66 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 66-70, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.66.70 © 2019 by the authors; licensee asian online journal publishing group leaf hexane extracts of two turkish fig (ficus carica l.) cultivars show cytotoxic effects on a human prostate cancer cell line olcay boyacioglu1 betül kara2 hilal can3 tugçe naile yerci4 sima yilmaz5 seda orenay boyacioglu6 ( corresponding author) 1,3,4,5department of food engineering, faculty of engineering, aydin adnan menderes university, efeler, aydin, 09010, turkey. 2department of food engineering, faculty of engineering, aydin adnan menderes university, efeler, aydin, 09010, turkey; molecular biotechnology program, institute of health sciences, aydin adnan menderes university, efeler, aydin, 09010, turkey. 6department of medical genetics, faculty of medicine, aydin adnan menderes university, efeler, aydin, 09010, turkey. abstract the world-famous turkish fig, grown in aydin, is also used for the treatment of various diseases with its leaves and latex. studies have shown that fig leaf has antioxidant, antiviral, antidiabetic, antimicrobial, and anticancer effects. the objective of the study is to examine the anticarcinogenic and antimicrobial effects of nonpolar extracts from the leaves of two fig cultivars (sari lop and aydin black) that are the most widely grown in aydin. the fig leaves collected in the summer of 2017 were dried in shade at room temperature and crumbled. the n-hexane extracts of the dried fig leaves obtained with manual soxhlet and semi-automated soxhlet apparatus were applied on pc3 human prostate cancer cell line for 24h. the antimicrobial activities of the extracts were examined on escherichia coli and bacillus cereus using disc and agar well diffusion methods. as a result, antimicrobial activity of the n-hexane extracts on the bacteria was not detected at the highest dose tested (100 mg/ml). the n-hexane extracts showed cytotoxic effect on pc3 cells in a dose-dependent fashion and caused close to 100% death at 1000 μg/ml. no significant difference was observed between the cytotoxic effects of n-hexane extracts of two fig cultivars and the extraction methods (p>0.05). keywords: ficus carica, fig leaf, hexane extract, prostate cancer, cytotoxicity, antimicrobial. citation | olcay boyacioglu; betül kara; hilal can; tugçe naile yerci; sima yilmaz; seda orenay boyacioglu (2019) leaf hexane extracts of two turkish fig (ficus carica l.) cultivars show cytotoxic effects on a human prostate cancer cell line. agriculture and food sciences research, 6(1): 66-70. history: received: 31 january 2019 revised: 7 march 2019 accepted: 17 april 2019 published: 11 june 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. bacterial strains were generously gifted by prof. dr. h. halil bıyık, and the pc3 cell line by prof. dr. william h. gmeiner. funding: this work was supported by the scientific and technological research council of turkey (tubitak), bideb 2209-a program, grant number 1919b011700595 to bk. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 67 2. materials and methods ................................................................................................................................................................... 67 3. results and discussion ................................................................................................................................................................... 68 4. conclusion ......................................................................................................................................................................................... 69 references .............................................................................................................................................................................................. 69 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.66.70&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.66.70&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/1837 https://orcid.org/0000-0003-0436-3020 https://orcid.org/0000-0001-8313-3340 https://orcid.org/0000-0003-2511-3565 https://orcid.org/0000-0003-0258-5164 https://orcid.org/0000-0001-7504-4161 https://orcid.org/0000-0003-1651-1940 agriculture and food sciences research, 2019, 6(1): 66-70 67 © 2019 by the authors; licensee asian online journal publishing group 1. introduction fig (ficus carica l.), a fruit tree of the moraceae family, is a small tree with a height ranging from 1 to 10 m. f. carica plant is grown mostly in the city of aydin in aegean region of turkey [1]. the fig is cultivated worldwide for its nutritive and medical properties. it is widely used in ayurveda, siddha, and homeopathy medical systems [2]. fruits, leaves, and roots of fig plants are some of the remedies people resort to in various illnesses. while fig fruit is used for the prevention of indigestion, anorexia, cardiovascular problems, and cancers; fig roots and leaves are used against indigestion and anorexia. in addition, fig leaf is also used against the anemia [3]. the total phenolic substance, organic acid content, and antioxidant potential in the fig leaf have been reported to be higher than in fig pulp and crusts [4]. it is claimed that f. carica is useful in treating liver and spleen disorders and gout disease. fig leaves were also tried for the treatment of jaundice. f. carica leaves boiled in hot water were reported to be used in kidneys and liver, in diabetes and calcification, and as well as in hemorrhoid treatment [5, 6]. f. carica leaves have also been used in the treatment of vitiligo due to the presence of furanocoumarins, psoralen, and daidzein [7]. the fig leaf is also reported to be used in asthma, cough, sexual disorders, diarrhea, hematuria, earache, toothache, migraine, eye problems, stomach problems, and scabies [8]. in traditional medicine, the use of f. carica leaf, fruit, and root is a reality due to the antibacterial, antifungal, antioxidant, and antiviral activities it has [3]. in 1998, when the aqueous decoction of fig leaf was administered to type i diabetic patients, hypoglycaemic effect was revealed as the postprandial (after the meal) glycemia decreased significantly in the patients [9]. following this, canal et al. showed that chloroform extract obtained from aqueous decoction of f. carica leaves was successfully used to decrease cholesterol levels and hyperglycemia on the rats with experimentally-induced diabetes [10]. it was reported that fig leaf methanol extract (500 mg/kg) decreased the aspartate aminotransferase, alanine aminotransferase, total serum bilirubin, and malondialdehyde levels, and showed liver protective effect by reducing lipid peroxidation in the liver [11]. this activity of the fig leaf extract is comparable to slymarine [12] which is known as a liver protective agent. the petroleum ether extract of f. carica leaves provided promising results in the treatment of rifampicininduced hepatic damage in rats [13]. a study conducted on wistar albino rats showed that the petroleum ether, chloroform, and ethanol extracts of f. carica leaves had anti-imflammatory properties [8]. in a recent study, ethyl acetate extract of fig leaf was shown to be controlling the glucose and fat levels in type 2 diabetic rats, thus being effective in controlling the carbohydrate metabolism [14]. while promising results were obtained with f. carica leaf extracts, ethanol extract of f. carica fruit (fig) up to 1000 ug/ml for 72 h did not show any toxicity on u87 glioblastoma cells [15]. in a study conducted in iran, the anti-angiogenic effects of fig leaf ethanolic extract was evaluated on human umbilical vein endothelial cells (huvec). after 24 hours of incubation with the extract at 25 μg/ml, a statistically significant cytotoxic effect on huvecs was reported compared to the control group [16]. the antimicrobial effects of f. carica leaf extracts have been examined in several studies. the methanol extract of fig leaves collected and dried in south korea was detected to have antimicrobial activity on oral bacteria [17]. in 2016, total phenolic and flavonoid contents of the methanol extracts from ten different varieties of f. carica in algeria were examined and antimicrobial effects of the extracts were determined by disk diffusion test. as a result, all extracts were rich in phenolic compounds and had antimicrobial effect against gram-positive and gramnegative bacteria. staphylococcus aureus and bacillus cereus were reported to be the most susceptible bacteria to f. carica extracts [18]. in another study, the ethanol extract of fig leaf showed growth inhibitory effect against the bacteria staphylococcus aureus, salmonella typhi, and the mold fusarium oxysporum. also, fig latex was tried in this study and reported to have a stronger effect. aspergillus niger and e. coli were more resistant to ethanolic extract and fig latex [19]. to the best of our knowledge, no results were reported on the essential oil and n-hexane extracts of fig leaves. the objectives of this study, were to assess the antibacterial and antiproliferative effects of nonpolar extracts from leaf of two turkish fig cultivars on two bacterial samples and on pc3 human prostate cancer cell line, respectively. 2. materials and methods 2.1. preparation of fig leaves the research material fig leaves were collected from the f. carica cultivar sari lop trees grown in the aydin adnan menderes university central campus (37°51'28.3"n 27°51'23.8"e) and f. carica cv. aydin black (also known as bursa black) leaves were collected from a fig orchard in gencelli village of kuyucak district in the city of aydin (37°56'04.5"n 28°39'28.9"e) in the summer of 2017. the collected leaves were visually inspected not to have damage due to sunburn, plant pests, or human handling. the leaves were shade-dried at room temperature for a week and crushed. the resulting leaf pieces were stored in dry and cool room conditions. 2.2. determination of water content of the fig leaves fresh, mature or young (leaf length < 10 cm) fig leaf pieces ranging between 4 and 10 gr was weighed into a pre-weighted crucible. after 3 hours at 105°c in lab oven, the crucible with the sample was cooled in a desiccator and weighed again to determine the water contents. 2.3. essential oil extraction essential oil was extracted from 100 gr of crushed dry fig leaves mixed with 800 ml of distilled water by 6-hr hydrodistillation using the clevenger-type apparatus. 2.4. hexane extraction methods in the first manual method, classical, glass soxhlet extraction system was used. dried and crushed (24 gr) fig leaf samples in the cellulose thimbles were extracted with 300 ml of n-hexane for 2.5 hours. the resulting extract agriculture and food sciences research, 2019, 6(1): 66-70 68 © 2019 by the authors; licensee asian online journal publishing group was filtered twice through whatman no. 1 filter paper and concentrated by rotary evaporator (büchi labortechnik ag, r-100, flawil, switzerland) at 55°c at about 70 bar vacuum pressure. the resulting solid extract mass was weighed, diluted in n-hexane to a concentration of 0.1 gr/ml (10%), and stored at -18°c. for the second extraction method, dried and crushed sari lop and aydin black fig leaves (a total of 24 gr in 3 cellulose thimbles) were extracted with a total of 150 ml of n-hexane in a semi-automated soxhlet device (velf scienfitica, ser 148/3, 20865 usmate, italy). the extraction program is composed of 40 min immersion, 80 min washing, and 3 min recovery. at the end of the extraction, the leftover n-hexane was removed and the resulting solid extract mass was stored as described above. 2.5. bacterial species antimicrobial assays were performed on e. coli atcc 35218 and b. cereus atcc 11778 (gifted by prof. dr. h. halil bıyık, aydın adnan menderes university). bacteria were cultured in tryptic soy broth (tsb) (merck) and tryptic soy agar (tsa) media consisting of tsb supplemented with 15% agar agar (sigma aldrich). 2.6. disc diffusion and agar well diffusion methods fifty μl of e. coli or b. cereus culture grown in tsb for 6-8 hours was transferred onto mueller hinton agar (mha) plates (merck) and spread over the entire surface to form a thin film layer. ten μl of the n-hexane extract of fig leaf dissolved in n-hexane or acetone at 0.1 gr/ml concentration was impregnated into sterile 6 mm antimicrobial susceptibility disks (oxoid) on the surface of mha plate, and the discs were pressed down gently to ensure complete contact with the agar surface. the n-hexane or acetone (10 μl) and a gentamicin (oxoid) disk were used as negative and positive controls, respectively. plates were incubated at 37°c for up to 48 h and the bacterial growth inhibition zones around the discs were observed and measured. agar well diffusion assay was used to increase the amount of extract tested. after the test bacteria were spread on the mha plates as described in the disc diffusion method, wells with 5 mm in diameter were formed on the agar under aseptic conditions. each well was filled with the n-hexane extract of fig leaf dissolved in n-hexane or acetone at 0.1 gr/ml concentration. plates were incubated and the bacterial growth inhibition zones around the discs were observed as described above. 2.7. prostate cancer cell line culture the anti-carcinogenic properties of the n-hexane extracts of the fig leaves were tested on pc3 human prostate cancer cell line (gifted by prof. dr. william h. gmeiner, wake forest university). cells were cultured in rpmi1640 (sigma-aldrich) supplemented with 10% fetal bovine serum (fbs) (biowest) in the presence of 5% co2 at 37°c. cells were routinely passaged using trypsin (sigma-aldrich) when they reached 80-90% confluence. 2.8. cytotoxicity mtt assay pc3 cells were seeded at a density of 3000 cells/well in 96-well plate and incubated in co2 incubator. the following day, hexane extracts were applied to the cells at 250, 500, 750, and 1000 μg/ml concentrations for 24-48 hours, and cell viabilities were measured colorimetrically by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (mtt) (sigma-aldrich). mtt was dissolved in sterile phosphate buffered saline (pbs) and further diluted in rpmi with 10% fbs to a final mtt concentration of 0.5 mg/ml. at the end of the fig leaf extract treatment, the used media was replaced with 120 μl of new media with mtt. after a minimum 3 hours of incubation, the violet colored formazan crystals inside the cells were observed under a microscope, and the mtt containing media was replaced with 100 μl of dimethyl sulfoxide (dmso) (merck). after the plate was shaken for 10 min to dissolve the formazan crystals, results were obtained with a spectrophotometer (thermo labsystems multiskan spectrum mic) at 570 nm. all experiments were performed in triplicates and the experiments were repeated 3 times. 3. results and discussion the water contents of fresh, mature fig leaves and the young leaves (leaf length < 10 cm) for both fig cultivars were found to be 70 and 78%, respectively. these values are correlated with the water content of mint leaves, which was reported as 70-80% [20]. essential oil extraction from both fig cultivar leaves was not possible as no essential oil was observed after the water distillation method using clevenger apparatus, possibly due to low amount of essential oil in the leaves. to check the equipment functionality, essential oil of thymus vulgaris l. was successfully extracted. a larger volumed extraction system might help in extracting essential oil from fig leaves. the n-hexane extraction yield rates of f. carica leaves in manual and semi-automated soxhlet methods were 1.55 and 2.30 mg/gr, respectively for the sari lop cultivar and 2.15 and 2.40 mg/gr, respectively for the aydin black cultivar. first, the f. carica leaf extracts prepared at 10% concentration in n-hexane were used in disc diffusion tests table 1. however, considering the low volume of extract used in discs and the fast vaporizing nature of n-hexane, no antibacterial effect of the extracts or the n-hexane negative control was observed. for this reason, the n-hexane extracts were solubilized in acetone at 10% concentration but no antimicrobial effect on the growth of e. coli and b. cereus was observed either table 1. the negative control acetone showed greater antimicrobial effect than all of the extracts. none of the fig leaf extracts obtained by semi-automated soxhlet system and dissolved in acetone had antimicrobial effect against b. cereus. similar results were obtained in well diffusion tests (results not shown). agriculture and food sciences research, 2019, 6(1): 66-70 69 © 2019 by the authors; licensee asian online journal publishing group table-1. result of disc diffusion tests using the n-hexane extract of fig leaves dissolved in n-hexane or acetone. extraction solvent reagents tested diameter of growth inhibition zone (mm) e. coli atcc 35218 b. cereus atcc 11778 none gentamicin (pos. control) 12 21 n-hexane n-hexane (neg. control) 6 6 sari lop m 6 6 sari lop s 6 6 aydin black m 6 6 aydin black s 6 6 acetone acetone (neg. control) 15 15 sari lop m 12 12 sari lop s 8 6 aydin black m 10 10 aydin black s 13 6 note: m: manual soxhlet; s: semi-automated soxhlet. however, a recent study conducted in algeria reported that methanol extracts of 10 different varieties of f. carica leaves showed antimicrobial properties against gram positive and gram negative bacteria [18]. type of extraction solvent used, geographic and climatic differences between turkey and algeria, and the type of the fig trees may be counted as the potential reasons for the difference in antimicrobial effectiveness between the studies. another study conducted in south korea with fig leaf methanol extract also reported antimicrobial effect against the oral bacteria [17]. in 1998, hypoglycemic effect of fig leaf was investigated in type i diabetic patients [9] and in 2000, a chloroform extract from f. carica leaves was used to reduce cholesterol levels in diabetic mice [21]. in this study, cytotoxic effects of n-hexane extract of fig leaves collected from two local cultivars (sari lop and aydin black) grown in aydin, turkey were investigated on the pc3 human prostate cancer cell line. fig leaf extracts were applied on cells at doses ranging from 0-1000 µg/ml for 24 h. results reveal that fig leaf extracts showed cytotoxic effect on pc3 cells in a dose-dependent manner figure 1. the leaf extracts started showing 50% cytotoxicity on pc3 cells after 24 h treatment at 250 µg/ml dose. almost complete cytotoxicity was observed on pc3 cells after 24 h treatment at 1000 µg/ml. no statistically significant difference was available between the cytotoxic effects of sari lop and aydin black cultivars (t-test, p>0.05). also, there was no significant difference in the comparison of the extraction type (manual soxhlet vs. semi-automated soxhlet device) (t-test, p>0.05). figure-1. treatment of n-hexane extracts of fig leaves on pc3 cells for 24 h. m: manual soxhlet; s: semi-automated soxhlet. the petroleum ether extract of f. carica leaf was successfully used in rats to protect the liver from damage by rifampicin [13]. similarly, hepatoprotective effects of fig leaf ethanol [22] and methanol [23] extracts were reported against hepatotoxicity caused by carbon tetrachloride. ethanol extract of fig leaf was reported to have a curing effect against both cellular and humoral antibody response in mice [24]. it was also indicated that fig leaf ethanol extract has immunity stimulating effect. additionally, the water extract of fig leaf is known to have toxicity against herpes simplex virus [25]. 4. conclusion in conclusion, f. carica fruit, latex, and leaf have been researched for their beneficial effects by various studies. this study revealed that 24 h treatment of n-hexane extract of two fig cultivars shows cytotoxic effect on pc3 human prostate cancer cell line. references [1] n. günal, "the culture of figs in turkic world," turkish stud, vol. 3, pp. 561–581, 2008. available at: https://doi.org/10.7827/turkishstudies.440. 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[25] g. wang, h. wang, y. song, c. jia, z. wang, and h. xu, "studies on anti-hsv effect of ficus carica leaves," zhong yao cai, vol. 27, pp. 754–756, 2004. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn(e) : 2411-6653 issn(p) : 2518-0193 vol. 3, no. 2, 49-52, 2016 http://www.asianonlinejournals.com/index.php/aesr 49 effect of foliar boron application on rice (oryza sativa l.) growth and final crop harvest sajed ali1 syed ali raza2 shahid javed butt3 zahid sarwar4 1,3university of management and technology, sialkot campus, sialkot, pakistan. 2,4directorate of land reclamation, irrigation department, lahore, pakistan. ( corresponding author) abstract boron (b) is an essential micro nutrient and its deficiency caused a reduction in final crop harvest and quality of the yield. a field experiment was conducted to evaluate the effect of foliar application of b on yield and yield components of rice in calcareous soils under agro-climatic conditions of lahore, pakistan. the experiment was laid out in randomized complete block design (rcbd) with six b foliar application rates (0, 5, 10, 15, 20 and 25 mg l-1). the experiment was replicated three times. the results illustrated a significant effect of b foliar application on number of grains panicle-1, number of filled grains and final grain yield. the highest grain yield (352 g m-2) was recorded in 20 mg l-1 foliar application, whereas an increase in b application to 25 mg l-1 reduces the final grain yield significantly (313 g m-2). detrimental effects of the highest b application on yield components were also observed. the decline in the quantity and quality rice yield resulted by increasing b application might be due to the toxic effect of higher concentration of b application. keywords: micronutrients, boron, rice, yield. contents 1. introduction ............................................................................................................................................................................... 50 2. materials and methods ............................................................................................................................................................. 50 3. results and discussion ............................................................................................................................................................... 50 4. conclusion .................................................................................................................................................................................. 51 references ...................................................................................................................................................................................... 51 citation | sajed ali; syed ali raza; shahid javed butt; zahid sarwar (2016). effect of foliar boron application on rice (oryza sativa l.) growth and final crop harvest. agriculture and food sciences research, 3(2): 49-52. doi: 10.20448/journal.512/2016.3.2/512.2.49.52 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: this work is licensed under a creative commons attribution 3.0 license contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 25 january 2016/ revised: 10 february 2016/ accepted: 16 june 2016/ published: 10 october 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://www.asianonlinejournals.com/index.php/aesr http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.2/512.2.49.52 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=sajed%20ali https://orcid.org/orcid-search/quick-search?searchquery=syed%20ali%20raza https://orcid.org/orcid-search/quick-search?searchquery=shahid%20javed%20butt https://orcid.org/orcid-search/quick-search?searchquery=zahid%20sarwar http://search.crossref.org/?q=10.20448/journal.512/2016.3.2/512.2.49.52 agriculture and food sciences research, 2016, 3(2): 49-52 50 1. introduction crop plants require a sufficient supply of essential mineral elements for optimal crop growth and final harvest in terms of crop yield. an insufficient availability of essential nutrients (micro and macro) may cause stunted growth and limits the crop productivity. boron is an essential micronutrient for plant growth [1] and has a key role in plant metabolism [2]. its deficiency are reported commonly among crops grown in soil having higher amounts of free carbonates, low organic matter, and high ph [3, 4]. crop removal from the field is the main driving force for decrease of soil nutrient levels because hardly any crop residue gets recycled back to the soil resulting in decreasing nutrient pool including b. in addition, abundant soil moisture (because of torrential monsoon rains during july august, in flooded rice fields etc.) causes b leaching, beyond the root zone [5]. on the other hand, the dry surface soil layers, because of dry spells during the growing season, inhibit root absorption of b [6, 7]. also, subsequent to green revolution, crop yields and crop intensification have risen and, hence, increased amounts of b are being removed from soils, year after year. hence, these essential elements can be supplied as fertilizers in both intensive and extensive agricultural systems. however, excess b can cause toxicity to both plants and animals. fertilizer use efficiency including foliar application methods can be enhanced agronomically, through improved fertilizermanagement practices by cultivating crops that acquire and/or utilize mineral elements more effectively [8, 9]. field research in pakistan has demonstrated yield increases due to b use on several crops, including wheat, rice, maize, groundnuts, tobacco, and potato. boron application increased the yield of wheat (up to 14%), rice (up to 19%) and maize (up to 20%) [10]. although monocotyledous crops are less sensitive to b deficiency as compared to dicotyledonous, cereals including maize, sorghum, wheat, rice and barley are also being affected by deficiency world scale [11, 12]. the total b concentration in soils ranges from 20-200 mg kg-1 [13]. most of it is unavailable to plants [14]. the available (hot water soluble) fraction generally ranges from 0.4-5.0 mg kg-1 [15]. boron availability decreases with increasing soil ph; thus it is often inadequately available in calcareous soils. boron uptake by plants correlates well with the level of hot water soluble and hcl extractable soil b [16]. keeping in view the deficiencies reported causing reduction in final crop harvest and less availability of b in calcareous soils, the objective of the present study was to evaluate the efficiency of boron foliar application on rice yield and yield components in calcareous soils of pakistan. 2. materials and methods a field experiment was conducted in field experimental area of directorate of land reclamation, canal bank moghalpura lahore, pakistan. the experiment was consisted of six boron application levels (0, 5, 10, 15, 20 and 25 mg l-1) and laid down in randomized complete block design and replicated thrice. the rice seedlings were transplanted in plots (40.5 m2) and there were twenty lines of rice seedling 9 m length. boron was applied as a solution of borax and was sprayed on rice at three growth stages including tillering stage, booting stage and milking stage. phosphorus (as diammonium phosphate) and nitrogen (as urea) was applied at the rates of 200 and 350 kg ha-1, respectively and were added to the all plots. a thinning was carried out (how many days after transplanting) to final plant density of 19 plants m-2 at seedling stage. weeds were mechanically controlled. grain yields were taken at maturity by harvesting the 1 m2 area of the two inner rows of each plot at the end of april. grain yield was adjusted to a 10% moisture basis. the following growth and yield component variables were recorded for each plot: plant height, fresh weight, number of tillers plant-1, number of panicles plant-1, panicle length number of grains panicle-1, grain yield, filled grains (%), , and 1000-seed weight. all data were analyzed by the glm procedure using the statistix (version 8.1 analytical software). means were compared using lsd test at 5% probability level. 3. results and discussion 3.1. yield and yield components results showed that b application had significant effects on grain yield, number of grains panicle-1 and filled grains (table 1). the highest rice grain yield (352 g m-2) was obtained in treatment with 20 mg l-1 solution of boron (fig. 1). increasing boron foliar application from 5 to 25 mg l-1 decreased grain yield (313 g m-2) significantly. one possible explanation for the decline in grain yield of rice at the highest b application is the antagonism relation between ca and b [17, 18]. it has been shown that the relative uptake rates of calcium significantly decreased in both shoot and root of plants as solution b concentrations was increased [19]. there are different reports on the effect of b on grain yield of rice. an increase in rice yield with increasing b has been reported [19, 20]. in contrast, it was observed that the grain yield of rice decreased as b fertilizes was increased [21]. table-1. mean square of analysis of variance for some measurements in this study sov d.f yield/g biomass/ kg no. of grains panicle-1 plant height/ cm panicle length/ mm filled grains/ % 1000 grains weight/g no. of tillers plant-1 no. of panicles plant-1 block 2 106.26ns 0.00277 ns 2.657 ns 12.4689 ns 1.88222 ns 3.8672 ns 0.8156 ns 0.2756 ns 0.2572 ns boron 5 8177.14** 0.06067ns 551.613** 66.5276ns 9.03156ns 35.8382** 13.3329ns 12.3206ns 12.4206ns error 10 20.29 0.00271 13.642 3.5936 2.08222 1.8499 2.1402 1.9822 0.4432 c.v (%) 1.64 2.41 2.97 1.83 6.63 1.92 6.83 9.47 4.86 source: unpublished data the highest number of grains per panicle and filled grains of rice (137.67 grains panicle-1 and 74 % respectively) resulted in treatment with 20.0 mg/l solution of boron (fig. 2 and 3). increasing boron fertilizer from 5.0 to 25.0 mg/l decreased number of grains per panicle (123.33 grains panicle-1) and filled grains (67.67 %) significantly. filled grains are affected by b deficiency as it has key role in pollen vitality [22]. agriculture and food sciences research, 2016, 3(2): 49-52 51 figure-1. effect of foliar application on rice yield figure-2. effect of foliar application on number of grains source: unpublished data source: unpublished data one possible explanation for the decline in number of grains per panicle and filled grains of rice at the highest b application is the antagonism relation between ca and b [17]. b application had no significant effect on 1000 grain weight (table 1). figure-3. effect of foliar application on number of filled grains figure-4. effect of foliar application on number of filled grains source: unpublished data source: unpublished data 3.2. biomass number of tillers per plant, panicle length and plant height b application had no significant effect on biomass, number of tillers plant-1, panicle length and plant height (table 1). there was slight increase in biomass production due to b application (fig. 4) but this increase was statistically non-significant. 20 mg/l gave maximum total biomass (2.36 kg m-2) whereas minimum biomass (1.96 kg m-2) was found in control treatment. these results are similar to those of khan, et al. [20] who obtained nonsignificant difference in number of plants m-2, total number of tiller plant-1 and number of fertile tillers plant-1 in response to applied fertilizer b, whereas, a significant improvement in number of grains per spike and 1000-grain weight was found when fertilizer b was sprayed on wheat foliage at three growth stages i.e. tillering, booting and milking. similarly, it was reported that in spite of increase in grain yield of rice, vegetative parameters as number of tillers plant-1, panicle length and plant height were not affected by b application [19, 23]. 4. conclusion the result of this study indicated that application of b had significant effect on yield and most components of yield of rice. application of 20 mg l-1 solution of b led to the highest grain yield of rice. application of b fertilizer more than 20 mg/l decreased grain yield and other yield components. this decline was significant when 25 mg/l b was applied. the decline in the yield parameters of rice might be due to the toxicity of high b application particularly in calcareous soil. the results showed that oryza sativa grown in a calcareous soil doesn’t need more than 20 mg l-1 b as foliar application. references [1] w. loomis and r. durst, chemistry and biology of boron vol. 3. oxford, england: biofactors, 1992. [2] d. g. blevins and k. m. lukaszewski, "boron in plant structure and function," annual review of plant biology, vol. 49, pp. 481500, 1998. [3] t. matoh and m. kobayashi, "boron and calcium, essential inorganic constituents of pectic polysaccharides in higher plant cell walls," journal of plant research, vol. 111, pp. 179-190, 1998. [4] l. bolaños, k. lukaszewski, i. bonilla, and d. blevins, "why boron?," plant physiology and biochemistry, vol. 42, pp. 907-912, 2004. [5] v. m. shorrocks, "the occurrence and correction of boron deficiency," plant and soil, vol. 193, pp. 121-148, 1997. [6] h. g. zheng, m. babu, r. chandra, m. s. pathan, l. ali, and n. huang, "quantitative trait loci for root-penetration ability and root thickness in rice: comparison of genetic backgrounds," genome, vol. 43, pp. 53-61, 2000. [7] c. plank and d. martens, "boron availability as influenced by application of fly ash to soil," soil science society of america journal, vol. 38, pp. 974-977, 1974. [8] z. rengel, g. batten, and d. d. crowley, "agronomic approaches for improving the micronutrient density in edible portions of field crops," field crops research, vol. 60, pp. 27-40, 1999. [9] y. zuo and f. zhang, "soil and crop management strategies to prevent iron deficiency in crops," plant and soil, vol. 339, pp. 83-95, 2011. [10] a. l. singh, r. s. jat, v. chaudhari, h. bariya, and s. j. sharma, "toxicities and tolerance of mineral elements boron, cobalt, molybdenum and nickel in crop plants," plant nutrition and abiotic stress tolerance ii plant stress, vol. 4, pp. 31-56, 2010. agriculture and food sciences research, 2016, 3(2): 49-52 52 [11] a. karim and j. vlamis, "micronutrient deficiency symptoms of rice grown in nutrient culture solutions," plant and soil, vol. 16, pp. 347-360, 1962. [12] a. rashid, m. yasin, m. ashraf, and r. mann, "boron deficiency in calcereous soil reduces rice yield and impairs grain quality," international rice research notes, vol. 29, pp. 58–60, 2004. [13] h. l. bohn, r. a. myer, and g. a. o'connor, soil chemistry, 2nd ed. new york: john wiley & sons, 2001. [14] x. yu and p. f. bell, "nutrient deficiency symptoms and boron uptake mechanisms of rice," journal of plant nutrition, vol. 21, pp. 2077-2088, 1998. [15] u. c. gupta, "boron nutrition of crops," adv. agron., vol. 31, pp. 273-307, 1979. [16] a. rashid, e. rafique, and n. bughio, "diagnosing boron deficiency in rapeseed and mustard by plant analysis and soil testing," communications in soil science & plant analysis, vol. 25, pp. 2883-2897, 1994. [17] u. c. gupta, factors affect1ng boron uptake by plants. boron and its role in crop production. boca raton, fl: crc press, 1993. [18] a. a. agboola and r. corey, "the relationship between soil ph, organic matter, available phosphorus, exchangeable potassium, calcium, magnesium, and nine elements in the maize tissue," soil science, vol. 115, pp. 367-375, 1973. [19] a. asad, r. bell, b. dell, and l. huang, "external boron requirements for canola (brassica napusl) in boron buffered solution culture," annals of botany, vol. 80, pp. 65-73, 1997. [20] r. khan, a. gurmani, a. gurmani, and m. zia, "effect of boron application on rice yield under wheat rice system," international journal of agriculture and biology pakistan, vol. 8, pp. 805-808, 2006. [21] m. john, h. chuah, and l. c. van, "boron response and toxicity as affected by soil properties and rates of boron," soil science, vol. 124, pp. 34-39, 1977. [22] o. garg, a. sharma, and g. r. kona, "effect of boron on the pollen vitality and yield of rice plants (oryza sativa l. var. jaya)," plant and soil, vol. 52, pp. 591-594, 1979. [23] m. shafiq and t. maqsood, "response of rice to model based applied boron fertilizer," j. agr. resour., vol. 48, pp. 303-314, 2010. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 52 agriculture and food sciences research vol. 5, no. 2, 52-56, 2018 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2018.52.52.56 growth and yield response of sweet potato (ipomoea batatas l. lam) varieties to lowland agro-ecology of raya azebo, ethiopia haileslassie gebremeskel1  kedir jaleto2 wakuma biratu3 haile abebe4 ( corresponding author) 1,2,3,4department of horticulture, ethiopian institute of agricultural research, mehoni agricultural research center, maichew, ethiopia abstract evaluation activity was conducted using four sweet potato varieties (awassa-83, kabode, kulfo and tulla) at mehoni agricultural research center (mearc) during 2015 and 2016 testing years. the treatments were arranged in randomized complete block design with three replications. the analysis of variance table indicated the existence of significant variation among sweet potato varieties for all the characters considered in this study in both 2015 and 2016 testing years except number of nodes/plant and tuber diameter during 2016 cropping year. from this investigation, significantly higher marketable yield (29.20 t ha-1) and total yield (33.42 t ha-1) were obtained from kulfo variety. overall, kulfo was found well adapted and produced higher yield in the testing location. hence, it is possible to use kulfo variety for the production of marketable and total tuber yield by farmers and investors in raya azebo, ethiopia. keywords: sweet potato, growth, tuber diameter, marketable yield, total yield. citation | haileslassie gebremeskel; kedir jaleto; wakuma biratu; haile abebe (2018). growth and yield response of sweet potato (ipomoea batatas l. lam) varieties to lowland agro-ecology of raya azebo, ethiopia. agriculture and food sciences research, 5(2): 52-56. history: received: 5 march 2018 revised: 7 april 2018 accepted: 9 april 2018 published: 12 april 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: our especial thank also go to tewodros abrha and halefom hailu for their help in field works. our sincere thanks also go to all who made input for this study. funding: the authors would like to acknowledge mehoni agricultural research center (mearc) and roots, vegetables and fruits research project for providing all the facilities and financial support during the entire experimentation. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 53 2. material and methods ..................................................................................................................................................................... 53 3. result and discussion ..................................................................................................................................................................... 53 4. conclusion ......................................................................................................................................................................................... 55 references .............................................................................................................................................................................................. 55 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.52.52.56&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=haileslassie%20gebremeskel https://orcid.org/orcid-search/quick-search?searchquery=kedir%20jaleto https://orcid.org/orcid-search/quick-search?searchquery=wakuma%20biratu https://orcid.org/orcid-search/quick-search?searchquery=haile%20abebe agriculture and food sciences research, 2018, 5(2): 52-56 53 1. introduction sweet potato (ipomoea batatas l. lam); 2n=6x=90) is a herbaceous dicotyledonous plant with creeping, perennial vines and adventitious roots. it belongs to family convovulaceae (morning glory flowers) and is hexaploid which is usually considered the only ipomoea species of economic importance [1]; [2]; [3]. sweet potato is originated from south america and it is cultivated in china, uganda, nigeria, indonesia, tanzania, vietnam, india and united states [4]. sweet potato is highly heterozygous cross pollinated crop in which many of the traits show continuous variation. it is known for its resistance to drought, vigorous early growth and low input requirements [5]; [6]. globally sweet potato is the seventh most important food crop after wheat, rice, maize, potato, barley and cassava and second most important root and tuber crop in the world after potato [3] however, in sub-saharan africa sweet potato is the third most important tuber crop after cassava and yam [7]. throughout the world 107.6 million tons of sweet potatoes are produced [8] whereas the world average storage root yield of sweet potato has been estimated to be 14.8 t ha-1 [9]. sweet potato has large, starchy, sweet-tasting and tuberous roots which adapts to tropical and warm temperate regions [10]. high dry matter content is the main characteristic preferred by consumers and processors of sweet potato [11]. storage-roots (root tubers) of sweet potato contain 30% dry matter that 70% of it starch, 5% sugar and 5% protein with vitamin a, c and b. especially orange colored sweet potatoes contain vitamin a (ßeta carotene) and vitamin c [12]. it is a relatively drought tolerant crop providing the highest dry matter content for human consumption [11]. sweet potato provides comparatively high calorie at 152 mj ha-1 day-1. other crops such as cassava, wheat, rice and maize provide 121, 135, 151 and 159 mj ha-1 day-1 calories, respectively [13]; [2]. in ethiopia, among the most important root and tuber crops, sweet potato is one of the major traditional food crops [14]. it is cultivated in ethiopia mostly for human consumption and as animal feed. it ranks third after enset (ensete ventricosum (welw.) cheesman,) and potato (solanum tuberosum l.) as the most important root crop produced in the country [15]. the ethiopian national average storage root yield of sweet potato is 8 t ha-1 while experimental storage roots yields ranging between 30 and 73 t ha-1 [16]; [17]. according to those authors yields obtained in ethiopia are generally low; however, there is good potential for the crop since climatic and soil factors are largely favourable. it is an attractive crop among ethiopian farmers due to its high productivity, universal uses, high caloric content and good taste, tolerant to adverse environmental conditions such as drought, low soil fertility, high rainfall and it requires very little labor and care compared to other crops [5]. according to these authors in ethiopian, sweet potato is commonly grown by farmers in complex, mixed cropping systems where they normally plant several varieties with different characteristics like that of yield, maturity, root size, shape and disease and drought tolerance. farmers may use the vines left in the fields to improve soil fertility and the crop is used in crop rotation. despite the various advantages of sweet potato have, research work on adaptability of the plant is very limited. in general, lack of information on performance evaluation and appropriate agronomic practices is considered to be among the major hindrance to embark on higher production and wisely utilization of this valuable plant in the country. thus, it is necessary to evaluate the adaptability that would enable to maximize growth and yield in order to diversify and popularize this economically important plant in study area. therefore, the objective of this study was to study the growth and yield response of sweet potato varieties to lowland agro-ecology of raya azebo. 2. material and methods five months old sweet potato mother plant introduced from hawassa agricultural research center (harc) was used as a planting material for this experiment. the experiment was carried out at mehoni agricultural research center (mearc), ethiopia in 2015 and 2016 cropping season. the center is situated at about 678 km north of the addis ababa. geographically it is located at 12° 41'50'' north latitude and 39° 42'08'' east longitude with an altitude of 1578 m.a.s.l. the site receives mean annual rainfall of 430-750 mm with an average minimum and maximum temperature of 18 and 25°c, respectively. the soil textural class of the experimental area is clay loam (vertisol) with ph of 7.9. the experiment consisted of four sweet potato varieties (awassa-83, kabode, kulfo and tulla) and it was arranged in randomized complete block design (rcbd) with three replications according to gomez and gomez [18]. thus, there were four treatments in each replication. the space between rows and plants was 75x30 cm having a plot size of 4.50 x 3.60 m (16.20 m2) and 72 plants per plot will be considered. the space between replications and plots will be 1.5 m and 1 m, respectively. plants in the four middle rows out of the six rows per plot were constitute the net plot to be used as the sampling units. eight plants from the middle rows were taken for sampling and data analysis. all appropriate agronomic practices such as weeding, watering and hoeing were conducted manually at the experimental field. data on emergence percentage, plant height, number of nodes/plant, internodes length, tuber length, tuber diameter, marketable yield/ha, unmarketable yield/ha and total tuber yield/ha were recorded when yellowing and falling of leaves and also cracking of the soil was observed in the experimental plots. statistical analysis of experimental data was subjected to analysis of variance (anova) using sas proc glm (2002) at p<0.05. differences between means were evaluated using least significant difference (lsd) test at p<0.05. 3. result and discussion 3.1. variation in growth and yield characters of sweet potato mean square from analysis of variance table for all the traits tested for two respective years are summarized table 1, 2 and 3. the performance of all tested varieties were found statistically different (p<0.05) in internode length and tuber length, highly significant (p<0.01) in number of nodes/plant and very highly significant (p<0.001) in emergence percentage, plant height, tuber diameter, marketable yield, unmarketable yield and total yield during 2015 cropping year. however in 2016, number of nodes/plant and tuber diameter did not significantly influenced (p>0.05) by varietal difference. whereas emergence percentage and plant height were significantly agriculture and food sciences research, 2018, 5(2): 52-56 54 affected (p<0.05) by sweet potato variety; which however, highly significant influence (p<0.01) was found in internodes length and tuber length and very highly significant (p<0.001) in marketable yield, unmarketable yield and total tuber yield of sweet potato varieties. this implies that the different genotypes were exerted significant influence on the growth and yield characters of sweet potato. table-1. mean square of growth and yield response of sweet potato during 2015 and 2016 testing years source of variation degree of freedom emergence percentage plant height(cm) number of nodes/plant 2015 2016 2015 2016 2015 2016 replication 2 48.08 10.76 0.002 0.01 9.59 14.07 variety 3 820.81*** 141.24* 0.068*** 0.05* 95.48** 15.99ns error 6 16.51 21.48 0.002 0.01 5.21 14.02 cv (%) 5.44 4.99 4.19 6.18 5.52 8.97 note: *= significant at p< 0.05; ** =significant at p<0.01 and ***= significant at p< 0.001 probability level. cv= coefficient of variation table-2. mean square of growth and yield response of sweet potato during 2015 and 2016 testing years source of variation degree of freedom internode length(cm) tuber length(cm) tuber diameter(cm) 2015 2016 2015 2016 2015 2016 replication 2 0.03 0.18 0.49 1.91 0.12 0.13 variety 3 0.91* 33.71** 8.16* 54.91** 6.57*** 1.02ns error 6 0.10 0.07 1.09 3.48 0.05 0.40 cv (%) 5.94 7.95 6.33 8.77 4.18 10.22 note: *= significant at p< 0.05; ** =significant at p<0.01 and ***= significant at p< 0.001 probability level. cv= coefficient of variation table-3. mean square of growth and yield response of sweet potato during 2015 and 2016 testing years source of variation degree of freedom marketable yield(t ha-1) unmarketable yield( t ha-1) total yield(t ha-1) 2015 2016 2015 2016 2015 2016 replication 2 115.10 22.77 0.64 24.89 114.52 176.27 variety 3 37526.51*** 31030.70*** 703.51*** 1141.79*** 48299.54*** 46648.58*** error 6 91.92 359.30 13.27 19.57 109.55 206.72 cv (%) 6.60 12.37 14.93 20.70 6.19 8.30 note: ns=not significant at p< 0.05, *= significant at p< 0.05; ** =significant at p<0.01 and *** =significant at p< 0.001 probability level. cv= coefficient of variation 3.2. performance variation in growth and yield of sweet potato the mean performances of sweet potato varieties are summarized in table 4, 5 and 6. comparing among the tested varieties the highest emergence percentage (98.61%) was found at variety kabode during 2016 testing year. while emergence percentage (94.91% and 94.41%) recorded at kulfo and tulla varieties, respectively were statistically at par with kabode during the same testing year. however, the lowest emergence percentage (50.31%) was obtained at tulla variety when the experiment was conducted in 2015 cropping year. the highest overall mean emergence percentage (90.46%) was recorded at kabode whereas the lowest value (72.61%) was recorded at tulla variety. significantly higher plant height (1.74 cm) and number of nodes/plant (48.67) were obtained from awassa-83 variety during 2016 and 2015 cropping year, respectively; which however, significantly lower value (1.02 cm) and 35.02 plant height and number of nodes/plant were recorded at variety awassa-83 and kabode varieties during 2015 testing year. kabode produced significantly higher internode length (5.65 cm) and tuber length (26.33 cm) during 2015 and 2016 cropping season; respectively. however, internode length obtained from kabode variety in 2015 was not statistically different with the value obtained from kulfo (5.46 cm) and tulla (5.39 cm) varieties. on the other hand, the lowest internode length (2.92 cm) and tuber length (14.03 cm) were found at kabode and awassa-83 varieties when the experiment was carried out during 2016 and 2015, respectively. this result in harmony with the findings of teshome, et al. [19]; yahay, et al. [20] in sweet potato. tuber diameter was significantly influenced in the first year; which however, it did not significantly affected by varietal difference in the second testing year. even though there was no significant effect on tuber diameter in the second cropping year kulfo produced slightly higher tuber diameter (6.96 cm) while the lowest value (3.48 cm) obtained from awasssa-83 in 2015 testing year. similar result with this finding [21] was also reported on sweet potato response to fertilizer trial. concerning the yield parameters, all the three yield characters were influenced by varietal difference in both testing years. kulfo produced significantly higher marketable yield (29.20 t ha-1) followed by same variety (26.57 t ha-1) in 2015 and 2016 cropping season, respectively. however, the lowest marketable yield (2.16 t ha-1) and (3.21 t ha-1) were obtained from awassa-83 variety while the experiment is implemented during 2015 and 2916, respectively. overall, mean marketable yield varied from 27.98 to 2.68 t ha-1 among the testing genotypes in both cropping seasons. unmarketable yield/ha was significantly influenced by genotypic difference in both testing years. the maximum unmarketable yield (4.71 t ha-1) was obtained at variety kulfo along with the testing year of 2016 followed by (4.22 t ha-1) at the same variety when the research experiment conducted in 2015 testing year; whereas, the minimum value (0.30 t ha-1) was obtained at awassa-83 variety in 2016 cropping year. the highest sweet potato total tuber yield (33.42 t ha-1) was obtained from kulfo variety in 2015 followed by (31.48 t ha-1) from the same variety during 2016; while the lowest total yield (2.58 t ha-1) was obtained from awassa-83 in 2016 testing year. from the mean comparison table kulfo variety was produced significantly higher (32.45 t ha-1) mean total tuber yield. these results are in agreement with findings of kathabwalika, et al. [22] in sweet potato. agriculture and food sciences research, 2018, 5(2): 52-56 55 table-4. mean performance of growth and yield of sweet potato varieties tested at raya azebo during 2015 and 2016 cropping season variety emergence percentage plant height (cm) number of nodes/plant 2015 2016 mean 2015 2016 mean 2015 2016 mean awassa-83 86.76a 82.87b 84.82 1.02c 1.74a 1.38 48.67a 42.08 45.38 kabode 82.31a 98.61a 90.46 1.06bc 1.59ab 1.33 35.02c 42.67 38.85 kulfo 79.43a 94.91a 87.17 1.15b 1.41b 1.28 40.00b 38.42 39.21 tulla 50.31b 94.91a 72.62 1.36a 1.57ab 1.47 41.67b 43.73 42.70 lsd (5%) 8.12 9.26 0.10 0.19 4.56 ns cv (%) 5.44 4.99 4.19 6.18 5.52 8.97 note: means followed by the same letter with in the same column are not significantly different at 5% level of probability. cv=coefficient of variation, lsd=list significant difference table-5. mean performance of growth and yield of sweet potato varieties tested at raya azebo during 2015 and 2016 cropping season internode length (cm) tuber length(cm) tuber diameter (cm) 2015 2016 mean 2015 2016 mean 2015 2016 mean awassa-83 4.42b 4.06a 4.24 14.03b 23.27a 18.65 3.48c 5.56 4.52 kabode 5.65a 2.92b 4.29 17.59a 26.33a 21.96 4.23b 5.83 5.03 kulfo 5.46a 3.36b 4.41 17.02a 17.64b 17.33 6.35a 6.96 6.66 tulla 5.39a 3.01b 4.20 17.13a 17.79b 17.46 6.39a 6.31 6.35 lsd (5%) 0.62 0.53 2.09 3.73 0.43 ns cv (%) 5.94 7.95 6.33 8.77 4.18 10.22 note: means followed by the same letter with in the same column are not significantly different at 5% level of probability. cv=coefficient of variation, lsd=list significant difference table-6. mean performance of growth and yield of sweet potato varieties tested at raya azebo during 2015 and 2016 cropping season variety marketable yield ( t ha-1) unmarketable yield(t ha-1) total yield (t ha-1) 2015 2016 mean 2015 2016 mean 2015 2016 mean awassa-83 2.16d 3.21d 26.83 0.56d 0.30c 4.26 2.71d 2.58d 26.45 kabode 11.90c 11.63c 117.64 2.09c 1.05c 15.70 13.99c 12.68c 133.34 kulfo 29.20a 26.75a 279.75 4.22a 4.71a 44.63 33.42a 31.48a 324.48 tulla 14.66b 19.73b 171.93 2.90b 2.47b 26.86 17.59b 22.53b 200.59 lsd (5%) 19.09 37.87 7.28 8.84 20.91 28.73 cv (%) 6.60 12.37 14.93 20.70 6.19 8.30 note: means followed by the same letter with in the same column are not significantly different at 5% level of probability. cv=coefficient of variation, lsd=list significant difference 4. conclusion the analysis of variance table indicated that all agronomic and yield characters had significantly affected by sweet potato genotypic difference in the 2015 cropping season; whereas, variety did not exert any significant influence on number of nodes/plant and tuber diameter traits. from this experimental study, significantly higher mean marketable yield (27.98 t ha-1), unmarketable yield (4.46 t ha-1) and total yield (32.45 t ha-1) were obtained from kulfo variety. generally, kulfo was found well performed and produced higher yield in the testing location. likewise, the values obtained in this experiment demonstrated comparable and even higher values in yield characters with the different reports. hence, it is possible to use kulfo cultivar for the production of marketable and total tuber yield in raya azebo, ethiopia. references [1] l. degrass, the yam a tropical root crop. the technical centre for agricultural and rural cooperation (cta). london: the macmillian press, 2000. 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[19] a. teshome, c. amenti, and t. kassaye, "yield and yield components of sweet potato as influenced by plant density. in adami tulu jido kombolcha district, central rift valley of ethiopia," american journal of experimental agriculture, vol. 1, pp. 40-48, 2011. view at google scholar | view at publisher [20] s. u. yahay, a. m. saad, s. g. mohammed, and s. o. afuape, "evaluating the performance of improved sweet potato (ipomoea batatas l. lam) advanced lines in kano, sudan savanna of nigeria," international journal of agronomy and agricultural research, vol. 7, pp. 52-60, 2015. view at google scholar [21] f. uwah, u. l. undie, n. m. john, and g. o. ukoha, "growth and yield response of improved sweet potato (ipomoea batatas (l.) lam) varieties to different rates of potassium fertilizer in calabar, nigeria," journal of agricultural science, vol. 5, pp. 61-69, 2013. view at google scholar | view at publisher [22] d. m. kathabwalika, e. h. c. chilembwe, v. m. mwale, d. kambewa, and j. p. njoloma, "plant growth and yield stability of orange fleshed sweet potato (ipomoea batatas) genotypes in three agro-ecological zones of malawi," international research journal of agricultural science and soil science, vol. 3, pp. 383-392, 2013. view at google scholar asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://scholar.google.com/scholar?hl=en&q=yield%20and%20yield%20components%20of%20sweet%20potato%20as%20influenced%20by%20plant%20density.%20in%20adami%20tulu%20jido%20kombolcha%20district,%20central%20rift%20valley%20of%20ethiopia http://dx.doi.org/10.9734/ajea/2011/173 https://scholar.google.com/scholar?hl=en&q=evaluating%20the%20performance%20of%20improved%20sweet%20potato%20(ipomoea%20batatas%20l.%20lam)%20advanced%20lines%20in%20kano,%20sudan%20savanna%20of%20nigeria https://scholar.google.com/scholar?hl=en&q=growth%20and%20yield%20response%20of%20improved%20sweet%20potato%20(ipomoea%20batatas%20(l.)%20lam)%20varieties%20to%20different%20rates%20of%20potassium%20fertilizer%20in%20calabar,%20nigeria http://dx.doi.org/10.5539/jas.v5n7p61 https://scholar.google.com/scholar?hl=en&q=plant%20growth%20and%20yield%20stability%20of%20orange%20fleshed%20sweet%20potato%20(ipomoea%20batatas)%20genotypes%20in%20three%20agro-ecological%20zones%20of%20malawi 64 agriculture and food sciences research vol. 5, no. 2, 64-67, 2018 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2018.52.64.67 influence of fertilizing with nitrogen fertilizer on the content of amino acids in sweet sorghum grain n.k.nokerbekova1  ye. t.suleimenov2 r.k. zhapayev3 ( corresponding author) 1phd doctoral student, kazakh national agrarian university, 050010, almaty, republic of kazakhstan 2candidates of agricultural sciences, kazakh research institute of agriculture and plant growing”, 040909, almaty region, republic of kazakhstan 3candidates of agricultural sciences international maize and wheat improvement center; cimmyt, 050035, almaty, republic of kazakhstan abstract three-year results of studies on the effect of fertilizing nitrogen fertilizer on the content of amino acids in the grain of sweet sorghum of kazakhstanskoe 16 and kazakhstanskoe 20 varieties on irrigated light chestnut soils of the southeast of kazakhstan are presented.nitrogen fertilizers are one of the effective means of increasing the protein content of sweet sorghum. the later nitrogen is introduced as a second fertilizer in a dose of n50 in the period of its vegetation, the more effective its effect on the increase in the protein content of the grain. the amount of nitrogen introduced into the soil is of great importance. the number of essential amino acids in sweet sorghum depended on the norms of applying nitrogen fertilizers. the higher is the norm of nitrogen, the more it accumulates in the grain of the protein. at the same time, when nitrogen was added in two terms, the number of such essential amino acids as valine, leucine, isoleucine, and threonine increased significantly, which increases the biological value of the total protein. therefore, it is possible not only to increase the yield, but also to increase the collection of protein from a unit of area by scientifically substantiating the use of nitrogen fertilizers for sweet sorghum. keywords: sweet sorghum, fertilizer, fertilizing, amino acids, variety, ground, soil supply. citation | n.k.nokerbekova; ye. t.suleimenov; r.k. zhapayev (2018). influence of fertilizing with nitrogen fertilizer on the content of amino acids in sweet sorghum grain. agriculture and food sciences research, 5(2): 64-67. history: received: 5 june 2018 revised: 9 august 2018 accepted: 12 september 2018 published: 15 october 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 65 2. methods ............................................................................................................................................................................................. 65 3. research results .............................................................................................................................................................................. 66 references .............................................................................................................................................................................................. 67 http://www.asianonlinejournals.com/index.php/aesr/article/view/1506 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.52.64.67&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.52.64.67&domain=pdf&date_stamp=2017-01-14 https://orcid.org/0000-0001-5761-8993 https://orcid.org/0000-0002-8224-0045 https://orcid.org/0000-0002-4272-7203 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2018, 5(2): 64-67 65 1. introduction the achievements of national and world science in recent years in fodder production convincingly show that it is possible to optimize the feeding of farm animals through the use of new higher-nutrient crops, one of which is sorghum. however, new varieties of sorghum can differ greatly from traditional sorghum and this can not be ignored when they are included in diets. generally, information on nutritional and chemical composition of feeds used in diets is taken from directories that do not take into account varietal characteristics, soil, climatic and other peculiarities of plant cultivation. therefore, the most reliable is the determination of feed nutrition before feeding animals under laboratory conditions [1]. amino acid nutrition takes a special place in the physiology of nutrition of highly productive animals. effective and cost-effective production of livestock products is inconceivable without the right combination of amino acids in the diet. on the one hand, the lack of one or the other amino acid interferes with the synthesis of protein and inhibits growth and productivity. on the other hand, excess nitrogen in the diet leads to additional energy consumption and increases the burden on the kidneys and the body as a whole. therefore, at the present time there is an increasing application of the concept of “ideal protein”, the essence of which is to give the animal as many amino acids as it needs, no more and no less. the quality of the fodder protein is determined from the point of view of the animals’ need for amino acids, firstly, the amino acid profile, which represents the ratio of essential amino acids in the protein and, secondly, their availability for metabolism and growth [2]. the content of protein and amino acids is one of the most important characteristics of the biological value of grain. the amino acid composition is used as a biochemical criterion for the biological value of fodder and food products (by the total content of essential amino acids). amino acids are the structural units of protein molecules involved in all processes occurring in the body of humans and animals. without proteins, life, growth and development of the body are impossible. proteins perform specific functions in the cell: enzymatic, construction, regulatory, etc.3% reduction of protein in the diet of the recommended standard causes a violation of the production of enzymes and, accordingly, the absorption of essential nutrients. deficiency of proteins stimulates the search for new varieties and hybrids with a high protein content, with a well-balanced amino acid composition that could be used in breeding programs to create new improved varieties. as the main protein-containing raw material, the leading role belongs to cereal crops. globally, about 70% of the needs of mankind in proteins are covered by grain either directly from food, or indirectly by feeding it to animals for meat production. consequently, plant storage proteins serve as the basis for human nutrition [3]. as known, proteins consist of amino acids, while some of them irreplaceable must necessarily come from feeds, since they are not synthesized in sufficient quantities in the animal’s body. the use of protein deteriorates, metabolism is dsturbed and productivity declines because of lack or absence of these amino acids in the rations of animals. critical essential amino acids are lysine and methionine. amino acids are the main subunits which build molecules of all protein substances. as the amount of protein increases, so does the content of amino acids. the sum of amino acids in the protein of sorghum grain is lower than in the protein of corn grain [4]. sorghum is a grain and fodder crop, anditis highly competitive with corn in terms of grain yield and green mass, and in some cases it exceeds it, especially in areas with unfavorable soil and climatic conditions. taking into account the prospects of using sorghum grain as a concentrated feed and source material for breeding, in recent years the chemical grain composition of local and selection sorghum varieties grown under irrigation conditions has been studied [5, 6]. the task of obtaining a high-quality, environmentally friendly and balanced agricultural products, in particular forage and food grains, has always been and remains relevant at all times [3.p 1]. the protein contains 20 amino acids: alanine, arginine, aspartic acid, valine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, oxyproline, ornithine, proline, serine, threonine, tryptophan, tyrosine, phenylalanine, cystine, cysteine and two amide asparagine and luteamine. for animals and humans, essential amino acids are valine, isoleucine, leucine, lysine, methionine, threonine, tryptophan and phenylalanine [7-9]. the purpose of our research was to study the effects of nitrogen fertilization on the amino acid composition of the grain protein of sweet sorghum varietiescultivated on light chestnut soils of the southeast of kazakhstan, with different supply of soils with mobile phosphorus. 2. methods field research was conducted in 2015-2017 on irrigated light chestnut soil in the foothills of the zailiyskiy alatau at a different level of soil supply with mobile phosphorus. agrochemical characteristics of light chestnut soils: humus content 2,34-2,54%, alkaline hydrolyzable nitrogen 81,2-97,7 mg/kg and exchange potassium 320-340 mg/kg soil. the experiment scheme included the following options: 1 control (without fertilizers); 2 n50 (fertilizing in the phase of 3-5 leaves); 3 n100 (fertilizingwith n50 in the phase of 3-5 leaves + n50 in the phase of 6-8 leaves). the experiments were based on two grounds of supply with mobile phosphorus according to machigin: (average 30-35 mg/kg soil) ground 1 and elevated (40-45 mg/kg soil) ground 2. nitrogen fertilizers were added in the form of ammonium nitrate (34%). the objects of research were varieties of sweet sorghum – kazakhstanskoe 16 and kazakhstanskoe 20. field experience was laid in 4-fold replication with the use of agricultural techniques adopted for this zone. accompanying analyzes of plant samples were carried out according to generally accepted procedures: total nitrogen, total phosphorus and total potassium in plants after wet ashing of plant material from one sample according to kjeldahl, colorimetrically, on a flame photometer. determination of proteinogenic amino acids was carried out by the method of capillary electrophoresis. statistical processing of the research results was carried out according to the methodology of dospehov [10]. using the agros program (version 2.09) according to martynov [11]. agriculture and food sciences research, 2018, 5(2): 64-67 66 3. research results by the conventional method, we have determined protein and a number of essential and other amino acids. the data of table. 1 show that the content of essential amino acids in sweet sorghum grain depended on the period of application of nitrogen fertilizers. the later nitrogen was introduced into the soil, the more amino acids accumulated in the grain. table-1. the content of essential amino acids of grain protein in varieties of sweet sorghum, depending on the norms of nitrogen fertilizers, g/100 g, average for 2015-2017 № aminoacids sweet sorghum varieties kazakhstanskoe 16 kazakhstanskoe 20 experiment variants experiment variants control n50 n100 control n50 n100 1 lysine 0,13±0,04 0,11±0,04 0,12±0,04 0,12±0,04 0,16±0,05 0,18±0,06 2 methionine 0,09±0,03 0,09±0,03 0,09±0,03 0,08±0,03 0,09±0,03 0,11±0,04 3 leucine + isoleucine 0,37±0,11 0,38±0,10 0,46±0,12 0,39±0,10 0,43±0,11 0,39±0,10 4 phenylalanine 0,21±0,06 0,25±0,07 0,35±0,11 0,22±0,07 0,35±0,10 0,30±0,09 5 threonine 0,18±0,07 0,15±0,06 0,21±0,09 0,18±0,07 0,20±0,08 0,24±0,10 6 valine 0,23±0,11 0,25±0,10 0,30±0,12 0,25±0,10 0,28±0,11 0,28±0,11 amount 1,21 1,23 1,53 1,24 1,51 1,50 as the results of the research showed, the introduced increasing rates of nitrogen fertilizers positively influenced both the irreplaceable and the remaining amino acid composition of sweet sorghum and, at the same time, it depended on the biological characteristics of varieties. the content of individual types of amino acids also varied depending on the norms of nitrogen fertilizers. thus, in the protein of the kazakhstanskoe 16 and the kazakhstanskoe 20 in the n50 variant of the increased ground, the methionine content was 0,09-0,09 g/100 g, threonine 0,15-0,20 g/100 g and valine 0, 25-0,28 g /100 g, and in the variant n100 methionine (0,09-0,11 g/100 g) and threonine (0,21-0,24 g/100g), that is, it was at the level of control option. the effect of fertilizers was on such kinds of amino acids as lysine, phenylalanine and valine, where the content increased on the average in varieties to 0,15; 0,32 and 0,29 g /100 g, respectively, against the variant without fertilizers 0,12; 0,22 and 0,24 g/100 g. it is known from the literature that the last indispensable amino acid is a precursor for the formation of phytohormone indolylacetic acid and vitamin pp, whose role in plant life is well known [12, 13]. the maximum value for the content of both control and fertilized variants was noted for leucine+isoleucine respectively, in the kazakhstanskoe 16 – 0,37 and 0,38–0,46 g/100g, and in the kazakhstanskoe 20 – 0,39 and 0,39– 0,43 g/100 g. thus, we can say that the variety kazakhstanskoe 20 is more responsive to the introduction of nitrogen fertilizers into the fertilizing, the synthesis of the amino acid is enhanced, and therefore the total amount of amino acids of the protein in the grain also increases. the ratio of amino acid in sweet sorghum grain depends on various environmental factors. the introduction of fertilizers leads to a change in the ratio of amino acids in the grain of sweet sorghum. in the variant n50of kazakhstanskoe 16 valine amplifies, and also the activity of glycine and methionine decreases. the total content of essential amino acids in kazakhstanskoe 20 was slightly higher in variant n50 than in grain of kazakhstanskoye 16, and in variant n100 it was the same (table 1). thus, the sum of essential amino acids in the kazakhstanskoe 16 in variants n50 and n100 was 1,23 and 1,53 g/100 g, respectively, with a control content of 1,21 g/100g, whereas in the kazakhstanskoe 20 it was 1,51 and 1,50 g/100 g with a control of 1,24 g/100 g. the content of the remaining amino acids of the protein in the varieties of kazakhstanskoe 16 and kazakhstanskoe 20 varies considerably in n50 and n100 variants of the raised ground, especially arginine (0,18; 0,34 and 0,33; 0,35 g /100 g), tyrosine (0,11; 0,16 and 0,14; 0,14 g /100 g), proline (0,48; 0,55 and 0,58; 0,46 g/100 g), serine (0,19; 0,23 and 0,21; 0,22 g /100 g), alanine (0,37; 0,43 and 0,43; 0,39 g/100 g), glycine (0,21; 0,20 and 0,20; 0,20 g/100 g), glutamic acid (1,20; 1,26 and 1,00; 0,80 g/100 g), aspartic acid (2,48, 2,61 and 2,79 ; 2,89 g /100 g) and cystine (0,17; 0,19 and 0,22; 0,21 g/100 g), respectively (table 2). table-2. the content of essential amino acids of grain protein in varieties of sweet sorghum, depending on the norms of nitrogen fertilizers, g/100g, average for2015-2017 № amino acids sweet sorghum variety kazakhstanskoe 16 kazakhstanskoe 20 experiment variants experiment variants control n50 n100 control n50 n100 7 arginine 0,28±0,11 0,18±0,07 0,34±0,13 0,14±0,06 0,33±0,13 0,35±0,14 8 tyrosine 0,17±0,05 0,11±0,03 0,16±0,05 0,13±0,04 0,14±0,04 0,14±0,04 9 proline 0,40±0,13 0,48±0,11 0,55±0,14 0,49±0,13 0,58±0,15 0,46±0,12 10 serine 0,18±0,07 0,19±0,05 0,23±0,06 0,21±0,05 0,21±0,06 0,22±0,06 11 alanine 0,37±0,13 0,37±0,10 0,43±0,11 0,39±0,10 0,43±0,11 0,39±0,10 12 glycine 0,20±0,07 0,21±0,07 0,20±0,07 0,18±0,06 0,20±0,07 0,20±0,07 13 glutamic acid 0,87±0,48 1,20±0,35 1,26±0,50 0,99±0,39 1,00±0,40 0,80±0,32 14 aspartic acid 2,49±1,16 2,48±0,87 2,61±1,04 2,12±0,85 2,79±1,12 2,89±1,16 15 cystine 0,19±0,09 0,17±0,08 0,19±0,09 0,17±0,08 0,22±0,11 0,21±0,11 amount 5,15 5,39 5,97 4,82 5,90 5,66 total amount 6,36 6,62 7,50 6,06 7,41 7,16 data on both the amount of the essential and the total amount of amino acids show that they depended on the agriculture and food sciences research, 2018, 5(2): 64-67 67 norms of nitrogen fertilizers and the biological characteristics of varieties. so, if the amount of the essential amino acids in the variety kazakhstanskoe 16 in variants n50 and n100 were respectively 5,39 and 5,97 g/100 g, then in the kazakhstanskoe 20 it was 5,90 and 5,66 g/100 g. the total amount of amino acids was, respectively, according to the variants of the experiment in the kazakhstanskoe 16 were 6,62 and 7,50 in the kazakhstanskoe 20 variety 7,41 and 7,16 g/100 g. at the same time, in the control variant, these indicators were respectively in the kazakhstanskoe 16, 5,15 and 6,36 for the kazakhstanskoe 20 4,82 and 6,06 g/100 g. to conclude, nitrogen fertilizers are one of the effective means of increasing the protein content of sweet sorghum. if you introduce nitrogen during its growing season later, the more effective its effect on the increase in the protein content of the grain. the amount of nitrogen introduced into the soil is of great importance. the higher the dose of nitrogen, the more it accumulates in the protein grain. at the same time, when nitrogen was added in two terms, the number of such essential amino acids as valine, leucine, isoleucine, and threonine increased significantly, which increases the biological value of the total protein. therefore, by scientifically substantiating the use of nitrogen fertilizers for sweet sorghum, it is possible not only to increase the yield, but also to increase the collection of protein from a unit of area. references [1] a. astashov, s. kononenko, and i. kononenko, "sorghum as an ingredient of combined fodder for broiler chickens," maize and sorghum, vol. 5, pp. 13-14, 2009. [2] i. s. kononenko, "sorghum for broilers," agrobusiness, vol. 2, pp. 42-43, 2010. [3] p. kondratenkoye, o. b. konstantinova, o. m. soboleva, y. a. izhmulkina, n. v. verbitskaya, and a. s. sukhikh, "the content of protein and amino acids in the grain of winter crops that grow in the forest-steppe of the south-east of western siberia," chemistry of plant raw materials, vol. 3, pp. 143-150, 2015. [4] e. i. adeyeye, "the intercorrelation of the amino acid quality between raw, steeped and germinated pearl millet (pennisetum typhoides) grains," pakistan journal of scientific and industrial research, vol. 52, pp. 122-129, 2009. [5] t. a. akmaliyev, efficiency of the use of sorghum and chickpeas in the feeding of milking cows. the author's abstract of the dis. ... candidate of agricultural sciences: ust-kinelsky, 2015. [6] a. v. alabushev, "unique possibilities of sorghum," agriculture, vol. 3, p. 19, 2000. [7] p. aleshinye and a. a. ponomarev, plant physiology. kolos: publishing house, 1979. [8] v. l. kretovich, exchange of nitrogen in plants. moscow: publishng house nauka, 1972. [9] v. l. kretovich, fundamentals of plant biochemistry. moscow: statepublishing house vysshaya shkola, 1961. [10] b. a. dospehov, methodology of field experience (with the basics of statistical processing of research results), 5th ed. moscow: agroprom publishing house. (textbooks and teaching aids for higher educational institutions), 1985. [11] s. p. martynov, statistical and biometric-genetic analysis in plant growing and breeding: the program package agros, version 2.09: user manual / tver, 1999. [12] s. boonchoo, s. fukai, and s. hetherington, "barley yield and grain protein concentration as affected by assimilate and nitrogen availability," australian journal of agricultural research, vol. 49, pp. 695-706, 1998.available at: https://doi.org/10.1071/a97103. [13] m. r. ebadi, j. pourreza, j. jamalian, m. a. edriss, a. h. samie, and s. a. mirhadi, "amino acid content and availability in low, medium and high tannin sorghum grain for poultry," international journal of poultry science, vol. 4, pp. 27-31, 2005.available at: https://doi.org/10.3923/ijps.2005.27.31. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 127 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 127-133, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.127.133 © 2019 by the authors; licensee asian online journal publishing group traditional potato food types preparation practices and farmer reaction in welmera and ada berga district, west shewa, ethiopia egata shunka tolessa1 gebremedhin weldegiorigis2 abebe chindi3 ( corresponding author) 1,2,3ethiopian institute of agricultural research, holetta research center, horticulture research division, addis abeba, ethiopia. abstract in most developing countries potatoes are consumed in fresh with some processing into chips and crisps. in ethiopia there are only three type of potato processed products such as boiled or cooked product, wot and deep fried types. principally, farmers in ethiopia produce potato for fresh marketing and tubers seed sale. to diversify potato utilization, training on potato processing was held in two kebeles of welmera and one kebele’s of ada berga district for selected potato producing farmers group and cooperative members to promote potato processing and increase farmer’s attitude towards potato production as well as to diversify the use of potato product and foods farmers use. the recipes done were potato pasta, potato kinche, potato quanta, potato starch, potato crisp and chips, genfo, shiro wot, chechebsa, enjera, bread, firfir (from mixture of meat and egg). these food types were found food which have good look, nutritious and pleasant. use of potato in different food items raise the appetite of people and diversify food farmers’ use. farmers appreciated the food types prepared from potato. they recommended processing and promoting it through mass media (tv), radio and exhibition in town administration. in addition, the farmers appreciated the effort holetta research center puts in providing new seed tubers together with its technology to producers and they asked to get continues support in the future time. keywords: potato, receipts, chips, crisp, genfo, enjera, bread and chechebsa. citation | egata shunka tolessa; gebremedhin weldegiorigis; abebe chindi (2019). traditional potato food types preparation practices and farmer reaction in welmera and ada berga district, west shewa, ethiopia. agriculture and food sciences research, 6(1): 127-133. history: received: 15 march 2019 revised: 17 april 2019 accepted: 21 may 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: the authors express their great thanks to aster kassa and fantaye balcha for their practical work support during farmers practicing. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 128 2. methodology ................................................................................................................................................................................... 128 3. result ................................................................................................................................................................................................ 129 4. discussions ...................................................................................................................................................................................... 133 references ............................................................................................................................................................................................ 133 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.127.133&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.127.133&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/0000-0002-1490-5826 agriculture and food sciences research, 2019, 6(1): 127-133 128 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study is contributing to the existing literature by showing ways and the possibility of eating potato in different food items. it helps the food science to look at another potato food preparation procedures. it also raises consumer interest and price for producers. in addition, it helps to diversify the ways of eating potato product in different food items. 1. introduction potato (solanum tuberosum l.) is highly productive and well suited for all type of consumer. the demand for potatoes is increasing due to expanded diet diversity, requests for prepared food items, and a need for reasonably priced foods [1]. potato consumption growth was also because of its wider range of climates adoption with broad range of cultures [2]. according to king and slavin [2] potatoes played vital role in preventing starvation and malnutrition worldwide and is a source of income for developing world. potatoes are packed with vitamins and minerals, although the nutritional content varied with variety and preparation method [3]. according to ryan [3] potatoes can be processed in too many food items employing many preparation ways, including boiled, baked and steamed. potato is eaten as boiled, baked, or fried and frequently served as a side dish or snack according to the description of atli [4]. he also added the common potato-based foods and food products embrace french fries, potato chips, and potato flour. in most developing countries potatoes are consumed in fresh with some processing into chips and crisps. potato can be processed into stable products which are used for food and industrial applications such as in textiles, starch, alcohol, glucose, paper and etc. utilization in ethiopia is restricted mainly in areas of production and in major cities where it is processed into chips and crisps. the usual mode of food preparation is by cooking such as boiling, roasting and deep frying. in ethiopia there are only three type of potato processed products such as boiled or cooked product, wot and deep fried types. but in the research areas it can be processed in to 36 types of food in ethiopia and more than 100 types of food in india (www.ficciagroindia.com/aic/post-harvest-mgmt/vegetables/potato.htm). processing is transformation of the raw tubers into sweeter and more palatable product to enhance consumption. it is attractive alternative to sale in the fresh form since the bulkiness and persheability of potatoes can lead to cause marketing problem. potato processing is very important to diversify use of potatoes and expand its market. thus, continuous evaluation of new germ plasm and improvement of existing varieties has to be incorporated in utilization component and variety development strategies as education; marketing, breeding, field management, and finally preparation for consumption enhances its contribution to human nutrition [5]. in most cases, farmers in ethiopia produce potato for fresh marketing and tubers seed sale. due to the persheability of the product, fresh marketing fetch low income as result of high supply during harvesting. this forces farmers to have low profit and even throwing of the product near the market as they have limited chance of selling and consuming it in relation to bulkiness and high weight as well as difficulty to transport twice. even in tuber seed sale the demand and supply are not fitting each other. there is chance of throwing these products in some years due to all cooperative and farmers group are producing the potato for this purpose regardless of the demand. due to these reasons and others, promotion of potato processing in to different food types is a substitute less strategy to solve shortage of market, increase profit and diversify potato use. based on these, training on potato processing was held in two kebeles of welmera district and one kebele’s of ada berga district for selected potato producing farmers group and cooperative members to promote potato processing and increase farmer’s attitude towards potato production as well as to diversify the use of potato product and foods farmers use. 2. methodology training was held in tellecho and burka welmera kebele of welmera district and bishan dimo kebele of ada berga district table 1 in 2014 to prepare different cultural food of ethiopian from gudanie potato variety. model potato producing farmers were selected by the help of wereda and chair man of the farmer group/ cooperative members. the selected farmers were called to the kebele and the training was started with adequate general discussion about potato production and use for human economy and health, nutritional value as whole. those recipes types done from raw potato such as: potato pasta, potato kinche, potato quanta, potato starch, potato crisp and chips were done by trainers in front of farmers to indicate procedures how to prepare and the type of tubers to be used. table-1.the group numbers and members of the trainees group. group number recipes prepared by the group district ada berga welmera burka welmera telecho 1 pasta and chechebsa 7 4 7 2 kinche and genfo 7 5 6 3 starch 6 5 7 4 crisp and shiro wot 7 5 6 5 chips and firfir with egg 7 5 7 6 quanta and firfir with meat 7 6 8 7 enjera and bread 4 3 4 total 45(male =23) 33(male=14) 45(male=24) source: training trainee group and group member, recipes the trainee practice. following same procedures the remaining recipes: genfo, shiro wot, chechebsa, enjera, bread, firfir (from mixture of meat and egg) were done from boiled or cooked potato tubers. in between the recipes section there were important questioning and answering in order trainees internalize what they were told about the steps to be followed to prepare each food type under issue. the next day the traineer was grouped in to 7 groups with each of agriculture and food sciences research, 2019, 6(1): 127-133 129 © 2019 by the authors; licensee asian online journal publishing group them 38mumbers and given one or two type of food type in order they practice how to do it themselves under the supervision of the trainers table 1. the groups were made to present what they prepared and on the last general dissuasions were made for receiving their reaction about the training. the total recipes and group as well as group members were listed in table 1 figure-1. food types done. source: photo taken from recipes made during training. each of these food types had their own procedures figure 1. the procedures for foods made from raw potato were similar in which all of them share selection of appropriate quality and quantity. then washing until the soil or mud completely removed and peeling the potatoes to remove outer hard cover. the other separate procedures were: potato pasta:-peeled long potato tubers were cut using knife in to thin and long form or in pasta shape and cooked in the same procedure of pasta. potato kinche: peeled potato was cut using knife in to pieces in similar size of kinche of barley, wheat or oat and the ways of preparation is similar to the kwon table kinche. potato quanta: peeled potato was cut using either knife or rubbing rough metal materials and dry in oven or sunlight until constant weight was achieved in order to pack it for future use. potato starch: peeled potato was rubbed with rubbing rough metal materials and socked in water in the bucket for ten minute for settling. it is schussed in another bucket and again socked for ten minute for schussing in the same manner as above. socking and schussing was continued until clear water drops from schussing. the schussed liquid was stayed for 24hours again for settling and the water was carefully removed next day and dry the white dough found below the settlement from these process which is called starch after drying. tuber characteristics selected for processing into chips and crisps were shallow eyes bearing tubers, high dry matter containing, and long shape for chips and round for crisps as well as low reducing sugar concentrations. crisp: peeled potato was cut in circular or in diameter in thinner depth which is washed by clean water and put for partial drying. then roasted in drier than chips in oil until golden color was developed. chips: peeled potato was cut in length or in longitudinally in not more than pencil thickness which is washed by clean water and put for partial drying. then roasted in oil until golden color was developed. procedures for those food types made from cooked potato were sharing same procedures in which all involve selection of needed quality and quantities of tubers, washing and boiling to the required extent. then the boiled and cooled potato tubers were peeled. genfo:peeled potato tubers were made cut in to pieces and grinded in to dough like structure which is placed in small boiled water and mixed and again cooked to 30 minute through continuous mixing. then ready for eating after taken out of stove or fire and placed in dish by deeping in central part for pouring boiled oil or butter together with salt, rade pepper and spices to get good flavor for eating. shiro wot: peeled potato tubers were made cut in to pieces and grinded in to dough like structure which is placed in cooked onion in oil and water, and mixed in order to again cooked to 30 minute through continuous mixing. then ready for eating after taken out of stove or fire to gather with enjera or bread after salting and spicing. chechebsa: peeled potato tubers were made cut in to pieces and grinded in to dough like structure which is mixed with dough made from durum wheat. as soon as possible mixing was completed; place all dough in baking materials through flattening for baking. it became ready for eating after cutting baked bread in to pieces and mixed with boiled oil or butter together with salt and spices enjera:-yeast and teff flour was mixed with water first day and next day peeled potato tubers were made cut in to pieces and grinded in to dough like structure which is mixed with mixed dough. after making thinner by boiled water, enjera was baked. bread: yeast and teff flour was mixed with water first day and next day peeled potato tubers were made cut in to pieces and grinded in to dough like structure which was mixed with mixed dough and baked in to bread. firfir: peeled potato was cut in to somewhat large size and added to cooked egg or meat which was eaten together with enjera or bread. 3. result the training was done on processing of potato in to thirteen well adapted and commonly used food types. these were kinche, pasta, starch, quanta, chips, kirps, genfo, chechebsa, shiro wot, firfir from meat and egg, enjera and bread figure 2. agriculture and food sciences research, 2019, 6(1): 127-133 130 © 2019 by the authors; licensee asian online journal publishing group figure-2. food types prepared in the training. source: photo taken from prepared recipes during taining. 1. kinche: is very important food in ethiopia which was consumed mostly as breakfast. it is previously made from barley, wheat and oat. in these training, it was done from potato and it was found very delicious food. the figure-3 below show the photo of kinche made from potato. figure-3. photo of kinche made from potato. source: taining photo graph. 2. pasta: is previously made from wheat and it is a common food in ethiopia. the past made during training from potato was found very nutritious and pleasant. figure 4 shows the exact photo of pasta of potato. figure-4. pasta done during training. source: photo graph of pasta taken during training. 3. starch: was made from potato during training. it is small in weight due to small amount of potato were used to prepare it. starch made from potato was similar to bulla of enset. it is used for enjera, bread and shiro wot, and typical food of ethiopian people called mucku. it can be packed in plastic bag and stored in dry and cool areas for long time. this type of processing is one way of preserving potato for long period of time. 4. quanta: was also done from potato during the training. previously quanta were done from meat as ethiopian cultural food preservation through drying the meat after cutting meat in to long thinner thickness by the help of air or smoking. this quanta from meat was used as food either directly or after processing it in to food type it fit. but, quanta of potato consumed after processing it and were a means of preservation for potato product/tuber which have short shelf life. this food was found very nutritious and pleasant. the food done during training from quanta of potato was shown below in figure 5. agriculture and food sciences research, 2019, 6(1): 127-133 131 © 2019 by the authors; licensee asian online journal publishing group figure-5. potato quanta food done during the training. source: photo graph of quanta taken during training. 5. chips: is the best well known processed food of potato worldwide. in ethiopia, the ways of processing potato in chips was not well known except recently started in big towns and hotels. in the trainings the chips done was very pleasant and easy to prepare by any individual household people in ethiopia. it can be also one area of work for source of income for low living farmers. figure 6 show the exact photo of chips prepared in the trainings. figure-6. chips made from potato in the training. source: photo graph of chips taken during training. 6. crisps: is the best well known processed food of potato worldwide. in similar manner as chips, in ethiopia the ways of processing potato in to crisps was not well known except recently started in big towns and hotels. in the trainings the crisps done was very pleasant and easy to prepare by any household individual people in ethiopia using available resource and knowledge. it can be also one area of work for source of income for low living farmers. chips and kirps are similar except tuber shape selected, ways of cutting potato and extent of frying. figure 7 show the exact photo of crisps prepared in the trainings. figure-7. crisps made from potato in the training. source: photo graph of crips taken during training. the others food types prepared were foods done from well cooked potato. these food types were the commonly eaten food in ethiopian country feeding habit in most cases. these foods were genfo, chechebsa, shiro wot, firfir from meat and egg, enjera and bread. 7. genfo: is commonly made in ethiopia from flour of teff, barley, wheat and maize. but in the training it was made from potato and it was found sweeter and pleasant. figure 8 indicates the exact photo of genfo made from potato during the training. agriculture and food sciences research, 2019, 6(1): 127-133 132 © 2019 by the authors; licensee asian online journal publishing group figure-8. genfo made from potato. source: photo graph of genfo taken during training. 8. chechebsa: is an important cultural food of ethiopian peoples. it is formerly made from cereals. in the training it was made from equal sized mixture of potato dough and durum wheat flour dough. this type of chechebsa was found sweeter and pleasant than commonly known one. figure 9 indicates the exact photo of chechebsa made from potato during the training. figure-9. chechebsa made from potato during the training. source: photo graph of chechebsa taken during training. 9. shiro wot: is commonly prepared from bean and pea in ethiopia. in the training it was made from potato in the manner of work procedure of shiro wot previously prepared from flour of bean and pea. it was found sweeter, pleasant and easy to work than the previous one. figure 10 indicates the exact photo of shiro wot made from potato during the training. figure-10. shiro wot made from potato during the training. source: photo graph of shiro wot taken during training. 10. fir fir from mixture of potato and meat, potato and egg: this type of food was made from well cooked potato which was peeled and sliced into small size which was mixed with either of cooked egg or cooked meat. this food type was found very advantageous in which potato increased amount of food and give good flavor. as potato addition increases dry matter the mixture can have importance for human body and economy. the biological value of mixture of egg and potato is higher than the egg alone. hence, potato can be supplement of meat and milk products for improving their taste, lowering energy intake and reducing food cost. figure 11 indicates the exact photo of this type of made from potato during the training. figure-11. meat and potato mixture, potato and egg mixture foods done during the training. source: photo graph of meat and egg mixed firifir taken during training. agriculture and food sciences research, 2019, 6(1): 127-133 133 © 2019 by the authors; licensee asian online journal publishing group 11. enjera and bread: commonly made from cereals especially durum wheat for bread and teff for enjera. in the training the two were made from potato dough and teff for enjera and durum wheat for bread by equal size mixture. they were found very nutritious and pleasant. figure 12 indicates the exact photo of enjera and bread made from potato during the training. figure-12. photo of bread and enjera made from potato during the training. source: photo graph of bread and enjera taken during training. 4. discussions food types made during training were found food which have good look, nutritious and pleasant which is similar with description given for potato in ryan [3] as described potatoes healthy, delicious and versatile. these food types made would require shorter procedures, time, cost and labor to prepare them when compared to other relevant locally known food types. use of potato in different food items raise the appetite of people and diversify food farmers’ use. it helps to get same product with different ingredients due to additives’ difference. it also facilitate utilization of potato product contribution to development as these product have high energy, useful protein which involved in cell growth and amendments, vitamins and minerals which promote health achievements of individual person and the country as the whole. because of the dry matter, edible energy and edible protein content of potato, it is found nutritionally superior vegetable as well as staple food not only in our country but also throughout the world which have to be converted in to individual house hold and country food achievements. being a short duration crop, it produces more quantity of dry matter, edible energy and edible protein in lesser duration of time than cereals like rice and wheat. hence, potato may prove to be a useful tool to achieve the nutritional security of the nation due to high return compared to other crops per unit area and time. the protein of potato has high biological value than proteins of cereals and even better than that of milk. this means it has higher amount of protein used for body growth and maintenance than others from taken in whole amount of protein [2]. based on variety or cultivar grown the biological value of potato is 90-100 while it was 100 for whole egg and 84 for soybean and 73 for legumes [6]. according to hampson [7] the protein value of potato is higher than protein value of any plant protein heavily eaten. the biological value of mixture of egg and potato is higher than the egg alone. hence, potato can be supplement of meat and milk products for improving their taste, lowering energy intake and reducing food cost. in nutritional point of view, potato is a wholesome food and deserves to be promoted as a potential high quality vegetable food crop in the country. farmers’ attitude about the training and their recommendation: farmers appreciated the food types prepared from potato during the training. they promised to use potato products as their back bone of their economy. in the past they were throwing potato tubers when they loss markets. due to this, their interest and attitude weakened to produce potato as they did not know different ways of eating it and the function of food prepared from potato as well. they recommended that if potato have these importances and can be eaten in these different food types, its processing has to be promoted through mass media (tv), radio and exhibition in town administration. in addition the farmers appreciated the effort holetta research center puts in providing new seed tubers together with its technology to producers and they asked to get continues support in the future time. references [1] food and agriculture organization, food and agriculture organization plant production and protection division. rome, italy: food and agriculture organization of the united nations, 2008. [2] j. c. king and j. l. slavin, "white potatoes, human health, and dietary guidance," advances in nutrition, vol. 4, pp. 393s-401s, 2013. available at: https://doi.org/10.3945/an.112.003525. [3] r. m. r. ryan, "7 health and nutrition benefits of potatoes. written by ryan raman, ms, rd. the epoch times." available: https://m.theepochtimes.com/7-health-and-nutrition-benefits-of-potatoes_2471127.html, 2018. [4] a. atli, "potatoes 101: nutrition facts and health effects, written by atli arnarson, phd." available: https://www.healthline.com/nutrition/foods/potatoes, 2019. [5] k. m. kolasa, "the potato and human nutrition," american potato journal vol. 70, pp. 375-384, 1993. [6] m. e. camire, s. kubow, and d. j. donnelly, "potatoes and human health," critical reviews in food science and nutrition, vol. 49, pp. 823-840, 2009. available at: https://doi.org/10.1080/10408390903041996. [7] c. hampson, "nutritional value of potatoes," nutrition bulletin, vol. 3, pp. 299–309, 1976. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.healthline.com/nutrition/foods/potatoes, 114 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 114-119, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.114.119 © 2019 by the authors; licensee asian online journal publishing group microbial loads of ogiri-ahuekere condiment produced from groundnut seed (arachis hypogaea linn) chukwu, m. n1 ezeagwula, c. g.2 nwakaudu, a. a3 oti, w.4 anyaogu, i5 ( corresponding author) 1department of food technology, abia state polytechnic, aba, abia state, nigeria. 2department of microbiology, abia state polytechnic, aba, abia state, nigeria. 3department of food science and technology, federal university of technology, owerri, imo state, nigeria. 4department of biochemistry, abia state polytechnic, aba, abia state, nigeria. 5department of hospitality and hotel management, abia state polytechnic, aba, abia state, nigeria. abstract microbial loads of ogiri-ahuekere condiment produced from groundnut seeds were examined. the groundnut seeds were sun-dried for 8 hours, dehulled and boiled for 8 hours using kerosene stove. the cooked cotyledons were milled manually into a paste and wrapped in small portions (30g) with blanched plantain leaves. the wrapped samples were fermented in a container for 1-10 day(s) while the unfermented cooked groundnut paste was used as a control. the freshly prepared samples of ogiri-ahuekere were used for microbial analysis and this action was carried out under sterile aseptic conditions. statistical analysis of the data was carried out using anova method with application of spss version 20. the significant difference between the mean values was determined by tukey’s test at 95% level of confidence. there was no growth in unfermented sample for tcc and tfmc while there was growth for tbc. there was significant increases in tcc which ranged from` 0.67-6.47 x 107 cfu/g, tbc (0.83-8.60 x 107 cfu/g) and tfmc (0.30 – 4.90 x 107 cfu/g). the results obtained from the study have shown the prevalence of bacteria throughout the period of fermentation in an increasing population. keywords: fermentation, coliform count, bacterial count, fungal count, colony, paste. citation | chukwu, m.n; ezeagwula, c.g.; nwakaudu, a.a; oti, w.; anyaogu, i (2019). microbial loads of ogiri-ahuekere condiment produced from groundnut seed (arachis hypogaea linn). agriculture and food sciences research, 6(1): 114-119. history: received: 7 march 2019 revised: 5 april 2019 accepted: 10 may 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 115 2. materials and methods ................................................................................................................................................................. 115 3. results and discussion ................................................................................................................................................................. 117 4. conclusion ....................................................................................................................................................................................... 118 5. recommendation............................................................................................................................................................................ 118 references ............................................................................................................................................................................................ 118 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.114.119&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.114.119&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/129 https://orcid.org/0000-0003-3371-7983 https://orcid.org/0000-0002-8249-6219 https://orcid.org/0000-0002-2714-0906 https://orcid.org/0000-0002-9251-9349 https://orcid.org/0000-0002-6237-4548 agriculture and food sciences research, 2019, 6(1): 114-119 115 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the study of the microbial loads of ogiri-ahuekere condiment showed the contamination of the condiment as well as the number of cells of different microbial counts. ogiri-ahuekere condiment will compete favourably in the market if processed or handled under sanitary conditions. 1. introduction the scientific classification of groundnut as kingdom (plantae), order (fabales), family (fabaleae), sub-family (faboideae), tribe (aeschynomenae), genus (arachis) and specie (hypogaea) [1]. groundnut (arachis hypogeae linn) is an important oil crop of brazil origin. it is specie in the legumes or “bean” family (fabaceae). it is also widely produced in guinea savannah ecological zone of nigeria. it is cultivated in tropical and warm temperate climate [2]. groundnut is an important oil crop of brazilian origin, cultivated in tropical and warm temperate climates. the crop is grown usually as a component of a variety of crop mixtures including sorghum, millet, cowpea and maize [3]. sahayaraj and martin [4] reported that groundnut is an important oil seed and cash crop accounting for more than one third (1/3) of the total oil seeds in india. beside income for farmers, it provides an inexpensive source of high quality dietary protein and oil [5]. groundnuts are particularly susceptible to contamination during growth and storage. poor storage of groundnut can lead to an infection by the mould fungus aspergillus flavus, releasing the toxic and highly carcinogenic substance called aflatoxin. the aflatoxin producing mould exists throughout the groundnut growing areas and may produce aflatoxin in the groundnuts when the conditions are favorable to fungal growth [6]. condiments are edible substances which are added to impart a particular flavor, enhance its flavor, and in some cultures to complement the dish. many condiments are packaged in single-sachets/packets e.g. mustard and ketchup. they are prepared from both plant and animal materials using processes in which micro-organisms play active roles in the physical, nutritional and sensory modification of the starting materials. the local condiment is an oily paste with strong putrid ammonical odor made from fermented vegetable protein [7]. condiments are prepared by traditional methods of uncontrolled solid substrate fermentation resulting in extensive hydrolysis of the protein and carbohydrate components. condiments are used as soup condiments and they generally have strong aroma [8]. condiments are excellent sources of proteins with essential amino acids, and also contain lipids, carbohydrates, essential fatty acids and vitamins [9]. many families in west africa often used fermented condiments as low cost meat substitute. fermented condiments improve nutritive values of foods as well as sensory properties as taste enhancers; contain antioxidants and neutraceuticals that promote health [10]. fermented condiments often have a stigma attached to them as they are considered to be food for the poor [11]. traditional diets in nigeria often lack variety and consist of large quantities of the staple foods (cassava, yam, maize) with supplements of plantain, cocoyam, rice and beans depending on their availability and seasons [12]. soups eaten with the staples are essential components of the diet and may contain a variety of seeds, nuts, pulses and leaves [13]. fermentation is one of the oldest and most economical methods of producing and preserving foods in developing countries [14]. fermentation remains of interest since they do not require refrigeration during distribution and storage [15]. apart from increasing the shelf life, and a reduction in the anti-nutritional factors, fermentation markedly improves the digestibility, nutritive value and flavour of the raw seeds [16]. when fermentation is involved in food processing, microorganisms are present. fermentation is an energy yielding metabolic process which involves the decomposition of substrate in the anaerobic condition. members of the fermenting organisms are important; for example, bacillus spp in ogiri preparation [1, 2]. fagbemi, et al. [17] described fermentation of condiments as a multi-step process which does not include a formal inoculation step. bacteria required for the fermentation appear to be incidental to both the raw and processed materials. indigenous flora is likely carried over from fermentation to fermentation sieves, trays and bags which are respectively used in condiment production. contamination of spores from the local environment may also contribute to the fermentation micro flora. it is likely that fermentation begins when the cooled, softened, dehulled oil seeds are packaged with leaves or other materials [18]. changes over the course of 6 days of fermentation period include a ph decrease for the first four days and an increase thereafter [17]. achi [14] reviewed some important microorganisms found in fermented condiments after different periods of fermentation at optimum conditions. ogbonna, et al. [19] determined the optimum ph for the growth of these bacteria as 7.5 while the optimum temperature is 33-40oc for 3 days. the methods employed in the manufacture of fermented condiment differ from one region to another because these processes are based on traditional systems. according to local custom, climate conditions and the type of substrates used, specific process variation occur [8]. in general, fermentation takes place under conditions which the producers have found to be favorable for appropriate growth and activity of microorganisms. this work is aimed at determining the microbial loads of ogiri-ahuekere condiment produced from groundnut seed using natural fermentation method. the production of ogiri-ahuekere will give value addition to groundnut and as source of income to both the farmers and the producers. this also could help in determining the sanitary condition of the condiment in the market. 2. materials and methods 2.1. collection of materials the groundnuts seeds were bought from a local market at aba, abia state, nigeria. other materials such as swab sticks, syringes, whatman filter papers no 1, foil, cotton wool, hand gloves, cover lips, distilled water and media produced by titan biotech ltd and micro-master ltd, india were purchased from rufus chemicals, aba. other utensils used were produced by search tech ltd, india and they include glass wares, pots, furnace, desiccators, plates, autoclave, dhp 9032 incubators, bunsen burner, olympus microscope and colony counter. the reagents used were of analytical grade and they include iodine solution, ethanol, safaranin solution, sulphuric acid, agriculture and food sciences research, 2019, 6(1): 114-119 116 © 2019 by the authors; licensee asian online journal publishing group methylene blue, starch, glucose, lactose, sucrose, fructose, maltose, boric acid, sodium hydroxide, bromocresol green and methyl red, edta, petroleum ether etc. reagents were produced by bdh chemicals ltd, poole england. 2.2. production of ogiri-ahuekere from raw groundnut seeds five hundred grams (500g) groundnut seeds were weighed and spread under the sun for easy removal of the seed coats (hulls). the hulls were removed by rubbing the seeds in-between the palms in accordance with the method described by chukwu, et al. [1]. the preparation of ogiri-ahuekere was prepared according to chukwu, et al. [1];chukwu, et al. [2]. figure 1 shows the flow diagram for the production of ogiri-ahuekere. figure-1. flow chart for the production of fermented ogiri–ahuekere samples. source: chukwu, et al. [8]. 2.3. microbial analysis of ogiri-ahuekere freshly fermented samples of ogiri-ahuekere were used for microbial analysis 2.4. media preparation the media used were nutrient agar (na), salmonella–shigella agar (ssa), macconkey agar (ma), mannitol agar, blood agar (ba) and potato–dextrose agar (pda). pda was used for fungi isolation while all the other agars were used for bacterial isolations. these agars were prepared as follows: 2.8g of nutrient agar was dissolved in 100ml of water; 11.1g of mannitol was dissolved in 100ml of water; 5.2g of macconkey agar was dissolved in 100ml of water; 6.3g of ssa was dissolved in 100ml of water; 2.8g of nutrient agar was dissolved in 2ml of blood to form the blood agar; and potato extract was made up to 100ml mark with distilled water. these agars were sterilized in the autoclave at 121oc/15 psi for 15minutes. the media were poured into sterile petri dishes on gelling. these were done in triplicates. 2.5. serial dilution nine folds serial dilutions were made using 10 test tubes placed on a test tube rack. one gram of the test sample was dissolved in 9ml of normal saline and diluents in a test tube and the content was thoroughly shaken. subsequent serial dilutions (10-2, 10-3, 10-4, 10-5, 10-6) were made from the solution by adding serially 1ml of the sample solution from proceeding concentration to 9ml of the diluents using sterile syringe. after dilution, 0.1ml of 10-5 dilution (105 tubes) was picked out and dropped into different plates of nutrient agar, macconkey agar, salmonella–shigella agar, mannitol agar, and blood agar for bacteria isolation and potato–dextrose agar for fungi agriculture and food sciences research, 2019, 6(1): 114-119 117 © 2019 by the authors; licensee asian online journal publishing group isolation. both pour plate and streak plate methods were employed. the plates were incubated for 24 hours at 37oc. plates were examined and the colony growths were counted [20]. 2.6. determination of microbial loads of ogiri-ahuekere samples the method of the icmsf [21] was employed. one gram (1g) of the prepared sample was dissolved in 9ml sterile distilled water (diluents) and mixed vigorously by shaking. about 1ml of the resultant mixture was aseptically transferred to 9ml sterile water in the test tube and thoroughly mixed. this action was carried out under sterile aseptic conditions. the dilution was continued serially until the sixth dilution was attained. the potato dextrose agar culture plates were incubated at room temperature for two days (48 hours) while the nutrient agar culture plates were incubated at 37oc for 24 hours in the incubator. all the plates were observed daily. the number of colonies formed in each culture plate at the end of incubation period was counted using the gallenkamp electronic colony method [21]. the total bacterial count was determined by counting the number of viable cells on nutrient agar for bacteria and potato dextrose for fungi plates respectively. thereafter, the number of viable cells was counted by using model tt201 colony counter. total coliform count was determined by introducing the sample inoculums in eosin methylene blue agar using pour plate method. the plates were incubated aerobically for 18-24 hours at 57oc. the total viable cells from primary plates were calculated using model tt201 colony counter. thereafter, a standard formula was used to calculate the number of the total coliform count [22]. the total microbial counts (cfu per gram) of food sample were calculated according to equation 1. 𝑇𝑀𝐶 = 𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑐𝑜𝑙𝑜𝑛𝑖𝑒𝑠 𝑜𝑏𝑠𝑒𝑟𝑣𝑒𝑑 × 𝑖𝑛𝑜𝑐𝑢𝑙𝑢𝑚 𝑝𝑙𝑎𝑡𝑒𝑑 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑖𝑛𝑜𝑐𝑢𝑙𝑢𝑚 ×𝑑𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑓𝑎𝑐𝑡𝑜𝑟 (1) 𝑇𝑀𝐶 = 𝑇𝑜𝑡𝑎𝑙 𝑣𝑖𝑎𝑏𝑙𝑒 𝑐𝑒𝑙𝑙𝑠 × 𝑖𝑛𝑜𝑐𝑢𝑙𝑢𝑚 𝑝𝑙𝑎𝑡𝑒𝑑 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑖𝑛𝑜𝑐𝑢𝑙𝑢𝑚 ×𝑑𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑓𝑎𝑐𝑡𝑜𝑟 (2) 3. results and discussion table 1 showed the average total microbial counts of ogiri-ahuekere produced from groundnut seeds at various fermentation time. the microbial counts included total coliform count, total bacteria count and total fungi/mould count, no growth in the unfermented ahuekere sample. 3.1. total coliform count (tcc) the tcc of unfermented ahuekere (0.00) was not significantly (p≤0.05) different from tcc of fermented ogiriahuekere sample from 1 day fermentation (0.67×107cfu/g). table showed that tcc of fermented ogiri-ahuekere samples from 1 and 2 day(s) were significantly the same while tcc of 2 days fermented ogiri-ahuekere sample (1.03×107cfu/g) was significantly different from tcc of unfermented ahuekere though similar to tcc (1.60×107cfu/g) of 3 days fermented ogiri-ahuekere sample. it was observed that the tcc (2.43×107cfu/g) of 5 days fermented ogiri-ahuekere sample was not significantly different from the tcc of 3, 4 and 6 days fermented ogiriahuekere samples (1.60, 1.93, and 3.20×107cfu/g respectively) while the tcc of 6 days fermented ogiri-ahuekere sample was significantly different of that of 4 days fermented ogiri-ahuekere sample. the total coliform count of 8 days fermented ogiri-ahuekere sample was 4.67×107cfu/g. the total coliform count (tcc) of 8 days fermented ogiriahuekere sample was significantly different from tcc of 6 and 9 days (5.70×107cfu/g) fermented ogiri-ahuekere samples, and it was also the same with tcc of 7 days fermented ogiri-ahuekere sample (3.87×107cfu/g); although, tcc of 6 and 7 days fermentation were significantly the same. however, the tcc of 9 and 10 days fermented ogiriahuekere samples were the same statistically but significantly different from the tcc from other samples. the table also illustrated how the total coliform count increased with the fermentation day [23]. 3.2. total bacteria count (tbc) there was also significant increase in the tbc with respect to the days of fermentation. the unfermented ahuekere sample had tbc (0.83 x 107 cfu/g) which was statistically the same with the tbc from 1 day fermented ogiri-ahuekere sample. the tbc (1.37, 1.70 and 2.33×107cfu/g respectively) from samples fermented for 1, 2 and 3 days were significantly (p≤0.05) similar but were significantly different from the tbc (2.80×107cfu/g) of 4 days fermented ogiri-ahuekere sample. it was observed that after 4 days fermentation, the tbc of the samples increased significantly throughout the period of fermentation table 1 the tbc of 5 days fermented ogiri-ahuekere sample was 3.47×107cfu/g which is significantly different from the tbc (4.40×107cfu/g) of 6 days ogiri-ahuekere samples. there were significant difference among the tbc of 5, 6, 7, 8, 9 and 10 days ogiri-ahuekere samples. this significant increase in tbc was due to the increase in fermentation time and similar results were reported by peter-ikechukwu, et al. [24]. 3.3. total fungi/mould counts (tfmc) total fungi/mould counts (tfmc) was also shown in table 1. the tfmc of all the ogiri-ahuekere samples increased significantly with increase in fermentation time. the unfermented ahuekere had no fungi growth. the following samples were statistically similar in tfmc: unfermented, 1 and 2 day(s) samples (0.00, 0.30 and 0.60×107cfu/g respectively); 3 and 4 days samples (1.00 and 1.33×107cfu/g respectively); 6 and 7 days samples (2.73 and 3.23×107cfu/g respectively); and 7 and 8 (3.23 and 3.33×107cfu/g respectively) days fermented samples respectively. however, the tfmc (2.03, 4.33 and 4.90×107cfu/g respectively) from samples fermented for 5, 9 and 10 days were significantly (p≤0.05) different from tfmc of the rest of the samples. sanni, et al. [25] and reported that some nigerian soup condiments have the microbial load within the range of 107 and 109 cfu/g as well as peterikechukwu, et al. [23]. agriculture and food sciences research, 2019, 6(1): 114-119 118 © 2019 by the authors; licensee asian online journal publishing group table-1. mean microbial count of ogiri-ahuekere fermented for 0-10 days. fermentation time (day) total count (x107cfu/g) tcc tbc tfmc 0 0.00a 0.83a 0.00a 1 0.67ab 1.37ab 0.30a 2 1.03bc 1.70bc 0.60ab 3 1.60cd 2.33cd 1.00bc 4 1.93d 2.80de 1.33c 5 2.43de 3.47e 2.03d 6 3.20ef 4.40f 2.73e 7 3.87fg 5.50g 3.23ef 8 4.67g 6.50h 3.33f 9 5.70h 7.30i 4.33g 10 6.47h 8.60j 4.90h lsd 1.014 0.843 0.531 note: means with the same superscripts are statistically the same but means with the different superscripts are significantly different from each other (p ≥ 0.05) in the column. where tcc = total coliform counts tbc = total bacteria counts tfmc = total fungi/mould counts note: the microbial load increased significantly from 2-10 days fermentation of groundnut seeds. 4. conclusion the results obtained from the study have shown the prevalence of bacteria throughout the period of fermentation in an increasing population. total counts of the microorganisms involved in fermentation process showed that contamination might have occurred either from processing equipment, the handlers or from the raw materials and the environment. fermentation time caused significant increase in the microbial loads of ogiriahuekere samples 5. recommendation fermented groundnut condiments could improve nutritive values of foods as well as sensory properties as taste enhancers; contain antioxidants and neutraceuticals that promote health; however it is recommended that further study on the toxicology of this product be carried out before it can wholly be accepted for human consumption. references [1] m. n. chukwu, c. s. nwakodo, q. alozie, and j. c. ndulaka, "comparative studies on organoleptic properties of ogiri-ahuekere and ogiri-egusi condiments," journal of food science research quality control, vol. 4, pp. 11-19, 2018a. 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[25] a. sanni, g. ayernor, e. sakyi-dawson, and s. sefa-dedeh, "aerobic spore-forming bacteria and chemical composition of some nigerian fermented soup condiments," plant foods for human nutrition, vol. 55, pp. 111-118, 2000. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 1, no. 1, 11-14, 2014 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 11 effect of drying methods on the nutritive value of some aquatic macrophytes in river rima, north western nigeria mamman, t 1* --ipinjolu, j.k 2 --bilyaminu g.d 3 1,2,3 department of fisheries and aquaculture, usmanu danfodiyo, university sokoto, nigeria abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ......................................................................................................................................................................... 12 2. materials and methods ........................................................................................................................................................ 12 3. results .................................................................................................................................................................................. 12 4. discussion ............................................................................................................................................................................. 13 5. conclusion ............................................................................................................................................................................ 13 references ................................................................................................................................................................................ 14 a comparative study on the effectiveness of different drying methods on the nutrient contents of water hyacinth (eichorniacrassipes) , water lilly(nymphae lotus) and water primrose (ludwigiahysopifollia ) leavewas carried out in order to ascertain their potential as nutrient supplements in fish feed formulation. whole components of each plant were weighted and 600g each of the leave were separately weighted in triplicates and was subjected to oven dying, sun drying and air drying. results of analysis indicated that oven dried leave of ludwigiahysopifolliahad significant (p< 0.05) high crude protein (15.13± 0.01) than sun dried and air dried (11.64±0.01 and 4.84±0.60), this follows the same trend with eichorniacrassipes and nymphaea lotus 12.52± 0.12 and with 11.76 ± 0.02 respectively. the ash, crude fiber and crude lipid contents exhibit no significant (p> 0.05) difference in the oven drying method of the three plants. moisture content differ significantly (p< 0.05) in eichorniacrassipes in all the drying methods, higher values nitrogen free extract was recorded in each drying method. the percentage composition of leaves shows that ludwigiahysopifollia has 64.74% eichorniacrassipes has 16.01% and nymphea lotus 55.95%. the study concluded that oven drying recorded the best effect on the nutrients contents of the plant leaves this grossly followed by air and sun drying method, and also the leaves of these macrophytes can be used to substitute conventional carbohydrate not protein supplements in fish feeds. further studies should be carried to assess the potential of these macrophytes subjected to the various drying methods in feeding cultured fish species. keywords: drying method, nutritive values, aquatic macrophyte. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2014, 1(1): 11-14 12 1. introduction nutrition is vital in fish farming because feed cost represent 40-50% of the total variable cost of production [13]. for several decades fishmeal has been used as the main sources of protein in fish feeds [4, 5], however, the periodically occurring low availability, competition and continuously fluctuating price of fish meal and other animal protein sources are affecting aquaculture feed production [6, 7]. as a result, a lot of attention has been focused on feedstuffs alternative to fish both from animal and plant sources [5]. in nigeria, most water bodies are flourished with large quantity of aquatic macrophytes as weed covering most part of water surface [8, 9]. studies into the possibility of converting these macrophytes into economically useful materials will help a lot in curbing their menace [8, 10]. in aquaculture, the increasing cost, and scarcity of fishmeal plus other ingredients rich in protein of both plant and animal sources have resulted in fish nutritionist seeking alternative sources from non-conventional feed ingredients such as macrophytes. the potential of fresh water vascular plants as feedstuffs has been emphasized [11]. a few species appear to be suitable raw materials for leaf protein extraction, which can be used as human and non-ruminant animal feed in the tropics. the use of various ingredient of plant origin with nutritive value in order to reduce variable cost of production has been documented. these includes plantain peel [12], calabash seed [13-15], moringa oleifera seed meal [16], calabash seed meal [17]. however, there is dearth of information on the nutritional values of these plants when subjected to various drying methods. therefore, the objective of this study is to assess the potential of some aquatic macrophytes subjected to different drying methods as valuable ingredient in fish diets. 2. materials and methods 2.1. study area the study was carried – out at the fish hatchery and agric chemical laboratory of the faculty of agriculture, usmanu danfodiyo university sokoto, nigeria. 2.2. samples collection all samples of the study was collected from river rima in kwalkwalawa village, along usmanu danfodiyo university road (main campus) sokoto. all the samples were collected and analyzed from october, 2012 to december, 2012. 2.3. processing of the samples weighed samples (600g) each of water hyacinth, water lily and water primrose were collected in triplicate and subjected to the different methods namely: air, sun and oven drying. 2.4. air drying samples samples were spread under shade in the fish hatchery. temperatures of the environment were recorded four times daily at 6.00am, 12.00pm, 4.00pm and 10.00pm respectively with mercury glass thermometer. 2.5. sun drying of samples samples were dried under the sun on concrete floor guarded against high wind. sun dried was carried out for 8 hours daily during which ambient temperature was recorded at 10.00am, 12.00pm, 4.00pm and 6.00pm, using mercury glass thermometer. 2.6. oven drying of samples samples were oven dried at the agric chemical laboratory using uniscopesm9053 laboratory oven at 65 0 c for forty eight hours. 2.7. assays moisture content was determined by drying in an oven 100-105 o c to constant weight [18]. the crude protein content was evaluated by digestion of the sample, nitrogen determination by a spectrophotometer method as described by devani, et al. [19] and the crude protein was obtained by multiplying the quantity of nitrogen by the coefficient 6.25. total lipids were determined by continuous extraction in a sox let apparatus for 8 hours using hexane as solvent. ashing was carried out by incinerating in a furnace at 550 0 c. crude fiber was determined by sequential hot digestion of the defatted sample with dilute acid and alkaline. total carbohydrate was determined by difference (100moisture, crude protein, ash, crude fiber and crude lipid) [18]. 2.8. statistical analysis data obtained were subjected to analysis of variance (anova) in a completely randomized design (crd) and, treatment means were separated for significant differences using duncan’s multiple range test [20]. the analyses were carried out using the computer software statistical package for the social sciences version 9.0 for windows [21] 3. results the proximate composition of water hyacinth (eichornia crassipes), water lily (nympae lotus) and water primrose (ludwigia hyssopfislia) subjected to various drying methods (air dried, ovum dried and sun dried) are shown in table 1. there were no significant differences (p> 0.05) between the moisture content of the air dried samples between the treatments. however, other methods (ovum dried and sun dried) differ significantly (p< 0.05). similarly, ash content follows the same trend, with higher values recorded in ovum dried method. the highest crude agriculture and food sciences research, 2014, 1(1): 11-14 13 protein was recorded in ovum dried method with no significant difference (p> 0.05) between the treatments; similarly the crude protein content of air dried did not differ significantly between water hyacinth and water lily but differ significantly (p< 0.05) with water primrose. water hyacinth recorded highest value of fiber content (5.2±0.12) in ovum dried method with no significant difference between the treatments; however other methods exhibit a significant difference. the highest value of crude lipid (4.51 ± 0.01) was recorded in ovum dried method for water primrose, however the value did not differ significantly (p> 0.05) with those of water lily but other methods differ significantly between the treatments. higher energy values were recorded in the air dried method for water primrose with no significant difference (p< 0.05) from sun dried method employed for water hyacinth and water lily respectively. table 2 shows the percentage composition of the whole plants used for the analysis. 4. discussion the highest moisture content was recorded air dried samples in all the treatments. this observation was in line with boyd [11]who reported that water hyacinth (eichornia crassiopes) when dried contain as much as 7% moisture, the author further stated that the plant contain higher moisture and fiber content. the higher moisture contents in air dried samples in all treatments could have been attributed to the temperature and relative humidity of the air and the duration of the dying. the moisture content of water lily (nymphae lotus) reported in the present study were similar to those of cratera religiosa reported in hassan, et al. [22]. lowest moisture content was recorded water primrose (ludwigia hyssopfislia) subjected to ovum drying method. similarly, the ash content reported in the study of the aquatic macrophytes with response to the dying methods employed where in conformity with boyd [11] and this therefore is indicating that the aquatic could beneficially serve as good sources of minerals for aquaculture. the crude protein contents reported in the present study for (eichornia crassiopes) is higher in all the drying methods than those reported in kusemiji and akingboju [23], this could have been resulted to the soil fertility of habitat in which the plant grow. the crude protein content for nympheae lotus in oven dried method was higher than the crude protein of the other drying methods, however the were higher than those reported in anjana and matai [24], lower than obtained in mohammed, et al. [9]. the reason this variation could have been influenced by processing method and techniques employed in the processes. oven dried method adopted for ludwigia hyssopfislia recorded the highest crude protein which was significantly different from the other dried product, this could be attributed to the combination of moderate temperature and duration of drying. a moderate temperature over short period of time result in considerable retention of nutrient such as vitamins and certain minerals like calcium, iron and zinc. the fiber contents reported in the present study with all the aquatic plants subjected the various drying method were within the 5% base line fiber content for monogastic animals including fish [25]. however, the fiber content reported in the study with regard to nympheae lotus was lower than reported in [9, 24]. higher crude lipid were recorded in ludwigia hyssopfislia exposed to various drying method, the values obtained than those reported in the leaves of eichornia crassiopes [26], also higher than those reported for the other plants in the present study. carbohydrate levels reported in the present finding was relative higher in all the macrophytes subjected to various drying method, this is a clear trend that the plants could serve betterment as energy sources. 5. conclusion the finding of the is revealing that water hyacinth (eichornia crassiopes), water lily (nymphae lotus) and water primrose (ludwigia hyssopfislia) could serve as good source of carbohydrate rather than protein, the processing methods were found to have effect on moisture, ash, crude protein, crude fiber, crude lipoid and carbohydrate in all the three macrophytes but protein retention was higher in oven dried samples. the study recommends oven dried methods in all the samples if it is economically justifiable, further studies should be carried to assess the potential of these macrophytes subjected to various drying method highlighted using cultured fish species. table-1. proximate composition of aquatic macrophytes on different drying methods (%) samples drying methods moisture ash crude protein crude fiber crude lipid nfe water hyacinth eichornia crassiopes air dried 7.05±0.03 a 14.33±0.17 b 12.36±0.02 b 4.50±0.29 b 2.01±0.06 b 60.06±0.01 b oven dried 6.91±0.01 b 15.37±0.13 a 12.52±0.02 a 5.20±0.12 a 2.50±0.03 b 58.05±0.02 c sun dried 6.60±0.06 c 14.47±0.03 b 11.08±0.01 c 4.25±0.01 b 2.52±0.02 a 61.00±0.50 b water lily nymphae lotus air dried 6.32±0.06 a 21.35±0.17 b 8.55±0.25 b 4.57±0.09 2.89±0.06 c 55.46±0.14 c oven dried 6.25±0.03 b 23.42±0.02 a 11.76±0.02 c 4.97±0.03 3.95±0.03 a 50.03±0.48 c sun dried 6.39±0.06 a 19.90±0.10 c 7.57±0.32 c 4.99±0.07 3.56±0.32 b 57.86±0.89 a water primrose ludwigia hyssopfislia air dried 6.20±0.03 a 8.22±0.15 b 4.84±0.60 c 2.52±0.01 4.03±0.03 b 74.02±0.11 a oven dried 5.96±0.02 b 3.34±0.17 b 15.13±0.01 a 3.20±0.11 a 4.51±0.01 a 67.94±0.31 b sun dried 6.12±0.06 a 8.51±0.01 a 11.64±0.01 b 3.10±0.01 a 4.49±0.01 b 66.29±0.04 b means in rows with the same letters are not significantly different (p >0.05) agriculture and food sciences research, 2014, 1(1): 11-14 14 table-2. percentage composition of whole plants part aquatic macrophytes components weight (g) composition (%) water hyacinth eichornia crassiopes leaves 600 16.01 stem 3081.0 82.20 root 67.03 1.79 total 3748.03 100 water lily nymphae lotus leaves 600 55.95 stem 290.886 27.12 root 181.609 16.93 total 1072.495 100 water primrose ludwigia hyssopfislia leaves 600 64.75 stem 326.648 35.25 root total 926.648 100 references [1] y. c. shang, "the role of aquaculture in the world fisheries," 1992. [2] s. craigs and i. a. helfrich, "understanding fish nutrition, feeds and feeding," virginia co -operation extension. publication no. 420-256, 2002. [3] a. a. eyo, e. a. falaye, and o. a. adetunji, "response of genetically improved hetrobranchus longifilis juveniles to different diets containing banished meal and extruded soybean meal," journal of applied science and environmental management, vol. 8, pp. 2933, 2004. [4] a. g. j. talon, "feed ingredient for warm water fish, fish meal and other processed feedstuff," fao fisheries circular no. 8856. fao rome italy, 1993. [5] d. m. s. el-saidy and m. m. a. gaber, "use of cottonseed meal as protein source for nile tilapia oreochromis niloticus," aquaculture, vol. 53, pp. 24-252, 2004. [6] c. lim and w. domonary, "evaluation of soya bean meal as a replacement marine animal protein in the diets of shrimps (pinnaeus vennamei)," aquaculture, vol. 87, pp. 53-65, 1990. [7] t. watanabe and j. pongmaneerat, "quality evaluation of some animal protein sources for rainbow trout (orcorrhynchus mykiss)," nippon suisan gakkaish, vol. 57, pp. 495-501. available www.nonnaturespecies.org, 1991. [8] i. g. mbagwu and h. j. adeniji, "the nutritional content of duck weed (lemma pausicustata) in the kainji lake area," aquatic botany, vol. 29, pp. 25-36, 1998. [9] h. a. mohammed, r. o. awodoyin, f. daddy, and g. o. adesina, "ethnobotyany of water lily (nympae lotus) among the riverrne community in nigeria, a case study of kainji lake basin," journal of biological and environment, vol. 5, pp. 52-55, 2008. [10] e. o. ita, "aquatic plants and wet land resources of nigeria," cifa occational paper, no. 21 fao rome, 1993. [11] c. e. boyd, "the nutritive value of three species of water weeds," economy botany, vol. 23, pp. 123-127, 1969. 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[19] m. b. devani, g. i. shishoo, s. a. shah, and b. n. suhagia, "microchemical methods. spectrophometric methods for microdetermination of nitrogen in kjeldahl digest," journal of association of analytical chemist, vol. 72, pp. 953-956, 1989. [20] r. g. d. steel and j. h. torrie, principles and procedures of statistics. a biometrical approach., 2nd ed., pp: 633,1980. [21] s.p.s.s 16.0, "statistical package for social package sciences." available http://www.winwrap.com, 2007. [22] i. g. hassan, j. k. umar, s. m. dangoggo, and m. j. ladan, "nutritional composition of ipomoea aquatic (water spinach) leaves," journal of applied science, vol. 7, pp. 804-807, 2007. [23] k. kusemiji and d. s. akingboju, "comparative growth of sarotheredon melanatheron (puppell) on formulated feed and water hyacinth diets," op. cit., pp. 196-20, 1988. [24] b. anjana and s. matai, "composition of indian aquatic plant in relation to utilization as animal forage," journal of aquatic plant management, vol. 28, pp. 69-73, 1990. [25] nrc, nutrients requirement of fish. committee on animal nutrition, national research council. washinton d.c.: national academy press. pp:114, 1993. [26] o. booy, m. wade, and v. white, "creeping water primrose," 2009. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. http://www.nonnaturespecies.org/ http://www.winwrap.com,/ 79 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 79-84, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.79.84 © 2019 by the authors; licensee asian online journal publishing group evaluation of extra-early maturing maize genotypes for grain yield and stability in the groundnut basin agro-ecological zone of senegal georgina lala ehemba1 yedomon ange bovys zoclanclounon2 moustapha gueye3 cyril diatta4 martial barnabe etienne diatta5 cheikh thiaw6 alfred kouly tine7 alpha bocar baldé8 ghislain kanfany9 ( corresponding author) 1national agricultural research center of bambey, senegalese institute of agricultural research, bp 211 bambey, senegal; regional study center for the improvement of drought adaptation, bp 3320, khombole road, thiès, senegal. 2regional study center for drought adaptation improvement, bp 3320, khombole road, thiès, senegal. 3,4,5,6,7,8national agricultural research center of bambey, senegalese institute of agricultural research, bp 211 bambey, senegal. 9national agricultural research center of bambey, senegalese institute of agricultural research, bp 211 bambey, senegal; west african center for crop improvement (wacci), university of ghana. abstract maize is a highly valuable staple food crop in sub-saharan africa regions. its production is hampered by both biotic and abiotic stress. early-maturing genetic resources appear as an excellent choice to manage adverse climatic change’s effects. this study aimed to find the best extra-early maize genotypes based on genotype by environment interactions in the groundnut agro-ecological zone of senegal. the trials were conducted in paoskoto, ndiedieng, nioro and keur sene during the rainy seasons of 2016 and 2017. six extra-early maize genotypes were evaluated following a randomized complete block design with three replications. data on grain yield were recorded and analysed using additive main effect and multiplicative interaction and genotype plus genotype by environment biplot methods. analysis of variance revealed a significant (p < 0.01) genotype by environment effect on grain yield. the sum of squares variation was higher (77.2%) explained by the environment effect. the genotype 2008-tzee-w-str showed the highest yield performance (3299 kg ha-1) and lowest yield stability index (3). this genotype can be suggested to farmers for its cultivation and a good candidate for early maturing maize breeding program in senegal. keywords: ammi, g x e, rainfed, early-maturation, yield, senegal, zea mays. citation | georgina lala ehemba; yedomon ange bovys zoclanclounon; moustapha gueye; cyril diatta; martial barnabe etienne diatta; cheikh thiaw; alfred kouly tine; alpha bocar baldé; ghislain kanfany (2019). evaluation of extra-early maturing maize genotypes for grain yield and stability in the groundnut basin agro-ecological zone of senegal. agriculture and food sciences research, 6(1): 79-84. history: received: 14 february 2019 revised: 18 march 2019 accepted: 22 april 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. the authors thank gratefully farmers for field facilities and their collaboration. funding: this study was supported by the west africa agricultural productivity program (waap). competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.79.84&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.79.84&domain=pdf&date_stamp=2017-01-14 https://orcid.org/0000-0002-2654-1417 https://orcid.org/0000-0003-2781-0778 https://orcid.org/0000-0001-6014-7846 https://orcid.org/0000-0002-4194-0897 https://orcid.org/0000-0001-5270-9423 https://orcid.org/0000-0003-1423-6375 https://orcid.org/0000-0002-5249-4677 https://orcid.org/0000-0002-3834-4584 https://orcid.org/0000-0002-0860-0532 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/122 agriculture and food sciences research, 2019, 6(1): 79-84 80 © 2019 by the authors; licensee asian online journal publishing group contents 1. introduction ...................................................................................................................................................................................... 80 2. materials and methods ................................................................................................................................................................... 80 3. results and discussion ................................................................................................................................................................... 81 4. conclusion ......................................................................................................................................................................................... 83 references .............................................................................................................................................................................................. 83 contribution of this paper to the literature this study contributes to existing literature by finding the best extra-early maize genotypes based on genotype by environment interactions in the groundnut agro-ecological zone of senegal. 1. introduction maize is one of the most important cereal grown in sub-sahara african countries, feeding more than 300 millions of people [1]. its grains are staple food crop and livestock feed [2]. in senegal, due to the rainfall distribution, maize production is restricted in the southern part of the groundnut basin, casamance and senegal oriental agro-ecological zones where annual rainfall may reach more than 800 mm per year. drought and low nitrogen soil, as well as parasitic weeds, seriously affect cereal production, particularly maize ones [3]. the invasion of the parasitic weed striga hermontica (del.) benth coupled with severe drought conditions can cause 50% of yield loss [4]. striga alfera willd., known as the pink-flowered was also reported destructive for maize production in senegal parker [5]. however, okuyama, et al. [6] reported that the potential of maize yield in senegal may be increased in the range of 6% on the basis of the current farming practices. in order to address drought stress issue in maize production in west african savannah zones, development of extra-early-maturing varieties have been implemented by the joint effort of the international institute of tropical agriculture and national agricultural research systems [4]. the basis of early flowering strategy as a drought escape mechanism enables the plant to complete its life cycle before drought stress exacerbation [7]. in addition, sowing date flexibility regarding non-desirable climatic events (for instance short period of rainfall and delay of the rains) is an important advantage of extra-early maturing varieties. thus, environmental conditions variation can influence maize growth and yield [8]. variation in phenotypic expression in a target environment for the same genotypes can be related to the genotype, the environment and genotype by environment interaction [9]. thereby, multi-environment trials constitute a relevant strategy for accurate identification of high-yielding and stable genotype [10]. yield stability and adaptation spectrum of genotypes over the years, in fluctuating environments, have become an important criteria for plant breeders [11]. numerous models were proposed to investigate genotype by environment interaction patterns, including additive main effect and multiplicative interaction (ammi) and genotype plus genotype by environment (gge) biplot. the ammi model deals with the traditional analysis of variance for additive main effects in combination with principal component analysis [12]; [13]. this model was widely used in the identification of superior genotypes of barley [14] cassava [15] wheat [16] and rice [17]. the gge biplot method proposed by yan and tinker [18] unravels both genotype and genotype by environment interactions based on gabriel [19] biplot methodology. gge biplot displays a powerful graphic on genotype stability and performance, genotype by environment interaction and mega-environment identification through “which on where” pattern graph [20]. this two-year study conducted in four locations in the groundnut basin agro-ecological of senegal, aimed to identify high yielding and stable genotypes using both ammi and gge biplot methods. 2. materials and methods 2.1. experimental sites experimentation was carried out during the rainy seasons of 2016 and 2017 in four sites (nioro, ndiedieng, paoskoto and keur sene) located in the “groundut basin” of senegal. details of testing environments are given in table 1. table-1. information on testing environments during rainy seasons 2016 and 2017. site soil type geographical position cropping season total rainfall (mm) temperature (°c) latitude (n) longitude (w) altitude (m.a.s.l) minimum maximum nioro sandy clay 13°43'48" 15°46'48" 28 2016 663 23.9 31.4 2017 1103 23.0 31.6 ndiedieng, sandy clay 13°34'12" 16°05'24" 18 2016 377 22.6 31.8 2017 257 24.5 35.7 paoskoto sandy clay 13°46'48" 15°47'24" 28 2016 1269 22.3 35.2 2017 1307 23.6 36.0 keur sene sandy clay 13°50'60" 16°02'24" 25 2016 330 23.3 35.6 2017 360 24.3 37.9 source: national agency of civil aviation and meteorology of senegal. 2.2. plant materials and test-environments six extra-early maturing maize genotypes table 2 were evaluated in a randomized complete block design with three replications in each of the environments. sowing was done with a spacing of 0.75 m between rows and 0.25 m between plants within a row in plots of 6 × 5.25 m2 area. each plot contained 8 rows. a basal 15n-15p-15k fertilizer at a rate of 200 kg ha-1 was applied before sowing. recommended cultural practices were applied for crop management. at the maturity stage, grains data were recorded and converted to kg ha-1 using the plot size as a factor. agriculture and food sciences research, 2019, 6(1): 79-84 81 © 2019 by the authors; licensee asian online journal publishing group table-2. information on extra-early maize genotypes used in this study. genotypes code pedigree/original name origin type g1 2008-tzee-w-str iita-ibadan opv g2 2009-tzee-or1-str iita-ibadan opv g3 tzee-wpop-strc5 iita-ibadan opv g4 tzee-y isra-senegal opv g5 tzee-w isra-senegal opv note: iita: international institute of tropical agriculture. opv: open pollinated variety. isra: institut sénégalais de recherches agricoles. 2.3. ammi model analysis of variance was performed using ammi model implemented in statistical software genstat [21]. assuming 𝜇 is the grand mean of extra early maize genotypes yield, 𝛼𝑔 is the genotype deviation of the grand mean, 𝛽𝑒 is the environment deviation, 𝜆𝑛 is singular value for interaction principal component (ipc) n and 𝜆𝑛 2 is the corresponding eigenvalue, 𝛾𝑔𝑛 is the eigenvector for genotype g and component n, 𝛿𝑒𝑛 is the eigenvector for environment e, 𝜌𝑒 is the residual, 𝜅𝑟(𝑒) is the block effect for replication r within environment e and 휀𝑔𝑒𝑟 is the error, the yield of genotype g in environment e for replication r (𝑌𝑔𝑒𝑟) is expressed by gauch [22] model: yger= μ + αg + β e + ∑ λn γ gn δen n + ρ e + κr(e)+ εger based on the relative contributions of the ipc 1 and 2 axis scores to genotype by environment interaction effect, the ammi stability value (asv) was computed using the following formula described by danquah, et al. [15]: asv = √[ ipc1sum of squares ipc2sum of squares (ipc1score)] 2 + (ipc2score) 2 using based-yield genotypes ranking and ranking based on ammi stability value, yield stability index was calculated as follows: yger= μ + αg + β e + ∑ λn γ gn δen n + ρ e + κr(e)+ εger lower is ammi stability value and ranking based on yield values, more stable and high yielding is the genotype [23]. 2.4. gge biplot analysis data recorded on grain yield were arranged in site-maize genotypes two-way table containing mean yields. biplot graphs were plotted using statistical software genstat 18 [21] following the gge biplot model defined by yan, et al. [24] as follows: y ij = ∑ λnξin η jn r n=1 where r is the number of principal components required to approximate the original data, λn is the singular value of principal components, ξin and ξjn are the ith genotype score and the jth environment score for principal components respectively and yij the response yield according to ith genotype in jth environment. 3. results and discussion 3.1. performance of the tested extra-early maize in different locations during the rainy season of 2016, g3 showed the highest yield (3893 kg.ha-1) at paoskoto, followed by g2 (3846 kg ha-1) at keur sene and g5 (3253 kg ha-1) at paoskoto table 3. table-3. mean grain yield (kg. ha-1) of maize genotypes in different environments during 2016 and 2017 crop seasons. year site g1 g2 g3 g4 g5 mean minimum maximum 2016 nioro 2959 2951 2524 2809 2684 2785 1920 3680 ndiedieng 2365 3484 3591 2187 1742 2674 747 4853 paoskoto 2951 2258 3893 2382 3253 2948 1173 4053 keur sene 3715 3846 3589 2382 1742 3788 2954 4652 2017 nioro 3790 3528 3259 2829 4206 3522 1723 4794 ndiedieng 3790 2957 3259 2952 2994 2998 2097 4085 paoskoto 2400 925 1849 2382 978 1387 587 2667 keur sene 4886 4737 4205 3988 4574 4478 3514 5333 source: field data. yields ranged from 747 kg ha-1 to 4853 kg ha-1 across environments during 2016 cropping season. in 2017, g1 and g2 showed the highest yields in keur sene with 4,886 kg ha-1 and 4737 kg ha-1, respectively. 3.2. additive main effect and multiplicative interaction analysis the ammi analysis of variance revealed a significant effect of genotype, environment and genotype by environment on yield variation table 4. agriculture and food sciences research, 2019, 6(1): 79-84 82 © 2019 by the authors; licensee asian online journal publishing group table-4. analysis of variance for ammi2 model of five extra early maize genotypes in eight environments. source of variation df ss ms tve (%) g × e (%) total 119 145733332 1224650 treatments 39 112590559 2886937*** genotype (g) 4 4941432 1235358** 4.38 environment (e) 7 86901293 12414470*** 77.18 g × e interaction 28 20747834 740994** 18.42 ipc1 10 9279579 927958** 44.72 ipc2 8 6508658 813582* 31.37 residual 10 49599596 495960 blocks within environments 16 11888665 743042* error 64 21254108 332095 note: *significant at 0.05 probability level. **significant at 0.01 probability level. ***significant at 0.001 1evel of probability. ns: non-significant. df: degree of freedom. ss: sum of square. ms: mean of square. tve (%): percentage relative to total sum of square. ipc: interaction principal component. g × e (%): percentage of sum of square variation explained by genotype by environment effect. environment factor explained the highest (77.2%) part of total variation followed by genotype by environment interaction (18.4%) and genotype (4.4%). the partitioning of the sum of square variances revealed a predominant effect of environment. this suggests that the environment influences range was wider than genotype. as found in several studies [25]; [26] the variability observed in environmental parameters such as rainfall, temperature and soil texture can be related to the strong environmental effect. ipc1 and ipc2 represent 44.7% and 31.4% of genotype by environment variation, respectively. the ipc 1 effect was highly greater (10 times) than the effect of the genotype and contributed to more than 40% of ge interaction variation. the ammi ipc1 and ipc2 scores were respectively positive and negative, reflecting location-year crossover pattern in genotype by environment interactions [27]. table-5. ranking of the tested genotypes based on asv and ysi. genotype mean rank (µ) ipc1 ipc2 asv asv rank (η) ysi (µ + η) ysi rank g1 3299 1 8.93 -12.45 17,81 2 3 1 g2 3086 3 -10.47 27.89 31,63 3 6 2ex g3 3209 2 -29.12 -19.04 45,68 5 7 4 g4 2704 5 4.13 11.03 12,50 1 6 2ex g5 3065 4 26.53 -7.42 38,54 4 8 5 source: field data. based on ammi stability values (asv), g4 appeared as the most stable genotype across environments. however, yield stability index (ysi) revealed that g1 was the best genotype followed by g2 and g3 table 5. 3.3. genotype plus genotype by environment biplot analysis the genotype by environment patterns were graphically presented in figure 1 and figure 2 using gge biplot model. principal components 1 and 2 encompass 40.75% and 25.10% of total variation respectively. the figure 1 represents the average-environment coordination (aec) view based on yield performance and stability. figure-1. the average-environment coordination view showing mean performance and stability of five extra early genotypes tested in eight environments. the genotypes: g1(2008-tzee-w-str), g2(2009tzee-or1-str), g3 (tzee-wpop-strc5), g4(tzee-y), g5(tzee-w). the years: 16 (2016), 17(2017). the locations: ks (keur sene), pa (paoskoto), nd (ndiedieng), ni (nioro). source: field data. agriculture and food sciences research, 2019, 6(1): 79-84 83 © 2019 by the authors; licensee asian online journal publishing group figure-2. the which-won-where gge biplot showing five extra early maize genotypes behaviour in eight environments. the genotypes: g1(2008-tzee-w-str), g2(2009-tzee-or1-str), g3(tzee-wpop-strc5), g4(tzee-y), g5(tzee-w). the years: 16 (2016), 17(2017). the locations: ks (keur sene), pa(paoskoto), nd (ndiedieng), ni (nioro). source: field data. the aec arrow indicates the highest genotype based on yield performance. the circle represents the ideal genotype, which combines high yield and stability. according to figure 1, the genotype g1 was the best genotype followed by g5. in contrast, g4 is the most unstable genotype across all environments. the which-won-where polygon view figure 2 highlighted the genotype by environment interaction pattern of this study. the genotypes located at the vertex of the polygon showed either best or poor performance in a related environment. the genotypes g1 and g5 had a higher grain yield than average in the environments ni17, ks17, nd17, pa17, ks16 and ni16. g3 was the best genotype in the environments pa16 and nd16. in contrast, g2 and g4 yields were lower than average across the tested environments. the orthogonal lines from the origin of biplot which cut the sides of the polygon allow the comparison between two adjacent genotypes in relationship with their respective environments. thus, g1 performed better in ni17, ks17, nd17 and pa17, whereas g3 was better in nd16 and pa16. the orthogonal lines in the polygon also revealed the presence of location-year crossover patterns. the environment couples pa16-pa17 and nd16-nd17 were located on opposite sides of orthogonal lines, indicating a location-year effect. gge biplot polygon view supports this crossover presence, denoting of adaptation of maize genotypes according to a target environment. various studies [28-30], performed on a wide range of crops, successfully showed the efficacy of ammi and gge biplot methods to figure out genotype adaptation patterns. 4. conclusion the present study demonstrated a high environment effect in ge variation despite the fact that the environments were located in the same agro-ecological zone. both ammi and gge biplot analysis showed that the genotype 2008-tzee-w-str had the highest grain yield and was more stable across environments. it can be recommended for cultivation in the groundnut basin of senegal and used in maize breeding program as a gene donor regarding yield and stability. references [1] y. beyene, k. semagn, j. crossa, s. mugo, g. n. atlin, a. tarekegne, b. meisel, p. sehabiague, b. s. vivek, and s. oikeh, "improving maize grain yield under drought stress and non-stress environments in sub-saharan africa using marker-assisted recurrent selection," crop science, vol. 56, pp. 344-353, 2016. available at: https://doi.org/10.2135/cropsci2015.02.0135. 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[30] e. s. fabio, t. a. volk, r. o. miller, m. j. serapiglia, h. g. gauch, k. c. van rees, r. d. hangs, b. y. amichev, y. a. kuzovkina, and m. labrecque, "genotype× environment interaction analysis of north american shrub willow yield trials confirms superior performance of triploid hybrids," gcb bioenergy, vol. 9, pp. 445-459, 2017. available at: https://doi.org/10.1111/gcbb.12344. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 47 agriculture and food sciences research vol. 5, no. 1, 47-51, 2018 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2018.51.47.51 evaluation of some plant extracts and powders in control of bean damping-off by sclerotinia sclerotiorum zahra ibrahim el-gali dept. of plant protection, faculty of agriculture, omer almuhktar university. el-beida, libya abstract the antifungal effect of three botanical powdered plants and their aqueous extracts against damping-off disease incidence of bean were evaluated under laboratory and field conditions. also this study investigated the phytochemical profile of these plants. powder of peganum harmala, urtica dioica and helichrysum stoechas leaves were used in the present work. the botanical plant extracts were applied as seed dressing, while the powders were applied as seed coating in vitro or were added to the cultivation soil in vivo before or after inoculation with sclerotinia sclerotiorum then sowing. all applied treatments reduced dead seedlings incidence comparing with untreated control. higher significant reduction in disease incidence was observed for h. stoechas treatment than that of the others plants. it is interesting to note that botanical plants extract gave a similar effect to the powder in reducing disease incidence either at preor post-emergence stages of bean growth. the content of the active biological compounds different between all plants. keywords: plant extract, powder, phytochemical, damping-off, sclerotinia sclerotiorum. citation | zahra ibrahim el-gali (2018). evaluation of some plant extracts and powders in control of bean damping-off by sclerotinia sclerotiorum. agriculture and food sciences research, 5(1): 47-51. history: received: 13 march 2018 revised: 7 april 2018 accepted: 9 april 2018 published: 12 april 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 48 2. materials and methods ................................................................................................................................................................... 48 3. results ................................................................................................................................................................................................ 49 4. discussion .......................................................................................................................................................................................... 50 5. conclusion ......................................................................................................................................................................................... 50 references .............................................................................................................................................................................................. 50 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.51.47.51&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=zahra%20ibrahim%20el-gali agriculture and food sciences research, 2018, 5(1): 47-51 48 1. introduction dry white bean (phaseolus vulgares l.) is one of the major protein yielding plant found not only in libya but throughout the world due to the presence of high calorie protein. sclerotinia sclerotiorum is an important fungal pathogen with cosmopolitan distribution that has a wide host range [1-3]. damping-off and root rot disease caused by s. sclerotiorum is a serious and persistent diseases problem of bean plants during growing season. control of such diseases mainly depend on fungicidal treatments [4]. however, fungicidal applications cause hazards to human health and increase environmental pollution. therefore, alternative treatments for control of plant diseases are needed [5]. botanical plants made into powder were widely used by ancient civilizations to improve the palatability of foods and beverages as well as for their preserving characteristics. various plant products like gum, oil, resins etc. are used as fungicidal agents [6-8]. the biological control of plant diseases may have minimum adverse effect on physiological processes of plant and less environmental hazards [9]. the objective of the research work performed was to study the antifungal activity of some botanical plant extracts and powders against damping-off disease incidence of bean in vivo and in vitro conditions. 2. materials and methods 2.1. source of pathogen s. sclerotiorum was isolated from dry bean seeds and identified in laboratory of phytopathology, faculty of agriculture, omer almukhtar university, libya. 2.2. plant materials 2.2.1. preparation of powders three different plant species, peganum harmala, urtica dioica and helichrysum stoechas known for their medicinal value in traditional medicine and generally available in wild in al-jabal al-akhdaer region were selected for the study (fig. 1). the apparently healthy leaves of these plants were collected in flowering time, then washed thoroughly with running tap water to remove the dust and dried. the dried leaves, were crushed into powder with the help of grinder. the powders were passed through sieve to get fine powder. the powders of different plant were stored in polythene bags for the study. 2.2.2. preparation of aqueous extracts fifty grams of each of above mentioned botanical plant materials in powder form was homogenized by laboratory blender in 50 ml distilled water for 10 min, then left in dark place for 24 h for tissue maceration. the extracts were filtered through thin cheesecloth sheets. the final extracts/supernatant were filtered through whatman no. 1 filter paper and sterilized by zeits filter (0.24 μl), then collected separately in sterile dark glass tight bottles. the obtained extracts served as the crude extract (100 % concentration) and kept in a refrigerator at 4°±1 until use. fig-1. types of used plants (source: the photograph was took by author's camera) 2.3. application of plant materials for damping-off control 2.3.1. laboratory experiment the plant materials were evaluated for bean damping-off control by using different methods of application. the wet method depends on soaking the bean seeds in the plant extract for 12, 24 and 36h., then left for 1h to dry, and in pure water for check treatment. the dry method depends on mixing the seed with the formula, carboxy methyl cellulose (0.1g cmc in 10ml water) as adhesive material. and were rolled on plant powder to help it for adhesion. for studying the disease in lab., petri plates with potato sucrose agar medium were inoculated with s. sclerotiorum and incubated at 23±2°c. when the entire plate was covered with the white mycelium, the mycelial mat was covered with sterilized soil, 1.0g thick. soaked and coated seeds were germinated in plates and incubated at room temperature, four replicates were used for each treatment. in control plates, the seeds without treatment were planted in plates medium. 2.3.2. field experiment under field conditions, plastic pots (15 cm diam.) were used. pots were sterilized by immersing in 5% formalin solution for 15 min., and left several days before being used, then filled with sterilized sand and clay soil (1:1 v/v). the soil had infestation of 5 fungus sclerotia and irrigated for one week before sowing. the seeds soaked were sown at rate of 5 seeds/ pot. the botanical plant powder tested was applied by two methods: a) soil mixed with powder at rate of 3g powder/ 300g and left two days before infestation with sclerotia and irrigated for one week. b) soil infested with sclerotia and irrigated for one week before powder added. surface sterilized seeds were planted at the rate of 5 seeds per pot in two applications. agriculture and food sciences research, 2018, 5(1): 47-51 49 2.4. disease assessments disease effects due to s. sclerotiorum in the different treatments were assessed at 10 days after planting as decayed seeds, death of seedlings (pre and post emergence) and seedling survival after 30 days. these parameters were calculated and expressed as percentages. 2.5. data analysis all experiments were conducted on the basis of completely randomized designs (crd). each treatment was replicated at least five times. for statistical analysis, data were subjected to the analysis of variance (anova) using co stat program. angular transformed values were used for data analysis. statistical comparisons among means were performed using duncan’s multiple range test (dmrt). fig-2. effect of plant extracts and powders on seed decay. (source: the photograph was took by author's camera) 2.6. phytochemical analysis phytochemical analysis of the extracts was done by following the methods described previously [10-14]. 3. results from observations in the test, after 10 days, all the seeds had not grown in the plate treated with p. harmala and in the control plates, and it appeared completely decay. the dressing with h. stoechas was protected the seed against fungus invasion. in the treatment plates, u. dioica powder was effect in reduce seed decay than plant extract, while extract and powder of h. stoechas was the best in control seed decay disease (fig. 2). the decrease in seeds rot was observed in seeds that were treated with extract or coated with powder of h. stoechas, at ratio of 20% and 6.7% compared with 100% in control treatment (table 1). h. stoechas was more effective than others plants against seeds decay. table-1. effect of seed treatment with plant preparation on the percentage of bean seed decay in vitro. treatments extract powder time (h) 12 24 36 p. harmala. 100 (100.0) a 100 (100.0) a 100 (100.0) a 93.3 (75.00) a u. dioica 86.7 (68.53) a 93.3 (75.00) a 100 (100.0) a 33.3 (35.24). b h. stoechas 20.0 (26.56) b 20.0 (26.56) b 20.0 (26.56) b 06.7 (15.00) b con. 93.3 (75.00) a 93.3 (75.00) a 100 (100.0) a 100 (100.0) a lsd at 5% ex.: 15.77 t: ns ex × t: ns 28.95 note: each value is mean of four replicates percentage of infection transformed data by the arcsine square before analysis. ns: non-significant results in table (2) indicate that the treatment with h. stoechas was significantly reduced the seed decay disease of bean plants comparing with treatments with p. harmala and u. dioica and untreated check control treatment. it recorded disease incidence (33.3, 28 and 4%) and seedling healthy (66.7, 72. 60%) in both extract and powder compared with 86.7% dead seedlings and 13.3% seedlings healthy in control treatment. in terms of effect of extracts and powders on disease in field the study recorded occurrence the reduction in seedlings death and increased seedlings survival after seeds treatment with extract or soil amended with powder before or after inoculation with fungal sclerotial. the h. stoechas weed was more effective followed by u. dioica. agriculture and food sciences research, 2018, 5(1): 47-51 50 table-2. effect of dressing bean seeds with plant preparation on percentage of bean damping-off disease treatment formula tested suspension powder before inoculation after inoculation dead seedlings seedlings healthy dead seedlings seedlings healthy dead seedlings seedlings healthy p. harmala. 60.0 (50.77) ab 40.0 (39.23) ab 48.0 (43.85) 52.0 (46.15) 88.0 (69.73) 12.0 (20.27) u. dioica 73.3 (58.89) ab 26.7 (31.11) bc 40.0 (39.23) 60.0 (50.77) 36.0 (36.87) 64.0 (53.13) h. stoechas 33.3 (35.21) b 66.7 (54.76) a 28.0 (31.95) 72.0 (58.05) 40.0 (39.23) 60.0 (50.77) con. 86.7 (68.70) a 13.3 (31.39) c 86.7 (68.70) 13.3 (31.39) 86.7 (68.70) 13.3 (31.39) lsd 5% 24.9 17.4 method: ns treatment: 16.79 m × t: ns method: ns treatment: 11.88 m × t: ns note: each value is mean of three replicates. percentage of infection transformed data by the arcsine square before analysis values followed by the same letter(s) are not significantly different at p= 0.05. ns: non-significant. phytochemical properties of extracts of p. harmala, u. dioica and h. stoechas are represented in table (3). flavonoids were not found in p. harmala and u. dioica but were found to be the major phytoconstituent along with glycosides, phenols, tannins, resins and terpenoids in h. stoechas. saponins were present as the major phytoconstituents irrespective of p. harmala and u. dioica . p. harmala extract was poorer in antifungal compounds. table-3. phytochemical constituents of aqueous extracts of different plants plants compound p. harmala. u. dioica h. stoechas alkaloids + anthraquinones flavonoids + glycosides + + phenols + + saponins + + tannins + + terpenoids + + phlobatanins resins + + note: +: found, -: not found 4. discussion naturally occurring products from plants have played an important role in the discovery of new therapeutic agents since ancient times. the use of plant extracts and phytochemicals, with known antimicrobial properties, may be of immense importance in therapeutic treatments. synthetic fungicides are currently used as the primary means for the control of plant diseases. however, the alternative control methods are needed because of the negative public perceptions about the use of synthetic chemicals, resistance to fungicides among fungal pathogens, and high development cost of new chemicals. the uses of plant derived products as diseases control agents have been studied, since they tend to have low mammalian toxicity, less environmental effects and wide public acceptance [5]. to develop environment-friendly alternatives to synthetic fungicides for the control of fungal plant diseases, the interest on essential oils and plant extracts has been increased [9]. the results obtained in the present work indicate that the powders were more effective than extracts. this results agreement with that obtained by elmougy and abdel-kader [15] and mokhtar, et al. [16]. h. stoechas extract or powder had antifungal activity against plant pathogenic fungus tested in the present study. numerous studies have been conducted in the past few years to prove such efficiency [17-20]. plant of the genes helichrysum are prolific producer secondery metabolites as antiviral, antifungal, antimicrobial and anti-inflammatory [20-23]. the efficiency of h. stoechas may be due to the high content of phenols, flavonoids, glycosides, tannins, terpenids and resins in leaves more than the others plants. so, it is interested to state that we can used h. stoechas extracts and/or powders less expensive for controlling plant diseases under field. 5. conclusion promising applicable technique could be suggested in the light of the results obtained. the use of soil drench treatment with botanical plants powder might be considered as safe, cheap and easily applied method for controlling such soil-borne plant pathogens as eco-friendly means, and considering the avoidance of environmental pollution and the side effect of pesticide references [1] m. malvárez, i. carbone, n. j. grünwald, k. v. subbarao, m. schafer, and l. m. kohn, "new populations of sclerotinia sclerotiorum from lettuce in california and peas and lentils in washington," phytopathology, vol. 97, pp. 470-483, 2007. view at google scholar | view at publisher [2] z. i. el-gali, "cultural, morphological and physiological studies on some isolates of sclerotinia sclerotiorum libyan," journal of plant protection research, vol. 1, pp. 11-16, 2010. https://scholar.google.com/scholar?hl=en&q=new%20populations%20of%20sclerotinia%20sclerotiorum%20from%20lettuce%20in%20california%20and%20peas%20and%20lentils%20in%20washington https://scholar.google.com/scholar?hl=en&q=new%20populations%20of%20sclerotinia%20sclerotiorum%20from%20lettuce%20in%20california%20and%20peas%20and%20lentils%20in%20washington http://dx.doi.org/10.1094/phyto-97-4-0470 agriculture and food sciences research, 2018, 5(1): 47-51 51 [3] m. m. abdel-kader, n. s. el-mougy, and s. m. lashin, "chemical and biological measures against sclerotinia spp. the causal of foliage blight disease of cucumber and pepper plants in egypta review," international journal of engineering & technology, vol. 3, pp. 302311, 2014. 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10.20448/journal.512.2019.61.41.49 © 2019 by the authors; licensee asian online journal publishing group production and adoption constraints of improved coffee varieties in jimma zone, southwest ethiopia samuel diro1 beza erko2 kalkidan fikirie3 ( corresponding author) 1holeta agricultural research center; agricultural economics research process, holeta; ethiopia. 2jimma agricultural research center; agricultural economics research process, jimma; ethiopia. 3melkassa agricultural research center, melkassa, ethiopia. abstract the study was aimed to identify major coffee production problems that affected coffee farmers’ production and productivity. it was conducted in four districts of jimma zone namely gera, manna, limu kosa and gomma. multistage sampling technique was employed to select the population for the study which involved both purposive and random sampling techniques. data was collected through structured questionnaire administered to sampled farmers from 285 coffee producing households. both descriptive and inferential statistics were used to analyze the gathered and cleaned data. kendall’s coefficient of concordance was used to rank the most important coffee production constraints. the study has identified three top constraints on coffee production. low and fluctuating coffee price, lack of coffee market information and lack of preferred coffee seed and seedling varieties were the major problems explored. strengthening cooperatives and unions, sustainable supply of improved coffee seeds and seedlings, and sustainable coffee seed system are suggested to tackle these coffee production problems. keywords: coffee, constraint, cooperative, kendall, market information, unions. citation | samuel diro; beza erko; kalkidan fikirie (2019). production and adoption constraints of improved coffee varieties in jimma zone, southwest ethiopia. agriculture and food sciences research, 6(1): 41-49. history: received: 14 january 2019 revised: 25 february 2019 accepted: 29 march 2019 published: 4 june 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: the authors would like to gratefully acknowledge jimma agricultural research center for the encouragement and support of various kinds. they are very grateful for district coffee experts of gomma, gera, limu kosa and manna for their dedication in facilitation and coordination. they also like to thank our collaborators including respondents (farmers), development agents and kebele officials for their respective contributions. funding: the authors would like to thank jimma agricultural research for the financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 42 2. materials and methods ................................................................................................................................................................... 42 3. result and discussion ..................................................................................................................................................................... 43 4. conclusions and recommendations ............................................................................................................................................. 48 references .............................................................................................................................................................................................. 49 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.41.49&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.41.49&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/1829 agriculture and food sciences research, 2019, 6(1): 41-49 42 © 2019 by the authors; licensee asian online journal publishing group 1. introduction world coffee production is increasing from year to year in spite of huge market volatility and environmental constraints [1]. arabica coffee has its center of origin in southwestern and southeastern ethiopia [2, 3]. ethiopia produces 9% of world’s arabica coffee with a value of 7.2 million 60kg bags annually. brazil and colombia ranked first and second with 57% and 22% of the total production, respectively [4]. in ethiopia, coffee is cultivated in four distinct production systems. garden coffee refers to the bulk of ethiopian coffee which represents more than 50% of total coffee coverage. it is grown by smallholder farmers intercropped with cereals, fruits and vegetables in the southern and eastern regions. the last production system is plantation which is grown on large state owned or commercial farms (represents 5% of production). the plantation production system that mainly observed in the southwestern part of the country under heavy shade and intensive management is based on improved varieties and agronomic practices [5, 6] . pokorná and smutka [7] reported that international coffee trade does not support the developing or least developed countries [8] pointed out lack of capital, poor extension service, poor market infrastructure, low and volatile coffee price, poor linkage to cooperatives as a major coffee production and marketing constraints. apart from these, disease and lack of pest control programs also results in decline in coffee production [9]. jimma agricultural research center has devoted considerable effort and resource, and developed several coffee technology packages. a number of coffee cultivars that combine high yield, disease resistance and quality character were developed by the center. in addition to these technologies, several recommendations have been developed on pest and disease management, agronomic and soil fertility management [10]. jimma zone is one of the major coffee producing areas of ethiopia. despite the dissemination of coffee improved technologies through different coffee extension approaches, utilization of the improved technologies is poor. intensive study on the constraining factors for coffee production has not been studied using different methodologies. this study was designed to explore constraints of coffee production. the result of the study could be helpful for coffee related biological and physiological researchers, academicians and policy makers. 1.1. objectives the overall objective of the study is identifying major constraints of coffee production in jimma zone. the specific objectives are: • to identify constraints that hinder coffee production activities on the study area. • to suggest policy options the way coffee production bottlenecks could be eliminated. 2. materials and methods 2.1. study area description the study was conducted in four districts of jimma zone namely gera, manna, limu kosa and gomma districts. gera district is found in the southwest of jimma zone. it shares border with chekorsa to the south east, with gomma to the east, with setema to the north east, with sigmo to the north west, with shebe sombo to the south and the south ethiopian people’s nations and nationalities to the west and south west. its absolute location ranges between 7027’ to 7055’ north latitude and 38001’ to 36024’ east longitude. tropical, semi tropical and temperate agro climates respectively shares 15%, 35% and 50% of the district’s total area. the mean annual temperature of the district ranges from 15-220c. the vast area of the district’s annual rainfall varies between 1300mm and 1700mm. coffee and teff are the major local cash crops in the district. limu kosa district extends between 7050’ to 8036’ north latitudes and 36044’ to 37029' east longitudes. it is bordered with limmu seka district in north and west shoa zone in north east, with tiro afeta in southeast, with manna and kersa districts in south, with buno bedele zone and gomma district in west. it is situated in the north central part of the zone. sub-tropical and temperate agro climates do respectively constitute 70% and 15% of the district’s areas. the remaining 15% of the district’s agro climate does have tropical climate. the mean annual temperature of the district ranges from 18-230c. the mean annual rainfall of the district ranges from 13002300mm. maize and coffee are the main crops grown in the district. gomma district extends between 7040’ to 8004’ north latitudes and 36017’ to 360 46' east longitudes. it is bordered with didesa district in north, with limmu kosa district in east, with manna district in southeast, with seka chekorsa in south and with gera district in west. it is situated in the central part of the zone. most part of the district belongs to subtropical and temperate agro climates. sub-tropical and temperate agro climates do respectively constitute 88% and 12% of the district’s area. the mean annual temperature of the district ranges between 150c and 220c. the vast area of the district’s annual rainfall varies between 1700mm and 2100 mm. maize and coffee are also the main crops grown in the district. manna district extends between 7038’ to 7054’ north latitudes and 36038’ to 36053' east longitudes. it is bordered with gomma and limmu kosa districts in north, with kersa district in east, with seka chekorsa district in south and with gomma district in west. it is also situated in the central part of the zone. sub-tropical and temperate agro climates do respectively constitute 80% and 20% of the district’s total areas. the vast part of the district does have with mean annual temperature ranges between 180c and 200c. the district has mean annual rainfall which lies between 1300 and 1700mm. maize and coffee are the main crops grown in the district [11]. agriculture and food sciences research, 2019, 6(1): 41-49 43 © 2019 by the authors; licensee asian online journal publishing group figure-1. map of the study districts. source: manipulated by the authors from arc gis (2017). 2.2. sampling procedure multistage sampling technique was employed to select the population for the study which involved both purposive and random sampling techniques. first, districts were purposively picked, and secondly kebeles were selected using random sampling method. finally, households were randomly chosen from the sampling frame exist at kebele level. a total of 205 households were selected for the study. 2.3. data collection and analysis data was collected through structured questionnaire administered to sampled farmers from march 2017 to april 2017. all demographic, socio-economic, coffee production and utilization, technology use, adoption pathway, constraints of production and technology adoption were collected. before the actual survey, the questionnaire was pretested in non-sampled villages. the pretest was not only used to test the appropriateness of the tool in collecting the required data but also to evaluate the trained enumerators on the capability of administering the questionnaire. information related to coffee production and utilization was gathered from the respondents. households’ socio demographic, institution and economic features were also collected. data were cleaned, organized and analyzed using stata version 14.2 software. both descriptive and inferential statistics were used to analyze the gathered and cleaned data. the kendall coefficient of concordance was used to assess the constraints against the production of coffee [12]. in our case, constraints were ranked from 1-10; 1 being the most constraining factor and 10 being the least constraining factor. 3. result and discussion 3.1. farmers’ demographic structure the study was conducted on four coffee potential districts of jimma zone in oromia regional state. total number of respondents interviewed was 205. out of the total respondents, 95.1% were male headed households and the rest were female headed. table-1. socio-demographic characteristics by districts. variables gomma (n=46) gera (n=50) limu kosa (n=71) manna (n=38) overall (n=205) gender in % male 95.7 98.0 94.4 92.1 95.1 female 4.30 2.0 5.6 7.9 4.9 marital status in % married living with spouse 95.6 98.0 91.6 92.2 94.2 married living without spouse 0.0 0.0 1.4 2.6 1.0 single/never married 0.0 0.0 1.4 0.0 0.4 divorced 2.2 0.0 1.4 2.6 1.5 widowed 2.2 2.0 4.2 2.6 2.9 occupation of the household head in % agriculture self employed 93.5 96.0 93.0 97.4 94.5 agriculture wage labor 4.3 0.0 1.4 0.0 1.5 non agriculture self employed 0.0 0.0 2.8 0.0 1.0 non-agricultural wage labor 2.2 2.0 1.4 2.6 2.0 domestic work 0.0 2.0 1.4 0.0 1.0 education level of the household head in % non educated 8.7 20.0 22.6 10.5 16.6 adult/religious education 2.2 18.0 2.8 5.3 6.8 primary education (1-8) 65.2 56.0 56.3 68.4 60.5 secondary education (9-12) 23.9 6.0 18.3 15.8 16.1 college education 0.0 0.0 0.0 0.0 0.0 source: survey result, 2017. agriculture and food sciences research, 2019, 6(1): 41-49 44 © 2019 by the authors; licensee asian online journal publishing group the marital status of the farmers showed that the majority of them were married and insignificant amount were widowed household heads. regarding occupation, most of respondents were engaged on full time agricultural work on own farm. out of the total respondents, the education level of more than halve of respondents were primary education and few were non-educated. gomma and manna districts have the least non educated respondents, and limu kosa have the highest non educated respondents table 1. the age of the respondents was examined as it is an important demographic factor to affect agricultural activities. the result showed non-significant difference among the districts in age of respondents. the mean age of the respondents was 47.08 years. family size affects agricultural productivity and production as it is the proxy for labour. the survey result revealed that large mean family size was seen at gomma district and the lowest mean family size existed at limu kosa district. the overall mean family size was 6.64. table 2 showed statistically significant difference between districts in number of family size of the household at 5% significance level. table-2. other socio demographic variables. variable gomma (n=46) gera (n=50) limu kosa (n=71) manna (n=38) overall (n=205) p-value mean s.e mean s.e mean s.e mean s.e mean s.e head age 45.46 1.59 41.56 1.35 47.77 1.48 44.24 1.17 47.08 0.79 0.143 family size 7.02 0.33 6.76 0.27 6.38 0.28 6.50 0.33 6.64 0.15 0.049** note: * indicate significance level at 5%. source: survey result, 2017. the age of respondents across gender revealed that female headed households have larger age than male headed households. however, male headed households have high mean family size than female headed households which is significant at 10% significance level table 3. table-3. socio demographic characteristics by gender. variables male headed household (n=195) female headed households (n=10) p-value mean s.e mean s.e age 44.96 0.80 47.50 3.89 0.488 family size 6.70 0.16 5.40 0.72 0.066* note: * indicate significance level at 10%. source: survey result, 2017. 3.2. land ownership and tenure arrangement land is the main irreplaceable factor of production in agricultural sector. the study result showed that the mean land size of the respondents is 2.14 hectares. however, large land size was seen at gera and limu kosa. the study showed statistically significant difference between the districts in land size at 10% significance level. coffee land holding of the study area revealed that gera and limu kosa district’s farmers holds as large coffee land. the small land size was observed at gomma district. however, there was no statistically significant difference between districts in terms of coffee land size table 4. the mean share of coffee land from total land is 69.6% which is high at manna district. the land covered by coffee at manna district is 77.4% which is by far higher than the rest of districts. table-4. land holding and share of coffee by location. description gomma (n=46) gera (n=50) limu kosa (n=71) manna (n=38) overall (n=205) p value mean s.e mean s.e mean s.e mean s.e mean s.e total land in hectares 1.94 0.32 2.39 0.25 2.40 0.24 1.55 0.18 2.14 0.13 0.087* coffee land in hectares 1.35 0.19 1.64 0.20 1.63 0.23 1.20 0.16 1.49 0.11 0.465 share of coffee in % 69.6 68.6 67.9 77.4 69.6 0.002*** note: ***, * indicate significance level at 1% and 10% respectively. source: survey result, 2017. land holding among gender also revealed that male headed households have large mean land holding as compared to the female counterparts though no statistically significant difference. on other hands, coffee land holding by gender has also been seen. the result pointed out that male headed households have large coffee land relative to female headed counterparts. despite the result, there is no significant difference between coffee lands among gender. the share of coffee land among male and female headed households showed coffee has covered 69.4% of male headed household’s land and 81.6% of the female headed counterparts table 5. table-5. land holding and share of coffee by gender. description male headed households (n=195) female headed households (n=10) p-value mean s.e mean s.e mean land in hectares 2.16 0.13 1.74 0.49 0.438 mean coffee land in hectares 1.50 0.11 1.42 0.52 0.886 share of coffee in % 69.4 81.6 0.995 source: survey result, 2017. agriculture and food sciences research, 2019, 6(1): 41-49 45 © 2019 by the authors; licensee asian online journal publishing group number of plots affects the resource allocation and utilization of the farmers which in turn affect the gross margin of a farm. the study was tried to assess the number of coffee plots that farmers own. the result of the study showed that the mean number of coffee plot is 2.33 across the study districts. however, coffee land fragmentation is high at gomma district and lower at manna district (table 6). table-6. coffee plot holding by location. gomma (n=46) gera (n=50) limu kosa (n=71) manna (n=38) overall (n=205) p-value mean s.e mean s.e mean s.e mean s.e mean s.e 2.46 0.18 2.18 0.16 2.46 0.15 2.09 0.17 2.33 0.08 0.310 source: survey result, 2017. 3.3. farmers’ coffee farm characteristics the study area is endowed with a good production environment for growing coffee with a combination of appropriate altitude, temperature, rainfall, soil type, and ph. totally 539 coffee plots of 205 farmers were investigated on the survey. farmers’ perception on coffee plots’ soil fertility showed that 37.85% of plots are good and 13.73% are poor in fertility. gera district has relatively higher proportion of coffee plots and gomma district has lower fertile plots according to farmers’ traditional evaluation and perception. the slope of the coffee plots was also seen on the survey. the descriptive result of the study showed that 32.84%, 48.98 and 18.18% of coffee plots have gentle, medium and steeply slope respectively. farmers’ evaluation of depth of the soil of the coffee plots showed that 50.1% has medium and 23% has deep soil. gomma district has relatively high proportion of deep soil and manna has the least. regarding soil color, the majority of the plots’ soil is red (42.49%). black and brown soil covers 38.03% and 19.48% of the total coffee plots table 7. table-7. coffee plots characteristics by location. coffee plots’ features gomma (n=137) gera (n=111) limu kosa (n=195) manna (n=96) overall (n=539) soil fertility in % from total plots good 34.30 48.65 34.87 36.46 37.85 medium 51.11 45.05 50.26 44.79 48.42 poor 14.59 6.30 14.87 18.75 13.73 slope in % from total plots gentle slope 33.58 31.53 29.23 40.63 32.84 medium slope 42.33 56.76 53.85 39.58 48.98 steeply slope 24.09 11.71 16.92 19.79 18.18 soil depth in % from total plots shallow 18.25 35.14 30.26 22.92 26.90 medium 51.82 43.24 48.72 58.33 50.10 deep 29.93 21.62 21.02 18.75 23.00 soil color in % from total plots black soil 41.61 40.54 35.38 35.41 38.03 brown soil 19.71 20.72 20.51 15.63 19.48 red soil 38.68 38.74 44.11 48.96 42.49 note: where n=total number of plots examined. source: survey result, 2017. the study was also tried to examine soil and water conservation methods and structures applied on farmers’ coffee land. accordingly, soil bunds and terrace were structures used by farmers relatively. about 77.5% of respondents do not used any conservation structures on their coffee land (figure 2). figure-2. soil and water conservation structures on coffee farms in %. source: survey result, 2017. 3.4. farmers’ awareness for improved coffee varieties awareness is the most crucial step for the adoption of agricultural technologies. farmers were asked whether they have information about coffee cultivars with high productivity, disease resistant and high sensory quality. accordingly, most of the farmers of each district knew the existence of improved coffee cultivars. agriculture and food sciences research, 2019, 6(1): 41-49 46 © 2019 by the authors; licensee asian online journal publishing group figure-3. awareness for improved coffee cultivars by location. source: survey result, 2017. the information for the cultivars also differs among gender. the result pointed out that 80% of male headed households have awareness about the cultivars. out of female headed households, 50% have information and 50% do not. the reason could be male headed households have more access to and socially delegated to meetings, training and farmers’ field days. 3.5. coffee productivity the overall objective of coffee production is yield. coffee yield differ from location to location. aside the coffee’s morphological and physiological characteristics, different socio economic and geographical features affect the productivity of coffee. despite the factors, high coffee yield was seen at manna district and lowest clean coffee productivity per hectares was seen at limu kosa. the mean overall coffee yield per hectare was 769 kg/ha which is higher than national average (710 kg/ha) by 8.3%. however, there was no significant productivity difference between study areas (p=0.768). figure-4. productivity by districts. source: survey result, 2017. due different socio economic factors, productivity may differ among gender groups. despite non-significant difference between the groups (p=0.520), male headed households’ productivity (842 kg/hectare) is higher than the female headed households’ productivity (766 kg/hectare). 3.6. source of planting material seed and seedlings are two planting materials for coffee that has been diffused to users. the main source of seed and seedlings is government extension services. there is no certified coffee seed supplier in ethiopia. however, jimma agricultural research center produces seed and disseminate to districts according to their request and agro ecology. the districts disseminate the seed to model farmers and farmers’ groups. figure-5. source of coffee planting materials in %. source: survey result, 2017. agriculture and food sciences research, 2019, 6(1): 41-49 47 © 2019 by the authors; licensee asian online journal publishing group the extension also prepares seedlings at government nurseries which could be sold to farmers at low price. accordingly, the result of the survey revealed that 46% and 61% of farmers got seed and seedlings respectively from government extension. own prepared, gifts and ngos are also the sources of the planting materials. research center also supplies seedlings in some cases like for the establishment of demonstration and scaling out of improved coffee technologies (figure 5). 3.7. sources of information information is the basic tool for the transfer of agricultural technologies. different information dissemination and awareness creation method has been modeled and used by researchers. training is one of the main methods among the models. the result revealed that 62% of respondents have got training on coffee production, postharvest handling and marketing. different bodies provided information for the farmers in the study area. the major one was government extension service which accounts for 61.1% of farmers. research center specifically jimma agricultural research center also contributes its share in providing information and knowledge for 19.1% of respondents (figure 6). figure-6. source of information about improved coffee technologies in %. source: survey result, 2017. this study identified the interrelationship between training and adoption of improved coffee technologies. the result showed that 61% of respondents who have participated on coffee related training have adopted the improved varieties. however, 39% of respondents who got training didn’t adopted improved coffee varieties figure 7. figure-7. descriptive relation of participation on coffee related training and adoption. source: survey result, 2017. different studies revealed that participation on farmers’ field days positively and significantly affects adoption of improved agricultural technologies. this study was also identified the descriptive impact of participation coffee field days on adoption. the result revealed 68% of respondents who have participated on farmers’ field days adopted the improved coffee varieties and 32% did not (figure 8). figure-8. descriptive relation of participation on coffee related field days and adoption. source: survey result, 2017. agriculture and food sciences research, 2019, 6(1): 41-49 48 © 2019 by the authors; licensee asian online journal publishing group 3.8. coffee produce utilization the utilization of coffee produce was also examined on the study area. the result of the study showed that the highest commercialization level was seen at gera district (90.8%) and relatively the lowest was recorded at manna district (86.5%). the mean commercialization level of coffee on the study area was 88.6%. when we see the consumption of coffee, the mean consumption of coffee was 6.8% and the highest coffee consumers were farmers of limu kosa district (7.8%). generally, there is no wider difference in coffee consumption among the districts (table 8). table-8. utilization of coffee produce. description gomma gera limu kosa manna total mean production (kg/household) 1828 1807 1453 1851 1697 mean coffee sold (kg) 1587 1641 1302 1601 1504 mean coffee for other uses (kg) 124 62 99.6 96.5 77 mean coffee consumed (kg) 117 110 114 125 116 sale % (commercialization) 86.8 90.8 89.6 86.5 88.6 consumption % 6.4 6.1 7.8 6.8 6.8 source: survey result. 3.9. perception on improved coffee technologies perception about the specific technology strongly affects farmers’ adoption decision [13]. the respondents were asked to give their level of agreement on perception statements comparing the improved varieties with the local varieties. the agreement levels were arranged on hedonic scale of 1 to 10 (1 indicating strong agreement and 10 indicating strong disagreement to the statement). accordingly, farmers strongly agree in high yield, vigorously and large canopy size and disease tolerance of improved coffee varieties relative to the local counterparts table 9. table-9. farmers’ perception of important varietal attributes. coffee characteristics mean rank global rank coffee improved varieties have high yield 3.52 1 coffee improved varieties are vigorous and have large canopy 5.54 2 coffee improved varieties are disease tolerant 5.71 3 coffee improved varieties matures early 5.86 4 coffee improved varieties have good berry size 6.06 5 coffee improved varieties are insect tolerant 6.23 6 coffee improved varieties have good berry color 6.76 7 coffee improved varieties are drought tolerant 6.92 8 coffee improved varieties are frost tolerant 7.16 9 coffee improved varieties have good sensory quality (taste) 7.25 10 coffee improved varieties are labor demanding 7.58 11 coffee improved varieties are input demanding 9.40 12 chi-square: 283.355’; kendall’s coefficient of concordance=0.187*** source: survey result. 3.10. constraints to improved coffee adoption agriculture is risky and uncertain sector of developing world economy. production, marketing, financial, human and institutional factors are the most commonly known risks in agriculture. coffee farmers were told ten major problems related to coffee production on their area to rank them based on their importance. the result of kendall’s coefficient of concordance summarized below showed that fluctuating coffee price is the most important problem ranked first. this was the problem raised by almost all farmers interviewed. the problem of coffee price is related to coffee global price which is set by giant coffee processing and marketing companies. the constraint ranked second was lack of reliable coffee market information. own observation in this regard also showed farmers sell their product to local traders without having any know how about update coffee price. the third important problem identified by the coffee farmers was lack of coffee seed and seedling varieties preferred table 10. table-10. ranking of coffee production constraint. constraints mean rank global rank coffee prices fluctuation 2.90 1 lack of coffee markets information 4.90 2 getting preferred coffee variety(seed/seedling) 4.93 3 prices of coffee seed/seedlings 5.10 4 getting required quality coffee seed/seedlings 5.26 5 timely availability of coffee seed/seedlings 5.31 6 getting required quantity of coffee seed/seedlings 5.40 7 availability of credit to buy improved coffee seed/seedlings 6.14 8 availability of herbicides 6.95 9 timely availability of coffee seed/seedlings 8.10 10 chi-square = 53.40; kendall’s coefficient of concordance=0.272*** source: survey result. 4. conclusions and recommendations the study was aimed to identify major coffee production problems that affected coffee farmers’ production and productivity. the result of the study identified three main coffee related problems raised and ranked by farmer namely coffee price fluctuation, lack of coffee market information and lack of improved coffee seed and seedlings. based on the above findings, the study has drawn the following implication. agriculture and food sciences research, 2019, 6(1): 41-49 49 © 2019 by the authors; licensee asian online journal publishing group • strengthening cooperatives and unions: cooperatives and unions on the study area collect both dry and red coffee in relatively attractive price. the outlet has reduced additional transaction cost and also eliminates/reduces unnecessary chain actors such as brokers or assemblers. farmers also collects dividend in proportion to the coffee they supplied to cooperatives. however, the main drawback of this market outlet is that they do not pay the money on the day farmer sale coffee. farmers stay for a weeks or even months to collect the money. this makes the farmers to not sell their coffee to cooperatives which in turn leads to sell for local traders and brokers that fluctuates coffee price significantly. therefore, strengthening the financial capacity of cooperatives and union should be a good option for farmers to sell their coffee product to these outlets so that coffee price could be modified and farmers could collect the margin they deserve. • supply of improved coffee seeds and seedlings: the study result identified lack of preferred coffee varieties’ planting materials as the main constraint on the study areas. encouraging smallholder farmers in supplying planting materials (seed/seedlings) to replace aged coffee for improved coffee technologies is a critical option to be given emphasis by stakeholders such as extension, research centers, universities and ngos. on other hands, government nurseries need to emphasize on seedling distribution to their maximum capacity. • sustainable seed system: there is no formally recognized enterprise which multiplies and supply coffee seed. its only research center that have limited seed multiplication sites which is incapable to satisfy huge and raising demand for improved coffee seed. there is high mismatch between coffee seed demand and supply. therefore, concerning bodies need to be concerned the way coffee seed sources would be established for each coffee producing area. references [1] icc international coffee council, "cc development strategy for coffee," presented at the international coffee council 105th session, london, 2010. [2] k. f. wiersum, "indigenous exploitation and management of tropical forest resources: an evolutionary continuum in forest-people interactions," agriculture ecosystem environment, vol. 63, pp. 1-16, 1997. available at: https://doi.org/10.1016/s01678809(96)01124-3. [3] dfsc and ipgri, forest genetic resources conservation and management: overview, concepts and some systematic approaches vol. 1. rome, italy: international plant genetic resources institute, 2001. [4] l. c. olmos, e. duque, and e. rodriguez, "state of the art of coffee drying technologies in colombia and their global development," espacios, vol. 38, pp. 27-37, 2017. [5] g. adugna, b. bellachew, t. shimber, e. taye, and t. kufa, coffee diversity and knowledge. addis ababa, ethiopia: ethiopian institute of agricultural research, 2008. [6] usaid, "ethiopian coffee industry value chain analysis. profiling the actors, their interactions, costs, constraints and opportunities. chemonics international inc. [accessed 18 june 2010]," 2010. [7] i. pokorná and l. smutka, "what is the structure of the coffee market: can the real poor benefit from the coffee trade? memeographed paper; electronic copy. czech university of life sciences, faculty of economics and management, department of economics. available: http://ssrn.com/abstract=1734902," 2011. [8] a. bizualem, g. degye, and s. zekarias, "analysis of marketed surplus of coffee by smallholder farmers in jimma zone, ethiopia," journal of biology, agriculture and healthcare, vol. 5, pp. 242-251, 2015. [9] g. dessalegn and s. kebebew, "evaluating coffee market structure and conduct in bench-maji zone, south west ethiopia," journal of agricultural economics, extension and rural development, vol. 2, pp. 156-163, 2014. [10] ectdma /ethiopian coffee and tea development and marketing authority, "coffee productivity and quality improvement package training manual (amharic); ministry of agriculture and rural development, addis ababa, ethiopia," 2016. [11] janrd/jimma zone agricultural and natural resource development, "profile and features of districts," unpublished report; jimma, ethiopia2016. [12] p. a. pinamang, n. a. eyram, a. nimo-wiredu, d. adogoba, b. nsiah-frimpong, j. haleegoah, a. adu-appiah, b. owusu-asante, k. adofo, and e. baafi, "root and tuber crops technologies adoption and impact study in ghana: the case of improved sweet potato technologies," final report to the west africa agricultural productivity program, ghana2017. [13] a. a. adesina and m. m. zinnah, "technology characteristics, farmers' perceptions and adoption decisions: a tobit model application in sierra leone," agricultural economics, vol. 9, pp. 297-311, 1993. available at: https://doi.org/10.1016/01695150(93)90019-9. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://ssrn.com/abstract=1734902, agriculture and food sciences research vol. 4, no. 1, 30-36, 2017 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2017.41.30.36 30 value addition in cassava processing: evidence from women in ilesa west local government area of osun state e.o. okebiorun1  n.a. jatto2 ( corresponding author) 1,2department of agric. economics, faculty of agriculture, usmanu danfodiyo university, sokoto, sokoto state, nigeria abstract to develop policies in addressing women problem in respect of their participation in cassava processing, the determinants and the level of their interest have to be adequately explored and understood. the study examined value addition to cassava in term of net return and factors influencing women interest in cassava processing in ilesa west local government area in osun state. a purposive sampling technique was used to select 4 wards and 40 respondents were randomly selected within the l.g.a. using a structured questionnaire. the result shows that rate of investment on cassava processing was ₦0.39 indicated that for every ₦1 spend on cassava processing; ₦0.39 will be realized as profit. it is evident from the study that cassava processing is a profitable agricultural activity in the study area. the socioeconomic variables such as age, years of experience, income and membership in cooperative society contributed significantly to women interest in cassava processing in the study area. years of experience and income are the main socio factor responsible for their interest in cassava processing. the major constraints to cassava processing in the area are heat from firewood. it is recommended that government through its research agencies should make available and affordable improved processing technology that is simpler to use for cassava processing more accessible. keywords: value addition, cassava processing, evidence, women interest, ilesa west. contents 1. introduction ...................................................................................................................................................................................... 31 2. methodology ..................................................................................................................................................................................... 31 3. results and discussions .................................................................................................................................................................. 32 4. conclusion and recommendation ................................................................................................................................................ 35 references .............................................................................................................................................................................................. 35 citation | e.o. okebiorun; n.a. jatto (2017). value addition in cassava processing: evidence from women in ilesa west local government area of osun state. agriculture and food sciences research, 4(1): 30-36. history: received: 30 october 2015 revised: 20 september 2017 accepted: 27 september 2017 published: 2 october 2017 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2017.41.30.36&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=e.o.%20okebiorun https://orcid.org/orcid-search/quick-search?searchquery=n.a.%20jatto agriculture and food sciences research, 2017, 4(1): 30-36 31 1. introduction women form the majority of rural areas and are responsible for about 80% of all food items produced [1]. when focusing on agriculture as an economic activity the role of women cannot be undervalued as they account for 49.6% percent of nigeria population[2]. even though women play a dominant role in agriculture, they are hardly trained and reached by extension agents with improved technologies [3]. many women believe that the agricultural industry is mostly for men. however, what most women do not realize is that the percentage of women in the agricultural industry continues to increase almost every day [4] cited in jatto, et al. [5]. along with raising children and tending to duties inside the home such as cooking and laundry, women also tend to chores outside. some duties include feeding animals, scooping manure and driving grain truck, these jobs are no longer seen only for man [2]. jatto, et al. [2] also suggested that women’s contribution to farm work is as high as the total farm task performed. the contribution of women ranges from such tasks as land clearing, land tilling, planting, weeding, fertilizer application to harvesting, food processing, threshing, milling and marketing as well as management of livestock [6].with these, most tasks are supposedly meant for man but the benefits gained by them are not commensurate to the hours women spend on it. many women believed that agricultural industry is still for men only, making them entrenched in a vicious cycle of poverty that placed them at less advantage of income and resource empowerment [6]. the national research centre for women in agriculture (nrecwa) [7] stated in their 2003-2004 report that women are faced with health hazards and about 50% comes from transplanting on farm activities, 50% in post harvest activities comes from threshing and 47%in livestock management comes from shed cleaning, these may be the part of reason while women’s participation in agriculture is impeded. policy issues in the past failed to address the unpleasant and deploring conditions under which women in the country contribute to agriculture; this perception arises because there is limited knowledge among the policymakers about the numerous contributions of women to agriculture especially in the area of food production in the country. efforts to add value to crops such as cassava require more attention now than ever before if nigeria’s agricultural sector is to evolve. aside from improving farmers’ incomes, value addition is one of the best ways to address the increasing level of unemployment in nigeria international institute of tropical agriculture [8]. abubakar [9] reported that value addition to cassava would help turn around the fortunes of farmers and it offers tremendous opportunities that would further broaden the country’s economic viability and improve its competitiveness particularly in the nonoil sector. cassava is a major source of energy with very high food security value similar to most cereal crops [10, 11]. cassava possesses special attributes, which include ability to make returns of root yield even at extreme stress conditions; tolerance to unfavorable conditions like low soil ph, marginal soil fertility, and drought; suitability to various farming and food systems in africa, minimal external input requirements, all year round availability as well as efficient production food energy. these attributes single cassava out as ideal crop for the tropics [12, 13]. alternative uses of cassava through value addition has resulted in emergence of wide food recipes from cassava through processing which involves conversion of edible food into another form more acceptable or more convenient to the consumer. some value added products from cassava include cassava flour, cassava starch, cassava bread, cassava chin-chin, cassava chips, cassava flakes, cassava odorless fufu, cassava doughnut, cassava cake, cassava biscuit and cassava salad cream. the value addition technology in cassava possesses enormous potentiality for increasing cassava consumption, diversifying its uses as well as using same to enhance livelihoods of farm families through providing opportunities for employment; micro agro enterprises development income and boosting economy of rural households [14]. in other to address women problems in respect of their participation in agricultural processing activities most especially cassava processing and their level of interest have to be properly understood. toward this end a study has been conducted to identify the value addition and factors influencing women to participate in cassava processing ilesa west local government area in osun state. 2. methodology the research was carried out in ilesha west local government area in osun state. the area lies between latitudes 70 39'n and 70 617'n and longitudes 40 43'e and 70 767'e. it covers an estimated landmass of about 75 square kilometers with rivers like adeti, oora and oromu traversing the local government area. its headquarters is located at ereja square (now at omi-aladiye, osogbo road) and is purely an urban area. the square is the headquarters of ijesa division. the local government area is unique in ijesa land as its headquarters is the seat of owa-obokun adimula, the paramount ruler of ijesa land. the area was carved out of the old ilesa local government area, which was split into two in november 1996. the population of the inhabitant of the area as projected from the provisional figures of the 2006 national population census by 2014 is 137,495 peoples. the annual mean temperature is about 850f (29.440c) and annual mean rainfall measures about 60 inches (1,524mm). the area interspersed with hills within the ranges of 800 to 950 feet above sea level and it is relatively plain in contradistinction to the neighboring ilesa east local government area. majority of the inhabitants in the area are farmers; this is due to the rich soil which support agricultural activities especially crop production (cassava, maize, soybeans, potatoes, cotton, etc.) and a handful of them are engaged in cash and food crop production, they also involved in livestock farming (poultry, cattle, sheep and goat, etc.). four wards were purposively selected from ilesa west local government area of osun state due to large population of cassava processor and 40 cassava processors (women) were randomly selected from the four wards at random. primary data was collected for the study, using a structured questionnaire, containing information on the socio-economic characteristic of cassava processors, value addition to cassava in term of net return by the cassava processors and constraint faced by the cassava processors. the analytical tools used to analyze the data generated from the study descriptive statistic and a binary logit regression to assess the factors that influenced women participation n cassava processing. agriculture and food sciences research, 2017, 4(1): 30-36 32 2.1. budgetary techniques this is to estimate cost and return of the cassava processing and to determine value added to cassava in term of net return. net return is the amount of money realized in a business after removing the cost involved in the business. this can therefore be stated mathematically as: nr = trtvc where: nr = net return at each level of actor along cassava value chain. (₦/month) tr = total revenue at each level of actor along cassava value chain. (₦/month) tvc = total variable cost at each level of actor along cassava value chain. (₦/month) roi = 𝐺−𝐶 𝐶 where: roi = rate of investment (₦/month) g = gain from production (₦/month) c = cost of production (₦/month) 2.2. binary logit model specification 1 𝑋ij , 𝐽 = 1 − 8 = 𝐹(𝑍𝑖) = 1 1 + 𝑒𝑧𝑖 = 𝑒𝑧𝑖 𝑒𝑧𝑖 + 1 ; 𝑖 = 1 − 𝑛 where, 𝑍𝑖 = 𝛼 + 𝛽1𝑋𝑖1 + 𝛽2𝑋𝑖2 + ⋯ + 𝛽8𝑋𝑖8 + 𝑈 1 = women interested in cassava processing 0 = not interested in cassava processing i = number of women sampled j = 1-8 are the socio economic characteristic of women interested defined as: x1 = processing experience (yrs.) x2 = education (yrs.) x3 = amount of cassava processed (kg/month) x4 = age (yrs.) x5 = income (n) x6 = dependency ratio x7= children (number) x8= participation in cooperative society (years) β1 β8 = logit coefficient α= constant term. u= error term which will be assumed to be normally distributed with zero mean and constant. 3. results and discussions the responses of the cassava actors in ilesa west local government area were analyzed using descriptive statistic. the results on socio-economic characteristic of the cassava processors are presented in table 1. the socio-economic characteristics of cassava processors have direct or indirect influence on cassava processing. the variables identified under this include; age, marital status, family size, level of education, year of experience and the main occupations of the cassava processors. table-1. distribution of the cassava processors according to age, gender, marital status and family status and family frequency percentage age range (years) ≤ 30 4 10 31 – 40 3 7.5 41 – 50 23 57.5 51 – 60 7 17.5 > 60 3 7.5 mean 45.6 marital status single 4 10 married 15 37.5 divorced 12 30 widowed 9 22.5 family size 1 – 5 24 60 6 – 10 15 37.5 11 – 15 1 2.5 source: field survey, 2014. the socio-economic characteristics of cassava farmers have direct or indirect influence on resources use efficiency in cassava production and are presented in table 1. the age of the respondents determine the effort and quality of labour he/she employs in any given area [15]. the results show that majority, 65% of the cassava processors were between the age group of 31 – 50 years old. this indicated that most of the cassava processors are in their active age years. this implies that most of the actors involved are likely to be physically fit to perform cassava processors activities. the mean age of cassava actors was 45.6 years. rathman, et al. [16] shows that 47.96 years is within the economically active age and as such will respond positively to any intervention aimed at improving their productive capacity. agriculture and food sciences research, 2017, 4(1): 30-36 33 marital status determines the family size of the cassava processors and consequently determines the number of people expected to work in processing activities. the marital status of cassava processors is presented in table 1. the result reveals that majority of the cassava farmers (37.5%) are married while 30% are divorced. this result agrees with the work of ofuoku [17] and fakoya, et al. [18] who noted that married people have the responsibilities of catering for the needs of their families and also engage in various economic activities that would serve as source of income. the household size of cassava farmers determines the contribution of the family members in cassava processing. this determines the number of family members in a household that could contribute to family labour. table 1 reveals that majority of the farmers (60%) have family sizes ranging from 1 to 5 individuals. the result of this finding is in line with the result of igodan, et al. [19]who reported that educated farmers tend to have smaller families. arthur [20] also observed that small family size enjoy better economic and social lives, which have great influence on better understanding of environmental conditions but reduced labour availability. table-2. distribution of cassava processors according to education status, source of capital, occupation and year of experience frequency percentage education status no formal 8 20 primary 19 47.5 secondary 10 25 tertiary 1 2.5 islamic 1 2.5 adult education 1 2.5 source of capital personal saving 21 52.5 friend & relative 12 30 money lender 5 12.5 bank 2 5 occupation primary 36 90 secondary 4 10 year of experience 1 -10 3 7.5 11 – 20 17 42.5 21 – 30 12 30 > 30 8 20 source: field survey, 2014. education plays a vital role in adoption of innovative practices among the cassava processors. education is seen as a means of acquiring knowledge on new technology being practiced to add value and increase efficiency. with high level of education, adoption of new technology becomes easy. table 2 revealed that 47.5% of the cassava processors have attained and obtained primary school certificate, while 25% attended secondary school. it was observed that farmers with primary and secondary school education dominated the cassava production activities. williams [21] and adesoji and farinde [22] noted that secondary education could equip farmers with some managerial skills in agri-business and help in understanding innovations. capital is one of the factors of production, without which the production will not be possible. the importance of capital in production is much particularly as related to smallholder farmers, as most farmers had little or no money to carry out important farm operations or purchase modern yield exchanging input in production at the required. capital also is important because it help the farmers to expand their activities. sources of capital of cassava processors are presented in table the result shows that 52.5% of processors source their capital from their personal savings to finance their activities, while 30% source their capital from friends and relatives. this result indicated that cassava processors do not have much access to capital from banks in the study area. the result agrees with that of alimi [23] who observed that food crop farmer’s major source of capital (working capital) was personal saving which was inadequate. occupation involves activity that is carried out by an individual to derive a certain benefit or to enable the individual sustains a living. the distribution of the cassava processors based on occupation is shown in table 2. the table shows that 90% of cassava processors consider their engagement into cassava processing activities as primary occupation while the remaining 10% involved in secondary occupation in casa of failure in one activity, they will easily change to the other. years of experience of the cassava processors, this refers to the number of years over which the cassava processors have been engaged in cassava processing activities. the more experienced one has, the higher the profit margin. also, the more the period a farmer spends in the practice, the more he/she will improve in managerial capability and overall efficiency. the results of the cassava processors based on the years of experience are represented in the table 2. the result indicates that most of the cassava processors (92.5%) have not less than 11 years of experience in their engagement into cassava processing activities. it is expected that the processors are aware of new processing technology, and this would likely help the processors in achieving higher level of productivity. agriculture and food sciences research, 2017, 4(1): 30-36 34 3.1. cost and returns structure of the cassava processors table-3. average cost and returns structure of the cassava processors / month item amount percentage quantity of cassava purchased (bag) 71.80 unite price of cassava (₦/bag) 1,139.00 total value of cassava purchased (₦) 81,780.20 66.55 unit cost of processing (₦/bag) 572.50 total cost of processing (₦) 41,105.50 33.45 total variable cost (₦) 122,885.70 100 unit price of cassava output (₦/bag) 2,372 total returns (₦) 170,309.60 net returns (₦) 47,423.9 net returns as a % of cost incurred 38.59 r o i (₦) 0.39 source: field survey, 2014 cost and returns structure of the cassava processors is presented in table 3. it can be observed that an average cassava processor utilized 71.8 bags of cassava per month in their processing activities. out of the total cost incurred by an average cassava processor per month, the greatest share (66.55%) was spent on the purchase of raw cassava and the remaining 33.45% was incurred as the cost of processing the cassava. an average cassava processor realizes a net return of ₦47,423.90 per month represent 38.59% of the total cost incurred per month. further analysis on rate of investment on cassava processing showed that for every ₦1 invested into cassava processing, ₦0.39 is realized as profit. this implies that cassava is a lucrative enterprise. 3.2. constraints faced by cassava processors table-4. constraints faced by cassava processors problems frequency percentage high cost of cassava 25 62.5 drainage problem 32 80 hazard faced 20 50 scarcity of firewood 29 72.5 low demand for gari in the market 21 52.5 shortage of laborers 18 45 note: * multiple responses. source: field survey, 2014. the constraints in cassava processing in the study area were found to be numerous, stating from high cost of cassava, water lodge, heat from firewood and scarcity of firewood, table 4 shows the gravity of these constraints provided by cassava processors. these constraints reported by the cassava producers are critical. the table shows that 80% of cassava processors encounter drainage problems as a result of poorly drained topographically and 72.5% of them find it difficult to get enough wood in preparing their product while 62.5% of them are being faced with the problem of high cost of cassava. the results are in line with abdulkarim, et al. [24] who argued that water logging and erosion are one of the factors affecting cassava-processing activities. table 4 shows the distribution of maximum likelihood estimate women interest in cassava processing in ilesa west local government. the table shows that number of children, dependency ratio and years spent on education are not significant, but age and membership in cooperative society had a significant negative coefficient but significant while years of experience and income had a positive coefficient and significant. it is being noted that a positive sign on a parameter indicated that higher values of variables tends to increase the likelihood of women processors in cassava production. similarly, a negative value of coefficient implied that higher values of the variables would reduce the probability women interest in cassava production. table-5. maximum likelihood estimate women interest in cassava processing in ilesa west local government variables coefficients std. error pvalue constant 25.8584 9.15772 0.00475*** age -0.424411 0.157892 0.00719*** number of children 0.228914 0.346629 0.50900 dependent ratio -0.830329 1.08079 0.44233 years of education 0.264233 0.208643 0.20537 years of experience 0.383089 0.166958 0.02176** income 0.0000036 0.00000166 0.03045** membership in cooperative society -6.03709 2.07297 0.00359*** mean dependent variable 0.50000 log-likelihood -13.02386 source: field survey, 2014. the result indicates that age, years of experience; income and membership in cooperative society might influence women interest in cassava processing. the variables age and membership in cooperative society had a negative coefficient and significant at 1%, while years of experience and income have a positive coefficient which are significant at 5% level. considering years of experience and income it might imply that for every one year experience gained might increase the probability to participation in cassava processing. this could be because they agriculture and food sciences research, 2017, 4(1): 30-36 35 are likely to be more sensitive to risk in their decisions and actions that may affect their productivity and might imply also that a woman cassava processor whose income has increased is due to her experience in the processing activities this is also supported by jatto, et al. [5]. while for every one-year increase in age and membership in cooperative society may decrease the likelihood of women participation in cassava processing. the log-likelihood (-13.02346) indicate that there is no close relationship within the variables and the omnibus test of the model which gives the overall fit test indicate a chi-square value of 84.8566which is significant (p<0.01) implies that the model as a whole fit significantly better. the predicted marginal probability of women interest in cassava processing as related to the significant socioeconomic variable is shown in table 5. the table showed that age and membership in cooperative society have a negative marginal probability. income and years of experience shows a positive marginal probability table-6. marginal effect of binary logit regression variables coefficient std. error marginal effect constant 24.1543 9.09713 age -0.392912 0.157012 -0.053984 years of experience 0.28416 0.107409 0.0390421 income 0.00000226 0.00000163 0.000000311 membership in cooperative society -4.47877 1.29146 -0.615358 source: field survey, 2014. the result shows that age and membership in cooperative society may not necessary be a factor contributing to women interest in cassava processing meaning that they may not be educated and may not belong to any cooperative society to be a cassava processor. years of experience have the highest extent of influence to which a unit change increases the probability of the women interest in cassava processing. the implication is that it increases the likelihood of more access to income. 4. conclusion and recommendation it is evident from the study that, cassava processors in the study area showed that cassava processing is profitable agricultural activity in the study area and majority of them having not less than 11 years of experience. the socioeconomic variables such as age, years of experience, income and membership in cooperative society contributed significantly to women interest in cassava processing in ilesa west local government area of osun state. years of experience and income are the main socio factor responsible for their participation in cassava processing. the major constraints to cassava processing in the area are heat from firewood. it is recommended that government through its research agencies should make available and affordable improved processing technology that are simpler to use and also make cassava processing more accessible. references [1] d. o. awotide, "assessment of women’s participation in cooperative societies and its determinants in yewa north local government area of ogun state, nigeria asian," journal of agriculture and rural development, vol. 2, pp. 344-350, 2012. view at google scholar [2] n. a. jatto, u. i. gunu, and a. alkali, "determining women interest in poultry egg farming in ilorin, kwara state," international journal of current research, vol. 5, pp. 065-067, 2013. 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[24] n. a. jatto, m. a. maikasuwa, u. b. kyiogwom, and i. muhammad, "evaluating the factors influencing poultry farmers’ participation in egg production: a binary logit regression model," european scientific journal, vol. 8, pp. 125-134, 2012. view at google scholar asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://scholar.google.com/scholar?hl=en&q=application%20of%20markov%20chain%20analysis%20in%20forecasting%20arable%20crop%20farm%20size https://scholar.google.com/scholar?hl=en&q=evaluating%20the%20factors%20influencing%20poultry%20farmers’%20participation%20in%20egg%20production:%20a%20binary%20logit%20regression%20model https://scholar.google.com/scholar?hl=en&q=evaluating%20the%20factors%20influencing%20poultry%20farmers’%20participation%20in%20egg%20production:%20a%20binary%20logit%20regression%20model 57 agriculture and food sciences research vol. 5, no. 2, 57-63, 2018 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2018.52.57.63 comparative evaluation of the nutritional, physical and sensory properties of beef, chicken and soy burgers adeniyi paulina o.1 obatolu veronica a.2 kehinde hammed a.3 1,2institute of agricultural research and training, obafemi awolowo university, moor plantation, apata, ibadan, nigeria 3federal college of agriculture, moor plantation, apata, ibadan, nigeria ( corresponding author) abstract the sporadic technological advancement witnessed worldwide since the beginning of the twenty first century has encouraged civilization and urbanization culminating into changes in lifestyle from active to sedentary globally. people now prefer instant foods and eating out in different food service units to home cooked meals. burger is a special delicacy which is seldom prepared at home but commonly purchased from restaurants. the need to encourage human consumption of soybean cannot be over emphasized due to the unrivalled nutritional and health benefits derivable from it especially in combating some communicable diseases, the epidemic of which is becoming alarming globally, hence, harnessing soybean potentials in the production of burger is of pertinent importance. this experimental study was therefore designed to compare the nutritional, physical and sensory properties of soy burger with those of the conventional beef and chicken burgers. soy cheese was prepared from soybeans using appropriate culinary procedure. equal portions of minced beef, minced chicken and soy cheese were processed into burgers using basic recipe. beef, chicken and soy burgers were subjected to proximate, thiamine, riboflavin, cholesterol, colour, texture and sensory evaluation using standard methods. mean data were compared using analysis of variance at p≤0.05. protein content (dry weight basis) for beef burger (bb), chicken burger (cb) and soy burger (sb) were 57.87, 50.36 and 53.89% respectively. in similar sequence fat content were 10.02(bb) 10.15(cb) and 8.58 (sb). sb was significantly higher than the other two samples in crude fibre, riboflavin and niacin while its cholesterol content was negligible. there was no significant difference between sb and cb in lightness and redness while the values for yellowness were 10.60, 25.12 and 18.25 for bb, cb and sb respectively. in the same vein there was no significant difference in chewiness, cohesiveness and springiness of the three burger samples. for overall acceptability bb was most acceptable followed by sb and then cb. the nutritional, physical and sensory properties of soy burger were comparable to that of beef and chicken burgers, hence, soy burger may compete favourably with the conventional types of burger and is hereby presented as a cholesterol-free alternative with higher health benefits suitable for all even vegans. household and commercial production and consumption of soy burger is hereby recommended. keywords: beef burger, chicken burger, soy burger. citation | adeniyi paulina o.; obatolu veronica a.; kehinde hammed a. (2018). comparative evaluation of the nutritional, physical and sensory properties of beef, chicken and soy burgers. agriculture and food sciences research, 5(2): 57-63. history: received: 23 april 2018 revised: 9 may 2018 accepted: 14 may 2018 published: 16 may 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 58 2. methodology ..................................................................................................................................................................................... 58 3. result .................................................................................................................................................................................................. 60 4. discussion .......................................................................................................................................................................................... 61 5. conclusion and recommendation ................................................................................................................................................ 62 references .............................................................................................................................................................................................. 62 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.52.57.63&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=adeniyi%20paulina%20o. https://orcid.org/orcid-search/quick-search?searchquery=obatolu%20veronica%20a. https://orcid.org/orcid-search/quick-search?searchquery=kehinde%20hammed%20a. agriculture and food sciences research, 2018, 5(2): 57-63 58 1. introduction the pros and cons of any concept commonly go hand in hand. if the pros outweigh the cons the concept may be considered desirable and helpful. the same is witnessed in the sporadic technological advancement which was being experienced since the beginning of the twenty first century. the affirmation of the very rapid advancement in technology in the 21st century has been reported [1]. it has enhanced good security while concomitantly threatening adequate security globally [1]. it has encouraged sedentary lifestyle which has increased the epidemic of some non communicable diseases which may be fondly referred to as ‘diseases of comfort’ the overall causes of death by the 22nd century [2]. sedentary lifestyle predisposes an individual to chronic diseases such as cancer, cardiovascular disease, type 2 diabetes mellitus and osteoporosis and may be regarded as ‘civilization diseases [3]. it is therefore imperative that feasible measures should be designed and actualized to cushion the adverse effect of technological advancement and civilization on human health and development. one of such measures is to influence food choices and habit towards the consumption of adequately healthy foods which are naturally endowed to combat or prevent these civilization diseases or diseases of comfort. soybean is of unrivalled nutritional and health benefits. it is the only plant source of complete protein and competes favourably with animal protein [4]. the ability of soybean consumption to prevent heart disease, breast cancer, prostate cancer and osteoporosis has been well affirmed by both old and recent scientific reports [5-9]. all these ascertained the ability of soybean in preventing the so called civilization diseases or diseases of comfort; hence, it is of pertinent importance to encourage the consumption of soybean globally. burger, the origin of which is traceable to germany and america, is now a special delicacy of international acceptance and reception, that is, it is now commonly consumed and enjoyed by people all over the world. harnessing the global acceptability of burger in encouraging the consumption of soybean is therefore a thoughtful, feasible and viable approach in combating the alarming epidemic of civilization diseases. burger is commonly prepared with minced beef or chicken but other meat sources such as turkey, ostrich, deer, buffalo, sheep, etc may be used [10]. a burger is a sandwich of cooked patties of minced meat, usually beef, inside a sliced bread roll or burger bun and may be accompanied with cheese, lettuce, onion, tomato, bacon and pickles while condiment such as mayonnaise, mustard, ketchup, barbecue sauce may be added. the meat patty may be cooked by pan roasting, pan frying, grilling or flame broiling [10]. meat-like product has been produced from tofu (soy cheese) using food extruder [11] a food processing invention which may be expensive and may not be affordable for household and small/medium scale commercial production and consumption. preparation of burger from soybean using tofu is an approach of producing meat-like product from soybean, the process of which is simple, affordable and easily accessible, hence, this study was designed to compare the nutritional, physical and sensory properties of soy burger with that of the conventional beef and chicken burgers with the view of producing soy burger that can compete favorably with beef and chicken burgers in these parameters so as to encourage the consumption of the wonder bean towards alleviating the incidence of civilization diseases globally. 2. methodology 2.1. collection of materials soybeans (tgx-1740) were collected from the seed store of the institute of agricultural research and training, obafemi awolowo university, apata, ibadan, nigeria. minced beef and chicken were purchased from shoprite supermarket, palm mall, ring road, ibadan. breadcrumbs was prepared from oven dried sliced bread (my joy, ibadan, nigeria). other ingredients were purchased from local market in apata, ibadan. 2.2. preparation of soy cheese soybeans were cleaned manually and soaked in water for16hours at ambient temperature after which the water was decanted and the beans were washed with clean water. this was then allowed to simmer in water for 5 minutes. the hot water was drained off and the beans were wet-milled using plate attrition mill (amuda plate mill, india). the paste was diluted with water and sieved using cheese cloth. the extract (soymilk) was allowed to boil and while still boiling sour steep water was added in ration 7:1 (v/v) for soymilk: steep water. the curd separated immediately and further boiling for 30 minutes was done after which this was poured in a cheese cloth and allowed to drain with a heavy load placed on it. 2.3. preparation of burger samples the three burger samples were prepared using basic recipe. ingredients used are expressed in table 1. table-1. basic ingredients for the preparation of beef, chicken and soy burgers ingredients beef burger chicken burger soy burger meat minced beef (500g) minced chicken(500g) soy cheese (500g) wheat flour 50g 50g 50g bread crumbs 50g 50g 50g bouillon cube (knorr) 1 cube 1 cube 1 cube bell pepper and onion paste 20ml 20ml 20ml salt 2g 2g 2g margarine (for greasing tray) 5g 5g 5g source: aidells [10] ingredients were gently mixed together and divided into small portions which were shaped into flat forms of about 1.5cm thickness. these were placed in greased aluminium trays and positioned under the grill and allowed to grill for 20 minutes for each side to give the three burger samples. these were the subjected to protein, fat, ash, fiber, carbohydrate, thiamine, riboflavin, niacin, cholesterol, color, texture and sensory evaluation. agriculture and food sciences research, 2018, 5(2): 57-63 59 2.4. moisture content determination this was determined using the air oven method [12]. a known weight of the sample (3g) was put in a washed, dried and cooled crucible and this was dried at 103oc until a constant weight was obtained. this was allowed to cool in a desiccator and the difference in weight was used to calculate the moisture content. 2.5. protein content determination the crude protein content was determined using the micro kjeldahl method as described by kirk and sawyer [12]. a tablet of kjeldahl catalyst was added to a known weight of the sample (0.2077g) in a long necked kjedahl flask. this was heated in a fume cupboard with 25cm3 of concentrated h2so4 until a clear solution was obtained. this was cooled, poured into a 10cm3 volumetric flask and made up to mark with distilled water after which 10ml of this was measured into a distillation set. 5cm3 of boric acid was pipette into a 100ml conical flask and placed at the receiving end of the distillation unit with the delivery tube completely dipped into the flask. 40% naoh was used to liberate ammonia out of the digest into the boric acid under alkaline condition and this was titrated against 0.1n hcl until the first permanent color change was observed. blank sample was run through the procedure and the titer value was used to correct the titer value for the test samples. the protein content was calculated thus: %n = molarity of hcl x (sample titreblank titre) x0.014 x df x100 weight of sample %n was converted to the percentage crude protein by multiplying by 6.25 2.6. crude fat content determination the fat content was determined using soxhlets extraction method as described by kirk and sawyer [12]. a known weight of the sample (2g) was put into a weighed filter paper and folded neatly. this was put inside a preweighed thimble (w1). the thimble with the sample (w2) was inserted into the soxhlets apparatus and extraction was carried out under reflux with petroleum ether (40oc – 60oc boiling range) or 6 hours. at the end of the extraction, the thimble was dried in the oven for about for about 30 minutes at 100oc to evaporate the solvent and thimble was cooled in a desiccator and later weighed (w3). crude fat content of the sample was calculated thus: % fat = loss in weight of sample x 100 = w2 – w3 x 100 original weight of the sample w2 – w1 2.7. ash content determination the ash content denotes the total amount of minerals present in the products. this was determined using the method as described by kirk and sawyer [12]. a known weight (1.5g) of finely ground sample was weighed into clean and dry previously weighed crucible with lid (w1). the sample was ignited over a low flame to char the organic matter with lid removed. the crucible was then placed in muffle furnace at 600oc for 6 hours until it was turned to ash completely. this was then transferred directly to desiccators to cool and was later weighed (w2). % ash = w2 – w1 x 100 weight of sample 2.8. crude fibre determination the crude fibre was determined using the method as described by kirk and sawyer [12]. two hundred milliliters (200ml) of freshly prepared 1.25%h2so4 was added to a known weight (3g) of the residue obtained from fat extraction and this was boiled for 30 minutes and then filtered after which the residue was washed until it was free from acid. the residue was transferred quantitatively into a digestion flask and 1.25%naoh was added after which this was boiled for 30 minutes. this was followed by filtration and the residue was then washed with methylated spirit and then petroleum ether to be free of alkali. this was then allowed to drain and the residue was transferred to a silica dish (previously ignited at 600oc and cooled). the dish and its content were dried to constant weight at 105oc. the organic matter of the residue was burnt by igniting for 30 minutes in a muffle furnace at 600oc). the residue was cooled and weighed while the loss on ignition was reported as crude fibre. 2.9. carbohydrate content determination this was calculated by difference of all other nutrients from 100. 2.10. thiamine, riboflavin and niacin determination these b vitamins content were determined using the method as described kirk and sawyer [12]. 2.11. thiamine determination method as described by kirk and sawyer [12] was used fifty milliliters (50ml) of 50% methanol and 50ml 0f 17% sodium carbonate was added to 1g of the sample in order to extract the vitamin. this was then filtered after which folins-denis reagent was added. this was allowed to cool until a bluish color was developed and absorbance was read in a spectrophotometer at 415nm. a standard curve was prepared using the data obtained with tannic acid in place of the sample and the values for the sample were extrapolated from this curve. 2.12. riboflavin determination the method as described by kirk and sawyer [12] was used. to 0.5g of the sample 30ml of dichloroethane and 30ml of 30% hcl were added. this was followed by the addition of 50ml of ammonium hydroxide solution after which filtration was carried out and later the absorbance was read at 415nm. a standard curve was constructed using the data obtained from the use of standard riboflavin in place of the sample and the curve was used to extrapolate the values for the samples. agriculture and food sciences research, 2018, 5(2): 57-63 60 2.13. niacin determination method as described by kirk and sawyer [12] was used. niacin was extracted by autoclaving the sample (1g) with 0.75g calcium hydroxide and 20ml deionised water at 121oc for 30 minutes. the mixture was diluted with 30 ml of water, mixed thoroughly and allowed to cool after which it was centrifuged at 0oc and 2500 rpm for 15 minutes. a 15ml sample of the supernatant was adjusted to ph 7 with aqueous oxalic acid. the resulting suspension was centrifuged at 2500 rpm for 10 minutes to precipitate the calcium oxalate and the absorbance was measured at 650nm. a standard curve was constructed using the absorbance readings obtained from the reference niacin solutions in place of the sample and this was used to extrapolate the niacin content of the samples. 2.14. cholesterol determination this was carried out using the liebermann-burchard reaction as described by kirk and sawyer [12]. a small quantity of the sample was added to 1ml of chloroform and this was stirred with magnetic stirrer. a combination of 40ml acetic anhydride and 2ml conc. h2so4 was added. this was later poured into distilled water and a green coloration was observed the absorbance of which was read with a spectrophotometer at 640nm. the concentration was extrapolated from a calibration curve. 2.15. color analysis the intensity of the color in the samples (lightness-l, redness-a, and yellowness-b) was determined using a colorimeter (chroma meter cr-410, japan). the samples were set in the sensor and the value was read from the reader. 2.16. texture analysis cylindrical samples of 10mm diameter and 12mm height were cut from the burger samples. the texture properties of the samples were determined using a texture analyzer (texvol tvt 300 ph, sweden). the texture parameters determined were: hardness, chewiness, cohesiveness and stringiness.. 2.17. sensory evaluation burger samples were subjected to sensory evaluation with a total of 20 trained taste panelists using a 7 point hedonic scale with 1 denoting ‘dislike extremely’ and 7 denoting ‘like extremely’. the following sensory properties were evaluated: texture, appearance, aroma, taste and overall acceptability. 3. result data are expressed in mean± standard deviation. table-2. proximate composition of beef, chicken and soy burgers in dry weight basis (dwb) samples protein (%) fat (%) ash (%) crude fiber (%) carbohydrate (%) bb 57.87a± 0.15 10.02a±0.15 5.11a±0.10 1.29c±0.58 25.71c±1.10 cb 50.36c± 1.25 10.15a±2.03 5.04a±0.15 1.80b±0.22 32.65a±0.25 sb 53.89b± 0.20 8.58b± 1.15 4.27b±0.21 2.88a±0.10 30.38b±0.36 note: data in the same column with different alphabets are significantly different (p≤0.05); bb-beef burger; cbchicken burger; sbsoy burger. bb was highest in protein content followed by sb and then cb. soy burger was least in ash and fat content but contained the highest quantity of crude fibre. table-3. vitamin and cholesterol composition of beef, chicken and soy burgers in dwb (mg/100g) samples thiamine riboflavin niacin cholesterol bb 0.98a±0.02 0.51b±0.05 1.04a±0.06 2.99b±0.15 cb 0.75b±0.15 0.33c±0.03 0.57c±0.10 3.58a±0.21 sb 0.13c±0.01 0.96a±0.06 0.85b±0.05 0.03c±0.01 note: data in the same column with different alphabets are significantly different (p≤0.05); bb-beef burger; cbchicken burger; sbsoy burger thiamine, riboflavin and niacin content of the three burger samples are significantly different. soy burger was highest in riboflavin and niacin content but contained the least quantity of thiamine. the cholesterol content of soy burger was negligible compared to that of beef and chicken burgers. table-4. colour properties of beef, chicken and soy burgers. samples l (lightness) a (redness) b (yellowness) bb 48.31b±0.39 18.45a±0.12 10.60c±0.80 cb 51.28a±1.28 10.58b±2.13 25.12a±0.59 sb 50.09a±1.34 9.54b±1.35 18.25b±0.34 note: data in the same column with different alphabets are significantly different(p≤0.05); bb-beef burger; cbchicken burger; sbsoy burger soy burger was not significantly different from chicken burger in lightness and redness while for yellowness beef burger had the least value followed by soy burger and then chicken burger. agriculture and food sciences research, 2018, 5(2): 57-63 61 table-5. texture properties of beef, chicken and soy burgers parameter bb cb sb hardness (n) 16.54a±0.64 17.71b±1.21 15.40c±1.14 chewiness (n.mm) 7.80a±1.21 7.12a±0.20 7.24a±0.68 cohesiveness 0.66a±1.23 0.62a±0.12 0.65a±0.24 springiness (mm) 0.81a±1.42 0.78a±0.45 0.72ab±0.25 note: data in the same row with different alphabets are significantly different (p≤0.05); bb-beef burger; cbchicken burger; sbsoy burger the three burger samples were not significantly different in chewiness, cohesiveness and but differed significantly in hardness and springiness. table-6. scores for the sensory properties of beef, chicken and soy burgers samples texture appearance aroma taste overall acceptability bb 6.43a±0.58 6.77a±0.06 6.80a±0.10 6.97a±0.06 6.90a±0.10 cb 5.60b±0.10 6.87a±0.12 6.10b±023 4.50c±0.10 5.07b±0.12 sb 6.07a±0.12 6.43a±0.06 6.15b±0.28 6.67b±0.12 6.37a±0.15 note: data in the same column with different alphabets are significantly different (p≤0.05); bb-beef burger; cbchicken burger; sbsoy burger soy burger was not significantly different from beef burger in texture and overall acceptability. in the same vein, soy burger was comparable to chicken burger in aroma while there existed no significant difference the appearance of the three burger samples. 4. discussion soy burger was significantly higher in protein content than chicken burger while beef burger had the highest value (table 2). since same ingredients were used in preparing the burgers, except the meat or meat alternative sources, the difference in composition may be attributed to the meat sources. raw lean beef was observed to contain 30.61% protein and 3.49% fat at 64.8% moisture which translates to 86.93% protein and 9.91% fat in dry weight basis [13]. in the same vein, lean chicken meat of the breast and thigh contained an average of 21.1% protein at 71.1% moisture content and this translates to 73.1% crude protein in dry weight basis [14]. soy bean on the other hand contained an average of 48.5% protein in high protein seeds and 41.5% in normal protein seeds at 15% moisture content thus translating to 57.05% protein in high protein seeds and 48.82% in normal protein seeds [15]. it can therefore be inferred from these that in dry weight basis the protein content of lean beef, chicken and soybean are 86.96, 73.01 and 57.05% respectively. however, in this study chicken burger had the least protein content while the highest value was observed in beef burger. this implies that tofu (soy cheese) is a rich in protein and this has been ascertained by past study [16] coupled with the fact that the soybean variety used in this study was of exceptionally high protein content. this may not be surprising since it is an improved variety. this may also be as a result of the lower protein content of the minced chicken used which may be traceable to genetic, dietary and other environmental factors. soy burger contained the least quantity of fat and ash while there were no significant differences in values for beef burger and chicken burger. the lower fat content of soy burger may be expected since this ultimately gave allowance for the high protein content as earlier discussed. soy burger was significantly higher than both beef and chicken burgers in crude fibre. this is quite expected since it is from plant source compared with the other two conventional burgers which are animal-based foods. chicken burger contained the highest quantity of carbohydrate followed by soy burger and then beef burger. the major common sources of carbohydrate in the burgers produced are wheat flour and bread crumbs. the proximate composition of beef burger observed in this study was in disparity with that of carvalho, et al. [17] who reported a protein, fat, ash and carbohydrate content of 40.23, 42.50, 8.94 and 8.23% respectively in dry weight basis. this may be as a result of variation in ingredients used. similarly, chicken burger was observed to contain in dry weight basis 29.91% protein, 52.61% fat and 7.48% ash [18]. the lower protein and higher fat content of this compared with that of the chicken burger observed in this study may be as a result of differences in the ingredients used. beef burger was highest in thiamine followed by chicken burger while soy burger was too low in this vitamin (table 3). on the other hand soy burger was significantly higher than beef and chicken burgers in riboflavin while the highest niacin content was observed in beef burger with chicken burger having the least value (table 3). the cholesterol content of soy burger was negligible compared with that of beef and chicken burgers. the differences in these nutritional composition, as earlier stated, was mainly attributed to the difference meat or meat alternative used since all other ingredients used were the same in type and quantity. beef was reported to contain 1.2mg/100g thiamine, 1.3mg/100g riboflavin and 16mg/100g niacin [19] chicken meat contained 0.09mg/100 thiamine, 0.16mg/100g riboflavin and 7mg/100g niacin [20]. the thiamine, riboflavin and niacin content of soybean were; 0.66, 0.22 and 2mg/100g respectively [20]. judging from these data, beef burger should contain the highest quantity of these vitamins followed by soy burger while chicken burger is expected to be higher in niacin than soy burger. however, the result observed in this study showed that beef burger contained highest quantity of thiamine and niacin only. this may be as a result of variation in the diet of the cattle used. unexpectedly, chicken burger was higher than soy burger in thiamine though the frmer contained the least quantity of riboflavin ad niacin as expected. the unexpectedly low thiamine content of soy burger may be as a result of loss of this vitamin with ‘whey’ water that was drained from the bean curds to obtain soy cheese. for color analysis soy burger was not significantly different from chicken burger in lightness and redness. beef burger had the highest value for redness (table 4) and least values for lightness and yellowness. this may be as a result of the higher myoglobin constituent of beef compared with chicken [21] and which is absent in soybean. this myoglobin commonly coverts to red oxymyoglobin on exposure to air and when cooked it is converted to brown metmyoglobin. agriculture and food sciences research, 2018, 5(2): 57-63 62 the color properties of beef burger observed in this study were different from that of past scientific report. carvalho, et al. [17] observed the lightness, redness and yellowness values for beef burger to be 44.7, 3.3 and 10.6 respectively. in the same vein, bestos, et al. [22] observed beef burger to have the following values for lightness, redness and yellowness respectively; 32.09, 5.70 and 4.46. the disparity of these from the observation in this study may be as a result of differences in ingredients used since only basic ingredients were used in this study in order to minimize the influence of confounding variables. variation in color analyzer used may also be responsible for this. on the other hand, the color properties of chicken burger in this study were in close proximity with that observed by uran and yilmaz [18] who reported values for lightness, redness and yellowness of chicken burger to be 50.29, 9.07 and 22.12 respectively. similarity in materials and method used as well as the colour analyzer may be responsible for this. all the three burger samples were similar in chewiness and cohesiveness (table 5). for chewiness, beef burger had the highest value followed by soy burger and then chicken burger while for cohesiveness followed similar trend. the three burgers produced were significantly different in hardness with chicken burger having the highest value followed by beef burger and then soy burger. springiness, which is the ability of the sample to resume its original shape when deforming force is exerted and removed, was observed to be highest in beef burger and least in soy burger. there existed no significant difference between beef burger and soy burger in this texture parameter. in overall, soy burger was comparable to beef and chicken burgers in texture parameters since there were close proximity of values even where significant difference existed. texture parameters from past findings are comparable to that observed in this study. carvalho, et al. [17] reported beef burger with hardness, chewiness, cohesiveness and springiness to be 15.38, 8.23, 0.69 and 0.78 respectively while in this study these parameters were 17.51, 6.58, 0.65 and 0.57 (table 5). the close proximity of the values observed in this study with that of the past study could be as a resultant effect of similarity in the methods used in the preparation of the beef burgers. for evaluation of sensory properties, the three burger samples did not differ significantly in appearance and were judged to be equally liked. for texture there was similarity in the likeness of beef and soy burgers and these were more preferred to chicken burger. the aroma of the three burger samples were liked but beef burger was most preferred while there existed no significant difference in chicken and soy burgers. the taste of beef and soy burgers were more preferred than that of chicken burger. in overall evaluation, beef and soy burgers were equally acceptable and more preferred to chicken burger. these show that soy burger can compete favorably in sensory properties with beef and chicken burgers 5. conclusion and recommendation soy burger is comparable to beef and chicken burgers in proximate composition and texture properties. it is higher in riboflavin and niacin than the two conventional burgers types with negligible cholesterol content. furthermore soy burger is similar to chicken burger in color attributes. it is more preferred than chicken burger in taste, texture and overall acceptability and is at par with beef burger in overall acceptability. soy burger can therefore compete favorably with beef and chicken burgers, hence, may be presented as a cholesterol-free healthier alternative suitable for all, even strict vegetarians. household and commercial production and consumption is hereby encouraged. references [1] a. mallik, "technology and security in the 21st century, a demen-side perspective in sipri (stockholm international peace research institute)," research report, no 20, oxford university press, new york, 2004. 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http://dx.doi.org/10.1111/j.1747-0080.2007.00197.x https://scholar.google.com/scholar?hl=en&q=effect%20of%20high%20pressure%20treatment%20on%20the%20colour%20of%20fresh%20and%20processed%20meats:%20a%20review http://dx.doi.org/10.1080/10408398.2017.1363712 https://scholar.google.com/scholar?hl=en&q=alternative%20fat%20substitute%20for%20beef%20burger:%20technological%20and%20sensory%20characteristics http://dx.doi.org/10.1007/s13197-013-1233-2 agriculture and food sciences research vol. 4, no. 1, 37-44, 2017 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2017.42.37.44 37 the role of institutional policies in promoting agribusiness development in rural china owusu samuel mensah1  zhuang jincai2 asare bediako isaac3 acheampong patrick4 ( corresponding author) 1,2,3,4school of management, jiangsu university, jiangsu, p.r. china abstract sustainable agricultural growth has become an area of interest for many researchers in the quest to increase food production in the midst of an escalating population. however, the evidence remains largely scanty, isolated and devoid of an in-depth analysis of how some economic policies promote agribusiness development in mainland china. using time series data from 1990 to 2013, this paper adopts semi-parametric quantile regression to study the complex relationship between institutional policies in rural areas and agribusiness development. the study revealed the role of family household in promoting agribusiness development at the higher quantiles of the distribution. moreover, government investment in rural health and education contributed significantly across the quantile distribution. the impact of research and development on agribusiness development is positively significant across the quantile points of the distribution within the study period. the result from the quantile graph clearly shows the disparities between ols and quantile regression coefficients. keywords: agribusiness development, institutional policies, quantile regression, rural china. citation | owusu samuel mensah; zhuang jincai; asare bediako isaac; acheampong patrick (2017). the role of institutional policies in promoting agribusiness development in rural china. agriculture and food sciences research, 4(1): 37-44. history: received: 29 september 2017 revised: 7 november 2017 accepted: 13 november 2017 published: 16 november 2017 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: for comments, suggestions and data collection, the authors would like to thank dr. emmanuel opoku marfo, li na, chen cong, li junqiang and liu jiayi. funding: national natural science foundation of china (nsfc) supported this research work, with grant number 7147310. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 38 2. related literature............................................................................................................................................................................ 38 3. data source and model specification .......................................................................................................................................... 39 4. conclusion ......................................................................................................................................................................................... 42 references .............................................................................................................................................................................................. 43 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2017.42.37.44&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=owusu%20samuel%20mensah https://orcid.org/orcid-search/quick-search?searchquery=zhuang%20jincai https://orcid.org/orcid-search/quick-search?searchquery=asare%20bediako%20isaac https://orcid.org/orcid-search/quick-search?searchquery=acheampong%20patrick agriculture and food sciences research, 2017, 4(1): 37-44 38 1. introduction the contribution of agriculture to the growth of the world’s economy has been a key policy issue in both less economically developed countries (ledc) and more economically developed countries (medc). currently, agriculture, which was previously perceived as a low-tech industry designed for small families and rural dwellers has changed dramatically due to liberalization of the economy and availability of agricultural markets [1, 2]. the contribution of agriculture sector to economic growth and development is manifested through the creation of agricultural and non-agricultural job opportunities and provision of food for both human and animal consumption [3-6]. currently, china’s agricultural sector is facing many challenges due to increasing urbanization and industrialization, which has decreased the productivity growth of agriculture. however, the large growth of human population and the need to increase food production has compelled the chinese government to introduce agricultural reform policies to remedy the situation [3, 7]. literature has revealed that the rural reforms instituted by china’s government have been the backbone of agricultural productivity growth [8-13]. according to pufahl and weiss [14] the impact of the central government programs on agricultural production and farm structure is a major issue in the international trade negotiation on agriculture. moreover, the introduction of the household responsibility system, adjustments of prices of major farm products, tax exemptions given to smallholder farmers, and public investments have contributed enormously to the growth of agricultural sector [12, 15-17]. for instance, with less than 8-10% of the world’s arable, china feeds about 20% of the world population. in 2008, the total grain production for the worldwide by china amounted to 18%, it then produced 27% of its meat and 43% of its poultry products. figure-1. agribusiness growth showing at different quantiles [18]. source: authors’ construct in addition, the domestic consumption of edible beans in china increased to 4.4 million tons in 2011, which was equivalent to 3.27kgper capital as compared to 9.9kgper capital in japan in 2011 [19]. the agricultural sector in mainland china, aside its continual food supply to the large populace, gives more than eight hundred million jobs to farmers dwelling in both rural and urban areas [20]. china’s agricultural sector has experienced rapid growth in other sectors such us livestock, fishery and forestry aside the crop-first agriculture. after 1978 reforms, the output growth of crop reduced from 82% in 1970 to 55% in 2010, whilst the fishery production also moved from 2% in 1979 to 10% in 2010 and forestry sector recorded 4% in 2010 [8]. for the past decade, there has been a rapid growth in the output of livestock and fishery production as compared to that of crop production due to dietary changes. the effect of the various institutional reforms in promoting china’s agriculture sector has been subjected to rigorous research [8, 12, 21-29]. in theory, the growth of agriculture sector is attributed to many factors, such as fertilizer application, the use of agricultural machinery, increase in agricultural and non-agricultural labor, and more importantly the introduction of household responsibility system, which was formally called production teams [30, 31]. the aim of this paper is to adopt flexible statistical techniques to investigate the effect of the various reforms on the growth of agribusiness in mainland china. surprisingly, few studies on this topic has employed semi-parametric regression model to study the complex relationship between rural policies and growth in agribusiness sector. however, we adopt quantile regression, which is the appropriate quantitative technique to measure the effect predictor variables on agricultural growth in rural china. the rest of the study is organized as follows. section 2 reviews the related literature. section 3 presents the source of data. section 4 describes methodology and model specification. section 5 summarizes empirical results and discussion of findings. section 6 provides concluding remarks. 2. related literature after successful rural reforms in china, the change from the commune system to the household responsibility system (hrs) in the late 1970s and 1980 has been a major contributor to agribusiness sector and agricultural productivity growth [17, 20, 25, 29]. according to huang, et al. [32] the implementation of household responsibility system increased agricultural production by 8.2% as compared to 2.7% before the reform period. agriculture and food sciences research, 2017, 4(1): 37-44 39 moreover, the study conducted by fan and pardey [28] also revealed that between the period of 1965 to 1993, the institutional reform policy contributed to the growth of agriculture by 17.6 percent. figure-2. the trend of family household and public investments in health, education and agricultural research from 1990-2013 [18]. source: authors’ construct moreover, government investment in agricultural research and development has resulted in the introduction of new varieties of crops, which has increased food production in rural china [33]. in addition, the massive economic reforms and rapid economic growth in china has increased government investment in the health sector, which has resulted in the provision of quality health services in rural china [34, 35]. according to literature, the monetary investments in education, which consist of in-school and out-of-school expenditure, which serve as a major aspect of human capital investment has improved the standard of education among farmers resulting into an increase in agricultural production [36]. most studies conducted in china’s agricultural sector have ascribed the rapid growth of agriculture to several factors. for example, mcmillan, et al. [37] investigated into the impact of china’s economic reforms and agricultural productivity growth. they established that price adjustment policies have increased the productivity growth of agriculture. wu, et al. [38] use a non-parametric malmquist procedure to investigate the temporal and spatial nature of agricultural productivity growth over the study period (1988-1995). the authors concluded that agricultural productivity growth in mainland china has increased through continuing innovation, the use of modern inputs and competitive market. in this line of research huang and ma [15] analyzed the determinants of public and farm household capital stock formation and their effects on the productivity growth of agriculture. by using cobb-douglas production function, the study reveals that capital investment in agriculture, private capital formation and technological changes have increased agricultural growth while fertilization application has had mild agricultural growth. [12] using province-level panel data investigated the effect of rural reforms on agricultural growth in rural china. by employing cobb-douglas production function, the author indicated that decollectivation and adjustment in state procurement prices have augmented the growth of agricultural production. lezin and long-bao [16] explored the impact of production variables used on agricultural productivity growth. however, using 1989-2002 data for zhejiang province and employing cobb-douglas production function, the study indicates that there is a strong correlation between the production variables and agricultural productivity growth. xu [17] used of cobb-douglas production function to synthesis the empirical literature on the subject chinese agricultural growth in post-reform era. the study reveals that there is a relationship between household responsibility system and agricultural productivity growth. li [39] adopted stochastic frontier analysis and employed 1995-2009 village-level data to investigate the effect of china’s agricultural reforms on productivity changes. the study disclosed that for the past fifteen years, chinese agricultural productivity has increased due to the institution of agricultural policies (such as tax exemptions, insurance for farmers), and technological changes. based on the related literature, an important aspect of measuring the productivity growth of agriculture, which is necessary but have been over-looked, is the estimation of the conditional median or the other quartiles of agricultural growth and not only the mere conditional mean. 3. data source and model specification 3.1. data source the data set consist of a country-data for 23 years observation over the period 1990-2013, which is obtained from china statistical yearbook ,various issues, 2015. the data on agribusiness development consists of a countryagriculture and food sciences research, 2017, 4(1): 37-44 40 data from the period of 1990 to 2013 obtained from china statistical yearbook, various issues, 2015. agribusiness performance (agb) is measured by total profit of agro-industries in rural china within the study period. family household refers to the total number of farmers in the various households from 1990 to 2013 in china. expenditure in education includes government appropriations for education, which consists of rural expenditure on education, taxes, and fees, collected for education by the government, which is measured in 10 000 yuan. agricultural research & development expenditure capture the public total expenditure on research and development in agricultural sector that is measured in 10 000 yuan. government expenditure in health is the previous and current expenditure by the government in the health of rural people measured in 100 million yuan. all the variables were converted into log form to avoid heteroskedasticitity, which will help us to estimate elasticity. table-1. variables definition variable definitions unit source agb agribusiness growth 100 million yuan china statistical yearbook hsd the total number of farmers in the various family households household china statistical yearbook edu government investment in rural education 10 000 yuan china statistical yearbook hlt government expenditure in rural health 100 million yuan china statistical yearbook r&d government expenditure in agricultural research and development 10 000 yuan china statistical yearbook note: all the variables are annual data from 1990-2013. data source: national bureau of statistics of china china statistical yearbook [18] 3.2. model estimation the quantile regression method proposed by koenker and bassett [40] performs regression analysis of sample data at different quantile points. thus conditional quantile estimates are performed by quantile regression with each nth quantile showing different behavior in the conditional distribution [41]. the quantile regression generalized median regression estimate to other quantiles. however, the use of traditional regression (ols) only pays attention to the mean effects, which may result in either over estimation or under estimation of coefficients and even fail to identify the actual relationships that exist between variables [42]. to investigate the role of institutional policies in promoting agribusiness development, the study adopts quantile regression (qr) over the ordinary least square model (ols) for the estimation. the least squares regression only measures how the changes in the vectors of the covariates affects the conditional mean function of . furthermore, ols regression is very sensitive to outliers, which may lead to inaccurate predictions due to the presence of multicollinearity, abnormal distribution and the independent nature of the residuals [43]. moreover, qr is invariant to monotonic transformation which is different from ols [40, 43]. in addition, qr is more robust to the non-normal error term and outliers in the model and takes into consideration the major effects of the covariates on the distribution of the dependent variable holistically and not only the conditional mean. however, this study employs the specifications of the previous literature on quantile regression. as stated by chen [44] the quantile regression generalizes the concept of a univariate quantile to a conditional given qauntile or many covariates and is modeled as follows; for a random variable y with probability distribution function ( ) ( )prf y ob y y=  , (1) the th quartile of y* is defined as the inverse function ( ) ( ) inf :q y f y =  , (2) where 0 1  . in particular, the median is ( )1/ 2q . according to chen [44] for random sample  1,..., ny y of y, it is known that the sample median minimizes the sum of absolute deviations 1 min d i r i y    − − . as compare to the general th sample quantile ( )  , which is analogue of ( )q  is donated by ( ) 1 min d i r i y     − − , where ( ) ( )( )0 ,0 1z z i z  = −    thus, ( ).i is the indicator function. chen [44] further argues that likewise the sample mean, which tries to minimize the sum of the squared residuals ( ) 2 1 1 ˆ arg min d i r y  − = − (3) is extended to linear conditional mean function as ( )e y x x x = = by solving ( ) 2 1 ˆ arg min d h i i i r y x   − = − (4) the linear conditional quantile function modeled as ( ) ( ) ,q x x x  = = is also estimated by solving ( ) ( ) 1 ˆ arg min d h i i i r y x    − = − (5) for any quantiles ( )0,1  .the, quantity ( )̂  represents the th regression quantiles. agriculture and food sciences research, 2017, 4(1): 37-44 41 the 0.5 = , denotes the median regression called the 1l regression, following chen [44]. 3.3. discussion of results the study, however, measured the effect of the dependent variables on the response variable by adopting quantile regression model. for effective comparison, we present the ols estimate, which serves as the baseline of mean effect and compare to the results of the selected quantiles in the conditional distribution of output produced in table 5. the estimation of quantile regression is reported in table 5 at 15th, 25th, 35th, 45th, 55th, 65th, 75th, 85th, and 95th quantiles. table 5 presents the estimation of a quantile regression model whereby family household and government investments in rural health, rural education and agricultural research and development are used to measure the agroindustry performance in china. from the results, the values of the parameters used for the estimation vary across ols and the selected quantiles both in magnitude and direction. for example, table 2 shows that the coefficient of family household in the 90th quantile is 24 times more than the ols estimate results. according to the coefficients of family household in table 2, the contribution of farmers in each household to the growth of agribusiness sector in the lower quantiles is negative. however, the coefficients of ols estimate to the lowest quantile (15th, 25th, 35th, 45th, and 55th quantile), are significant at the 10 % level. moreover, the influence of individual-based farming system in the various households on agro-industry growth increases at the higher quantile and significant at the lower quantile (fig. 3). this implies that an increase in farmers in the family household corresponds with the agribusiness development in the higher quantile of the distribution. in our opinion, this due to the technological changes in the agricultural sector, whereby farmers use simple farm tools to boost food production. in addition, land fragmentations, which is a major problem facing smallholder farmer, and increase in off-farm employment among the youth and the able-bodied people in the rural areas are contributing factors contributing to the poor performance of family household to the agricultural growth in the lower quantiles and ols estimate results. from table 2, the ols results indicate that a unit increase in family household reduces agricultural productivity growth by 2.62 percent. furthermore, the results of the study indicate some variations across selected percentiles in the conditional distribution of agribusiness growth. this shows the heterogeneity in the effect of rural health investments in agribusiness performance. the coefficient is significant and has a positive impact across the selected quantiles and ols estimates at 1% and 10% levels, whereby, the magnitude increases and then decreases toward the higher quantile (fig. 3). the results imply that, all things being equal, improve health status among rural farmers in china contributes greatly to the development in the agribusiness sector at both the ols estimate and the selected quantiles. this is due to increasing government spending in rural health, as a means of developing the human capital in the rural areas to increase food production. according to sckokai and moro [45] effective government policies promote agribusiness sector and agricultural growth and more importantly, create agricultural and non-agricultural jobs that increase farmers’ income level. report from chappell and lavalle [3] indicates that life expectancy increased from 67.77% in 1981 to 74.83% in 2010 and, there was a dropped in infant mortality rate (imr) from 32.89% in 1990 to 13.93% in 2012. table-2. institutional policies and agribusiness development: ols versus quantile regressions. (dependent variable is total profit of agro-industries from 1990 to 2013) coefficients selected quantiles ols q (.15) q (.25) q (.35) q (.45) q (.55) q (.65) q (.75) q (.85) q (.95) constant 29.29* 22.93 26.35* 36.42* 45.28 48.78 5.65 13.22 7.93 5.65 (1.48) (0.82) (0.87) (1.09) (1.38) (1.71) (0.11) (0.34) (0.17) (0.11) hsd -2.62 -1.92* -2.31* -3.35* -4.28* 4.63* 0.11 0.93 0.35 0.11 (1.29) (0.67) (0.67) (0.97) (1.26) (1.55) (0.02) (0.23) (0.07) (0.02) hlt 0.89*** 0.59*** 0.71*** 0.76** 0.88** 0.83** 0.87* 0.65* 0.75* 0.88* (4.32) (1.92) (1.96) (2.03) (2.01) 1.72 (1.12) (1.16) (1.01) (1.12) edu -0.07 0.17 0.11 0.1 0.42 0.94 0.24 0.36 0.08** 0.24** (0.33) (0.58) (0.33) (0.310 (0.12) (0.25) (0.43) (0.08) (0.16) (0.43) r&d 0.03* 0.01* 0.03 0.02** 0.26 0.02 0.02* 0.04** 0.02* 0.02* (1.48) (1.15) (2.56) (1.94) (2.43) (1.99) (0.68) (1.79) (0.88) (0.68) observation 23 23 23 23 23 23 23 23 23 23 r -square 0.9843 pseudo 0.9295 0.9157 0.9055 0.899 0.8943 0.889 0.8823 0.8789 0.8903 note: t statistics are in parentheses. *p< 0.10, **p< 0.05, ***p< 0.01 the per-capita health spending of urban area as compared to rural health investment was less than 2% in early 1990s, but increased to 2.67% in 2010. the 2.67% increased in health expenditure recorded in 2010 is attributed to the implementation of the new rural cooperate medical scheme (nrcms) and the current reforms [46-48]. this led to rapid increase in agro-industry performance in the agribusiness distribution. in addition to these, the contribution of rural investment in education to agro-industry performance is statistically insignificant across the selected quantiles but recorded positively at the ols estimate. table 2 clearly shows that the coefficients of the number of variables used differ across ols and the quantile levels. as demonstrated in figure 3, the impact of investment in rural education in agribusiness development across different quantiles is positive and its influence under 55th quantile is higher than the estimates above 90th quantile. the results of the ols estimate suggest that a percentage increase in rural investment in education decreases agribusiness performance by 0.07 percent. this supports why the study adopted quantile regression model to measure the effect at the various quintile points other than using only ols model. however, the contribution of public investment in education to the agro-industries performance across the selected quantiles recorded positively. for instance, from table 2 the output agriculture and food sciences research, 2017, 4(1): 37-44 42 growth at 55th quantile is greater than those in15th, 25th, 35th, 45th, 65th, 75th, 8th, and 95th quantiles. from figure 3, results for both family household and rural education give clear indications of the changes in the coefficients throughout the distribution of agribusiness development. the quantile regression shows significantly negative effect of household at 15th to 45th quantiles, however, the impact is positive at the upper quantiles of the conditional distribution. although, the results of family household contradicts the results of various studies, we will not make any policy conclusion from the results, but has to be thorough investigated. figure-3. the effects of family household (hsd), rural investment in education (edu), health (hlt) and agricultural r&d (rd) agribusiness development (abd): ols versus quantile regression. notes: the dashed lines represent ols parameter estimate, and the dark shaded areas are confidence intervals for the estimation of quantile regression parameters. data source: national bureau of statistics of china china statistical yearbook [18] in our view, the growth in the various quantiles from table 2 is due to improvement in the educational system in rural areas, which has gone a long way to improve the skills of agricultural entrepreneures in rural china. finally, for agricultural r&d, the results indicate that the impact of research and development on agribusiness performance is positively significant across the quantiles at 1%, 5% and 10% level. however, the effect on agribusiness development differs slightly, despite its positive effect on at all selected quantiles. for instance, the coefficient of r&d at 25th quantile is the same as compare to the coefficient in ols regression. according to ols results, an increase in entrepreneurial knowledge increases agribusiness growth by 0.03 % within the study period. moreover, the parameter estimate of the ols regression is quite lower than the estimate results in the 45th quantile but higher than the values for 15th, 35th, 55th, 65th, 85th, and 95th quantiles. this confirms study conducted by audretsch, et al. [49] which maintained that investment in new knowledge promotes productivity growth. the contribution of r&d to agribusiness development may attribute to high investment in research and development. according to liu, et al. [50] patent applications and grants in the agricultural sector has increased over the past decades, which has promoted agricultural production. 4. conclusion using time series data in pr china from 1990-2013, this paper measures the linkage between institutional policies and agribusiness development in china. that is the impact of government investment in education, health research and development and the introduction family households on agribusiness sector. the study however, adopted ols mean regression and compared with quantile regression because ols regression model only expresses the conditional mean in a linear form. the results indicate that the impact of rural investments on agribusiness development in china is quite heterogeneous. for instance, effects family household on growth of agribusiness sector across the selected quantiles are negative but significant at 15th, 25th, 35th, 45th, and 55th quantiles. however, agribusiness development continuously increases at 85th and 95th quantiles. the influence of rural investment in health on the agribusiness sector increases monotonically across the quantile distribution. for instance, the output growth in 15th and 75th quantiles are lower than 25th, 35th, and 85th quantiles but at the 45th, 55th, and 95th quantiles, health influence on agro-industry performance is stronger. this is due to the desire of the government to ensure the well-being of the people and promote human capital development through provision of effective health care system. moreover, the contribution of educational investment to agribusiness development at 55th quantile of the distribution is greater than the effect at the higher quantiles of the distribution. the difference in the output growth at different quantiles may be attributed to the inability of smallholder farmers to identify the various opportunities in the farming business and have access to the technological market. the reason is that most farmers in the rural areas still use traditional farm tools for farming activities and engage in subsistence farming. based on agriculture and food sciences research, 2017, 4(1): 37-44 43 the results from the quantile regression, we suggest that policy makers could be misled by using simple results achieved by ols regression to determine the effect of institutional policies on agribusiness development. this is because simple aggregations achieved by simple regression fail to indicate the existence of predictability in the exogenous variables. references [1] j. hansen, f. tuan, and a. somwaru, "do china's agricultural policies matter for world commodity markets?," china agricultural economic review, vol. 3, pp. 6-25, 2011. view at google scholar | view at publisher [2] g. mcelwee, "developing entrepreneurial skills of farmers. a literature review of entrepreneurship in agriculture." retrieved from http://www.esofarmers.org/documents/esofliteraturereview_000.pdf, 2005. 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shelmith waruguru mwaniki4 1,2,3department of plant science and crop protection faculty of agriculture; university of nairobi 4kenya agricultural and livestock research organization (kalro-kabete) ( corresponding author) abstract banana weevil is a pest with a great impact on banana production in the world. the objective of this study was to evaluate the pathogenicity of the kenyan entomopathogenic nematodes (epns) of genus steinernema against the weevil cosmopolites sordidus (germar) under laboratory conditions. adult weevils were trapped from banana fields in maragua, central kenya and larvae extracted from the infested banana rhizomes. the adults were treated with three kenyan epns steinernema weiseri, steinernema yirgalemense and the new steinernema spp and with steinernema carpocapsae all strain as a standard and plain distilled water as a control in five replicates. the treatments were 500ijs, 750ijs and 1000ijs per adult on petri dishes and 1000ijs, 3000ijs and 5000ijs per adult on pseudo-stems. the treatments for larvae in petri dishes were 300ijs, 400ijs and 500ijs. the adults were not susceptible to all the nematodes at all doses and preparations while larvae were highly susceptible to the four test epns at all concentrations. there was a significant difference at p<0.05 in the mortality of larvae between nematode steinernema carpocapsae and steinernema weiseri. a significant difference was observed between nematode steinernema carpocapsae and steinernema yirgalemense. there was also a significant difference between the new steinernema spp and steinernema weiseri and finally in the two nematodes the new steinernema spp and steinernema yirgalemense. all the test nematodes caused over 90 % larval mortality within 48 hours. the mean percent mortality of larva increased with nematode concentration for all the test nematodes. the results indicated that the four test epns significantly (p<0.05) caused high mortality within 24-48 hrs. the findings indicate that the banana weevil larvae are susceptible to the local entomopathogenic nematodes and potentially useful in the management of the banana weevil. it is however recommended that more research be conducted on the formulation and application technology to enhance their effectiveness in the field. keywords: banana, cosmopolites sordidus, management, entomopathogenic nematodes, steinernema spp. contents 1. introduction ............................................................................................................................................................................... 30 2. materials and methods ............................................................................................................................................................. 30 3. results ........................................................................................................................................................................................ 31 4. discussion ................................................................................................................................................................................... 34 5. conclusions and recommendations ........................................................................................................................................ 35 references ...................................................................................................................................................................................... 35 citation | moses mwaniki ndiritu; dora kilalo; john wangai kimenju; shelmith waruguru mwaniki (2016). pathogenicity of selected kenyan entomopathogenic nematodes of genus steinernema against banana weevil (cosmopolites sordidus). agriculture and food sciences research, 3(1): 2936. doi: 10.20448/journal.512/2016.3.1/512.1.29.36 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: contribution/acknowledgement: this work is licensed under a creative commons attribution 3.0 license all authors contributed to the conception and design of the study. we are grateful to the university of nairobi for offering us the opportunity to pursue the study and to the national council for science and technology for supporting the research work. we appreciate the assistance of the agriculture extension officer and farmers of maragua region of murang’a county for their support. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 11 may 2016/ revised: 30 may 2016/ accepted: 6 june 2016/ published: 10 june 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.29.36 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=moses%20mwaniki%20ndiritu https://orcid.org/orcid-search/quick-search?searchquery=dora%20kilalo https://orcid.org/orcid-search/quick-search?searchquery=john%20wangai%20kimenju https://orcid.org/orcid-search/quick-search?searchquery=shelmith%20waruguru%20mwaniki http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.29.36 agriculture and food sciences research, 2016, 3(1): 29-36 30 1. introduction entomopathogenic nematodes (epns) of the families steinernematidae and heterorhabditidae have been exploited for several decades as biological tools against many important insect pests in the world [1]; [2]. this has been made possible due to major advances in understanding the natural behavior of these nematodes [3]. they have also been studied intensively because of their ability to cause natural mortality of soil dwelling arthropods hence potential as biocontrol agents [4]. use of epn was part of integrated population suppression of pine weevil in the united kingdom [5]. they have been reported to control sweet potato weevils in india and kenya [6]. in addition, use of s.carpocapsae as a biological control against flat-headed root dwelling weevils in roots of apricot trees achieved 95% control [7]. many researchers in the world are working on these important biological controls [8]. of these, research on status of commercially available epns have been carried out intensively in north american countries and europe while in asian countries including china, korea and india the much stressed research work is on the use of epns to control insect pests and plant pathogens [8]. for most african countries epn research is still being carried out and in some countries non-existent. in developing countries more emphasis and interest is in the mutualistic relationship between the epns and bacteria hence the need to use them as biological agents for soil pests [8]. the epns are also commercially available in many parts of the world [9]. studies on the occurrence of epns in africa have been reported. the first record of both families was in a survey done in nigeria where h. bacteriophora and s. feltiae were reported [10]. a number of surveys have been documented showing new species and strains isolated from african countries. these have been found to have widespread abundance and are associated with types of habitats in south africa, kenya, ethiopia and egypt [11]; [12]; [13]; [14]; [15]; [16]; [17]. the epns were first reported in kenya in a survey conducted in the central highlands and coastal areas of kenya where a total of 154 nematode isolates among them the new species steinernema karii was identified [18]; [19]. further surveys in the rift valley yielded 12 nematode isolates (mwaniki, pers com.). currently 33 nematode isolates are maintained in three laboratories at kenya agricultural and livestock research organization (mwea, thika and kabete). the most studied genera are those that are useful in the control of insect pests, the steirnematidae and heterorhabitidae [20]. they have been identified in kenya with different species described in central highlands, rift valley and coastal areas [21]. the banana weevil is the major banana pest in wherever bananas are grown. it originated from south east asia [22] and it is now found in all banana growing regions including the new world, afro tropics, and oriental and australasian regions [23]. it came into the african continent from the south east asia through infested planting material and has since established in banana production areas within the continent such as nigeria, rwanda, democratic republic of congo, uganda, tanzania and kenya [22]. it is a major pest in east africa [24]. banana weevil is a coleopteran insect [23]. the adult weevil is nocturnal and very susceptible to desiccation and it rarely flies [22]. the newly emerged adult is red brown but turns black two to three days later [23]. it measures about 12mm, hard shelled and it has a pronounced snout. the larva is the most destructive stage [22]. they develop from the eggs and get into the corms and sometimes in the pseudo-stems making numerous tunnels. the tunneling interferes with root initiation and development, nutrient and water uptake there by weakening the plant leading to production of a bunch with less weight or eventual death [24]. this study was undertaken to evaluate the pathogenicity potential of the kenyan entomopathogenic nematodes (epns) of genus steinernema against the weevil (cosmopolites sordidus) (germar) under the laboratory conditions. 2. materials and methods 2.1. entomopathogenic nematodes laboratory investigations were carried out in the entomology laboratory of kenya agricultural and livestock research organization (kalro) kabete in nairobi. entomopathogenic nematodes were obtained from the entomology laboratory. multiplication of the nematodes was done by the in-vivo method where the insect-bait technique with galleria mellonella larvae described by poinar [25] and modified by woodring and kaya [26]; parra [27]. four nematode species were tested against the adult weevil and the larval stage of the banana weevil. these were the new steinernema sub spp, steinernema carpocapsae all strain, steinernema weiseri and steinernema yirgalemense. three selected epns steinernema sub spp, steinernema weiseri and steinernema yirgalemense had been isolated from kenyan soils. steinernema carpocapsae all strain was used for comparison purposes as it is a widely tested species. infective juveniles were kept in aqueous suspension in plastic containers with perforated lids at room temperatures and used in the experiment seven days after harvesting. 2.2. banana weevils adult of c. sordidus were trapped using pseudo-stem traps placed in banana orchards in maragua (0°46′59″ s and 37°07′59″ e) in central kenya. they were transferred to the laboratory, and maintained in perforated plastic buckets with moistened soil and pieces of banana pseudo-stems and corms. they were covered with opaque clothing and kept at room temperature. they remained until their use in experiment. every week, there was exchange of soil and both pseudo-stem pieces and corms. in addition, banana corms heavily infested with weevil larvae were obtained from banana orchards from maragua and transferred in plastic bags to the laboratory. weevil larvae were extracted the following day by opening up the corms that were characterized by holes due to infestation. they were carefully handled to avoid any injury or mortality before the assay was carried out. the extracted larvae were removed and immediately inoculated with nematode treatments. 2.3. pathogenicity tests procedures for inoculation and incubation were the same as those used for entomopathogenic nematode multiplication with galleria mellonella larvae [25] modified by woodring and kaya [26]; parra [27]. the only agriculture and food sciences research, 2016, 3(1): 29-36 31 difference was that this time adult and larval stages of the banana weevil were used instead of the normal last instar stage of wax moth. the design for all the bioassay experiments was completely randomized with five treatments, each treatment had five replicates and the experimental unit consisted of six petri dishes 9cm in diameter with 3 banana weevil adults or larvae. in the first bioassay experiment adult weevils were used as the target insect pests to test the laboratory pathogenicity of the selected entomopathogenic nematodes. four entomopathogenic nematodes were used as described above. five adult weevils were introduced in sterilized petri dishes per nematode species. filter papers were lined on the surface of petri dish to drain the excess water that carried the nematode suspension. inoculation was done by introducing aqueous suspension containing nematodes through sterilized pipette that was specific for each nematode treatment. each nematode treatment was replicated five times. three different nematode concentrations were used and they included: 500 ijs, 750 ijs and 1000 ijs per adult weevil in the petri dish and a control that contained plain distilled water only. this first experiment was repeated using the same procedure. after the inoculation, each petri dish was sealed with parafilm. the contents were then transferred to a dark chamber at room temperature. observations for mortality were made at 24 hours, 48 hours, 72 hours and 96 hours. the weevils were dissected under a light microscope to confirm nematode infection. a second bioassay with adult weevils was conducted using the same procedure but this time with a much higher dose of entomopathogenic nematodes. the dosage levels were 1000 ijs, 3000 ijs and 5000 ijs per adult weevil and plain distilled water was used as a control. observations were done as before. this bioassay was also done twice. a third bioassay with the adult weevil was conducted by using pseudo-stems which were laced with epns. small blocks of pseudo-stem of the same size were cut from a fresh banana pseudo-stem. each nematode treatment was applied to the small blocks using a hand pump. this was done all around to maximize the introduction of nematodes. sterilized plastic containers were used in place of petri dishes. they were lined up with filter papers at the bottom so as to drain excess water. the same nematode concentration was applied as in the second weevil bioassay. only one weevil was introduced per treatment with five replications. plain distilled water was used as the control per concentration level. the last bioassay involved the use of banana weevil larvae. the procedure was similar to the first bioassay that was conducted using the adult weevil. however, the number of larvae introduced per petri dish was three and the concentrations were 300, 400 and 500ijs per larval stage. nematode suspensions were introduced into the petri dishes through a sterilized pipette. plain distilled water was used as a control. each concentration level was replicated five times. the petri dishes were then closed and sealed with parafilm. they were then incubated at room temperature in dark chambers. the experiment was done twice. observations for mortality were made every 24 hours up to 96hours. a dead treatment was still with no response after brushing. cadavers infected with the nematodes were pale in color. cadavers were dissected under a light microscope to confirm presence of epns. the number of the dead larvae was recorded. 2.4. statistical analysis the mortality data was subjected to one and two way analysis of variance (anova). mean separation of the treatments that were significantly different at p<0.05 was done using the turkey-cramers comparison test. the analysis was done to compare the pathogenicity of the selected local entomopathogenic nematodes and test their effectiveness against banana weevil adult and larval stage. 3. results 3.1. pathogenicity of different entomopathogenic nematodes on the adult stage of the banana weevil under laboratory conditions adult weevils were not susceptible to all the four entomopathogenic nematodes at all concentrations in all the test arenas and time periods in the three bioassay experiments. 3.2. comparison of pathogenicity and virulence of selected entomopathogenic nematodes against the larvae of banana weevil under laboratory conditions all the test nematodes were pathogenic to the larvae of cosmopolites sordidus. the dead larvae got a soft consistency and a yellow-brown to black colour (pale), a characteristic of larvae killed by the bacteria of the genus xenorhabdus associated with steinernema species [28]. the mean mortality for each test epn differed though not significant in both the experiments which were carried out under the same conditions. the mean mortality for s. yirgalemense spp ranged from (2.00-2.20), new steinernema spp (2.40-2.55), s.weiseri spp (2.20-2.33) and steinernema carpocapsae spp (2.60-2.73). there was no mortality recorded in the control treatment in both experiments (figures 1 and 2). agriculture and food sciences research, 2016, 3(1): 29-36 32 (means with the same letter are not significantly different (p<0.05, turkey’s multiple range test). s.e.m -standard error of mean) figure-1. mean mortality of banana weevil larvae following 96hr of exposure to infective juveniles of selected epns in the one-on-one assay. source: kalrokabete there was a significant difference (p<0.05) in mortality of weevil larvae attributed to the three nematode species. the mean mortality of banana weevil larvae caused by s.carpocapsae spp was significantly higher (p<0.05) compared to s.weiseri spp and s.yirgalemense spp (figure 1). more so, a significant difference at (p<0.05) was observed between nematode new steinernema spp and s.weiseri spp; new steinernema spp and s.yirgalemense spp (figure 1). however, no significant difference (p>0.05) was observed between nematode s.carpocapsae spp and new steinernema spp (figures 1 & 2) s.weiseri spp and s.yirgalemense spp (figures 1) s.carpocapsae spp and s.weiseri spp; new steinernema spp and s.weiseri spp (figure 2). (means with the same letter are not significantly different (p<0.05, turkey’s multiple range test). s.e.m -standard error of mean) figure-2. mean mortality of banana weevil larvae following 96hr of exposure to infective juveniles of selected epns the one-on-one assay. source: kalrokabete s.carpocapsae spp was found to be the most pathogenic and virulent among the tested epns in both larval assays compared to other test nematodes as shown in figures 1 and 2 above. 3.3. comparison of banana weevil larvae mortality recorded at different time periods in an assay under laboratory conditions there were significant differences (p<0.05) between the exposure times of larvae to the test entomopathogenic nematodes as shown in figures 3 and 4 below. (means with the same letter are not significantly different (p<0.05, turkey’s multiple range test). s.e.m -standard error of mean) figure-3. mean mortality of banana weevil larvae recorded after exposure to infective juveniles of selected epns for a variable time periods. source: kalrokabete agriculture and food sciences research, 2016, 3(1): 29-36 33 (means with the same letter are not significantly different (p<0.05, turkey’s multiple range test). s.e.m -standard error of mean) figure-4. mean mortality of banana weevil larvae recorded after exposure to infective juveniles of selected epns for a variable time periods. source: kalrokabete all the nematodes tested caused a high mortality of banana weevil larvae within the first 24 hrs of exposure (figures 3 and 4) but except for s.carpocapsae (figure 3). the new steinernema spp caused significantly (p<0.05) higher mortality within 24hrs than the rest of the test nematodes. it took only 48 hrs for the new steinernema spp to cause more than 90% larval mortality. s.weiseri spp and s.yirgalemense spp took the longest time period to cause more than 90% larval mortality. however, at 48 hrs exposure time s.carpocapsae spp achieved a significantly higher (p< 0.05) mean mortality compared to other test nematodes (figures 3 and 4). 3.4. effect of ijs concentrations on the mortality of banana weevil larvae in a dose-response study under laboratory conditions in this study, the results indicated that the mean mortality of banana weevil larvae increased with increasing nematode concentration (300ijs, 400ijs and 500ijs) as shown in figures 5 and 6 below. (means with the same letter are not significantly different (p<0.05, turkey’s multiple range test). s.e.m -standard error of mean) figure-5. mean mortality of banana weevil larvae following inoculation with different concentrations of infective juveniles of the selected epns in a dose-response assay. source: kalrokabete (means with the same letter are not significantly different (p<0.05, turkey’s multiple range test). s.e.m -standard error of mean) figure-6. mean mortality of banana weevil larvae following inoculation with different concentrations of infective juveniles of the selected epns in a dose-response assay. source: kalrokabete agriculture and food sciences research, 2016, 3(1): 29-36 34 there was a significant difference (p<0.05) in larvae mortality between nematode concentrations 300ijs and 500ijs for the two nematodes new steinernema spp and s. weiseri. however, no significant (p>0.05) differences were observed between nematode concentrations 300 ijs and 400 ijs in all the test nematodes. no mortality was observed in control treatment. test nematodes s.yirgalemense spp and new steinernema spp did not show significant difference in all the three nematode concentrations in both bioassays. however, the trend in all the test nematodes was evident that after 96 hr of exposure the 500 ijs concentration caused the highest larval mortality (figures 5 and 6). 4. discussion in this study, it was evident that all epns tested were pathogenic to cosmopolites sordidus larvae and they caused different mortality rates under laboratory conditions. however, none of the test nematodes affected the adult stage of the banana weevil. although high concentrations of the test epns were used, the adult stage was not susceptible. this can probably be attributed to the heavily sclerotized body and pronounced long snout of the adult weevil [23] making it difficult for the ijs to penetrate inside the weevil. infective juveniles penetrate the host through the natural openings i.e. spiracles, mouth and anus or in some cases directly through the cuticle of certain insects [29]; [30]. according to trevellow and bedding [31] resistance is almost certainly due to difficulty of nematode entering the adult weevil than from establishment once infection is successful. once inside the insect host, they work symbiotically with bacteria carried in their guts. the latter multiply releasing a number of virulence factors. these are toxin complexes, hydrolytic enzymes, hemolysins and anti-microbial compounds that cause mortality of insects within 2448 hours [29]; [31]. a study done by sirjusignh, et al. [32] demonstrated similar results that local caribbean epns were not effective on the adult weevil under laboratory conditions. furthermore, the introduction of fresh pseudo-stem laced with nematode at different dose rates was to maximize the contact between the adult weevil and the nematodes in the process of feeding. adults are attracted to the volatiles emanating from the fresh or decomposing pseudo-stems [33]. however, no mortality was recorded. in tanzania, a study carried out by mwaitulo, et al. [15] reported similar results of resistance of the adult weevil to locally isolated epns from banana fields in three regions of morogoro, mbeya and pwani. all test epns caused larval mortality. these results are consistent with a study conducted in tanzania by mwaitulo, et al. [15] showed that all local epn isolates were pathogenic to banana weevil larvae hence causing mortality. the test nematodes caused different mortality rates of the weevil larvae. the dead larvae got a soft consistency and a yellow to brown to black colour, characteristic of larvae killed by bacteria of the genus xenorhabdus [28]. s. carpocapsae caused higher mean mortality compared to the rest of the test nematodes. this can be attributed to the specificity in pathogenicity and virulence of individual epn isolate. in a study by koppernhoofer and fuzy [34] noted that differences in the pathogenicity of infective juveniles can be attributed to their foraging strategy, the responsiveness of the host immune system, the pathogenicity of the symbiotic bacteria and the number of bacterial cells transported by dauers. this explains why in this particular study, the test nematodes caused different mortality rates. a report by koppernhoofer and kaya [35] indicated that steinernema dauers can carry different amounts of bacterial cells in their intestines. for instance, the species s. scapterisci contains a small amount of bacterial cells compared to s.carpocapsae that has a large number of bacterial cells, causing higher pathogenicity but at the expense of lower survival in the environment [36]. all tested nematodes caused more than 90% mortality of the larvae. in a study conducted in canary island, a 100% mortality banana weevil larva was reported using indigenous epn (heterorhabditis and steinernema species). previous authors have reported susceptibility to epns among larvae of other beetles such as diaprepes abbreviatus (coleoptera: curculionidae) [37] capnodis tenebrionis [38] and premnotrypes sutulicallus [39]. the mortality of banana weevil larvae in this study could be attributed to the physical appearance (soft and fleshy cuticle, abdominal segments) that enabled epns to penetrate easily alongside the mouth and anus. previous histopathological studies have shown that ijs enters the larvae through the cuticle and less often through the anus and mouth [40]. from the results in this study all the test nematodes caused a significant high mean mortality rate of larvae within the first 24-48 hours of exposure. this demonstrates the high virulence and infectivity potential of the local epns tested against the banana weevil larval. the mortality was confirmed to be due to epn infection through dissection of the cadavers to check for the presence of ijs in the haemocoel. entomopathogenic nematodes kill the host by inducing septicaemia within 24-48 hours of infection [41]. a dose-response assay showed no significant difference in larval mortality between 300ijs and 400ijs concentrations. 500 ijs concentration for all the test nematodes caused larval mortality and was different to 300ijs. this can probably be attributed to the numbers. it is said there is strength in numbers and hence the higher concentration of ijs showed high virulence and pathogenicity differences in the epn species, humoral and immune response of the host [41]. although there was no consistency across all the three ijs concentrations, there was a positive relationship that weevil larval mortality increased with increasing concentrations. larval mortality observed for s.carpocapsae and s. yirgalemense did not differ in the mortality rates caused to the larvae based on concentration. the rest of the tested nematodes did not differ in causing mortality at the two concentration levels of 300ijs and 500ijs. this indicates the importance of dosage rates. the results partially concur with a study conducted by mwaitulo, et al. [15] who reported that the banana weevil mortality increased significantly with increasing native nematode dosage. however, these same results differ with a study conducted by padilla-cubas, et al. [42] that showed an increase in ijs dosage does not necessarily cause an increase of banana larvae mortality. a dosemortality response was observed by schmitt [43]. previous studies have reported significant differences based on epn agriculture and food sciences research, 2016, 3(1): 29-36 35 concentration and larvae stage mortality in other insect pest orders. a study conducted in turkey showed that native epn ijs concentration (0,100, 500 and 1000ijs) had differences in causing mortality of p. operculella larvae [44]. 5. conclusions and recommendations the tested kenyan epns were able to parasitize cosmopolites sordidus larvae but not the adult under laboratory conditions. all the nematodes tested caused above 90% banana weevil larval mortality. nematode species and the nematode concentration levels showed differences in efficiency against the weevil larvae. in addition all tested nematodes caused high mortality within 24-48 hrs of exposure indicating a high susceptibility of banana weevil larvae to the tested nematodes. these tested nematodes have potential for field management of the banana weevil since epns are known to search and locate susceptible hosts in cryptic habitats like within the banana pseudo-stem and corms. larvae are found within the small holes on the surface of the rhizome, which are in contact with exterior. they are also found living in long galleries inside the rhizomes which are ideal conditions for epns survival and infection of hosts. the larval stage is the most destructive stage of banana weevil and if targeted for control through epn application would lead to a reduction of banana weevil damage hence higher banana yields. the most effective epn was s.carpocapsae and can be used at 500 ijs concentration level in the laboratory to kill the weevil larvae future work is necessary to test more native nematodes for efficacy, appropriate formulations and application techniques in banana fields to enable the adoption epns against the banana weevil larvae in the field. in addition, further research in epn storage is needed for longer nematode life span. better storage and application technology of epns will enhance the adoption of epns for the management of banana weevil larvae and in the integration of entomopathogenic nematodes in pest management programs. references [1] r. georgis, a. m. koppernhoofer, and l. a. lacey, "successes and failures in the use of parasitic nematodes for pest control," journal on biological control, vol. 38, pp. 103–123, 2006. 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[44] i. kepenecki, a. tulek, m. alkan, and s. hazir, "biological control potential of native entomopathogenic nematodes against potato tuber moth (pthorimea operculella (zeller) (lepidoptera: gelechiidae) in turkey," pakistan journal of zoology, vol. 45, pp. 14151422, 2013. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 3, no. 1, 1-11, 2016 http://www.asianonlinejournals.com/index.php/aesr 1 sweet potato agronomy research in ethiopia: summary of past findings and future research directions daniel markos1  gobeze loha2 1,2 awassa agricultural research center, awassa, ethiopia ( corresponding author) abstract sweet potato (ipomoea batatas (l.) lam) is economically important food crop in ethiopia. since its introduction, numerous agronomic research activities were carried out in agricultural research centers, non-governmental organizations and universities. the objective of this piece of work is to document available research findings in a usable manner and present it to first national sweet potato workshop held between 6-7 june 2013, sari head quarter, awassa, ethiopia. agronomic research has been carried out on preparation of planting materials, curing vine cuttings, planting methods, depth of planting, plant density, planting time, cropping systems, and etc. results showed that sweet potato crop has a potential of 50 to 60 t/ha in ethiopia and the length of vines to be used for planting shall be 30 to 40 cm. weeding trials elaborated that weeding twice on 30-40das and 70das regardless of the study sites would suffice for the crop. planting date experiments justified early planting with the onset of rainfall in non irrigated fields. however, the results of population density and fertilizer regimes varied across locations and varieties. thus generation and promotion of site specific recommendations are of paramount importance for root crops like sweet potato. keywords: ipomoea batatas, ethiopia, agronomy, cultural practices, varieties and research findings. contents 1. introduction ...................................................................................................................................................................................2 2. materials and methods .................................................................................................................................................................2 3. results and discussion .................................................................................................................................................................2 4. conclusion .....................................................................................................................................................................................9 5. future research directions .......................................................................................................................................................10 references ........................................................................................................................................................................................10 citation | daniel markos; gobeze loha (2016). sweet potato agronomy research in ethiopia: summary of past findings and future research directions. agriculture and food sciences research, 3(1): 1-11. doi: 10.20448/journal.512/2016.3.1/512.1.1.11 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: contribution/acknowledgement: this work is licensed under a creative commons attribution 3.0 license all authors contributed to the conception and design of the study. especially thanks go to awassa, areka, adami tulu and melkasa research centers and haramaya and wolita sodo universities for giving due attention to sweet potato research at national level. heart felt thanks also go to cip for sponsoring the first sweet potato national workshop held on 6-7 june 2013 at sari (southern agricultural research institute) head quarter and also the organizing committee for giving us this opportunity to present the findings. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 12 january 2016/ revised: 23 january 2016/ accepted: 29 january 2016/ published: 4 february 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://creativecommons.org/licenses/by/3.0/ http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.1.11 https://orcid.org/orcid-search/quick-search?searchquery=daniel markos https://orcid.org/orcid-search/quick-search?searchquery=gobeze loha http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.1.11 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.1.11 https://orcid.org/orcid-search/quick-search?searchquery=daniel markos https://orcid.org/orcid-search/quick-search?searchquery=gobeze loha http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.1.11 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.1.11 https://orcid.org/orcid-search/quick-search?searchquery=daniel markos https://orcid.org/orcid-search/quick-search?searchquery=gobeze loha http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.1.11 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.1.11 https://orcid.org/orcid-search/quick-search?searchquery=daniel markos https://orcid.org/orcid-search/quick-search?searchquery=gobeze loha http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.1.11 agriculture and food sciences research, 2016, 3(1):1-11 2 1. introduction sweet potato (ipomoea batatas (l.) lam) is a herbaceous dicotyledonous plant with creeping, perennial vines and adventitious roots. it belongs to family convovulaceae (morning glory flowers) and is hexaploid, and usually considered the only ipomoea species of economic importance. it has large, starchy, sweet-tasting and tuberous roots. it adapts tropical and warm temperate regions. it is a highly heterozygous cross pollinated crop in which many of the triats show continuous variation. this crop is known for its resistance to drought, vigorous early growth and low input requirements. it also does well in areas of high rainfall and it requires very little labor and care compared to other crops. because it readily produces adventitious roots and has trailing vines, sweet potato can colonize soils easily. sweet potato is grown in several agro-ecological zones and usually plays significant roles in the farming and food systems. sweet potato is commonly grown by farmers in complex, mixed cropping systems where they normally plant several varieties with different characteristics (yield, maturity, palatability, time to maturity, root size and shape, root colour, storability in the ground, pest and disease tolerance, drought tolerance, and sweetness) in a single plot. farmers may use the vines left in the fields to improve soil fertility, and the crop is used in crop rotation. since it has a short growing period, it stores well in the soil and performs well in marginal lands. it is recognized as ideal crop for food security. the yellow and orange-fleshed sweet potato varieties are also known as a good source of vitamin a that is frequently lacking in diets of most african farming communities. however, most varieties in subsaharan africa are white-fleshed, low yielding and lacking beta-carotene, the precursor of vitamin a that was found vital to pregnant women and children. also, sweet potato is widely used as animal feed (assefa, et al. [1]; engida, et al. [2]). globally, sweet potato is the seventh most important food crop and second most important tuber crop in the world after irish potato. it ranks seventh among all food crops worldwide, with an annual production of 124 million tons. among the root and root crops, it ranks third in acreage (9.1 million ha) behind irish potato and cassava. in africa, sweet potato is the second most important root crop after cassava and production is concentrated in the east african countries around lake victoria (dantata, et al. [3]; ndole, et al. [4]). sweet potato productivity is limited by both abiotic and biotic constraints, leading to poor yields at farm level. they include low soil fertility and drought, shortage of improved varieties, shortage of planting materials, pests and diseases particularly viruses, post-harvest problems such as storage, and market availability and demand as well as low socioeconomic status in some communities. as a food security crop, it can be harvested piecemeal as needed, thus offering a flexible source of food and income to rural households that are mostly vulnerable to crop failure and consequently fluctuating cash income. in addition to being drought tolerant and having a wide ecological adaptation, it has a short maturity period of three to five months. sweet potato has several advantages within the context of african cropping systems: i) it produces food in a relatively short time, ii) it gives reliable yields in sub-optimal growth conditions, iii) it requires lower labor inputs (appropriate for vulnerable households) than other staples, vi) it serves as an alternative food source for urban populations, facing increasing prices of cereals and v) it provides a potential option to reduce vitamin a deficiency (andreas, et al. [5]; getahun and tenaw [6]). sweet potato has a potential of giving over and above 50 to 60 tons/ha in ethiopian conditions; however, yield obtained from farmer’s field is by far lower than 6 to 8 tons. thus the yields are ten times lower than the potential sought. this huge variation is attributed to biotic and abiotic stresses, lack of improved varieties, and weak attitude of people toward sweet potato, inefficient means of sweet potato technology transfer, inadequate, etc set of package recommendations. 1.1. objectives to summarize research findings in cultural practices, cropping systems and physiology of sweet potato in ethiopia. to recommend appropriate site specific management practices in sweet potato agronomy for future use to set future research directions in sweet potato management 2. materials and methods identification of major growing areas in the country was carried out initially from secondary sources. nextly available published and unpublished research findings and best practices were sought from universities, research centers and non-governmental organizations like farm africa, jica and etc. working in major sweet potato growing areas like sidama, wolaita, harargie, bako, etc... personal communication with growers, seed multipliers and researchers who made a research in the past was carried out to confirm their findings. available information was assembled into categories of cultural practices and cropping systems by pictures, tables and summary notes. however, by no means the results are complete and exclusive of all observations and achievements in the country. finally results were summarized in usable form by providing appropriate citation for future referencing. 3. results and discussion 3.1. preparation of vines for planting results of various studies showed that sweet potatoes are propagated using seeds, vines and roots in ethiopia. the use of seed is mainly for breeding purpose and not for commercial production. if the crop is to be grown from the root, then sets should be chosen from healthy robust marketable tubers. the most common method of planting sweet potatoes in commercial production is exclusively using vines mainly because vines are free from soil borne diseases, provide greater yield besides provision of uniform roots. the crop has a great deal of foliage; however, it has low multiplication rate compared to cereals. the vines are those parts of sweet potato that do not die after the roots are mature. however, there is shortage of preservation methods of these planting materials over the long dry season. hence there is inability to meet large and regular demand for planting coming from governmental and nongovernmental organization particularly in food insecure periods of the year. preparation of planting material involves http://en.wikipedia.org/wiki/starch http://en.wikipedia.org/wiki/tuberous_root agriculture and food sciences research, 2016, 3(1):1-11 3 removing leaves, cutting the vines to appropriate size and curing them accordingly. these reduce moisture loss and bulkiness (fig 1). fig-1. preparation and packing of sweet potato planting materials to compare and evaluate the yield performance different parts of the sweet potato vines used as planting material (top, middle and basal parts), experiments were conducted at awassa and areka. although it was not statistically significant, the result indicated that top vine part gave higher root yield compared to the middle and basal portion of the vine cuttings. however, in case of low moisture conditions, the top portion easily wilts, as it is delicate. top part is relatively free from weevil and other diseases compared to basal parts (crct (crop science research department) [7]). therefore, it was recommended to plant top and middle portion of sweet potato compared to basal portion. vine pre rooting treatments were carried out to solve problem of establishment and yield obtained from vine parts. they have to be rooted under ideal condition before transferring to the field. some farmers employ pre rooting to replace the dead vines in the field and minimize the time gap with the first planted crops (geleta [8]). table-1. mean tuber yield (tha-1) of pre rooted vines tested for three years pre rooting time (days) 1990 1991 1992 mean 0 4.5a 9.1b 8.8c 7.4b 3 4.5a 15.9a 12.2b 10.8a 6 4.2a 16.0a 15.8a 12.0a 9 4.9a 16.6a 14.5a 12.0a vine pre rooted for 6 and 9 days gave better root yield than 0 and 3 days. both pre-rooting days were found to be consistent and better in root yield in all years (table 1). 3.2. curing of vine cuttings an experiment was conducted at awassa to compare establishment and root yield of sweet potato obtained from vine cuttings which were stored under shade for different length of time (0, 2, 4, 6, 8 and 10 days). the three years result of the experiment indicated that storing vines for two days could enhance better root initiation, increase vine establishment and root yield of sweet potato compared to others levels of vine curing. vines stored for longer period dried off prior planting. hence vine storage for two days before planting is recommended for better establishment and root yield (crct (crop science research department) [7]). 3.3. length of vine cuttings based on the studies carried out a vine having 30 to 40 cm length with 3 to 4 buds on the cutting are preferred for better yield and ease of management in comparison to using whole vine. 3.4. depth of planting comparisons were made on depth of planting the vine cuttings. results confirmed that burying two-third of the vine would suffice for better establishment and subsequent yield compared to other levels of burying the sweet potato cuttings. 3.5. vine treatment with insecticides the older sweet potato vines and roots harbor sweet potato weevil (cylas puncticollis). therefore it is recommended that all the cuttings should be dipped in the fentain or fentrotion insecticide solutions at the recommended rate just before planting in order to kill the egg or adult weevil (endale, et al. [9]). 3.6. land preparation for root production according to the studies carried out by national research coordination unit, land for sweet potato production should be ploughed thoroughly to 15-20 cm depth and could be prepared using either tractor drown implements, oxen drown plows or by hand tools. the land prepared could be flat as in wolaita area and could be of ridges as in hararige. in fact in research centers like that of awassa, ridges are prepared by tractor drown implements. series of studies has been carried out to explore the comparative advantage of these land preparation methods with consideration of different varieties. the researchers concluded that since the varieties had no significant response to methods of land preparation, any of the cost effective methods could be employed (table 2 and 3). however, flat planting is beneficial in areas where the amount of rainfall is sufficient and the soil is more of sandy loam. in areka, where the rainfall was higher and water-holding capacity of soil is good, flat planting produced significantly higher agriculture and food sciences research, 2016, 3(1):1-11 4 root yield of sweet potato. but, generally in moisture stressed areas, tie ridges are the most universally recommended methods of growing sweet potato (crct (crop science research department) [7]). table-2. effects of land preparation methods on root yield (t/ha) of sweet potato varieties in awassa land preparation methods koka -6 koka-12 1987 1988 1989 mean 1987 1988 1989 mean flat 15.7 16.2 12.9 14.9 17.3 14.7 7.1 13.0 open ridge 14.8 16.7 12.9 14.8 18.1 17.2 8.2 14.5 tied ridge 16.3 14.8 12.7 14.6 17.5 18.2 7.0 14.2 source: endale, et al. [9] table-3. effects of land preparation methods on root yield (t/ha) of sweet potato varieties in areka land preparation methods koka -6 koka-12 1987 1988 1989 mean 1987 1988 1989 mean flat 9.5 11.2 10.4 10.4 12.4 7.0 9.1 9.5 open ridge 11.4 10.8 10.6 10.9 11.5 8.7 6.8 9.0 tied ridge 9.3 9.5 11.4 10.1 9.9 18.2 6.7 11.6 source: endale, et al. [9] 3.7. land preparation methods and production of herbage for animal feed research conducted at afar (melkaworer agricultural research center) using koka-12 variety consisted of three planting methods (ridge, flat, and sunken), and three vine harvesting times (45, 75 and 105 days after planting including one control treatment without vine harvesting). the experiment was laid out in a randomized complete block design in a factorial arrangement and replicated four times. the results revealed that planting sweet potato on ridges and harvesting the vines 105 days after planting (when about 60% of the growth phase of the plant was completed) led to optimum production of herbage for fodder without compromising yield of tuberous roots. thus planting sweet potato on ridges and harvesting the vines when about 60% of the growth phase of the plant was completed enhanced the production of herbage for animal feed without compromising yield of tuberous roots to be used for human food. therefore, pastoralists as well as sedentary smallholder farmers in ethiopia who produce sweet potato using furrow irrigation should plant the crop on ridges and harvest the vines for animal feed at a later stage of growth by which time tuberous roots may have grown and bulked sufficiently. the farmers should also plant several adaptable cultivars having similar or different maturity times to spread vine harvests throughout the season for ensuring sustained availability of food for humans and fodder for animals (mohammed, et al. [10]). 3.8. ridge height and furrow width according to findings, ridge width and height happened to be researchable. research recommendations showed that medium to high ridges (25 to 36 cm) are advantageous on poorly drained soils when there are heavy rains, and flat or low ridges are practiced where drainage is good and rainfall is not very high (endale, et al. [9]). in tractor plown farms, furrows are spaced 75 cm apart and plants are placed 30cm away from one another. 3.9. planting date studies planting date experiments were carried out in awassa, areka and bako using varieties called koka 6 and koka 12. results indicated that planting could be carried out between june 4 to july 7 in bako areas. this holds true to similar agro ecologies to that of bako [11]. in awassa, early planting on 13 may 1987, 4 may 1988 and 16 may 1989 gave highest mean root yield of 28.4t/ha, 39.2t/ha and 30t/ha, respectively. thus in awassa and similar agroecological areas the onset of rainy season particularly mid may to early june was recommended as optimum unlike early june planting of bako (crct (crop science research department) [7]). 3.10. planting positions studies on planting positions were carried in areka and awassa from 1987-1989 comparing inclined (slant), vertical, u-shaped and horizontal planting positions in relation to different varieties. results indicated that none of the varieties responded significantly (p<0.05) to varying positions of planting both in areka and awassa (table 4 and 5). at awassa, planting cuttings at slant (inclined) position gave higher root yield compared to horizontal plantings. this was due to more vigorous establishment and better contact of soil and the planting material. however, in dry season horizontal planting is advantageous compared to slant planting. similarly during wet seasons, slant planting is preferred compared to horizontal planting in awassa. in harargie vines are planted in inclined positions where plantings are done on ridges. in wolaita, vine cuttings are buried horizontally in furrows when the land is prepared by oxen drown implements. in other parts of the country vertical and inclined positions predominate when planting is done on flat or ridges. table-4. effects of planting positions on root yield (t/ha) of sweet potato varieties in awassa planting positions koka -6 koka-12 1987 1988 1989 mean 1987 1988 1989 mean inclined 22.2 18.7 15.9 18.9 28.3 15.5 15.9 16.6 vertical 19.2 15.3 15.5 16.6 23.0 18.1 6.8 16.6 u-shaped 21.3 19.2 11.3 17.2 25.2 15.1 10.1 16.9 horizontal 24.9 9.9 7.3 14.0 15.3 11.5 6.3 11.1 source: endale, et al. [9] agriculture and food sciences research, 2016, 3(1):1-11 5 table-5. effects of planting positions on root yield (t/ha) of sweet potato varieties in areka planting positions koka -6 koka-12 1987 1988 1989 mean 1987 1988 1989 mean inclined 9.2 6.4 9.0 8.2 13.2 5.5 9.3 9.3 vertical 10.3 6.0 7.6 7.9 12.4 6.8 8.8 9.3 u-shaped 9.7 6.0 9.7 8.5 13.3 5.1 9.5 9.3 horizontal 7.5 5.8 6.6 6.6 15.5 1.3 6.8 7.8 source: endale, et al. [9] 3.11. plant density experiments in jido combolcha as shown in the table below (table 6) values of total tuber yield of balella variety with spacing 20 cm x 80 cm performed better (644 ± 105 q ha -1 ) than the rest of the treatments 20 cm x 60 cm and 50 cm x 60 cm, with the yield value of 590 ± 104 q ha -1 and 522 ± 137 q ha -1 , respectively. but the net marketable yield obtained at spacing combination of 20 cm x 60 cm (590 ± 104 q ha -1 ) was by far better followed by 20 cm x 80 cm and 50 cm x 60 cm that gave average yield of 583 ± 82 q ha -1 and 463±93 q ha -1 , respectively. table-6. result of mean and standard error comparison of balella variety on farm in jido combolcha variable root diameter (cm) root length (cm) green top (kg) marketable root (q/ha) unmarketable root (q/h) total yield (q/ha) df 2 2 2 2 2 2 20 x 100 7. 2±.66a 15±3.4ab 3e4±8e3ab 438±110abc 42±21a 479±116abc 20 x 60 5.5±.25bcd 13±.62ab 2e4±1e4ab 590±104a 0±0 a 590±104a 20 x 80 4.9±.1d 14±2.2ab 3e4±2e4ab 583±82a 61±54 a 644±105a 30 x 100 6.1±.25abcd 15±2.2ab 4e4±4e4a 384±221bcd 65±89 a 449±309abc 30 x 60 5±1.1d 12±2.6ab 2e4±9e3ab 332±74bcd 12±21 a 344±63bc 30 x 80 5.6±.63bcd 14±2.2ab 2e4±1e3ab 288±1.3cd 12±21 a 300±110bc 40 x 100 7±1.5ab 15±1.4ab 1e4±5e3ab 285±43cd 57±81 a 323±68bc 40 x 60 5.3±.5d 14±1.2ab 1e4±5e3ab 324±36bcd 12±20 a 336±27bc 40 x 80 6±1.4abcd 14±1.8ab 1e4±5e3ab 278±54cd 22±20 a 299±57bc 50 x 100 5.4±0.29cd 16±3.6a 4e4±4e4ab 294±77bcd 28±17 a 322±83bc 50 x 60 6.9±.17abc 13±2.8ab 2e4±1e4ab 463±93ab 88±124 a 522±137ab 50 x 80 6.1±1.3abcd 15±.91ab 1e4±8e3ab 233±47d 28±32 a 260±38c cv% 13.51 15.13 66.55 24.3 136.9 28.12 *mean of the same letter across the column indicates non-significant difference among the treatments (p<0.05) (teshome, et al. [12]) it is clear form table 7 that the general yield performance of the bareda (tubers horizontal vines spreading) variety was much lower than that of yield obtained by balella (tubers vertical vines erect) in jido combolcha area. in general 20 cm x 60 cm spacing gave total yield of 409±257 q/ha followed by 20 cm x 100 cm and 30 cm x 60 cm that gave 347±139 q/ha and 294 ± 63 q/ha, respectively. among all the treatments 40 cm x100 cm gave the lowest performance (129±69 q/ha) followed by 50 cm x 100 cm (144 ±34 q/ha) and 30 cm x 80 cm (185±111 q/ha). it is indicative of the fact that if spacing between plant and rows increases beyond provision of enough nutrients, the net number of vine decreases resulting into lower yield, since the net number of vine is positively correlated with yield obtained at the end of the day. therefore, 20 cm x 60 cm between two consecutive plants and rows should be adopted regardless of tuber and vine orientation for those farmers involved in sweet potato production in jido combolcha and similar agro-ecologies (frg [13]). table-7. result of mean and standard error comparison of bareda variety on farm in jido combolcha variable root diameter (cm) root length (cm) green top (kg) marketable root (q/ha) unmarketable root (q/h) total yield (q/ha) df 3 3 3 3 3 3 20 x 100 4.3±1a 19±1.3 a 3e4±3e3bcd 322±115ab 25±27a 347±139ab 20 x 60 4.2±0.91a 19±1.9 a 3e4±2e4ab 396±267a 13±11 a 409±257a 20 x 80 4.4±102a 17±2.6 a 4e4±1e4a 220±3.7abc 41±50 a 188±167ab 30 x 100 4.8±1.4a 20±2.0 a 3e4±2e4bcd 270±164abc 17±4.9 a 281±160ab 30 x 60 3.6±1.2a 19±3.2 a 2e4±3e3bcd 290±71abc 4.6±8 a 294±63ab 30 x 80 4±1.4a 21±1.2a 2e4±1e4bcd 176±103bc 8.3±12 a 185±111ab 40 x 100 5.5±1.2a 20±5.2 a 2e4±1e4d 110±40c 19±29 a 129±69b 40 x 60 5.3±0.66a 21±309a 3e4±2e4abc 323±136ab 6.9±12 a 330±132ab 40 x 80 5.1±1.4a 19±2.6 a 2e4±1e4dc 220±131abc 13±12 a 233±130ab 50 x 100 5.7±1.4a 17±0.81 a 1e4±7e3d 130±36bc 14±4.1 a 144±34b 50 x 60 4±0.85a 20±4.6 a 2e4±4e3d 170±74bc 3.7±6.4 a 174±71ab 50 x 80 5±2.2a 19±0.6 a 9 1e4±9e3d 154±93bc 27±19 a 181±109ab cv% 14.2 12.81 31.9 42.86 138.16 50.14 (teshome, et al. [14]) 3.12. plant density experiments in bako areas the population study at bako was carried out during 1978 to 1982 using a cultivar white star (table 8). the row spacing employed were 80, 100 and 120 cm whereas plant spacing included 30, 40 and 50 cm with population ranging from 41, 700 to 16,700 in a hectare of land. the results indicated that marketable and total yield increased as plant densities were decreased where as unmarketable yield was higher due to either higher or lower plant densities. unmarketablity was mainly due to oversized and undersized roots. undersized roots increased as spacing between agriculture and food sciences research, 2016, 3(1):1-11 6 rows and plants decreases. an increase in oversized roots was observed with wider spacing. 100cm x 30 cm was recommended for the cultivar white star around bako and similar area. table-8. effects of plant density in mean root yield (q/ha) at bako, 19781982 row x plant spacing (cm 2 ) number of plants/ha (1000) marketable root yield (kg/ha) total root yield (kg/ha) 80 x 30 41.7 206.28 319.54 80 x 40 31.3 193.15 309.19 80 x 50 25.0 187.70 299.16 100 x 30 33.3 209.80 322.12 100 x 40 25.0 174.59 289.94 100 x 50 20.0 157.39 272.62 120 x 30 28.6 188.79 302.25 120 x 40 21.4 158.81 269.28 120 x 50 16.7 157.84 265.53 3.13. plant density experiments in awassa and areka areas plant density recommendations varied in sari mandate areas with variety and location (table 9). in areka, different varieties responded differently as shown in table below. however, at awassa 60 cm x 30 cm recommendation was adopted after a three-year plant population experiment for a white fleshed variety called awassa-83. table-9. recommendations of plant density in sari mandate areas variety location results sp-1499 areka 100 cm x 25 cm sp-koka 12 areka 70 cm x 20 cm sp-2498 areka 100 cm x 20cm awassa-83 awassa 60cm x 30 cm source: crct (crop science research department) [7] 3.14. weeding weeding trials revealed that the crop is highly susceptible to weeds at early stage of growth as most other root and tuber crops. so a grower shall induce a weeding practice staring the 7 th day after planting. however, the crop starts suppressing annual broadleaved weeds starting 45 th day of planting. that is because the crop is known for its allelophatic effects at full growth. there was an increase in yield per unit area as plant density was increased from 5 to 10 plants m -2 . however, there was not significant variation in yield (kg/ha) due to 7 and 10, and 7 to 12.5 plants m 2 in tis 1499 and koka 6 (intermediate), respectively (table 10). the yield of tis 2498 did not show significant variation among plant densities despite the high yield from 10 plants m -2 . the results showed that a population of 10 plants m -2 produced significantly high tuber yield although it was not significantly different from 7 plants m -2 . effect of weeding on tuber yield was dictated by variation in variety and plant density. weeding of tis 1499 (erect and early) and koka 6 twice increased the yield by 22.5 and 23.9%, respectively, while weeding practice did not have a significant effect on tuber yield of tis 2498 (long vine, spreading and early). although weeding once and twice produced more as plant density was increased (up to 10 plants m 2 ), further increase reduced the yield. however, weeding twice increased tuber yield by 23.6, 18.2, 15.4, and 5.1% for the 5, 7, 10 and 12.5 plants m -2 , respectively, compared with weeding once. the variation in root yield was not statistically significant in both weeding practices due to the range of 7 to 12.5 plants m -2 . generally speaking plant density of 25x10 3 or less revealed poor ground cover percentages; thus lower stand densities are of less value to the producers. varieties with erect and intermediate growth habits can be weeded twice if high population is maintained. growth with spreading canopy structure and plant density could be used as means to reduce weed infestation. this will save farmers time and labor and thus, breeders should focus in developing cultivars with spreading canopy structure with high yield. table-10. weeding frequency x variety x plant density on tuber yield of sweet potato (tons/ha) at areka variable weeding frequency one time (30 40 das) two times (30 40 and 70 das) variety tis 1499 16.96bc 20.78a koka 12 6 15.45cd 19.15ab tis 2498 12.53de 11.82e plant density (no.ha-1 ) 50 × 103 11.60c 14.34bc 70 × 103 16.19ab 19.14a 100 × 103 17.16ab 19.81a 125 × 103 14.96bc 15.72ab (source: tenaw, et al. [15]) das = days after sprouting, same letter across a column and row for each factor shows no significant difference at 5% probability level sweet potato used to be grown without any fertilizer amendment despite its requirement and mining from the soil. however studies confirmed that use of either fym or chemical fertilizers or both would increase yield. fertilizer studies were carried out in sandy loam soils of adami tulu consisting of five levels of fym (0, 5, 10, 15, agriculture and food sciences research, 2016, 3(1):1-11 7 20 t ha -1 ) and three levels of p (0, 90,180 kg p2o5 ha -1 ). it was laid out as a randomized complete block design in a factorial arrangement, and replicated three times (table 10). a sweet potato cultivar known in the area called balella was used for the study. the result indicated that the main effect of fym significantly (p < 0.05) affected total tuberous root yield, tuberous root dry weight, fresh total biomass and dry harvest index. however, the main effect of p had no significant influence on all parameters studied (table 11). moreover, the interaction effects of fym and p significantly (p< 0.05) affected marketable root yield, specific gravity and total dry biomass yield. combined application of 20 t farmyard manure ha -1 and 180 kg ha -1 p2o5 resulted in production of highest marketable yield (32.56 t ha -1 ). the lowest marketable yield (8.8 t ha -1 ) was obtained at the application of 0 t ha -1 fym combined with 180 kg p2o5 ha -1 . thus, most of the yield and yield estimate parameters were enhanced in response to the application of fym. for the two commonly used sweet potato quality parameters (dry matter and specific gravity), dry matter was not significantly responsive to both fym and p while the highest specific gravity (1.09) was obtained at the combined application of 10 t fym ha -1 and 0 kg p2o5 ha -1 . tuberous root dry weight was correlated negatively (r = 0.32*) with shoot dry weight and was statistically significant (p < 0.05). but it was correlated positively with yield and yield estimate parameters like tuberous root fresh weight (r=0.965**), marketable tuberous root weight (r = 0.842**), unmarketable tuberous root weight (r = 0.639**), average tuberous root number per plant (r = 0.582**) and average tuberous root length (r = 0.492**) at (p<0.01) indicating the existence of close associations among those parameters. therefore, it could be concluded that tuberous root yield of sweet potato was significantly enhanced in response to the application of farmyard manure, indicating that enriching the soil of the area with organic matter through use of organic fertilizers holds the key for maximizing the yield of the sweet potato crop in the study area. these studies showed that higher nitrogen application would increase vine growth and suppress root growth. table-11. farmyard manure x phosphorus effect on marketable tuberous root weight (t ha-1) of sweet potato fym ( t ha -1 ) p2o5 (kg ha -1 ) 0 90 180 0 15.89 12.41 8.80 5 19.01 22.65 9.72 10 18.67 9.01 32.04 15 14.20 11.17 12.47 20 24.72 22.07 32.56 f-test * lsd (p*fym) 15.41 cv (%) 32.70 *= significant at p < 0.05probablity level; cv = coefficient of variation according to sweet potato without fym was not foreseeable around sandy loam soils of adami tulu with chemical fertilizers alone. if a grower can have fym as high as 5t/ha or 10t/ha then 90 kg ha -1 or 180 kg ha -1 would suffice to obtain agronomic optimum sweet potato root yield of 22.65 tha -1 and 32.04 tha -1 respectively (table 11 and 12). table-12. unmarketable and total root yield of sweet potato as affected by fym and p application treatment unmarketable tuberous root yield (tha -1 ) total tuberous root yield (tha -1 ) fym( t ha-1) 0 10.00 22.37b 5 9.70 26.83b 10 9.39 29.30b 15 10.75 23.30b 20 15.91 42.89a f-test ns * lsd 6.54 12.48 p2o5(kg ha-1) 0 9.59 28.09 90 11.49 26.94 180 12.39 31.48 f-test ns ns lsd 5.05 9.67 cv (%) 31.13 23.25 *=significant at p<0.05 probability levels; ns= non-significant; cv= coefficient of variation; fym=farmyard manure a study was undertaken to assess the effects of combined application of inorganic (np) and organic (fym) fertilizers on root and biomass yield of sweet potato (ipomoea batatas (l.) lam). the field experiment was conducted from june-dec. 2008 at delbo watershed, two n levels (23 and 46 kg/ha) and two p levels (20 and 40 kg/ha) were considered as inorganic fertilizers along with one unfertilized control treatment. the findings of this study clearly showed that yield and yield components of sweet potato at delbo watershed can be enhanced by combining fym and inorganic fertilizers. in this study the highest yield (24.12t/ha) was achieved using 46kg/ha n and 5t/ha fym. accordingly, the economic analysis showed that the highest net benefit of 37,880 birr was obtained from 46kg/ha n and 5t/ha fym. considering the scarcity associated with inorganic fertilizer and limited amount of fym, 23 n kg/ha and 2.5 t/ha fym with a net benefit of 29,635 birr, could also alternatively be recommended for use by sweet potato producers in the area. poor farmers who cannot afford fertilizer would be encouraged to use 2.5t/ha fym (gezahegn and andergachew [16]). in fact, horticulture development department of ministry of agriculture [17] recommended use of 100-200kg/ha dap or equivalent animal manure in previous years. if ridges agriculture and food sciences research, 2016, 3(1):1-11 8 are to be made, it is recommended to apply farmyard manure prior ridging. in fact, at the level of awassa agricultural research center 50q fym/ha was applied during ridging, 25kg /ha dap was applied during planting and 50kg/ha urea applied after a month of planting. 3.15. irrigation practices even though this crop is said to be drought tolerant, research results showed needs for sufficient moisture at early stage especially during the first six weeks to secure healthy stand establishment. this was mainly because sweet potato vines are succulent and fragile, and if no sufficient moisture is supplied it dries up soon. if there is no rain in early parts of planting, supplementary irrigation of 2mm of water per day is required even in the main rainy season. to meet this requirement, irrigating in alternative days is vital. moisture stress during growth significantly reduces storage root yield. as a crop grows on, irrigating with 4-5mm water would suffice for a given week. because sweet potato crop needs sandy and well-drained soil; and if the soil has high moisture content, planting in raised bed is preferable (moard [18]). with growing interest of sweet potato as emergency food security crop in the country, there were increased pressure from concerned bodies to supply sweet potato cuttings in off seasons. obviously the production of vines under such situations required application of irrigation water pumped from springs, rivers or lakes (fig 2). there was a practice of applying water to field capacity once in a week time in such cases in majority of sweet potato fields. yield and quality of sweet potato in non-irrigated experiments were extremely low compared with that of irrigated experiments. fig-2. furrow irrigation practices in sweet potato seed multiplication schemes 3.16. intercropping three sweet potato cultivars (koka-12, koka-14 and alemaya) were grown under three sorghum varieties (kobomash76, 76t1 no.23, hybrid sorghum) were tested under rainfed and irrigated conditions at melkasa and melka werer, respectively. all pure sweet potato stands gave higher yield compared to intercrops. sorghum grain yield of pure stands was lower when compared to intercrops. the results also indicated that koka 12 and kobomash 76 were relatively compatible (sirak [11]). results further depicted that sweet potato plants are sensitive to shade leading to low yields. the crop should be grown under full sunlight. tesfa, et al. [19] also noted that maize intercropped with sweet potato showed reduced levels of leaf blight and common rust intensity when both crops were planted at the same time. 3.17. relaying and double cropping therefore, it is only with erect leaved varieties that intercropping is recommended. however, all the varieties could be grown with relay crops, double crops or rotations with maize or sorghum (table 13). thus the crop can take advantage of residual moisture and fertility. results showed that in areas where sweet potato was grown previously, long season maize varieties could be grown successfully. table-13. double cropping of maize varieties after cereal, pulse and root crops at awassa (1993, 1994 and 1996) precursor crop yield (q/ha) of katumani yield (q/ha) of a 511 no fertilizer 46/46kgha -1 np mean no fertilizer 46/46kgha -1 np mean katumani 25.00 26.04 25.52 a-511 5.52 5.52 5.10 tef 10.83 10.83 5.52 18.75 19.79 19.27 haricot bean (hb) 9.53 9.53 12.29 15.63 25.00 22.04 irish potato (ip) 14.47 14.47 13.75 18.75 26.04 22.40 sweet potato (sp) 7.08 7.08 6.65 17.71 26.04 21.88 9.49 8.66 9.08 19.17 24.58 21.88 source: (tesfa, et al. [19]) a double cropping experiment of maize grown after different crops in the same year was executed from 1993 to 1996. the objective of the experiments was to properly utilize the available moisture, enhance productivity and fill food deficit periods. hb, ip and sp were found to be good precursors for a-511. teff and irish potato happened to be good pre cursors for katumani under unfertilized conditions. haricot bean would become good precursors only after fertilized plots. by double cropping the productivity of land was increased by 50 and 70% as compared to single crops under unfertilized and fertilized conditions (table 12). in another study carried out in awassa by tolesa, et al. [20] farmers were advised to grow either teff, haricot bean or sweet potato by harvesting at green ear or dry stage of maize. the land equivalent ratio analysis revealed agriculture and food sciences research, 2016, 3(1):1-11 9 that the advantage of relay cropping is higher, 71% when maize was harvested green and 54.9% when maize was harvested at dry grain stage. the researchers obtained the highest gross return of 8912 br./ha when maize was harvested at green ear stage and 5683 br./ha when maize harvested dry grain and when sweet potato was relayed into maize at 50% flowering. this manifested that harvesting maize for green ears and relaying sweet potato into maize are economically attractive. thus the appropriate time of relay planting sweet potato is at 50% flowering of maize for both harvesting systems (table 14 and 15). table-14. effect of relay cropping sweet potato on maize green ear harvest and ler at bako, 1990-1992 treatments maize ears/ha sweet potato yield (q/ha) gross returnbr./ha ler maize + 50% mf 36310.82 67.47 3911.99 1.27 maize +sp 15 da 50% mf 36310.82 47.13 7372.81 1.16 maize +sp 30 da 50% mf 40958.60 8.86 6480.39 1.07 sole mz 5967.08 1.00 mz-maize, spsweet potato, mf-mid flowering, da-days after table-15. effects of relaying sweet potato after maize at bako, 1990-1992 treatments sp yield (q/ha) sp yield reduction relative to sole (%) @ 50% mf 67.5 64.2 15 da 50% mf 47.1 75.0 30 da 50% mf 8.9 95.3 sole sp 188.5 0 lsd5% 26.6 lsd5% 37.3 cv (%) 27.1 3.18. rotation cropping according to results of numerous year crop rotation study, maize, sweet potato and common bean coming in the sequence in the three year crop rotation scheme was productive compared to other cropping sequences. 3.19. growing sweet potato near shrub and tree crops as leaves, barks, roots, twigs, exudates and branches release phenolic allello chemicals, eucalyptus spp. were generally recognized with their allelophatic effects on growth of various crops. this negative effect coupled with its fierce competition for sunlight, moisture and nutrients posed great threat to crop production and have been known as major court case in many places. however, driven by expansion of construction economy the reality of eucalyptus spp. expansion in most agro-ecologies of ethiopia has become a reality. so serious studies were carried out to identify tolerant crops to the cumulative effect of eucalyptus and identify a mechanism for amelioration of the challenge. results showed that a distance of some 5 meters should be left in lowlands to produce crops like teff, sweet potato and common beans; however, growing crops like wheat and barley is possible with eucalyptus in highland areas where moisture is not limiting. in fact, cutting ditches was recommended to break root exudation and completion for moisture and nutrients (fig 3). fig-3. ditches dug between eucalyptus hedge row and adjacent farmland there are also growing trends of growing sweet potato under other shrub crops like coffee and enset in some parts of the country. however, the production is maintained until canopy closure of coffee and enset, and is meant to fill food gaps of some forthcoming months. 4. conclusion sweet potato that was once known as poor man’s crop has become a food security crop all over the country with numerous potentials for income generation and export. although it required low inputs, it matured in short period of time and provided reliable yield in ethiopian conditions. consequently the crop rightly obtained national focus due to the various organizations in the ethiopian agricultural research system which contributed much to the generation of crop management options. sweet potato crop has a potential of 50 to 60 t/ha in ethiopia; however, farmers are obtaining 6 to 8 tons/ha only. the yield is ten times lower than the potential. this yield gap would be attributed to a number of factors like absence of high yielding improved and virus free planting materials, appropriate insect pest management and use of improved crop management options. based on research findings to date, vines taken from agriculture and food sciences research, 2016, 3(1):1-11 10 upper or middle portion of the plant chosen for sweet potato planting should be of 30 to 40 cm length having 3 to 4 buds, shall be stored for two days under shade and should be buried in the soils until two-third of the cutting. the land prepared could be flat as in wolaita area and could be of ridges as in hararige depending on soil texture, moisture regime and tillage implements. when planting dates are considered, in awassa and similar agro-ecological areas the onset of rainy season particularly mid may to early june was recommended as optimum unlike early june planting of bako. planting position study confirmed that planting cuttings at slant (inclined) position gave higher root yield (p<0.05) compared to horizontal plantings in areas like awassa. in harargie vines are planted in inclined positions where plantings are done on ridges. in wolaita, vine cuttings are buried horizontally in furrows when the land is prepared by oxen drown implements. plant density experiments in jido combolcha revealed that net marketable yield of balella variety obtained at spacing combination of 20 cm x 60 cm (590 ± 104 q ha -1 ) was by far better followed by 20 cm x 80 cm and 50 cm x 60 cm that gave average yield of 583 ± 82 q ha -1 and 463±93 q ha -1 , respectively. according to similar studies employing cultivar whitestar 100cm x 30 cm plant spacing was recommended for the bako and similar agro-ecologies. the plant density study at areka showed that sp-1499 should be grown with 100 cm x 25 cm, sp-koka 12 with 70 cm x 20 cm, and sp-2498 with 100 cm x 20cm. however, similar studies employing cultivar awassa-83 ended up in recommending 60cm x 30 cm for awassa and similar agro-ecologies. for a sweet potato crop grown with recommended plant density, varieties (eg. tis 2498) with long spreading vines could be weeded once on 30-40das but varieties with erect vines (e.g.tis 1499) should be weeded twice on 30-40das and 70das regardless of the study sites. in one fertilizer study carried out in adami tulu combined application of 20t farmyard manure ha -1 and 180 kg ha -1 p2o5 resulted in production of highest marketable yield (32.56tha -1 ). in another fertilizer study carried out at delbo watershed, the highest root yield (24.12t/ha) was achieved using 46kg/ha n and 5t/ha fym. if rains stop during early or late growth of sweet potato, supplementary irrigation of 2mm/day during planting (for a week or so) and 4-5mm/week during vegetative growth was recommended. in intercropping study carried out at melkasa and melka worer using three sweet potato varieties called koka-12, koka-14 and alemaya and three sorghum varieties called kobomash76, 76t1 no.23 and hybrid sorghum, sorghum grain yield of pure stands was lower when compared to intercrops and koka 12 and kobomash 76 were relatively compatible. based on relay and double cropping trial carried out at awassa, in places where sweet potato was grown previously, long season maize varieties could be grown successfully and sweet potato could act as precursor for maize varieties. based on green ear and dry harvest analysis, best maize and sweet potato compatibility was seen when sweet potato was relay cropped at 50% flowering of maize. whenever sweet potato is grown near eucalyptus tree, ditches should be cut with 60 cm depth and 40 cm width to prevent root exudates of allelophatic effect. totally there is a great deal of concern on production, research and technology dissemination; however, there is also a need to create sustainable and site specific integrated nutrient management options, appropriate irrigation schedules, generation of ways of conserving planting materials in dry seasons and development of compatible multiple cropping options for sweet potato in the country. 5. future research directions 1. as we are in the era of global climate change and soil degradation, adopting and generating eco friendly site specific sustainable crop production technologies through well designed research projects is necessary 2. information on productivity and on suitable soil fertility management practices for sustained crop production for the major soils of ethiopia is still very limited. there are limited suitable options for improving soil fertility management (which is essential to increase crop and farm productivity) in a crop like sweet potato that is considered a low-input and subsistence crop and where there are few incentives to use labor and resourceintensive technologies. sweet potato comprehensive integrated soil fertility management (isfm) studies for early, medium and late maturing varieties preferred in seed multiplication and root production schemes for dominant soils found in major agro-ecologies is recommended as future research strategy. 3. drought management is an important factor for increasing sweet potato productivity, given that sweet potato yields can be severely affected by limited water availability. so sweet potato irrigation studies aimed at economizing water use while optimizing vine and root production shall be sought in future. 4. design and assessment of intercropping options and crop rotations that could extend the supply period. 5. development of alternatives for conserving planting materials and for having planting material on a timely manner, particularly in drought prone areas. references [1] t. assefa, a. teshome, t. engida, and t. tesfaye, "summary of progress on orange-fleshed sweetpotato research and development in ethiopia," in proceedings of the 13th istrc symposium, 2007, pp. 728 –731. [2] t. engida, e. v. devakara, and d. nigussie, "correlation and path analysis in sweet potato and their implications for clonal selection," journal of agronomy, vol. 5, pp. 391-395, 2006. [3] i. j. dantata, f. e. babatunde, s. mustapha, and a. s. fagam, "influence of variety and plant spacing on tuber size, tuber shape and fresh marketable yield of sweet potato in bauchi nigeria," biol. environ. sci. j. trop., vol. 7, pp. 140-144, 2010. [4] p. j. ndole, t. mcharo, e. e. carey, s. t. gichuki, c. ndinya, and j. malinga, "participatory on-farm selection of sweet potato varieties in western kenya," african crop science, vol. 1, pp. 41-48, 2001. [5] o. andreas, k. regina, l. berga, o. oscar, k. jürgen, and l. john, "challenge theme papers 5. integrated crop management. cip. social sciences working paper 2009-1, 130 unleashing the potential of sweet potato in sub saharan africa," 2009. [6] d. getahun and w. tenaw, initial results of informal survey. areka mixed farming zone, wolayita district, sidamo region. working paper no. 11. addis ababa, ethiopia: institute of agricultural research, 1990. [7] crct (crop science research department), available technologies and constraints in technology dissemination. awassa, ethiopia: crop science research department of sari, 2005. [8] l. geleta, "establishment and root yield of sweet potato as affected by pre rooting of vine portions," in proceedings of the sixth annual conference of the crop science society of ethiopia, seble, may 1994, addis ababa, ethiopia, 1994, pp. 116-119. [9] t. endale, b. terefe, d. mulgeta, and l. geleta, "improvement studies in enset and sweet potato. in: horticulture research and development in ethiopia," in proceedings of the 2nd national horticultural workshop of ethiopia, 1-2 december 1992, addis ababa, ethiopia, 1992. agriculture and food sciences research, 2016, 3(1):1-11 11 [10] a. mohammed, r. nigussie-dechassa, and a. bekele, "effect of planting methods and vine harvesting on shoot and tuberous root yields of sweet potato [ipomoea batatas (l.) lam.] in the afar region of ethiopia," african journal of agricultural research, vol. 7, pp. 1129-1141, 2012. [11] b. sirak, "review of sweet potato and other minor root crops research in ethiopia," in proceedings of first ethiopian horticulture workshop, 20-22 february 1985, w. godfrey-sam-aggrey and bereke-tsehai tuku eds, 1985, pp. 313-322. [12] a. teshome, c. amenti, t. kassaye, t. fiseha, and a. geremew, "yield and yield components of sweet potato as influenced by plant density: in adami tulu jido kombolcha district, central rift valley of ethiopia," american journal of experimental agriculture, vol. 1, pp. 40-48, 2011. [13] frg, frg research inventory. ethiopia: melkasa, 2009. [14] a. teshome, d. nigussie, and a. yibekal, "sweet potato growth parameters as affected by farmyard manure and phosphorus application at adami tulu, central rift valley of ethiopia," agricultural science research journal, vol. 2, pp. 1 12, 2012. [15] w. tenaw, m. waga, and h. legesse, "growth habit, plant density and weed control on weed and root yield of sweet potato (ipomoea batatas l.) areka, southern ethiopia," journal of horticulture and forestry, vol. 3, pp. 251-258, 2011. [16] g. gezahegn and g. andergachew, combined effects of inorganic (np) and organic (fym) fertilizers on: root and biomass yield of sweet potato (ipomoea batatas l.) at delbo watershed in wolaita. southern ethiopia: lap lambert academic publishing, 2011. [17] horticulture development department of minstry of agriculture, practical guide to horticulture in ethiopia. addis ababa, ethiopia: ministry of agriculture, 1976. [18] moard, "crop variety register, animal and plant health regulatory directorate, issue no. 12, june 2009, addis ababa, ethiopia," 2000. [19] b. tesfa, d. tolessa, g. setegn, t. tamado, g. negash, and w. tenaw, "development of appropriate cropping systems for various maize producing regions of ethiopia," proceedings of thesecond national maize workshop of ethiopia12-16 november 2001 addis ababa, ethiopia, 2001. [20] d. tolesa, g. gemechu, and l. melakesilase, "relay cropping of different crops in short cycle maize guto at bako," in proceedings of the sixth annual conference of the crop science society of ethiopia, in: seble may 1994. addis ababa, ethiopia, 1994, pp. 75-79. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research vol. 4, no. 2, 45-57, 2017 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2017.42.45.57 45 determinants of nutritional status of under-five children in ethiopia: with particular reference to anelmoworeda, hadiya zone, southern nations, nationalities and peoples region nigussie temesgen1+ abebe haile2 ( (+ corresponding author) 1,2addis ababa university, college development studies, centre for food security studies addis ababa, ethiopia abstract malnutrition is one of leading problem which affect children well-being and growth in the study area as well as in ethiopia. however, insufficient scientific knowledge limits to understand the factors of nutritional status at community level for adequate intervention. therefore, this study was undertaken to investigate the determinants of nutritional status of under-five children in anlemo woreda to tackle the malnutrition problems. a purposive sampling followed by random sampling procedure was employed to draw 330 sample households from four kebeles. a household survey was undertaken using structured questionnaires. different characteristics of the households were investigated; anthropometric measurement of children was also measured. the data were analyzed using spss.v.22, for descriptive statistics, stata14 for multivariate probit regression analysis and who anthrop v.3.2.2 software to convert raw nutritional data into zscores. the result of multivariate probit analyses revealed that age of household head, water source, land size, complementary feeding, antenatal care, toilet, time elapse to fetch water and livestock ownership were significant and associated with child stunting. sex of child, toilet, income, and source water was found to be significant and associated with child wasting, and child age, family size, livestock ownership, income and water source were significant and associated with underweight. the findings of this study revealed that the prevalence of stunting of children is 42.4%, underweight 16.1% and wasting 9.7% respectively. the prevalence of stunting and wasting among children aged 6-59 months is relatively high but underweight is lower than national and regional prevalence. malnutrition puts substantial burden on under five children in anlemoworeda. local government of the hadiya zone and anlemoworeda health institution should design effective nutritional implementation on child stunting and wasting is a vital task for the community to reduce malnutrition. keywords: nutritional status, under-five children, determinants, anlemoworeda. citation | nigussietemesgen; abebe haile (2017). determinants of nutritional status of under-five children in ethiopia: with particular reference to anelmoworeda, hadiya zone, southern nations, nationalities and peoples region. agriculture and food sciences research, 4(2): 45-57. history: received: 12 october 2017 revised: 22 november 2017 accepted: 29 november 2017 published: 6 december 2017 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 46 2. materials and methods ................................................................................................................................................................... 46 3. results and discussions .................................................................................................................................................................. 47 4. descriptive statistic and results .................................................................................................................................................. 47 5. discussion .......................................................................................................................................................................................... 54 6. conclusion and recommendations ............................................................................................................................................... 55 references .............................................................................................................................................................................................. 56 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2017.42.45.57&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=nigussie%20temesgen https://orcid.org/orcid-search/quick-search?searchquery=abebe%20haile agriculture and food sciences research, 2017, 4(2): 45-57 46 1. introduction malnutrition is a public health problem which causes the deaths of 3.5 million underfive children peryear in the world. it is the third source disease for children. even though, childhood malnutrition had shown a trend of relative decline in 1990s at a global level; however, its prevalence in africa had tremendously increased during the aforementioned decade [1]. malnutrition is nothing, but a disorder resulting from deficiency or excess of one or more essential nutrient in developing world. usually it is described as under nutrition whereby the existence of varying degrees of deficiencies in essential nutrients. true, malnutrition clearly identified in children where the bodies of children respond to malnutrition and/or under nutrition by stunting (low height for age), wasting (low weight for height) and/or underweight. malnutrition on the one hand, weakens the immunization of a child against both infectious and non-infectious diseases; and it reduces appetite and hence, prevents the body for normal absorption of food and worsening malnutrition on the other [2]. as far as nutritional status is concerned, it is the weakness/strength of the body resulting from the intake, absorption and utilization of food. it is, therefore, determined by a complex interaction between internal factors such as age, sex, nutrition, behavior, physical activity and diseases and the external environmental factors such as food safety, cultural, social and economic circumstances [3]. more than half of child mortality is associated with malnutrition worldwide. according to the lancet series studied in 2008, 20 per cent of children below the age five in low and middleincome countries are underweight. globally, 10%; or 55 million children are wasted and out of these 19 million are severely wasted. about 32%; or 178 million of under-five children in the world are stunted. among the 178 million children under five 90% of them live in 36 countries including ethiopia. the ethiopian demographic and health survey shows that 53% of underfive mortality is attributed to malnutrition; thirty-eight percent of children younger than five years are stunted; 9.9% are wasted; and 23.6% are underweight [4]. according to the world bank, adequate nutrition is the fundamental right of every human being; and malnutrition is not just a stark manifestation of poverty, it is rather non-income face of poverty and it helps perpetuate poverty (2012). even though its patterns and prevalence vary significantly malnutrition is a global issue. it is not only a problem at a country level, but is also common among different regions within a given country [5]. ethiopia is a case in point: prolonged malnourishment results in improper physical and mental growth to children and hence, has negative impact for the well-being of children. thus; malnutrition is a menace for children’s development particularly in the developing nations [6]. under nutrition remains a devastating than one problem in many developing countries affecting over 815 million children causing more -half of child deaths [7]. the levels of mortality are also worsened particularly by poverty, inadequate maternal education, lack of potable water and sanitation, high fertility and inadequate birth spacing. lancet study shows 195 million under-five children are affected by malnutrition; 90% of them live in subsaharan africa and south asia [8]. ethiopia is one of the world nations with the highest under-five mortality rates. at least 53% of death of children can be attributed directly or indirectly to malnutrition. the ethiopian demographic health survey (edhs) report shows that nearly one in two children is malnourished i.e., 38.4% of ethiopian under five children are being stunted, 9.9% wasted, and 23.6% underweight. according to the estimates, one in every 17 ethiopian children dies before the first birthday, and one in every 11 children dies before the fifth birthday [4]. several factors which are associated with malnutrition have been identified, including poor breastfeeding and child feeding practices, lack of access to enough nutritious food, low levels of parental education and belonging to the low-income group [9]. according to edhs [4] nutritional status varies greatly by region among the ethiopian regional national states, where the highest rates of malnutrition found in amhara, tigray, snnpr and oromia regions, 46.3 %, 39.3%, 38.6% and 36.5%, respectively. addis ababa has the lowest rate, which is 14.6%. according to anlemo woreda agricultural and natural resource development office [10] the woreda under study is one of food insecure and targeted in productive safety net program since 2005. thus, all the kebeles in the woreda are beneficiaries of psnp program. the above source also indicated that this area is not self-sufficient in crop and livestock production, though they are the sources of improving food security status. according to anlemo woreda health and development office [11] both chronic and acute malnutrition problems are exist in anlemo and children were affected by malnutrition. besides, sufficient scientific knowledge is hardly available to understand the factors of nutritional status at community level for adequate intervention. therefore, this study is designed to investigate the prevalence of malnutrition and associated factors among children aged 6-59 months, which can be used as a reference in priority setting, to bridge the knowledge gap between the community and designing effective nutritional intervention programs within the study area. 2. materials and methods 2.1. study design and sampling the research design employed in this study was cross-sectional survey design which incorporates anthropometric measurement of 330 children and structured questionnaire to care-givers and mothers of the children under study to find out the situation of households in anlemoworeda. the sample size is determined by assuming the prevalence rate of malnutrition to be 50% in the survey area, 95% confidence interval (ci), 8% margin of error (d) and 10% non-response rate is added to the total calculated sample size, un economic and social affairs division (2005). calculation of sample size for infinite population: sample size (n) = z2 ×p (1-p)/d2 calculation of sample size for finite population: according to anlemoworeda health office we found that the total number of children of 6 -59 months are 13,626.thus we apply finite population sample formula to obtain new sample size to conduct survey in the study woreda therefore, new sample size (nss) = n / [1+ { (n-1) / top}] agriculture and food sciences research, 2017, 4(2): 45-57 47 where: new ss = new sample size for finite population. n = sample size in infinite population. top = total number of population (in this case total number of population is number of 6-59 months age children in the woreda). new sample size obtain as = n / [1+ {(n-1) / top}] = 150 / [1+ {(150-1) / 13626}] = 299.9 i.e., 300 thus calculated sample size is adjusted for non-response rate as 10%, the sample size is calculated to be 330. 2.2. ethical consideration the ethical approval was obtained from ephi ethical review committee. the ethical clearance was submitted to the snnpr, health office and following the zone, woreda and kebeles health offices/centers had received the ethical clearance copies. thelactating mothers were informed about the data collection and its contribution for their future nutritional benefits for the woreda and the country as a whole. the verbal consent was obtained from each participant it was assured that the information obtained from them was maintained confidential. 3. results and discussions this chapter presents the results, discussion and interpretation of the study findings. both descriptive statistics and multivariate probity regression analysis were employed to identify the relationship between explanatory variables and dependent variables. multivariate probity regression was used to identify the determinant factors that affect the nutritional status of under-five children while who, anthrop v 3.2.2 software was employed to converting direct measurement of child in to z-scores and to measure the prevalence of stunting (low-height-for age (haz), underweight (low-weight-for age (waz) and wasting (low-weight-for height (whz). 4. descriptive statistic and results 4.1. demographic socioeconomic characteristics of households table 4.1.indicateshousehold characteristics, from 312 (94.5%) child mothers are engaged with male headed, 18 (5.5%) of the household are female headed. among the household heads 314 (95.2%) are married, 6 (1.8 %) is divorced, 6 (1.8%) is widowed and 4 (1.2%) are engaged in polygamy. the educational status of the child mothers is low among sample households. at least 108 (32.7%) of mothers cannot read and write 107 (32.4%) of mothers can read and write 58 (17.6%) have 1-4 grade education levels and only 26 (7.9%) have 5-8 grade education 25 (7.6%) have 9-10grade education and only 6 (1.8%) percent have above grade 11 education levels. the family size of the sample households 41 (12.4%) had less than or equal to three family 173 (52.4%) had 4-6 family, 112 (30.6%) have 7-10 and 4 (4.5%) more than 11 family size. table-4.1. demographic and socio economic characteristic of the respondents distribution frequency and percentage in anlemoworeda, 2017 (n=330). source: own field survey, 2017 from the table 4.2 shows the age of household headed 3 (0.9%) equal to 20 years, 284 (86.1 %) 21-40 years, 42 (12.7%) 41-60 and only 1 (0.3 %) greater than 61year. with respect to livelihood bases 133 (40.3 %) are crop production 156 (47.3%) mixed farming and 41 (12.4%) other. variables number percent household type child mothers engaged with male headed 312 94.5 female headed only 18 5.5 total 330 100 marital status married 314 95.2 divorced 6 1.8 widowed 6 1.8 engaged in polygamy 4 1.2 total 330 100 family size ≤ 3 41 12.4 4-6 173 52.4 7-10 112 34 >11 4 1.2 total 330 100 education status of mothers cannot read &write 108 32.7 can read & write 107 30.5 1-4 grade 58 17.5 5-8 grade 26 7.9 9-10 grade 25 7.6 above grade 10 6 1.8 total 330 100 agriculture and food sciences research, 2017, 4(2): 45-57 48 table-4.2.demographic and socio economic characteristic of the respondents distribution frequency and percentage in anlemoworeda, 2017 (n=330). variables number percent age of household headed ≤20 3 0.9 21-40 284 86.1 41-60 42 12.7 >61 1 0.3 total 330 100 livelihood base crop production 133 40.3 mixed farming 156 47.3 other 41 12.4 total 330 100 source: own field survey, 2017 from table 4.3 it is found that the majorityofthehouseholds257 (77.9 %) in anlemoworeda produce their own food, while 45 (13.6%) purchase from market, 20 (6.1%) from shared production and 8 (2.4 %) food aid. regarding to size of land 186 (56.4%) households have less than or equal to 0.5 hectare 55 (16.7%) have 0.51-0.75 hectare, 71 (21.5%) have 0.76-1 hectare and only 18 (5.4%) households have greater than 1 hectare of land. with respect to income 176 (53.3%) of mothers have less than 500 birr and 124 (37.6 %) have got 501-1000 birr 29 (8.8%) had more than 1001-2000 birr and only 1 (0.3%) had greater than 2001 etb (ethiopian birr). table-4.3. socio-economic characteristic of the respondents in anlemo woreda,2017 (n=330). variables number percent land size in hectare (ha) ≤ 0.5 ha 186 56.4 0.51-0.75 ha 55 16.7 0.76-1 ha 71 21.5 >1 ha 18 5.4 total 330 100 source of food for household produce own food 257 77.9 purchase from market 45 13.3 food aid 8 2.4 shared production 24 6.1 total 330 100 income (etb) ≤500 176 53.3 501-1000 124 37.6 1001-2000 29 8.8 >2001 1 0.3 total 330 100 source: own field survey, 2017 etb= ethiopian birr 4.2. water, sanitation and hygiene table 4.4 revealed the largest proportion of the sampled household use unprotected wells, spring and surface water 181(54.8%) while relatively fewer people about 149 (45.2 %) use public taps and protected wells. anlemoworeda has limited or no access to safe drinking water. waste disposal methods of the household 10 (3%) by burying 11 (3.4%) throwing to nearby road 72 (21.8) keeping in compound for fertilizer and 237 (71.8%) by burning. the toilet/latrine access about 117 (35.5%) have pit latrine with slap 203 (61.5%) have open pit/without slap and 10 (3%) do not have latrine. table-4.4. environmental and sanitation associated factors in anlemoworeda, 2017(n=330). variables number percent source of water/access public-tap 132 40 unprotected dug well 75 22.8 protected spring 17 5.2 unprotected spring 86 26 surface water 20 6 total 330 100 latrine pit latrine with slap 117 35.5 open pit/without slap 203 61.5 no facility/bush 10 3 total 330 100 waste disposal methods by burying 10 3 as fertilizer 72 21.8 throwing to nearby roads 11 3.4 by burning 237 71.8 total 330 100 source: own field survey, 2017 agriculture and food sciences research, 2017, 4(2): 45-57 49 table 4.5: indicates that the housing condition of the respondent the floor of dwelling 316 (95.8%) are not cemented and only 14 (4.2%) were cemented floor, 132 (40%) were corrugated roof and 198 (60%) were thatched roof and 319 (96.7%) wall of dwelling were wood and mud only 11 (3.3%) stone and cement wall. table-4.5. households environmental associated factors in anlemoworeda, 2017(n=330). variables number percent housing condition (floor) earth and cow dung 316 95.8 cemented 14 4.2 total 330 100 roof of dwelling thatched 198 60 corrugated 132 40 total 330 100 wall of dwelling wood and mud 319 96.7 stone ad cement 11 3.3 total 330 100 source: own field survey, 2017 4.3. child care and health services as table 4.6 shows the age a child started complementary feeding over eighty percent 271 (82.1%) of the children start complementary feeding at the age of six month and the remaining 59 (17.9%) start at 7-12 months. child these had got immunization or received vaccination were only 3 (0.9 %) bcg 3 (0.9 %) polio 3 (0.9%) dpt and 314 (95.7%) of children took bcg, polio, dpt and 10 (2.6%) of children were not vaccinated. the sources of information for mothers of children how to feed her child and nutrition were 57 (17.3) percent of mothers’ from radio 265 (80.5%) from health extension workers and 8 (2.2%) from relatives. based on antenatal care 320 (97%) of child mothers or care takers attended antenatal care and only 10 (3%) do not follow the antenatal care. with respect to colostrum 279 (84.5%) of child was depleted the first breast milk and 51(15.5%) did not get the first breast milk, and 316 (95.8%) of children got vitamin-a supplement and only 14 (4.2%) do not supplemented by vitamin-a. with respect to child diarrhea 66 (20%) were sick by diarrhea and the remaining 264 (80%) were not sick by diarrhea. exclusive breast feeding for six month 327 (99.1%) of mothers feed their child for 6 months and only 3 (0.9%) do not feed. table-4.6. child caring, and health services, in anlemoworeda, 2017 (n=330). variable number percent age complementary feed start 6 months 271 82.1 >7 months 59 17.9 total 330 100 immunization/vaccination of child not vaccinated 10 2.6 bcg 3 0.9 polio 3 0.9 dpt 3 0.9 bcg,polio & dpt 314 94.7 total 330 100 source of information for mothers on child nutrition and feeding radio 57 17.3 health extension workers 265 80.5 relatives/friends 8 2.2 total 330 100 colostrum feeding/depleting yes 279 84.5 no 51 15.5 total 330 100 vitamin-a supplement yes 316 95.8 no 14 4.2 total 330 100 antenatal care yes 320 97 no 10 3 total 330 100 diarrhea illness yes 66 20 no 264 80 total 330 100 exclusive breast feeding for 6 months yes 327 99.1 no 3 0.9 total 330 100 source: own field survey, 2017 agriculture and food sciences research, 2017, 4(2): 45-57 50 4.4. anthropometric prevalence of stunting, wasting and underweight anthropometry: is the measurement of human body used to assess the nutritional status of individuals and population groups. this s tudy has three observation or dependent variables in identifying the key risk factors associated with malnutrition among children aged6-59 months in the study area: stunting, wasting and underweight. stunting (low-height-for-age haz) is an indicator of chronic or long-term nutritional status of children. it was computed or estimated by comparing the height-for-age of a child with a reference population [12]. the z-score was computed for each measure to see how far a child is from the median height/weight of the reference distribution for children of the same height/weight, taking in to consideration the standard deviation of the reference distribution. the analysis of the three anthropometric indices height-for-age, weight-for-age and weight-for-height in the study area: showing z-scores and their corresponding means and standard deviations for haz, waz and whz were, the mean z-scores show that stunting is the greater of the malnutrition problems with a z-score of -1.43 and a standard deviation of 1.21 followed by wasting with a z-score of -0.2 and standard deviation 1.39 and a z-score of -0.91 and a standard deviation of 1.11 is underweight. during calculations of z-score using the who anthrop v.3.2.2 software, and hence, z-score below –4sd and above 4sd were excluded from the analysis. a positive zscore means that an individual’s measurements are higher than the reference mean and a negative z-score means that the measurements are lower than the reference mean. the prevalence of stunting (low-height-age) was 42.4%, underweight (low weight-for-age) 16.1% and wasting (low-weight-for-height) 9.7% (see figure 4.1 below). figure-4.1. nutritional status of children in anlemoworeda, 2017 (n=330) source: own field survey, 2017 according to edhs [4] the national figure of stunting is 38.4% wasting 9.9% underweight 23.6%. however, the finding of this study revealed that stunting 42.4% which is slightly higher than the edhs prevalence figure as well as regional 38.6% and wasting 9.7% is nearly similar with national prevalence 9.9% but higher than regional 6% respectively. but underweight 16.1% is lower than the national 23.6 % and regional prevalence 21.1 % respectively. similarly according to dejene and ayele [13] study result of hawassa zuria revealed that 45.8% stunting, 31.9% underweight and 23.9% wasted. the child characteristics show that 168 (50.9%) of children in the sample are female and 162 (49.1 %) are male the average age of children in the sample is 27.8 months. the result of survey shows 18.2 percent of children in the sample were ill by diarrhea in the last two weeks preceding the survey (see table 4.8 below). table-4.8. distribution of children by sex and age in anlemoworeda, 2017 (n=330). age group/months male female total 6-11 25 (7.6%) 21 (6.4%) 46 (13.9%) 12-23 36 (10.9%) 41 (12.4%) 77 (23.3%) 24-35 49 (14.8%) 39 (11.8%) 88 (26.7%) 36-47 28 (8.5%) 41 (12.4%) 69 (20.9%) 48-59 24 (7.3%) 26 (7.9%) 50 (15.2%) total 162 (49.1%) 168 (50.9%) 330 (100% note: height –forage z-scores boy and girls (n=330) source: own field survey, 2017 agriculture and food sciences research, 2017, 4(2): 45-57 51 figure-4.2. shows height for age z-score distribution of children as compared to the standard reference population by sex indicates that height for age z-score below -2sd 48.8% male with mean -1.69 , sd 1.06 and 36.3% female mean -1.17 and sd 1.28. as indicated in the figure stunting in male children is higher than the female children. source: own field survey, 2017 figure-4.3. shows weight for height distribution as compared to standard population reference by sex of under-five children with z-score below -2sd 8 % male with mean -0.12, sd 1.1 and 11.3%% female with mean-0.28 and sd 1.42. source: own field survey, 2017 figure-4.4. shows that weight for age distribution as compared to standard population reference by sex of under-five children with z-score. the result indicates that 18.5% male children and 13.7% of female are underweight. the anthropometric figure revealed that male children are more underweight than the female children and z-score are negative which indicates less than the standard population references. source: own field survey, 2017 nutritional status of under-five children and factors associated in anlemoworeda, 2017 results sex of combined (n=330) agriculture and food sciences research, 2017, 4(2): 45-57 52 figure-4.5. height-for-age distribution as compared to standard population reference by sex combined with z-scorebelowthe-2sd 42.4% source: own field survey, 2017 the above result of anthropometric indicate that children stunting in anlemoworeda is 42.4% which is higher than the national and regional prevalence and the graph is deviated to negative z-score or less than the reference population. figure-4.6. shows weight-for-age nutritional status of children in reference population of combined sex indicates that 16.1% of children were below 2sd which means less than the reference population and deviated to negative z-score. source: own field survey, 2017 4.5. muac result table 4.9 shows that mid upper arm circumference; in this survey muac used only for emergence case/ sam: severe acute malnutrition triangulation; at the time of survey 6 children were severely acute malnourished and recommended to health post in order to treat with (otp) and corn soya blend (csb) to recovery from severe malnutrition. figure-4.7. shows weight-for-height 9.7%childrenwerebelowthe-2sdz-scores source: own field survey, 2017 agriculture and food sciences research, 2017, 4(2): 45-57 53 table-4.9. muac results in anlemoworeda, 2017. normal (>12.5cm) mam (11.5 < 12.5cm) sam (<11.5cm) total 293 26 6 330 source: own field survey, 2017 4.6. dietary diversity food groups/ dietary diversity/: proportion of children 6-59 months of age who receive foods 4 or more groups: where 7 food groups used including grains/ root& tubers, legumes and nut, milk and milk-products, meat, eggs, vitamin–a rich fruits and vegetables and other fruits and vegetables. three hundred thirty households survey were applied for this study the respondent answer yes takes the value 1 and no 0. in terms of dietary score, grain, root and tubers were the most popular food group consumed by children, followed by other fruits and vegetable and legumes. the eggs and meat was least consumed food group in the study area. the result of figure 4.8 indicates that 27(8.2%) children had high diversity more than 6 food groups and 86 (26.1%) had medium dietary diversity 4-5 food groups and 217 (65.7%) of children had low dietary diversity less than or equal to 3 food groups out of seven food groups. figure-4.8. food groups consumed by children and dietary diversity in the last 24-hours, anlemoworeda, 2017 (n=330) (using the fao [14] and fanta [15] guide line) source: own field survey, 2017 4.7. multivariate probit regression analysis table 4.10 presents results from the multivariate probity regression analysis of haz, waz and whz. the model statistics show that the r-squared ranges from1percent in the haz model 1 to 7 percent in the waz model. the f-statistics show that the hypothesis that all the slope coefficients are equal to zero, except the constant, in each model is rejected at the 1percent significance level. all child characteristics: sex of the child, age of the child and complementary feeding of the child, are statistically significant in all the three models. the relationship between age of child and nutritional status of children is curvilinear with the coefficient of the age of a child being negative and that of age squared being positive and statistically significant at 5percent levels. this indicates malnutrition (stunting) in children worsens with age, sex or being maleness and femaleness are significant with stunting. age of the household headed: the analysis of this study raveled that age of household is positively associated with child stunting. the age of households increase children more likely to be stunted. educational status of mothers: children from educated mothers were less likely stunted as compared with illiterate mothers. the association of maternal education with chronic malnutrition of children in his study is consistent with m a n y o f the previous studies on the subject. education of women exerts impacts on health and nutritional status of children since it provides the mother with the necessary skills for childcare, increases awareness of nutritional needs and preference of modern health facilities as well as change of traditional beliefs about diseases causation [1]. income are positively associated with child nutritional status (underweight and wasting) which means the house hold those receive better income have more probability to averse the risk of child underweight and wasting as compared with household with less income. land-size: households with more land size are less likely to have children who are stunted, underweight and wasted. livestock ownership (tlu) is positively associated and statistically significant in the models (haz, and waz) this indicates that households with more live-stocks children are less likely to be stunted, and underweight. family size: the negatively significant relationships among household size and underweight could be explained by the fact that the family meal is distributed among large numbers of household members resulting to inadequate diet for an extended period eventually causing malnutrition which is similar with the findings of other study conducted in tanzania or kenya [16]. water source and sanitation: statistically significant and associated in all three models (haz, waz and whz) households that use water from unprotected/unsafe sources like dug well, river, spring children who were more likely wasted and underweight or in other words, child who had access to potable clean water were less exposed to illness of diarrhea in comparison to the child who get unprotected water [17]. health care: the result raveled that in the households where the mother made more antenatal visits the child was less wasted and underweight. vaccination reduces the incidence of wasting and underweight. time elapse to fetch water: when the child mothers went long distance to fetch water and took more time and create addition work load for mother/women/, this reduce child care and feeding and increase child stunting. toilet/latrine: households these have latrine without slab children were more likely stunted, underweight and wasted due to sanitation problem as compared to households these have latrine with slab. agriculture and food sciences research, 2017, 4(2): 45-57 54 sure regression results indicate that stunting and underweight of children were complementary and occurs together (positively related), but stunting and wasting are supplementary (negatively related) they are substitutable. this revealed that a child who stunted had a probability of underweight also. underweight and wasting are complementary occurs together and positively related, that means children those who are underweight had a probability of wasting. multivariate probity regression estimation results indicate probability of stunting, wasting and underweight multivariate probity (msl, # draws) =5, number of obs = 330, wald chi2 (45) = 847.74, log pseudo like hood = -423.34337, prob> chi2 = 0.0000 table-4.10. multivariate probity regression analysis estimation results in anlemoworeda, 2017. variables stunting (haz) underweight (waz) wasting (whz) coef. std.err z coef. std.err z coef. std.err z agehh 0.352 0.212 1.66* -0.363 0.243 -1.49 -0.743 0.337 -2.20 hhsize -0.188 0.115 -1.64* 0.091 0.122 0.75 0.403 0.147 2.73** edulevel -0.033 0.052 -0.64 -0.047 0.062 -0.75 -0.153 0.061 -2.51* childagm 0.008 0.005 1.57* 0.008 0.005 1.46* 0.005 0.006 0.93 sexochild -0.347 0.145 -2.39*** -0.120 0.170 -0.71 0.272 0.187 1.45* maincome -0.028 0.116 -0.25 0.163 0.139 1.17* 0.186 0.139 1.34* landsize -0.104 0.070 -1.47* 0.045 0.70 -0.65 0.027 0.036 0.32 antnatal 0.006 0.436 0.02 -0.285 0.599 -0.48 -0.782 0.328 -2.38* exclusive -4.332 0.389 -11.12 -3.227 0.370 -8.72 -2.692 0.468 -5.74 compf 0.108 0.191 0.56** -0.294 0.241 -1.22 -0.340 0.248 -1.37 watersourc 0.078 0.041 1.91** 0.111 0.048 2.28* -0.003 0.052 -0.07* toilt 0.034 0.136 0.25 -0.036 0.140 -0.26 0.011 0.200 0.06* wasteds 0.024 0.053 0.46 -0.046 0.068 -0.68 -0.075 0.069 -1.09 fetchwt 0.122 0.120 1.02* -0.345 0.138 -0.25 -0.009 0.136 -0.07 tlu 0.076 0.047 1.60* 0.004 0.052 0.08* -0.102 0.059 -1.71* cons 3.077 1.112 2.77 3.406 1.381 2.47 3.446 1.570 2.19 /atrho21 0.635 0.111 5.6 /atrho31 -0.224 0.124 -1.81 atrho32 0.529 0.130 4.07 rho21 0.561 0.076 7.34 rho31 -0.220 0.117 -1.87 rho32 0.484 0.099 4.88 note: likelihood ratio test of rho21=rho31=rho32=0 chi2 (3) = 73.5964 pro>chi2=0.0000 above table result shows that the probability of occurrence of stunting and underweight is complementary and positively related while stunting and wasting are negatively related. underweight and wasting are also complementary and positively related. figure-4.9. indicates that the kernel density estimate indicates that the error term is less and data is similar with the normal distribution estimate. source: own field survey, 2017 focus group discussion (fgd): were held in 4 groups each holds 8 members, which includes health extension workers, child mothers and care givers were the members of group discussion, key findings from focus group discussion and associated factors that cause child malnutrition were, shortage of crop land, low income, low knowledge on child feeding and care, rain fall variability cause shortage food, lack of clean water, poor sanitation, large family number insufficient knowledge on complementary food preparation for their children. 5. discussion in ethiopia as well as in the study area and in other developing countries, malnutrition in under-five children is a major public health problem. the findings of this study revealed that more than forty two percent of children aged between 6-59 months were affected by stunting. the prevalence of stunting, underweight and wasting among children ages 6-59 months in anlemo woreda were about 42.4%, 16.1% and 9.7% respectively. 5.1. stunting the study finding to the national prevalence for public health significance, is greater than 38.4% for stunting, it is higher than the national figure which needs the attention of all concerned body. edhs [4] shows that the prevalence rate of stunting, amhara, tigray, snnpr and oromia, regions were found as 46.3 %, 39.6%, 38.6 % and agriculture and food sciences research, 2017, 4(2): 45-57 55 36.5 % respectively. stunting in this study when compared as to the above regions was less than amhara, but higher than tigray, snnpr, oromia regions. this may be due to sample size differences and also the woreda under study is food insecure and psnp beneficiary since, 2005. the study result revealed a higher prevalence of stunting in boys 48.8% as compared to girls 36.3%. female children were 1.3 times less likely to be stunted compared to male children. this result was similar to edhs [4] and other studies done in ethiopia 41%, 35% respectively. the cause of this discrepancy in sex is no well established in the literature, but it is believed that boys are more influenced by environmental stress than girls [18]. child health and care practice in this study 84.5% of children feed colostrums 15.5 % of children were not depilated the first milk/colostrums. 99.1% of children were exclusively breast feed until six months and 82.1 % of children were start complementary feed at the age of six months and 17.9% of children start seven and above seven months. 5.2. underweight the finding of this study shows 16.1% of the children age 6-59 months were affected by underweight. this finding was lower than the national prevalence figure 23.6% as well as the regional 21.1%.this might be due to difference in sample size and area coverage. age of children from 24-35 months was about 3.3 times more likely to be affected by underweight than children age 6-11 months. similarly children aged 36-47 months were about 2 times more likely to be affected by underweight. edhs [19] show that the proportion of underweight children was highest in the age group 24-35 months 34% and the lowest among those less than six months 10%. 5.3. wasting the finding of this study revealed 9.7% of children age 6-59 months was affected by wasting which is similar with 2016 edhs national figure 9.9 % as well as snnpr 6%. unlike stunting younger children were more likely to be wasted than older children. this might be due to the increased susceptibility of younger children to illness/infection such as diarrheal disease. 5.4. the result of multivariate probity regression model of analysis revealed that age of the household head, child age, antenatal care, complementary feeding, water source, and toilet, time elapse to fetch water and livestock ownership are significant and associated with child stunting, while family size, child age, income, livestock ownership and water source are significant and associated with child underweight. child age, family size, sex of child, income and latrine/toilet are significant and associated with child wasting. income are positively associated with child nutritional status (underweight and wasting) which means the house hold those receive better income have more probability to averse the risk of child underweight and wasting as compared with household with less income. land-size: households with more land size are less likely to have children who are stunted, underweight and wasted similar with harold [20]. livestock ownership (tlu) is positively associated and statistically significant in the models (haz, and waz) this indicates that households with more live-stocks children are less likely to be stunted, and underweight which is in line with study of unicef [21]. family size: the positively significant relationships among household size and wasting, and underweight could be explained by the fact that the family meal is distributed among large numbers of household members resulting to inadequate diet for an extended period eventually causing malnutrition which is similar with the findings of other study conducted in tanzania or kenya [16]. an increase in household members has shown a strong negative influence on children weight for age. that is, as the size of household members changes the likelihood of weight-for-age decreases or in other words increases the risk of underweight. this might be because of depletion of resources due to the large family size that decreases dietary intake which deteriorates health conditions. water source and sanitation: positively associated and statistically significant in all three models (haz, waz and whz) households that use water from unprotected/unsafe sources like dug well, river, spring children who were more likely wasted and underweight or in other words, child who had access to protected /clean water were less exposed to illness of diarrhea in comparison to the child who get unprotected water in line with the study of ephraim and harold [22]. antenatal care: the result raveled that in the households where the mother made more antenatal visits the child was less wasted and underweight. vaccination reduces the incidence of wasting and underweight which is in line with the study of kibebew [23]. time elapse to fetch water: when the child mothers went long distance to fetch water and took more time and create addition work load for mother/women/, this reduce child care and feeding and increase child stunting in line with the study of young live ethiopia. toilet/latrine: household these have latrine with slab children were less likely underweight and wasted as compared to no latrine.sure regression result indicate that the interaction between the three outcome variables (stunting wasting and underweight). stunting and underweight are complementary (positively related) which means they occurs together while stunting and wasting are supplementary which means they are substitutable. wasting and underweight are complementary they occurs together. 6. conclusion and recommendations 6.1. conclusion the aim of the study is to investigate the determinants of nutritional status of under-five children in anlemoworeda selecting 330 children within among households using random sampling method. the findings of the study indicate that nutritional status of under-five children especially stunting and wasting is highly prevalent. agriculture and food sciences research, 2017, 4(2): 45-57 56 the prevalence of stunting is 42.4% in the area under study, which is higher than the 2016 edhs national and regional prevalence figures, which are 38.4% and 38.6%, respectively. nevertheless, wasting in the study woreda is 9.7%, which is nearly similar to the national prevalence, i.e., 9.9%, though it is higher than the prevalence of the region, which is 6%. to determine the factors associated with nutritional status of under-five children, multivariate probity regression analysis is applied thus, these factors are categorized as demographic, socio-economic, child healthcaring and environmental factors. the result of multivariate probity regression analysis indicated that among the risk associated factors: age of household head, child age, complementary-feeding, ownership of livestock, latrine, source of water, land-size and time elapse to fetch water are significant and associated with stunting of children aged 6-59 months. whereas, family size, child age in month, sex, livestock ownership, water source and income are associated with child underweight and wasting. the study reveals that male children are significantly at higher risk of stunting, and underweight than their female counterparts. thus, in order to tackle these problems efforts should be done to improve sources of water. parental education is also crucial to prevent and control childhood illness. 6.2. recommendations child malnutrition is one of the major problems in the study area that affect the well-being and proper growth of under-five children. identifying and understanding the factors that determine stunting, underweight and wasting in different setting provide valuable information for policy makers and researchers; and the community at large; and practitioners who work in the area as well. therefore, based on the findings of the study, the following recommendations are forwarded: the local government of the hadiya zone, and anlemoworeda health institution should design and implement effective child nutritional and health programs for the community to reduce the identified malnutrition problems, which indicate 42.4% stunting and 9.7 % wasting. about 54.8% of the rural households get water from unprotected sources such as dug well, spring river/surface water which results in child diarrhea and illness. therefore, local government should facilitate clean water supply for rural households/community. the educational status of the child mothers 32.7% were illiterate, therefore should be promoted as it has been proved that it is the key to reduce both chronic and acute malnutrition problems in children (stunting, underweight and wasting) and to find sustainable solutions to malnutrition. for the community: more than sixty one percent of the households had open latrine without slab, measures aiming at promoting sanitation through encouraging people to own improved latrines among rural households should be devised to increase the level of sanitation and hence reduce diseases such as diarrhea and its consequences on children health. children are the most vulnerable group of the society, child age specific attention should be given a due attention while feeding and care, and producing in their backyards traditional nutritious foods. the availability of food alone was not sufficient to address the nutritional security of children therefore guiding and creating child feeding awareness to rural households/mothers are critical. child breast feeding after six months of age requires integration with appropriate complementary feeding. responsible ngos that works on health and nutrition like unicef shall expected to give due emphasis to minimize the mentioned gap by addressing exposed children. lastly, for researcher: stunting in male child is higher than female children however the rationale behind this was not clearly understood or well established it requires further research investigation references [1] b. gezae and r. nigatu, "nutritional status of children under five years of age in shire indaselassie, north ethiopia: examining the prevalence and risk factors," kontakt, vol. 16, pp. e161-e170, 2014. view at google scholar | view at publisher [2] g. world health organization, "who child 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[23] a. kibebew, "infant and young child feeding practices among mothers living in harar, ethiopia," harar bulletin of health sciences, vol. 4, pp. 66-78, 2012. view at google scholar asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://scholar.google.com/scholar?hl=en&q=prevalence%20of%20malnutrition%20and%20associated%20factors%20among%20children%20age%206-59%20months%20at%20lalibala%20town%20administration,%20north%20wollo%20zone,%20anrs,%20northern%20ethiopia https://scholar.google.com/scholar?hl=en&q=prevalence%20of%20malnutrition%20and%20associated%20factors%20among%20children%20age%206-59%20months%20at%20lalibala%20town%20administration,%20north%20wollo%20zone,%20anrs,%20northern%20ethiopia https://scholar.google.com/scholar?hl=en&q=infant%20and%20young%20child%20feeding%20practices%20among%20mothers%20living%20in%20harar,%20ethiopia agriculture and food sciences research vol. 4, no. 1, 20-23, 2017 issn(e) 2411-6653/issn(p) 2518-0193 doi: 10.20448/journal.512.2017.41.20.23 20 biocidal properties of cymbopogon citratus extracts on termite (microcerotermes beesoni) olotuah, o.f1  dawodu, e.o2 ( corresponding author) 1department of plant science and biotechnology, adekunle ajasin university, akungba-akoko, ondo state, nigeria 2department of agricultural education, college of education, ikere-ekiti, ekiti state, nigeria abstract the aim of this experiment was to provide a biological approach to termite control through the use of extracts of roots and shoots of lemon grass (cymbopogon citratus). this was a laboratory experiment that involved various treatments through in-vitro contact applications of plant extracts and a pre-screening experiment on solutions for lethal properties against termites. the experiment was conducted at the laboratories of the department of plant science and biotechnology and the chemical sciences of adekunle ajasin university, akungba-akoko, ondo state, nigeria between august, 2005 and march, 2006. the roots and shoots of c. citratus were obtained from its tuft, sundried for 4 weeks and chopped into match-stick sizes. extracts were obtained from the plant through non-polar (n-hexane) and polar (ethanol) solvents. in the laboratory experiments, the efficacy of the obtained oil extracts in controlling termites were determined through contact tests on termite castes and mortality rate recorded. this was also compared to a control contact invitro experiments where water, concentrated n-hexane and ethanol were determined for efficacy in termite control. means of the results obtained were separated using tukey's honestly significant test. analysis showed that there was no significant different at p < 0.05 between n-hexane shoots extract and n-hexane roots extract at 60seconds treatment applications. observations showed that extracts obtained from all the parts were effective in the control of the different castes of termites with the n-hexane roots extract showing the fastest efficacy within 50seconds. keywords: termites, pest control, cymbopogon citratus, lemon grass, plant extract. citation | olotuah, o.f; dawodu, e.o (2017). biocidal properties of cymbopogon citratus extracts on termite (microcerotermes beesoni). agriculture and food sciences research, 4(1): 20-23. history: received: 12 may 2017 revised: 8 june 2017 accepted: 23 june 2017 published: 5 july 2017 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: i wish to acknowledge the laboratory officers that assisted in the preparation of the extracts as well as my undergraduate project students that were on the field with me during sample collection. the work was solely done by the author as no other person contributed meaningfully to the manuscript. funding: this project was self-sponsored. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 21 2. material and methods ..................................................................................................................................................................... 21 3. results and discussion ................................................................................................................................................................... 22 4. conclusion ......................................................................................................................................................................................... 23 references .............................................................................................................................................................................................. 23 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2017.41.20.23&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=olotuah,%20o.f https://orcid.org/orcid-search/quick-search?searchquery=dawodu,%20e.o agriculture and food sciences research, 2017, 4(1): 20-23 21 1. introduction lemon grass is an aromatic plant belonging to the poaceae family akhila [1]. it is a tall, clumped perennial grass growing to a height of 1 m. the leaf-blade is linear, tapered at both ends and can grow to a length of 50 cm and width of 1.5 cm. conversely, flowering has never been observed under cultivation due to rapid harvesting time. the rhizome produces new suckers that extend vertically as tillers to form dense clumps. lemongrass can tolerate a wide range of soils and climatic conditions. however, vigorous growth is obtained on well-drained sandy loam soil with high fertility and exposed to sunlight. the insecticidal activities of various extracts of the plant on vectors of medical or veterinary interests or nonagricultural pests was reported ratnadass and wink [2] and which also encompasses mosquitoes, mites, cockroaches, houseflies, termites and water snails and a host of human or cattle parasites. the anticoagulant activity of the oil extract of lemongrass was detected while trying to examine its pesticidal properties. due to the environmental concern and expensive synthetic insecticides, the demand for alternative sources of insect pest control had increased. also, the pervasive use of these insecticides in granaries of small-scale farmers has also led to a number of shortcomings such as elimination of non-target species, user hazards, found residues, evolution of resistance to the chemicals, high cost of the chemicals and the destruction of the balance of the ecosystem. conversely, termites (odontotermes sp.) had been reported to reduce crop yield and destroy stored grains. they are reported to be serious pests of various crops (wheat, sugarcane, groundnut and paddy) and cause significant yield losses in annual and perennial crops and damage wooden structures in buildings, especially in semi-arid and sub-humid tropics. the control of termites had initially centered on the use of synthetic termiticides but observed hazards from residue had informed the adoption of chloropyriphos in the treatment of soil against subterranean termite, but this application was not devoid of hazard to human and livestock as its chronic exposure had been linked to neurological effects and developmental disorders. presently, continuous use of synthetic termiticides for soil as well as crop treatment has been allowed for the present time because of the lack of any effective substitute. although, researches are on-going for a quest for an effective formulation, which can reduce the damage by termites and also be environmental friendly. as an alternative to synthetic insecticides, several biodegradable botanicals had also be employed in pest management and had been proven to be reliable. sequel to this, this experiment was focused on the adoption of botanicals in pest control and it had centered primarily on the control of termites using extracts of cymbopogon citratus obtained through polar and non-polar solvents in a laboratory contact experiment. provide a factual background, clearly defined problem, proposed solution, a brief literature survey and the scope and justification of the work done. 2. material and methods the materials used during this experiment were: lemongrass roots and shoots, ethanol, n-hexane, cutlass, spade, hand trowel, petri-dishes, syringe, stop watch, jar bottle, cover bowl, rotary evaporator, foil paper, filter paper and knife and matured shoot and root of cymbopogon citratus were collected from the demonstration plot of adekunle ajasin university, akungba-akoko, ondo state, nigeria. the research project was conducted at the department of plant science and biotechnology laboratory, adekunle ajasin university akungba akoko. during extract preparation, obtained plant parts were washed and 400g of the fresh shoots was chopped into match-stick sizes and stepped in 700ml of ethanol and n-hexane into a 3litres of rubber bucket with a closed lid. this was kept intact with a paper tape securely wrapped around the cover of the bucket to prevent evaporation. this set up lasted for 7days using ethanol and n-hexane as the solvents in each experimental setup and then mixed with electric shaker for 2minutes and repeated 3times daily. it was then filtered using vacuum filtration through the buchner funnel. the content of this bucket was immediately poured to the rotary evaporator and the temperature set at the boiling point of each of the solvents under reduced pressure to obtain the extract from the fresh shoots. the extract was preserved in a covered conical flask in the laboratory at ambient temperature and used throughout the experiment. this was similarly repeated for the fresh roots. 2.1. collection of termites collection of termites was done early in the morning from the termite hill on the field within the premises of adekunle ajasin university, akungba akoko, ondo state, nigeria. the termite samples were collected from their natural habitat (termitarium) and transferred to mc cartney bottle which contained part of their termitarium. this was kept on the cabinet in the laboratory at ambient temperature throughout the experiment. 2.2. in-vitro experiments there were two in-vitro experiments as stated below. 2.2.1. control in-vitro experiment involving the use of solvents the mortality experiment was performed by applying at different time intervals of 10seconds, 20seconds, 30seconds, 40seconds, 50seconds and 60seconds, 5ml each of water, n-hexane and ethanol differently on 15 workers and 15 soldiers of termite in a petri dish and placed in a growth chamber at ambient temperature. the experiment had three replicates. 2.2.2. in-vitro experiments involving the use of plant extracts these were performed by applying 5ml of all the extracts of shoot and root differently prepared with n-hexane and ethanol with a syringe on the termite caste which contained 15 workers and 15 soldiers in a petri dish and placed in a growth chamber at ambient temperature. the extracts were tested for mortality on the selected castes of termites containing 15 soldiers and 15 workers and the experiment had three replicates. 5ml of the extracts was applied to the termite castle at different time interval 10seconds, 20seconds, 30seconds, 40seconds, 50seconds and 60seconds. the results obtained on the mortality rate at the different time intervals were cumulated and means agriculture and food sciences research, 2017, 4(1): 20-23 22 separated using tukey’s honestly significant test analysis. this experiment had three replicates and a control containing the castes of termites but without any treatment application. 3. results and discussion it was observed that the application of the different solvents had no effect on the termites from 10 to 60 seconds of treatment application. this is presented in table 1. table-1. cumulative means of mortality rate of termite caste under different applications of solvents solvent time of application 10 sec 20 sec 30 sec 40 sec 50 sec 60 sec water 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a n-hexane 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a ethanol 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a 0.00±0.0a note: *means sharing the same superscript along columns are not significantly different at p < 0.05 using tukey´s honestly significant test. conversely, after 10seconds of application of all the extracts, n-hexane root extract recorded the highest number of mortality. while the least mortality was observed in ethanol shoot extract and followed by ethanol root extract. there were significant differences among all the plant extracts used at 10seconds of treatment application. after 20seconds of application of all the extracts, the least mortality was observed in ethanol shoot extract replicate while the highest mortality was observed in n-hexane root extract replicate. this was followed by nhexane roots extract. there was no significant difference between ethanol shoots and ethanol plant extracts at 20seconds of treatment application. after 30 seconds of application, the extract proved to be more effective than the preceding treatments as higher mortality rate was observed. n-hexane root extract recorded highest mortality rate and the lowest mortality in ethanol shoot extract. at 30seconds of treatment application of all the extracts, there were significant differences among all the plant extracts. after 40seconds of application, it was observed that n-hexane roots extract recorded the highest mortality rate which was followed by n-hexane shoot plant extracts. the least mortality rate was observed in ethanol shoots extract but there was significant difference among all the plant extracts used at 40seconds of treatment application. between 50seconds and 60seconds of treatment application, all the extracts had almost been completely eliminated except in ethanol shoot extract. the highest mortality was eventually observed in n-hexane root extract, in which complete mortality was observed within 50seconds. there was no significant difference between n-hexane roots and shoots extracts at 60seconds of treatment application. this observation is presented in table 2. table-2. cumulative mean of mortality rate of termite caste under different application regimes extract type time of application 10 sec 10 sec 10 sec 10 sec 10 sec 10 sec n-hexane shoot 9.00±1.6c 11.66±0.7b 18.33±0.7c 22.33±0.9c 26.33±0.7c 30.00±0.0c n-hexane root 10.33±0.2c 13.66±0.9c 20.33±0.9d 25.33±0.5d 28.00±1.2d 30.00±0.0c ethanol shoot 4.00±1.2a 7.66±0.1a 11.33±0.6a 14.66±0.9a 15.33±0.9a 22.33±0.1a ethanol root 5.66±0.1b 7.00±0.7a 13.66±1.5b 16.66±1.2b 22.33±0.9b 24.33±0.9b note: *means sharing the same superscript along columns are not significantly different at p < 0.05 using tukey´s honestly significant test. pesticides had earlier been reported to be useful in the increase of farm yield products and farmers had depended heavily on the synthetic pesticides to control insects in their crops, which till today remains one of the most commonly used method in controlling insects. the use of synthetic pesticides in agriculture comes with a cost for the environment and the health of animals and humans. sequel to the attendant problems associated with the use of synthetic pesticides, the use of biodegradable botanicals are now preferred being that botanicals are readily available from local plants with little or no processing. zhu, et al. [3] opined that they should have low toxicity to non-target organisms, especially humans and beneficial insects. consequently, in the determination of the efficiency of extracts, the norm of non-contributory lethal tendencies of the solutions was also observed in the control experiments. this had convincingly proven that the solvents were exclusive of the lethal effects observed in the extracts in termite control. similar views on the adoptive use of botanicals in pest management, were shared by keital [4] who reported that pulses stored in gunny bags treated with aqueous extract from leaves of melia azadirachta, hyptis suaveolens, tuber of cyprus rotundus and chrysopogon zizanioides were effectively protected against crop pests. this use of botanicals unequivocally corroborates the use of chrysopogon zizanioides in this research. in addition, keital [4] reported that seeds treated with botanical oil extracts did not lose their viability and also established the powder made from essential oil at different basics provided complete protection against m. beesoni and did not show significant effect on the potent on seed germination rate. this is an assertion of the use of morphological parts plants as being not restricted to any specific part. this also substantiates the use of the shoot and root of cymbopogon citratus and the observed efficacy in termite control. although kim [5] had proven the potential insecticidal activity of extract from cinnamomum cassia bark and oil, horse radish oil, mustard (brassica juneea) oil against callosobruchus chinensis, within one day after eucalyptus seed powder treatment caused the death of emerging adult of c. chinensis. on the control of termites rahman, et al. [6] had made a reported on the toxicological study on termites. this was in support of the use of plant materials as surface protectant against termite’s population growth by reducing the seed damage rate. in a similar observation, isman [7] had previously reported that volatile oil extract from azadirachta indica at high concentration could effectively protect agricultural crops, forestry from microcerotermes beesoni. these are contentions that corroborate the use of cymbopogon citratus in termite control in this research. in the preparation of natural pesticides, different solvents could be used. report from moein and farrag [8] had specified the use of n-hexane and ethanol, and had also drawn a comparison on agriculture and food sciences research, 2017, 4(1): 20-23 23 their adoptive uses. the report of 0.5% concentration of extract inducing 50% mortality that later dropped to 14.28% with ethanol is in agreement with the methodology adopted in this research on the use of the solvents and the subsequent reduction in efficacy with ethanol. the efficacy of lemon grass is consequently in accordance with the report of zue, et al. [9] on the use of lemongrass plant extracts as anti-termitic and repellent ingredient against termites where also a better performance was obtained in comparison to a synthetic termicide. the use of lemongrass extract in this research has provided safer alternative against the use of chemical termicides because of its low toxicity to non-target organisms, especially humans and beneficial insects and as reported by zue, et al. [9] and also in agreement with ratnadass and wink [2] that various extracts of the lemongrass have been reported to have insecticidal activities on vectors of medical or interests or non-agricultural pests which encompasses mosquitoes, mites, cockroaches, houseflies, termites and water snails and a host of human or cattle parasites. all the plant extracts recorded high mortality rate of termites based on the allocated time, therefore, in corroboration of cunningham, et al. [10] assertion, cymbopogon citratus could be used effectively as a substitute for synthetic pesticides as its use is not accompanied with deleterious environmental and public health effects by consuming foods cultivated with agrochemicals, by exposure during application and by consumption of intoxicated species. 4. conclusion the results from this study affirms the adoptive use of cymbopogon citratus extracts in the control of termites. consequently, its choice of solvent for formulation has a reduced effect on its efficacy. thus, the cultivation of cymbopogon citratus could be promoted as well as its use as edges at homes and fields in order to control termites. references [1] a. akhila, chemistry and biogenesis of essential oil from the genus cymbopogon. essential oil bearing plants. the genus cymbopogon. edited by: anand akhila. boca raton, fl: crc press taylor & francis group, 2010. [2] a. ratnadass and m. wink, "the phorbol ester fraction from lemon grass oil: potential and limits for crop protection against insect pests," international journal of molecular sciences, vol. 13, pp. 16157-16171, 2012. view at google scholar | view at publisher [3] b. c. zhu, g. henderson, f. chen, h. fei, and r. a. laine, "evaluation of vetiver oil and seven insect active essential oils against the formosan subterranean termite," journal of chemistry, vol. 27, pp. 1617-1625, 2001. view at google scholar [4] s. w. keital, "efficacy of essential oil of ocimum basilicum l. insecticidal fumigant l. applied as an insecticidal fumigant and powder to control callosobruchus maculatus [coleoptera; bruchidea]," journal of stored products research, vol. 37, pp. 339-349, 2001. view at google scholar | view at publisher [5] s. i. kim, "insecticidal activities of aromatic plant extracts and essential oils against sitophilus oryzea and callosobruchus chinensis," journal of stored products 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google scholar | view at publisher [10] a. a. cunningham, p. daszak, and j. p. rodriquez, "pathogen pollution: defining a parasitological threat to biodiversity conservation," journal of parasitology, vol. 89, pp. s78-s83, 2003. view at google scholar https://scholar.google.com/scholar?hl=en&q=the%20phorbol%20ester%20fraction%20from%20lemon%20grass%20oil:%20potential%20and%20limits%20for%20crop%20protection%20against%20insect%20pests http://dx.doi.org/10.3390/ijms131216157 https://scholar.google.com/scholar?hl=en&q=evaluation%20of%20vetiver%20oil%20and%20seven%20insect%20active%20essential%20oils%20against%20the%20formosan%20subterranean%20termite https://scholar.google.com/scholar?hl=en&q=efficacy%20of%20essential%20oil%20of%20ocimum%20basilicum%20l.%20insecticidal%20fumigant%20l.%20applied%20as%20an%20insecticidal%20fumigant%20and%20powder%20to%20control%20callosobruchus%20maculatus%20%5bcoleoptera;%20bruchidea%5d http://dx.doi.org/10.1016/s0022-474x(00)00034-5 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http://dx.doi.org/10.1007/s10340-008-0239-4 http://dx.doi.org/10.1007/s10340-008-0239-4 https://scholar.google.com/scholar?hl=en&q=pathogen%20pollution:%20defining%20a%20parasitological%20threat%20to%20biodiversity%20conservation agriculture and food sciences research issn: 2411-6653 vol. 2, no. 2, 51-55, 2015 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 51 effects of drying methods on physico-chemical properties and antioxidant activity of shiitake mushrooms (lentinus edodes) jiang-lian duan 1 --jian-guo xu 2* 1,2 college of food science, shanxi normal university, linfen, shanxi, china abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ............................................................................................................................................................................... 52 2. materials and methods ............................................................................................................................................................. 52 3. results ........................................................................................................................................................................................ 53 4. conclusions ................................................................................................................................................................................ 54 references ...................................................................................................................................................................................... 54 the effects of different drying methods including natural air-drying (nd), hot air drying (hd), vacuum drying (vd), microwave drying (md), infra-red drying (id), and freeze drying (fd) on physicochemical properties and antioxidant activities of shiitake mushroom were investigated in this paper. the results showed that drying methods had obvious effects on the color, proximate chemical compositions, total phenolics, antioxidant activity of shiitake mushroom. the fd and nd products exhibited the best rehydration and shrinkage ratio; the effect of fd on color was the least, followed by nd, while md and id influenced significantly the color of mushroom; six drying methods had no effects on the content of ash, crude fat and carbohydrate of shiitake mushroom. drying resulted in the decrease of total phenolic content and antioxidant activity of shiitake mushroom. among them, the mushroom dried with fd had no obvious changes compared with fresh mushroom. keywords: lentinus edodes, drying, physical property, chemical property, phenolics, antioxidant activity. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(2): 51-55 52 1. introduction in recent years, mushrooms are favorable all over the world not only because of their texture and flavour, their chemical, nutritional and functional properties such as antiallergic, antiatherogenic, antihypoglycemic properties are also well documented [1, 2]. shiitake mushroom (lentinus edodes) is a good source of nutrition because of its higher protein, dietary fibers and important mineral contents [3]. besides, shiitake mushroom contains many different phytochemical contents such as phenolic compounds, tocopherols, ascorbic acid and carotenoids [4]. therefore, mushroom (l. edodes) is a healthy food in our daily diet. however, the shelf life of shiitake mushroom is short because of its high respiration rate, tendency to turn brown and having no physical protection to avoid water loss or microbial attack [5]. as a result, it is necessary to find a method to extend their shelf life. drying is the most common method for preserving mushrooms, but different drying methods have different effects on the quality of the products because of differences in characteristics and mechanism of drying and materials [6-8]. the effects of different drying methods including natural air-drying (nd), hot air drying (hd), vacuum drying (vd), microwave drying (md), infra-red drying (id), and freeze drying (fd) on physico-chemical properties and antioxidant activities of shiitake mushroom are investigated in this paper, which would provide some foundational information for the developing and application of shiitake mushroom. 2. materials and methods 2.1. raw materials fresh mushrooms (lentinus edodes) were purchased form local market in linfen, china on may 8, 2014. the mushrooms with homogeneous color and size were selected and stored in a refrigerator maintained at 4 o c. the average initial moisture content of these mushroom samples was 88.4 ± 1.2% (w/w), as determined using a hot air oven at 105 o c. 2.2. sample preparation and drying the middle sections of mushrooms were sliced and then were divided into seven portions (accurately 250 g of each) at random. one of the samples was stored at 4 o c for further use, and other portions were dried by different methods in optimised conditions until the final moisture content was less than 10 % (w/w), respectively. detailed procedures for each drying method are described below: (a) natural air-drying (nd) at 23–30 °c by sunshine and flowing air; (b) hot air drying (hd) at 45 °c; (c) microwave drying (md) in a microwave oven at 240 w; (d) vacuum drying (vd) in a vacuum drying oven at 45 °c and vacuum of 0.08 mpa; (e) infra-red drying (id) in an ir moisture analyzer at 45 °c; (f) freeze drying (fd) in a freeze drier at 20 °c with the condenser temperature and chamber vacuum at -55 °c and 30 pa. the dried samples were packaged in aluminum foil bags for further quality analysis. 2.3. rehydration ratio (rr) the rr of dried mushroom was determined by immersing the dried samples in distilled water (50 ml of water per gram of mushroom) at room temperature for 30 min. following this, the samples were taken out from the water, and the excess water was removed from the surface using a dry blotting paper and the mass of the samples was measured. the rr was calculated according to the formula [9] rr= (w1-w0)/wo, where w1 and wo are the mass values of the rehydrated and dried samples, respectively. the rr values were determined in triplicate. 2.4. shrinkage ratio the volume of the test sample was measured using an excluding method, and the clean sea sand was selected as filling material [10]. the mean particle diameter of sand is about 0.6 mm. five replicates were performed for each sample and mean value was calculated. the shrinkage ratio was calculated as: rs (%)=100×(v0−v1)/v0, where rs is the shrinkage ratio of the sample, v0 is the initial volume of the sample before drying, v1 is the volume of the dried sample. 2.5. surface colour measurement the colour of sample was measured through the cie l*a*b* system using a cr-330 minolta colorimeter (minolta, ramsey, nj) calibrated with a white standard tile. the results were expressed as hunter colour values of l*, a* and b*, where l* was used to denote lightness, a* redness and greenness, and b* yellowness and blueness. hunter values of the fresh and dried samples were measured in triplicate. 2.6. chemical analysis the crude protein, fat, fibre and ash were determined following the aoac procedures [11]. the crude protein content (n×4.38) of the samples was estimated by the macro-kjeldahl method; the crude fat was determined by extracting a known weight of the sample with petroleum ether, using a soxhlet apparatus; the crude fibre was estimated by acid/alkaline hydrolysis of insoluble residues; the ash content was determined by incineration at 600±15 °c; the carbohydrate content was calculated by subtracting the other proximate compounds from 1 g dry sample. the content of all compositions are expressed as grams per 100 gram dry matter basis (dm). 2.7. antioxidant activity 2.7.1. determination of total phenolic content (tpc) the ground samples are blended with methanol and shaken at 25 °c at 150 rpm for 2 h, and then the homogenates are centrifuged for 15 min at 4 o c and 5 000 g. after centrifugation, the supernatants are vacuumevaporated to dryness at 40 o c, redissolved in methanol at 10 mg/ml, and stored at 4 o c for further use. agriculture and food sciences research, 2015, 2(2): 51-55 53 the tpc was determined based on the folin-ciocalteu colourimetric method as described by hu and xu [12]. briefly, an aliquot (0.5 ml) of the suitable diluted extracts, 2.5 ml of deionized water and 0.5 ml of 1.0 m folinciocalteu reagent were mixed within 10 ml volumetric flasks and vortexed. after 8 min, 1.5 ml of 7.5% sodium carbonate solution was added and mixed thoroughly. the absorbance of the reaction mixtures was measured using a spectrophotometer at 765 nm wavelength after incubation for 2 h at room temperature. extraction solvent was used as the blank and gallic acid (ga) was used for calibration of standard curve. phenolic content was expressed as milligrams of gallic acid equivalents (gae) per 100 g of dm. 2.7.2. antioxidant activity by dpph radical scavenging assay dpph radical scavenging activity was determined according to the method of hu and xu [12] with some modifications. briefly, each of sample solutions (1 mg/ml in methanol) was serially diluted to various concentrations in methanol respectively, and then a 0.5 ml of samples was mixed with 2.5 ml of 60 μm dpph dissolved in methanol. the mixture was shaken vigorously and left to stand for 30 min in the dark, and the absorbance was measured at 517 nm against a solvent blank. the scavenging rate on dpph radicals was calculated according to the formula, scavenging rate (%) = [(ao-a1)/ao]×100, where ao is the absorbance of the control solution, a1 is the absorbance in the presence of samples in dpph solution. 2.8. statistical analysis all results are expressed as mean ± sd (n=3). in order to evaluate the significant differences between sample means, one-way analysis of variance (anova) and duncan’s test were performed with significant level being considered at p < 0.05. 3. results 3.1. the effects of drying methods on rehydration ratio colour, rehydration and shrinkage ratio are three important factors to evaluate appearance and texture of dehydrated products. the rehydration capacities of mushroom samples dried with different drying methods are presented in fig. 1. among all the treatments, md got the lowest rehydration radio which reaches at 5.2-fold of dried body, whereas fd and nd got the highest rehydration radio which reaches at 8.7 and 8.4-fold of dried body respectively. this may be due to the changes in the structure and texture of the samples during different drying processing [13, 14]. we postulated that fd and nd provides enough interval time, leading the internal moisture diffuse to the surface which avoids the shrinking of surface. for all samples, rehydration radio is fast at the beginning of re-watering process and falling to equilibrium state at the end, which was been reported by other authors [15, 16]. 3.2. the effects of drying methods on shrinkage ratio fig. 1 showed effects of different drying methods on the shrinkage ratio of mushroom samples, and the influence of drying methods on the shrinkage ratio was evident. contrary to the results of rehydration ratio, shrinkage ratio of mushroom dried with fd and nd were the smallest, and were 14.5% and 15.2%, respectively. the shrinkage ratio of other samples ranged from 18.5% to 28.6%. this is because much more moisture of samples was removed rapidly at higher temperature, which resulted in greater changes of shrinkage ratio under the same drying condition. therefore, temperature of drying may be the main factor affecting the shrinkage ratio of dried mushroom. the changes in shrinkage ratio also directly affect rehydration ratio of samples. 3.3. the effects of drying methods on color the effects of drying methods on color of dried mushroom are shown in table 1. cie l*, a*, and b* values were significantly different among drying methods. as an objective evaluation of dehydrated products, cie l* values appear to be important and sensitive to color evaluation, which can be an indicator of lightness of color. the cie a* value indicates the redness (positive a*) and greenness (negative a*), while the cie b* value indicates the yellowness. the cie l*, a*, and b* values of fresh mushroom were 26.33, 0.95, 1.13 respectively. however, after drying cie l*, a*, and b* values ranged from 31.56 to 42.34, from 1.08 to 3.86, and from 2.51 to 6.31, respectively. these changes in the cie l*, a*, and b* values implied that drying treatment had obvious effects on the color of dried mushroom. among them, the effect of fd on color was the least, followed by nd, while md and id influenced significantly the color of dried mushroom. the phenomenon can be attributed to the enzymatic browning which may be the main reason for brown stain of the products [17, 18]. 3.4. the effects of drying methods on proximate chemical compositions the effects of drying methods on the chemical compositions of dried shiitake mushroom are shown in table 2. based on dry weight, it can be easily seen that the crude ash ranged from 6.3 to 6.6 g/100 g dw, crude fat ranged from 4.2 to 4.8 g/100 g dw, and the carbohydrate ranged from 54.1 to 54.7 g/100 g dw, which indicate that six drying methods had no effects on the content of ash, crude fat and carbohydrate of shiitake mushroom. the content of crude fiber of fresh mushroom was 11.2 g/100 g dw, and it increased and ranged from 11.5 to 16.1 g/100 g dw after drying, especially md and id. the protein are found in high levels and varied between 18.5 to 20.6 g/100 g dw. nd and fd had no obvious effect on protein content compared with fresh mushroom, while other drying methods can result in decrease of protein. overall, fd is the best drying method which basically retains the proximate chemical compositions of fresh mushroom, followed by nd. 3.5. the effects of drying methods on phenolics the effects of drying methods on phenolic content of shiitake mushroom are shown in fig. 2. the results showed that the drying methods had a satisfactory effect on the phenolic content of shiitake mushroom. the total phenolic of agriculture and food sciences research, 2015, 2(2): 51-55 54 fresh shiitake was 35.3 mg/100 g. a reduction in total phenolic contents of dried samples was found. during fd, the loss of phenolics was the lowest, followed by vd, nd, md, hd, and id. investigates its reason, the loss may be come from enzymatic and non enzymatic reaction of phenolic compounds during the drying process [19]. the internal temperature of materials was very high although the heating time was short during microwave drying process, while the heating time was longer during hot air drying. therefore, the higher drying temperature and the longer drying time lead to oxidation of phenolic compounds. 3.6. the effects of drying methods on antioxidant activities the effects of drying methods on the antioxidant activities of shiitake mushroom are shown in fig. 2. the results showed that the drying methods had obvious effects on the antioxidant activities of shiitake mushroom, and the effect was similar to tpc. the drying resulted in the decrease of scavenging activity on dpph radicals. the scavenging rate of fresh shiitake on dpph radicals was 75.4%, while the scavenging activity of fd products on dpph radicals was the near to fresh shiitake with 70.5%, followed by vd, nd, md, id and hd. in addition, we found that the content of phenolics was highly associated with antioxidant activity, indicating that the phenolic compounds contributed significantly to the antioxidant activity of shiitake mushroom, which was in agreement with the previous studies [19, 20]. 4. conclusions in conclusion, this work showed that drying methods had obvious effects on the color, proximate chemical compositions, total phenolics, antioxidant activity of mushroom. the fd and nd products exhibited the best rehydration and shrinkage ratio; the effect of fd on color was the least, followed by nd; six drying methods had no effects on the content of ash, crude fat and carbohydrate of shiitake mushroom. drying resulted in the decrease of total phenolics, antioxidant activity of shiitake mushroom. among them, the mushroom dried with fd had no obvious changes compared with fresh mushroom. references [1] p. kalač, "chemical composition and nutritional value of european species of wild growing mushrooms: a review," food chem., vol. 113, pp. 9-16, 2009. 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[20] a. wojdyło, a. figiel, and j. oszmlański, "effect of drying methods with the application of vacuum microwaves on the bioactive compounds, color, and antioxidant activity of strawberry fruits," j. agric. food chem., vol. 57, pp. 1337–1343, 2009. agriculture and food sciences research, 2015, 2(2): 51-55 55 table-1. the color of fresh and dried shiitake mushroom l* a* b* fresh 26.33 ± 0.55 f 0.95 ± 0.05 d 1.13 ± 0.06 c nd 33.32 ± 1.46 de 1.13 ± 0.08 cd 2.56 ± 0.14 b hd 35.11 ± 1.65 cd 1.85 ± 0.52 cd 2.64 ± 0.52 b vd 37.27 ± 1.25 bc 2.18 ± 0.34 bc 2.93 ± 0.31 b id 40.55 ± 1.54 ab 3.25 ± 0.25 ab 5.84 ± 0.55 a md 42.34 ± 1.13 a 3.86 ± 0.84 a 6.31 ± 0.24 a fd 31.56 ± 0.82 e 1.08 ± 0.10 cd 2.15 ± 0.13 b numbers represent mean values of three independent replicates ± sd. different letters indicate statistically significant differences between the means (p < 0.05) for each parameter. table-2. proximate compositions (g/100 g dm) of fresh and dried shiitake mushroom crude protein crude fat carbohydrate crude fiber crude ash fresh 20.2 ± 0.4 a 4.5 ± 0.2 ab 57.7 ± 1.8 a 11.2 ± 0.6 de 6.3 ± 0.5 a nd 20.4 ± 0.2 a 4.4 ± 0.1 ab 56.2 ± 0.9 a 12.5 ± 0.3 d 6.4 ± 0.8 a hd 18.5 ± 0.1 c 4.7 ± 0.2 a 55.8 ± 1.4 a 14.5 ± 0.2 c 6.4 ± 0.6 a vd 19.4 ± 0.1 b 4.7 ± 0.1 a 54.7 ± 1.3 a 14.8 ± 0.5 bc 6.3 ± 0.5 a id 18.9 ± 0.2 bc 4.8 ± 0.3 a 54.1 ± 0.8 a 15.6 ± 0.1 ab 6.5 ± 0.4 a md 18.9 ± 0.3 bc 4.2 ± 0.4 ab 54.2 ± 1.4 a 16.1 ± 0.4 a 6.6 ± 0.2 a fd 20.6 ± 0.2 a 4.6 ± 0.2 ab 56.9 ± 1.2 a 11.5 ± 0.3 e 6.3 ± 0.5 a numbers represent mean values of three independent replicates ± sd. different letters indicate statistically significant differences between the means (p < 0.05) for each parameter. fig-1. effect of drying methods on rehydration and shrinkage ratio fig-2. the effects of drying methods on phenolic content and antioxidant activity of shiitake mushroom . views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. agriculture and food sciences research issn(e) : 2411-6653 issn(p) : 2518-0193 vol. 3, no. 1, 45-48, 2016 http://www.asianonlinejournals.com/index.php/aesr 45 effect of feeding regimen on relationship between carcass measurements, slaughter weight and hot carcass weight of sudanese desert goats m.o.mudalal1 i. bushara2  dafalla m. mekki3 elgazafey besheer4 s.a.babiker5 1,2,3department of animal production. faculty of natural resources and environmental studies, university of kordofan el-obied, sudan. 4faculty of medicine, university of eldaein, sudan. 5faculty of animal production, university of khartoum, khartoum, sudan ( corresponding author) abstract the study was conducted to determine the effect of feeding diets, on growth performance and relationship between carcass measurements, slaughter weight and hot carcass weight, of 72 intact male desert bucks divided randomly into three groups; each group (24 males), the animals were slaughtered according to halal standard procedures. the results obtained indicated that correlation coefficients between slaughters weight and carcass measurements were significantly higher (p <0.01) and positive. in free grazing group correlation coefficients between hot carcass weight and carcass measurements were significantly (p <0.01) and positive. also the relationship between slaughters weight and carcass measurements was significant and positive except internal chest length. in the group that grazing and supplemented with diets showed positive and significantly (p <0.01) correlation coefficients between hot carcass weight and carcass measurements and between slaughters weight and carcass measurements were significant and positive except carcass length. the zero grazing groups obtained non not significant correlation coefficients between hot carcass weight and carcass measurements except slaughter weight and external chest length. keywords: goat, carcass measurements, hot carcass and slaughter weights. contents 1. introduction ............................................................................................................................................................................... 46 2. materials and methods ............................................................................................................................................................. 46 3. results and discussions ............................................................................................................................................................ 46 4. conclusions ................................................................................................................................................................................ 47 references ...................................................................................................................................................................................... 47 citation | m.o.mudalal; i. bushara; dafalla m. mekki; elgazafey besheer; s.a.babiker (2016). effect of feeding regimen on relationship between carcass measurements, slaughter weight and hot carcass weight of sudanese desert goats. agriculture and food sciences research, 3(1): 45-48. doi: 10.20448/journal.512/2016.3.1/512.1.45.48 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: contribution/acknowledgement: this work is licensed under a creative commons attribution 3.0 license all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 25 may 2016/ revised: 15 june 2016/ accepted: 20 june 2016/ published: 2 july 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.45.48 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=m.o.mudalal https://orcid.org/orcid-search/quick-search?searchquery=i.%20bushara https://orcid.org/orcid-search/quick-search?searchquery=dafalla%20m.%20mekki https://orcid.org/orcid-search/quick-search?searchquery=elgazafey%20besheer https://orcid.org/orcid-search/quick-search?searchquery=s.a.babiker http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.45.48 agriculture and food sciences research, 2016, 3(1): 45-48 46 1. introduction the goat population of sudan is mostly located in smallholder farming areas, and their function is the production of meat. the traditional production system is extensive, with grazing on natural grassland with herbs, shrubs and forest and with little or no feed supplementation and this is mainly based on natural, unimproved savannahs, which lead to poor animal performance [1, 2]. goats are prominent browsers for herbs [3]. if herbs used in the diets of animals could have significant impact on feed intake, meat quality and composition [4-6]. the presence of high levels of herbs in feed have been reported to be responsible for decreased feed intake [6, 7] growth rate, feeding efficiency and live-weight gain [6, 8]. the carcass characteristics and meat quality are important criteria for consumers when it comes to making purchasing decisions. carcass classification for conformation and fatness are the main parameters for determining the marker price. the linear carcass measurements, included length of carcass, depth of chest, length of leg and width of leg [9]. many authors had showed that nutrition had significant effect on slaughter weight, hot carcass weight and fore and hind quarter [10]. therefore, the producers are encouraged to continue producing according to the traditional systems and methods because the products are well accepted to the consumers. the feeding trials are ongoing in order to recommend supplementation strategies for the use of relatively expensive concentrates with tropical forage for feeding local goats raised for meat. the aim of the pilot study was to evaluate the effect of feeding regimen and castration on relationship between carcass measurements, slaughter weight and hot carcass weight. 2. materials and methods 2.1. study area this study was conducted at el-obeid agricultural research station, north kordofan state, sudan. 2.2. experimental animal’s management seventy two male desert goats weighing 12.1 kg (aged 4-5 months) were divided randomly into three groups; each group content 24 males. group one (free grazing only) on some dry grasses like eragrotis turmula, zornia glochiidiata and cenphrus spp and browse the leaves of ziziphus spina chritis, balanite aegyptiaca and calotropis purocera. group two (grazing + concentrate supplement) and group three (concentrate supplement only) ad libitum fed the experimental diet. the experimental diet given at a rate of 324g/ head/ day. ingredients and chemical composition of experimental diet (table 1). live weight and external body measurements of each animal were recorded at the start of experiment, then weekly during the experiment, using spring balance. the animals were weighed in the morning at weekly interval for 12 weeks. table-1. ingredients % and chemical composition of the experimental feed stuffs. components % chemical composition % sorghum grains 30 dry matter (dm %) 98.6 wheat bran 38 organic matter (om %) 87.7 ground nut cake 20 crude protein (cp %) 22.0 ground nut hay 10 crude fiber (cf %) 25.0 sodium chloride 1 ether extract (ee %) 12.0 limestone 1 nitrogen free extract (nfe %) 31.0 ash (%) 08.0 energy density (me(mj/ kg dm) 12.0 note: ** calculate as in maff [11]. 2.3. slaughter procedure and data collection twelve animals from each group were slaughtered at 90 weeks. the animals were slaughtered according to halal standard procedures. the external and internal offals including gut content were weighed. hot carcass, carcass halves were weight; the carcass was split along midline. the left half carcass were done according owen [12] and cut into wholesale cuts, each cut was dissected according to cuthbertson, et al. [13] into lean, fat, and bone. 2.4. statistical analysis the data were statistically analyzed according to factorial experimental randomized design as a 3×2 (management vs sex). multiple range tests were used to compare the means using [14]. 3. results and discussions 3.1. effect of free grazing on slaughter weights and carcass characteristics there is positive significant correlation between slaughters weight and carcass measurements table (2). the highest correlation coefficients (r =0.73, (p< 0.001) was found between slaughters weight and proximal hind leg length and between hot carcass and slaughter weight(r =0.63(p< 0.05). the slaughter weight and distal foreleg length exert lowest relationship and between hot carcass weight and internal chest length(r =0.56, (p< 0.05) . this result was in agreement with the findings of jibir, et al. [15] and alexandre, et al. [1] who reported that the there is positive relationship between carcass measurements, slaughter weight and hot carcass weight were significant positive for animal depending on natural grazing. agriculture and food sciences research, 2016, 3(1): 45-48 47 table-2. correlation coefficients between slaughter weights, hot carcass weight and carcass measurements (free grazing) slaughter weight carcass length distal foreleg length proximal foreleg length distal hind leg length proximal hind leg length internal chest length external chest length hot carcass weight (kg) 0.91*** 0.60* 0.78*** 0.61* 0.64** 0.73*** 0.56* 0.73*** slaughter weight (kg) 0.45ns 0.63* 0.68** 0.64* 0.73*** 0.67** 0.68** note: **** ,** and * significant at p<0.001, p<0.01 and p<0.05 respectively, ns= non significant 3.2. effect of free grazing plus supplementation on slaughter weights, hot carcass weight and carcass measurements the linear carcass measurements are indices of skeletal development and indirectly help to determine carcass conformation. diet had significant and positive (p > 0.01) effect on relationship exert between slaughters weight and carcass measurements except internal chest length table (3). the significant and highest positive correlation coefficients were found between slaughter weight and external chest length(r =0.85, (p< 0.001), between hot carcass weight and carcass measurements and between hot carcass weight and slaughter weight(r =0.96, (p<0.001) respectively. the values increased (p<0.01) progressively with inclusion of concentrate in the diet. the lowest correlation coefficients (r =0.65, (p< 0.01) between slaughter weight and distal foreleg length. and between hot carcass weight and distal foreleg (r =0.65, (p< 0.05). generally the increase in weights of the main carcass cuts was related to the carcass weight increase due to availability of energy and protein in diets. diet and slaughter weight may be the main causes of the differences with the results obtained by owen [12] and abdullah and musallam [16]. the increase in weights of between hot carcass and slaughter weight with internal chest length was related to be not vary significantly. this results is online with studied of oman, et al. [17] who reported that fed goats produced heavier slaughters weight and carcasses measurements than pasture goats. table-3. correlation coefficients between slaughter weights, hot carcass weight and carcass measurements (grazing + supplemented group) slaughter weight carcass length distal foreleg length proximal foreleg length internal chest length external chest length hot carcass weight (kg) 0.96*** 0.83*** 0.65* 0.80*** 0.55ns 0.85*** slaughter weight (kg) 0.84*** 0.64* 0.82*** 0.51 ns 0.79** note: **** ,** and * significant at p<0.001, p<0.01 and p<0.05 respectively, ns= non significant 3.3. effect of zero grazing on slaughter weights and carcass characteristics feed intake reported in (table 4) followed the same trend as all correlation coefficients between slaughters weight and carcass measurements were significant and positive. whoever, the highest correlation coefficients (r =0.87, (p< 0.001) exert between slaughter weight and external chest length, and between hot carcass weight and slaughter weight (r =0.96, (p< 0.001) the lowest correlation coefficients (r =0. 58, (p< 0.05) between slaughter weight and distal hind leg length, and between hot carcass weight(r =0. 60, (p< 0.05). all correlation coefficients between hot carcass weight and carcass measurements were not significantly except slaughter weight and external chest length which agree with simela, et al. [18]. this is consistent with the results reported by peña, et al. [19] and marichal, et al. [20] who reported that the correlation coefficients between carcass measurements, slaughter weight and hot carcass weight were positive and significant. the variation between these results and that obtained by other authors may be due to differences in weight at slaughter and mature size of breed used. table-4. correlation coefficients between slaughter weights, hot carcass weight and carcass measurements (zero grazing) slaughter weight carcass length distal foreleg length internal chest length external chest length hot carcass weight (kg) 0.96*** 0.52 ns 0.50 ns 0.35 ns 0.60* slaughter weight (kg) 0.54 ns 0.58* 0.76** 0.87*** note: ****, ** and * significant at p<0.001, p<0.01 and p<0.05 respectively, ns= non significant. 4. conclusions improvement in the plane of nutrition by supplementation of concentrates to grazing goats can promote faster daily live weight gain and increase slaughters weight and carcass measurements then have good and positive relationships between them, and thus enable the attainment of acceptable market weight at earlier age. references [1] l. alexandre, a. liméa, o. fanchonne, n. coppry, mandonnet, and m. boval, "effect of forage feeding on goat meat production: carcass characteristics and composition of creole kids reared either at pasture or indoors in the humid tropics," asian-aust. j. anim. sci., vol. 22, pp. 1140 – 1150, 2009. 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[20] a. marichal, n. castro, j. capote, m. j. zamorano, and a. argüello, "effects of live weight at slaughter (6, 10 and 25 kg) on kid carcass and meat quality," livestock production science, vol. 83, p. 247−256, 2003. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 1 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 1-8, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.1.8 © 2019 by the authors; licensee asian online journal publishing group multivariate analysis of genetic variation in rapeseed (brassica napus l.) elora parvin1 firoz mahmud2 md. shahidur rashid bhuiyan3 md. maksudul haque4 ( corresponding author) 1scientific officer, bangladesh institute of research and training on applied nutrition (birtan), dhaka, bangladesh 2,3professor, department of genetics and plant breeding, sher-e-bangla agricultural university, dhaka-1207, bangladesh 4senior scientific officer, bangladesh institute of research and training on applied nutrition (birtan), dhaka, bangladesh abstract designing breeding programs for rapeseed (brassica napus l.) cultivars with developed seed and oil yields needs information about the genetic variability of characters. in this study, 40 rapeseed genotypes were evaluated for genetic variation and relationships between 12 agro-morphological characters. brassica species represent a broad range of crops. this redirects the high degree of genetic diversity and allied phenotypic plasticity. significant differences among the clusters were observed. the first two components with eigen value were greater than unity contributed a total of 42.77% variation to the divergence. the genotypes were grouped into five clusters. cluster v contained the maximum number of genotypes (11) and cluster iii contained the lowest (4). the highest inter-cluster distance was found between cluster ii and cluster v and the lowest between cluster ii and cluster iv. the highest intra-cluster distance was noticed for cluster i and the lowest for cluster v. considering diversity pattern and other agronomic performance lines nap0837, nap-0733-1, nap-2066, nap-9901, nap-108 and bari-8 could be considered apposite parents. this information is suitable for hybrid breeding and encouraging breeders to exchange their germplasm as to enlarge the genetic diversity of breeding accessions. keywords: multivariate analysis, genetic variation, rapeseed. citation | elora parvin; firoz mahmud; md. shahidur rashid bhuiyan; md. maksudul haque (2019). multivariate analysis of genetic variation in rapeseed (brassica napus l.). agriculture and food sciences research, 6(1): 1-8. history: received: 18 october 2018 revised: 23 november 2018 accepted: 4 january 2019 published: 2 february 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ......................................................................................................................................................................................... 2 2. materials and methods ...................................................................................................................................................................... 2 3. result and discussion ........................................................................................................................................................................ 2 4. conclusion ............................................................................................................................................................................................ 7 references ................................................................................................................................................................................................. 8 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.1.8&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.1.8&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/1667 https://orcid.org/0000-0003-0324-6834 https://orcid.org/0000-0001-9491-2753 agriculture and food sciences research, 2019, 6(1): 1-8 2 © 2019 by the authors; licensee asian online journal publishing group 1. introduction rapeseed (brassica napus l., 2n=38) is an important oil seed crop belonging to the family cruciferae. the seeds of mustard and rapeseed contain 42% oil and 25% protein. oilseed rape (brassica napus l.) is third most important oil crop in the world. today oilseed rape (b. napus ssp. napus) is the most important source of vegetable oil in europe and the second most important oilseed crop in the world after soybean. however, its limited geographic range and intensive breeding has led to a fairly thin genetic basis in present breeding material. the gene pool of elite oilseed rape breeding material has been more eroded by an emphasis on specific oil and seed quality characters. as a consequence, genetic variability in this vital crop is restricted with regard to several characters of importance for breeding purposes. oil and fat are not only the basis of energy but they also contain fat-soluble vitamins a, d, e and k. the oil cake holds proteins of high biological value and applicable quantities of calcium and phosphorus and is used as a very good animal feed as well as fertilizer for many crops. rapeseed laterally with mustard is currently ranked as the world’s third vital edible oil crop in terms of area and production after soybean and cotton. major producing areas include canada, china, northern europe, and the indian sub-continent. the rapeseed and mustard grown in bangladesh comprise three species viz. b. campestris, b. juncea, and the newly introduced b. napus. these crops have the largest zone and production among the oil crops grown in bangladesh. genetic diversity is vital to develop cultivars with improved yields, wider adaptation, desirable qualities, and pest and disease resistance. inclusion of other diverse parents (within a limit) in hybridization is supposed to raise the chance of obtaining supreme heterosis and give a wide spectrum of variability in segregating groups. variability and genetic diversity are the ultimate law of plant breeding which is a major tool being used in parent selection for effective hybridization program [1]. however, its partial geographic choice and intensive breeding has run to a moderately narrow genetic basis in present breeding material. the gene pool of best oilseed rape breeding material has been further worn by an emphasis on explicit oil and seed quality characters. as a consequence, genetic inconsistency in this vital crop is delimited with regard to many types of value for breeding purposes. approximations of genetic diversity in a crop species can contribution in the valuation of germplasm collections as potential gene pools to improve the concert of cultivars. genotyping accessions and assessing the level of genetic diversity inside a germplasm group can be of use to plant breeders in a number of ways. with this aim, an attempt was made in the present study to analyse genetic diversity among 22 advanced genotypes of b. napus. 2. materials and methods twenty two progressive genotypes of b. napus were grown in randomized complete block design with three replications. the advanced lines of brassica napus l. for the trial were collected from the department of genetics and plant breeding, sher-e-bangla agricultural university (sau) and three released varieties collected from bangladesh agricultural research institute (bari) as presented in table 1. the plot size was 3 m length with two rows. row to row and plant to plant distances were 25 cm and 15 cm, respectively. seeds were sown in lines in the trial plots. observations were recorded on 10 randomly chosen plants from each plot. data were collected on days to 1st flowering, days to 50% flowering, days to 80% flowering, days to maturity, plant height (cm), number of primary branches per plant, number of secondary branches per plant, siliqua/plant, siliqua/length (cm), number of seeds per siliqua, 1000 seeds weight (g) and seed yield per plant (g). multivariate analysis viz., principal component analysis (pca), principal coordinate analysis (pco), cluster analysis (clu) was done by using genstat 5 release 4.1 (pc/windos nt) software program [2]. table-1. list of the 40 brassica napus l. genotypes used in the experiment with their sources genotype no. name/acc no. source genotype no. name/acc no. source g1. nap-0717-2 bari g21. nap-10014 bari g2. nap-0733-1 bari g22. nap-10012 bari g3. nap-0762 bari g23. nap-0130 bari g4. nap-08-4 bari g24. nap-2012 bari g5. nap-0837 bari g25. nap-2013 bari g6. nap-0865 bari g26. nap-2022 bari g7. nap-0869 bari g27. nap-9906 bari g8. nap-0876 bari g28. nap-9908 bari g9. nap-0885 bari g29. nap-248 bari g10. nap-205 bari g30. nap-2001 bari g11. bari-8 bari g31. nap-9901 bari g12. bari-13 bari g32. nap-9904 bari g13. nap-10007 bari g33. nap-9905 bari g14. nap-10009 bari g34. nap-2057 bari g15. nap-10015 bari g35. nap-2037 bari g16. nap-10017 bari g36. nap-206 bari g17. nap-10019 bari g37. nap-2066 bari g18. nap-10020 bari g38. nap-179 bari g19. nap-1005 bari g39. nap-94006 bari g20. nap-1007 bari g40. nap-108 bari 3. result and discussion 3.1. diversity of the brassica napus l lines genetic diversity analysis involves several steps, i.e., estimation of distance between the varieties, clustering and analysis of inter-cluster distance. therefore, more than one multivariate technique was required to represent the results more clearly and it was obvious from the results of many researchers brassica napus l multivariate techniques were used. agriculture and food sciences research, 2019, 6(1): 1-8 3 © 2019 by the authors; licensee asian online journal publishing group 3.2. construction of scatter diagram based on the values of principal component scores 2 and 1 obtained from the main component analysis, a two dimensional scatter figure (z1-z2) using component mark 1 as x-axis and component score 2 as y-axis was constructed, which has been presented in figure 1. the position of the genotypes in the scatter graph was apparently distributed into five groups, which designated that there existed considerable diversity among the genotypes. figure-1. scattered distribution of 40 brassica napus l genotypes on principal component score superimposed with clustering 3.3. principal component analysis (pca) d2 analysis through non-hierarchical grouping has taken care of simultaneous disparity in all the characters under study. the scattering of genotypes in different clusters of the d2 analysis has tailed similar trend of the z1 and z2 vectors of the main component analysis. the d2 and principal component analysis were found to be another method in giving the information about the support of characters towards divergence of rapeseed and mustard. generally genetic diversity is related with geographical diversity but the former is not essentially directly related with geographical circulation. the genotype within the same cluster while formed specific clusters but were collected or originated from different places which indicated the topographical distribution and genetic separation did not follow the same trend. in the current study pattern of clustering revealed that genotypes originating from the similar country did not form a single cluster. the genotypes originating from dissimilar countries were grouped in the similar cluster (table 3). this indicates that topographical diversity was not related to genetic diversity, which might be due to nonstop exchange of genetic materials amongst the countries of the world. most important components were computed from the correlation matrix and genotype scores obtained from first apparatus and ensuing mechanism with dormant roots greater than the unity donation of the different morphological characters towards divergence were discussed from the dormant vectors of the first two most important component. the major component analysis yielded eigen values of each major component axes with the first axes entirely bookkeeping for the variation among the genotypes, while three of these with eigen values above unity accounted for 42.77%. the first two major axes accounted for 57.07% of the total variation among the 12 characters recounting 40 lines (table 2). based on major component axes i and ii, a two dimensional chart (z1z2) of the cultivars are offered in (figure 1). the scattered diagram discovered that apparently there were mainly five clusters. in vector i (zi) obtained from pca, the significant characters dependable for genetic divergence in the axis of discrimination were days to 50% flowering (5.137). plant height (0.756), secondary branches/plant (0.578) and siliqua/plant (0.429). in vector ii (z2), the second axis of discrimination, primary branches per plant (0.711), number of seeds per siliqua (0.031), siliqua per plant (0.429) and thousand seeds weight were imperative because all these characters had optimistic signs. on the other hand days to maturity, primary branches per plant, siliqua length, seeds per siliqua, thousand seeds weight and seed yield per plant in the first axis of delineation and days to maturity, siliqua length and seed yield per plant in the second axis of delineation had a minor role in the genetic divergence because they had productive signs. leaf length and siliqua per plant in both the vectors had constructive signs, which indicated they were the significant component characters having higher involvement to the genetic divergence among the materials calculated. it was distincted that three methods gave similar results. but factorial makes a distinction and mahalanobis’s d2 distance methods mandatory collecting data plant by plant, while the pca method obligatory taking data by plots. agriculture and food sciences research, 2019, 6(1): 1-8 4 © 2019 by the authors; licensee asian online journal publishing group table-2. eigen values and percentage of variation in respect of twelve characters in brassica napus l principal component axis eigen values % of total variation accounted for cumulative percent i 5.132 42.77 42.77 ii 1.716 14.3 57.07 iii 1.452 12.1 69.17 iv 0.965 8.04 77.21 v 0.756 6.3 83.51 vi 0.711 5.93 89.44 vii 0.578 4.81 94.25 viii 0.429 3.58 97.83 ix 0.228 1.9 99.73 x 0.031 0.26 99.99 xi 0 0 99.99 xii 0 0 99.99 among five clusters, cluster i was unruffled of eight lines: 2,3,4,5,25,34,35 and 38. from the clustering mean value (table 3), it was pragmatic that cluster i fashioned the highest mean for days to first flowering (58.54 days), days to 50% flowering (60.75 days), days to 80% flowering (63.50 days), days to maturity (105.14 days), plant height (107.99 cm), number of primary branch per plant (3.49), number of secondary branch per plant (3.49), siliqua per plant (146.19), siliqua per length (7.49 cm), seed per siliqua (22.99), thousand seed weight (3.39 gm) and seed weight per plant (11.72 gm) (table 3). cluster ii was composed of eight lines; 6,7,8,9,10,14,15 and 22. these genotypes formed the highest mean for seed weight per plant (12.35 gm). cluster iii was constituted of four lines; 1,17,19, and 27. the genotypes of this cluster formed the highest mean for siliqua per length (7.62 cm), seed per siliqua (24.27) and lowest value for number of secondary branch per plant (3.93) and siliqua per plant (130.25). cluster iv constituted of nine genotypes, lines; 1,12,13,16,18,20,21,23 and 24. the line produced the highest mean for siliqua per length (7.62 cm), yield per plant (11.89 gm) and lowest value for plant height (92.55 cm). cluster v constituted of eleven genotypes; line 26,28,29,30,31,32,33,36,37,39 and 40. the genotypes of this cluster produced the highest mean values for plant height (118.63 cm), number of secondary branch per plant (4.82), siliqua per plant (181.39). jagadev, et al. [3] showed utmost genetic distance in between cluster iii and iv suggesting wide diversity in rapeseed and mustard. table-3. distribution of 40 brassica napusl genotypes in five different clusters cluster total no. of line genotype number genotype designation i 8 2,3,4,5,25,34,35,38 nap-0733-1, nap-0762, nap-08-4, nap-0837, nap2013, nap-2057, nap-2037, nap-179 ii 8 6,7,8,9,10,14,15,22 nap-0865, nap-0869, nap-0876, nap-0885, nap205, nap-10009, nap-10015, nap-10012 iii 4 1,17,19,27 nap-0717-2, nap-10019, nap-1005, nap-9906 iv 9 11,12,13,16,18,20,21,23,24 bari-8, bari-13, nap-10007,nap-10017, nap10020, nap-1007, nap-10014, nap-0130, nap-2012 v 11 26,28,29,30,31,32,33,36,37,39,40 nap-2022, nap-9908, nap-248, nap-2001, nap9901, nap-9904, nap-9905, nap-206, nap-2066, nap-94006, nap-108 table-4. cluster means for twelve characters of brassica napus l characters cluster i ii iii iv v days to 1st flowering 58.54 32.58 39.58 42.89 57.82 days to 50% flowering 60.75 34.58 41.58 44.89 59.82 days to 80% flowering 63.50 37.25 44.25 47.56 62.55 days to maturity 105.14 85.48 88.38 90.81 102.85 plant height(cm) 107.99 90.39 98.69 92.55 118.63 no. of primary branch/plant 3.49 2.97 3.12 3.25 3.45 no. of secondary branch/plant 4.20 3.96 3.93 4.16 4.82 siliqua/plant 146.19 157.48 130.25 173.14 181.39 siliqua/length (cm) 7.49 7.43 7.62 7.62 7.49 seed/siliqua 22.99 21.76 24.27 22.62 22.99 seed wt (1000) gm 3.39 3.33 3.46 3.63 3.53 wt of plant seed (gm) 11.72 12.35 11.46 11.89 11.68 3.4. canonical variety analysis canonical variety analysis was performed to calculate the inter-cluster mahalanobis's values. statistical distances indicate the index of genetic diversity in the midst of the clusters. the average intra and inter-cluster distance (d2) values were offered in table 5. consequences indicated that the highest inter-cluster distance was observed between ii and v (8.145), followed by between i and ii (7.767), iii and v (7.434).the lowest inter-cluster distance was observed between the cluster ii and iv (3.917) followed by i and v (3.958), ii and iii (5.308) suggesting a close relationship among these clusters (figure 2). the inter-cluster distances were larger than the intra-cluster distances suggesting wider genetic diversity among the genotypes of dissimilar groups (figure 2). uddin [4] obtained superior inter-cluster distances in the multivariate analysis in mustard. khan [5] reported alike results in raya. subordinate intra-cluster distances in all the eight clusters indicated the genotypes within the same cluster were strongly allied. conversely, the utmost inter-cluster distance was recorded between cluster ii and v (8.145) followed by between i and ii (7.767), iii and v (7.434). genotypes from these clusters if implicated in hybridization might agriculture and food sciences research, 2019, 6(1): 1-8 5 © 2019 by the authors; licensee asian online journal publishing group fabricate a wide range of segregating population, as genetic variation was very discrete among these groups. the inter-cluster divergence varied from 0.304 to 0.420 utmost being from cluster v which comprised of eleven cultivars of diverse origin, while the minimum distance was pragmatic in cluster iii which comprised only four lines (table 5). alike results were obtained by anand and rawat [6] in brown mustard. verma and sachan [7] observed no parallelism between geographic and genetic diversity. gupta, et al. [8] showed no correlation between geographic and genetic diversity. mitra and saini [9] reported no substantiation for any correlation between genetic divergence and geographic diversity. chatterjee and khare [10] studied a downbeat relationship between geographic and genetic diversity. consequences obtained from different multivariate techniques were superimposed in figure 1 from which it might be accomplished that all the techniques gave more or less similar results and one technique supplemented and deep-rooted the results of another one. the clustering blueprint of the lines bare that varieties/lines originating from the same places did not form a single cluster because of direct selection pressure. it has been observed that geographic diversity is not always allied to genetic diversity and therefore, it is not ample as an index of genetic diversity. kumar, et al. [11] premeditated that genetic drift and selection in different environment could cause greater diversity than geographic distance. additionally, there is a complimentary replace of seed material among different region as a upshot, the characters assemblage that might be associated with fastidious region in nature loose their individuality under human obstruction and however, in some cases effect of geographic origin predisposed clustering that is why geographic allocation was not the sole criterion of genetic diversity. the free cluster of the lines suggested confidence upon directional selection pressure practical for realizing maximum yield in different regions; the nicely evolved homeostatic campaign would favour fidelity of the associated characters. this would suggest that it was not necessary to choose diverse parents for diverse geographic regions for hybridization. table-5. average intra and inter-cluster distances (d2) for 40 brassica napusl lines cluster i ii iii iv v i 0.420 ii 7.767 0.384 iii 5.331 5.308 0.414 iv 6.563 3.917 5.728 0.398 v 3.958 8.145 7.434 6.116 0.304 note: *bold figures denotes intra-cluster distance. figure-2. diagram showing inter-cluster (outside the circle) and intra-cluster (inside the circle) distances of 40 genotypes of brassica napus l plate-1. photograph showing seed type of the different genotypes of cluster i agriculture and food sciences research, 2019, 6(1): 1-8 6 © 2019 by the authors; licensee asian online journal publishing group plate-2. photograph showing plant type of the different genotypes of cluster ii plate-3. photograph showing plant type of the different genotypes of cluster iii plate-4. photograph showing plant type of the genotype of cluster iv agriculture and food sciences research, 2019, 6(1): 1-8 7 © 2019 by the authors; licensee asian online journal publishing group plate-5. photograph showing plant type of the genotype of cluster v 3.5. non – hierarchical clustering the multiplication from co-variance matrix gave non-hierarchical clustering among 40 genotypes. by application of nonhierarchical clustering using covariance matrix, the 40 brassica napus l genotypes were grouped into five different clusters. these results confirmed the clustering pattern of the lines according to the principal component analysis. so, the results obtained through pca were confirmed by non-hierarchical clustering. compositions of unlike clusters with their corresponding lines integrated in cluster were presented in table 3. cluster v had utmost eleven lines followed by cluster i, ii, iv, and iii, which had eight, eight, nine and four genotype correspondingly. cluster iv self-possessed of five lines; 1,17,19,27. verma and sachan [7] reported 12 clusters; gupta, et al. [8] five clusters; khan [5] seven clusters and srivastava and singh [12] six clusters in rapeseed and mustard. these results deep-rooted the clustering pattern of the genotypes according to the major component analysis. from the clustering mean value (table 4), pragmatic that cluster i produced the highest mean for days to first flowering (58.54 days), days to 50% flowering (60.75 days), days to 80% flowering (63.50 days), days to maturity (105.14 days), plant height (107.99 cm), number of primary branch per plant (3.49), number of secondary branch per plant (3.49), siliqua per plant (146.19), siliqua per length (7.49 cm), seed per siliqua (22.99), thousand seed weight (3.39 gm) and seed weight per plant (11.72 gm). photograph showing seed, siliqua and plant of the different genotypes of this cluster has been offered in plate 1. cluster ii was self-possessed of eight lines; 6,7,8,9,10,14,15 and 22. these genotypes fashioned the highest mean for seed weight per plant (12.35 gm). photograph showing eed, siliqua and plant of the different genotypes of this cluster has been presented in plate 2. cluster iii was constituted of four lines; 1,17,19, and 27. the genotypes of this cluster fashioned the highest mean for siliqua per length (7.62 cm), seed per siliqua (24.27) and lowest value for number of secondary branch per plant (3.93) and siliqua per plant (130.25). photograph screening seed, siliqua and plant of the different genotypes of this cluster has been existing in plate 3. cluster iv constituted of nine genotypes, lines; 11,12,13,16,18,20,21,23 and 24. the line formed the highest mean for siliqua per length (7.62 cm), yield per plant (11.89 gm) and lowest value for plant height (92.55 cm). photograph screening seed, siliqua and plant of the different genotypes of this cluster has been offered in plate 4. cluster v constituted of eleven genotypes; lines; 26,28,29,30,31,32,33,36,37,39 and 40. the genotypes of this cluster formed the highest mean values for plant height (118.63 cm), number of secondary branch per plant (4.82), siliqua per plant (181.39). photograph viewing seed, siliqua and plant of the different genotypes of this cluster has been presented in plate 5. from the class mean value it was observed that all the cluster mean values were more or less alike. the utmost range of variability was pragmatic for yield (11.46 g to 12.35 g) among all the characters in five clusters. cluster ii integrated mainly early flowering and early maturing genotypes with high yield. to develop high yielding varieties/lines, genotypes of this group could be used in hybridization program. 3.6. selection of genotypes for future hybridization genotypically far-away parents are competent to fabricate higher heterosis. consequences of the present studies indicated noteworthy variation among the genotypes for all the characters studied. number of siliqua per plant, siliqua length and seed weight per plant contributed maximum towards yield perfection. forty brassica napus l genotypes formed five different clusters. pca and cluster analysis gave alike results. by and large, diversity was influenced by the morphological characters, but not by the distribution of the genotypes, which indicated the consequence of consumer preference and growers aptness. considering diversity pattern and other agronomic performance lines nap-0837, nap-0733-1, nap-2066, nap-9901, nap-108 and bari-8 could be measured suitable genotypes for well-organized hybridization in future. concerning of such diverse lines in crossing curriculum could produce enviable segregants. so, more or less divergent genotypes are suggested to use as parents in future hybridization program. 4. conclusion assortment of genetically diverse parents is the most important assignment for any plant breeding actions. for that reason, taking into consideration the extent of genetic distance, contribution of character towards divergence, magnitude of cluster mean and agronomic performance the following genotypes were promising: in view of diversity pattern and other agronomic performance lines nap-0837, nap-0733-1, nap-2066, nap-9901, nap-108 and bari-8 might be recommended for future hybridization program. agriculture and food sciences research, 2019, 6(1): 1-8 8 © 2019 by the authors; licensee asian online journal publishing group references [1] g. bhatt, "comparison of various methods of selecting parents for hybridization in common bread wheat (triticum aestivum l.)," australian journal of agricultural research, vol. 24, pp. 457-464, 1973. available at: https://doi.org/10.1071/ar9730457. [2] p. digby, m. galway, and p. lane, genstat. a second course. oxford: oxford science publications, 1989. [3] p. jagadev, k. samal, and d. lenka, "genetic divergence in rape mustard," the indian journal of genetics and plant breeding, vol. 51, pp. 465-467, 1991. [4] m. j. uddin, "genetic divergence in mustard," bangladesh journal of plant breeding and genetics, vol. 7, pp. 23-27, 1994. [5] m. n. khan, "multivariate analysis in raya," applied biological research, vol. 2, pp. 169-171, 2000. [6] i. anand and d. rawat, "genetic diversity, combining ability and heterosis in brown mustard," the indian journal of genetics and plant breeding, vol. 44, pp. 226-234, 1984. [7] s. verma and j. sachan, "genetic divergence in indian mustard (brassica juncea (l.) czern & coss.)," crop research (hisar), vol. 19, pp. 271-276, 2000. [8] m. gupta, k. s. labana, and s. s. badwal, "correlation and path co¬efficient of metric traits contributing towards oil yield in indian mustard," presented at the international rapeseed congress, pozan, poland, 107(en), 1987. [9] j. mitra and h. saini, "genetic divergence for yield and its components in toria (brassica campestris var. toria)," international journal of tropical agriculture, vol. 16, pp. 243-246, 1998. [10] a. chatterjee and d. khare, "multivariate analysis in niger," research and development reporter, vol. 8, pp. 111-114, 1991. [11] c. h. m. v. kumar, v. arunachalam, and p. s. k. rao, "ideotype and relationship between morpho-physiological characters and yield in indian mustard (b. juncea)," indian journal of agricultural science, vol. 66, pp. 14-17, 1996. [12] m. k. srivastava and r. p. singh, "genetic divergence in indian mustard," crop research hisar, vol. 20, pp. 555-557, 2000. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research vol. 4, no. 1, 8-19, 2017 issn(e) 2411-6653/issn(p) 2518-0193 doi: 10.20448/journal.512.2017.41.8.19 8 growth performance and apparent nutrient digestibility coefficients of weaned rabbits fed diets containing different forms of cocoa pod husk meal p. o. ozung1  o. o. kennedy oko2 e. a. agiang3 p. o. eburu4 e. i. evans5 c. e. ewa6 ( corresponding author) 1,2,3,4department of animal science, university of calabar, calabar, nigeria 5department of animal science, akwa ibom state university, obio akpa campus, uyo, nigeria 6department of animal breeding & physiology, michael okpara university of agriculture, umudike, abia state, nigeria abstract the growth performance and apparent nutrient digestibility co – efficients of weaned rabbits fed corn – soybean meal basal diets containing different forms of cocoa pod husk meal were determined. a total of 60 mixed breed rabbits of both sexes (30 bucks and 30 does) with average initial weight of 606.42±1.30g were used in a nine week feeding trial using a completely randomized design. twelve is nitrogenous and is caloric diets were formulated containing sundried (t1 t2, t3 & t4), fermented (t5 t6, t7 & t8), and hot water treated (t9 t10, t11 & t12) cocoa pod husk meal (cphm) each at 0, 12.5, 25 and 37.5% levels of inclusion, respectively. diet with 0% cphm served as the control in the different forms of the cocoa pod husk meal based diets. growth performance parameters (feed intake, weight gain and feed conversion ratio) were determined, while apparent nutrient digestibility coefficients were determined at the 9th week using the direct in vivo method. the digestibility co efficients were calculated using appropriate formula. results of the study revealed that the final weight, total weight gain and average daily feed intake recorded significant (p<0.05) differences between dietary treatments. when compared with values in the control, diets with the highest inclusion level (37.5%) cphm had the least total weight gain with the sundried, fermented and hot – water treated groups having 557.50, 570.83 and 390.00 g/rabbit respectively. the average daily feed intake declined with increasing levels of sundried cphm (i.e. 66.94, 62.56, 59.27 and 52.98 g/rabbit/day, respectively). the hot – water treated cphm diets recorded the worst fcr beyond 12.5% level when compared with other forms of cphm. some of the apparent nutrient digestibility coefficients for ash and nfe recorded significant (p<0.05) differences between dietary treatments, while crude protein, crude fibre and ether extract digestibility coefficients were statistically similar across dietary treatments in all the three forms of cphm. the sundried cphm had the least digestible crude protein (75.07%) at the highest inclusion level (37.5%). the study concludes that the three different forms of cocoa pod husk meal in the following order of superiority (fermented, hot – water treated and sundried) can be included in diets meant for rabbits at levels not exceeding 25% for optimum growth performance and apparent nutrient digestibility co efficients. keywords: growth, digestibility, rabbits, cocoa citation | p. o. ozung; o. o. kennedy oko; e. a. agiang; p. o. eburu; e. i. evans; c. e. ewa (2017). growth performance and apparent nutrient digestibility co efficients of weaned rabbits fed diets containing different forms of cocoa pod husk meal. agriculture and food sciences research, 4(1): 8-19. history: received: 19 april 2017 revised: 8 june 2017 accepted: 14 june 2017 published: 20 june 2017 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2017.41.8.19&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2017, 4(1): 8-19 9 contents 1. introduction ......................................................................................................................................................................................... 9 2. materials and methods ...................................................................................................................................................................... 9 3. results ................................................................................................................................................................................................ 11 4. discussion .......................................................................................................................................................................................... 12 5. conclusion ......................................................................................................................................................................................... 14 6. recommendations ............................................................................................................................................................................ 14 references .............................................................................................................................................................................................. 14 1. introduction extant records show that livestock husbandry occupies about 30 percent of the total arable land in the world [1]. but with the rapid exhaustion of limited animal products, increasing population and better life styles, it is now imperative to explore other aspects of livestock production to bridge the gap of protein intake from animal origin. according to fao [2] from year 2000 to 2020, the projected world requirement for animal protein would increase tremendously due to population explosion. traditionally, ruminants and monogastrics (poultry and pigs) are the main sources of domestic meat. however, rabbits and other micro livestock species (grasscutters, snails and quails) are emerging in many developing countries like nigeria as potential sources of animal protein. the inadequacy of animal protein coupled with stiff competition between man and animals for agricultural produce call for enhancing the diversification and productivity of livestock and this has culminated in resurgence of interest in rabbit production [3]. the renewed interest in rabbit production by researchers and farmers in nigeria has necessitated research into alternative feed resources that are readily available, under utilized agro by products or agro industrial wastes to substitute or supplement the highly demanded conventional cereals [4]. a major aspect of livestock production that constitutes huge cost and demands urgent attention is the utilization of quality animal feeds. the quest for non conventional feed resources is imperative, due to the competitive pressure on grains and conventional seed meals which have outweighed production [5]. the immediate challenge therefore is for animal scientists/ researchers and feed mill operators to promote the utilization of cheap and affordable alternative feed resources that are rich in nutrients and are well adapted to tropical climatic conditions [2]. literature reports abound on the use of agro industrial by – products and farm waste materials in livestock nutrition. they include wheat and rice offals as well as maize bran and brewers dried grains (bdg) or yeast. according to atteh and opawande [6] bdg can replace groundnut cake in diets meant for finishing broilers. rice milling wastes have been utilized in diets meant for cockerel finishers [7] while afolayan [8] utilized corn bran meal at different levels in broiler chicken diets. however, the prices of most of these by – products have escalated as a result of high demand; thereby necessitating the quest for cheaper alternatives. there is the need to include alternative feedstuffs common in the tropics in rabbit diets, for the purposes of promoting optimum performance characteristics and reduction in cost of production and to make rabbit farming profit oriented to the teaming unemployed youth and rural poor farmers in nigeria [9]. the domestic rabbit is a pseudo ruminant (monogastric – herbivore or hind gut fermentor) that feeds on forages, grains/ concentrates and hay [10]. most diets for rabbits consist of ingredients from plant sources [1113] composite mixture of table scraps [14] and agro by products [15-17]. one of the promising agro by products that can be utilized in rabbit diet formulations is the cocoa pod husk. according to olubamiwa and akinwale [18] cocoa pod husk meal (cphm) can partially replace maize in layers diet at 25 percent level with no adverse effects on egg parameters. untreated cphm has been added in the diets of growing swine up to 300 g/kg without deleterious signs on body characteristics [19]. the practice of incorporating cocoa pod husk meal in various animal diets has been reported [13, 20-25]. accordingly, smith, et al. [26] reported that sundried cocoa beans contain 0.70 percent sugar, 0.70 percent starch, 1.70 percent theobromine (an alkaloid), 6.70 percent protein and 50 – 55 percent pale yellow, non drying fat known as cocoa butter. the cocoa bean testa has been used successfully in feeding trials for small ruminants without adverse effects on performance characteristics [27]. cocoa pod husks constitute 75 percent of the entire cocoa fruit on fresh weight basis fagbenro [28]. adomako [29] reported that cocoa beans account for less than 2.55 percent of the whole fruit. according to donkor, et al. [22] cocoa pod husks contain 6 7 percent crude protein, 9 10 percent total ash, 1 – 8 percent crude fat (ether extract) and 23 – 33 percent crude fibre. also findings by ozung, et al. [13] showed that cocoa pod husk meal has between 7.70 and 9.37 percent crude protein, ether extract (8.83 – 11.68%) and ash (9.30 – 10.85%). sobamiwa and longe [30] reported that metabolizable energy of cocoa pod husk is moderate and ranges from 2000 – 2100 kcal/kg; which is comparable to that of palm kernel cake, soybean meal, rice bran and brewers dried grain [31]. the processed cocoa pod husks have been reported to have low theobromine content. the crude fibre is easily digestible [32]. it is reported that cocoa pod husk is digestible by all classes of livestock, especially ruminants. however, the high crude fibre content (21.49 – 34.82 percent) hinders its effective utilization by monogastrics [33]. this constraint calls for the processing of the cocoa pod husks by various methods (fermentation, hot – water treatment, urea, enzyme, fungal treatment and microbial detheobromination), so as to promote digestibility and biodegradability in animals. other researchers have carried out nutritional and growth performance studies with cocoa pod husk meal on poultry and small ruminants, but there is paucity of research findings on the effect of sundried, fermented and hot – water treated cocoa pod husk meal on the growth performance and apparent nutrient digestibility co efficients of rabbits, hence the objectives of this study. 2. materials and methods 2.1. location of the study the study was carried out at the rabbitry unit of the teaching and research farm, department of animal science, university of calabar, calabar, cross river state, nigeria. according to the geonames geographical database, calabar is located at 4.95170 latitude and 8.3220 longitude (in decimal degrees) with an average elevation/ altitude of 42 metres [34]. while, akpan, et al. [35] reported that calabar is located at latitude 30n and longitude 70e, with a land mass of 233.2 sq. miles (604 km2). the annual rainfall ranges from 3000 – 3500 mm agriculture and food sciences research, 2017, 4(1): 8-19 10 (average of 1,830mm) per annum and the average daily temperature is 250c/770f which increases to 300c (860f) in august. the relative humidity is between 70 – 80 percent. 2.2. collection, processing and proximate analysis of cocoa pod husk meal (cphm) freshly broken composite cocoa pod husks (derived from improved/hybrid cocoa varieties crinc1 – 8, wacri 11 hybrids and f3 amazon) were obtained from the fermentation units of the cocoa research institute of nigeria (crin) sub station at ajassor, ikom lga of cross river state. the broken pods were washed and sun dried for two weeks, bulked and milled with hammer mill to produce cocoa pod husk meal (cphm) [36]. the resultant meal was shared into three (3) portions: the sundried cphm (scphm), fermented cphm (fcphm) and hot water treated cphm (hcphm), respectively. cphm for the fermented treatment was thoroughly mixed with 60 percent water, relative to its weight as ascertained by bello, et al. [37] and bagged using an air tight polythene bag. this was allowed to stay for three (3) days under room temperature, thereafter, it was opened and shade dried for five (5) days; before being packed, bagged and stored in a cool dry place until it was used. the final portion of cphm was treated with hot water that was boiled to 1000c for 15 minutes [38-40] which was later drained, shade dried and stored for later use. 2.3. experimental diets twelve (12) experimental isonitrogenous and isocaloric diets for the growth phase were formulated in line with the nutrient needs of rabbits as recommended by aduku and olukosi [10]. each processed form of cphm was included at 0, 12.50, 25.00 and 37.50 percent levels for t1, t2, t3, and t4 (sundried cphm), t5, t6, t7, t8 (fermented cphm) and t9, t10, t11, t12 (hot – water treated cphm), respectively in the experimental diets. diet without cphm (0 percent) served as control in the experiment. the choice of these levels was based on earlier reports on the use of much lower levels of cphm for poultry, pigs and rabbits without adverse and significant effects on growth performance characteristics [36, 39, 40]. feedstuff purchase/choice and procurement of cocoa pod husks as well as methods of processing cphm and ration formulation gave primary consideration to least cost and maximum biological returns [41]. the proximate composition of the experimental diets was done based on the procedures outlined by a.o. a.c [42]. the gross composition of experimental diets is presented in table 1. 2.4. experimental rabbits and management sixty (60) weaned mixed breed rabbits (chinchilla x new zealand x californian) between 5 and 6 weeks old of both sexes (30 bucks and 30 does), (average initial body weight of 606.42±1.30g) were used in this study. the rabbits were purchased from a reputable rabbitry (domino farms, use offot) in uyo, akwa ibom state. they were managed based on standard experimental procedures. on arrival at the rabbitry facility, the animals were provided with anti – stress vitalyte (anidone – vita dox) at 0.50g per 75 litres of chlorine free water. concrete drinking troughs and fabricated feeding troughs (empty beverage cans nailed to the wooden board) were provided in each cage. the rabbits adjusted for two weeks before the actual commencement of the feeding trial and within this period; they were placed on commercial pelleted grower mash and given prophylactic treatment against bacterial and viral infections using a broad spectrum antibiotic – oxytetracycline hydrochloride eq.50mg (soluble powder 5 percent w/w) at recommended dosage of one teaspoonful per 4 litres of chlorine – free water. the animals were thereafter screened against ecto and endo parasites via subcutaneous injection of ivermectin (kepromec) at the recommended level (0.20 ml per rabbit). thereafter, the animals were subjected to nine (9) weeks (63 days) initial feeding trial during which their pre pubertal growth performance characteristics and apparent nutrient digestibility co –efficients were evaluated. 2.5. housing and equipment the experimental animals were accommodated individually in double tier wooden hutches (with wire mesh floor) measuring 65 × 65 × 65 cm (l × h× w) and raised 25 cm from the ground and placed in a standard rabbitry with half walls to allow for cross ventilation. 2.6. experimental design animals were randomly distributed to the test diets using a simple completely randomized design (crd) with three processed forms of cphm. they were twelve (12) dietary treatments with five (5) rabbits per treatment (each rabbit served as a replicate). the rabbits were assigned to the various treatments after equalizing for body weights. 2.7. feeding of animals/ feed intake respective rabbits were given weighed amount of feed weight daily in the morning hours (07:00 – 08:00 am) and adjusted weekly to make provision for the weekly weight change, while water was given without restriction. the left over feed samples were collected into marked envelopes and weighed daily with a precision electronic balance; model hk dc-320as with minimum and maximum sensitivities of 0.01 and 320g, respectively. the feed intake was computed by difference. fresh aspilia africana leaves (supplementary forage) were harvested daily, air – dried and fed to the rabbits in the evening period (17:00 – 18:00 hours) at a uniform amount of 30 g in all the treatments. 2.8. weighing of animals at the beginning of the trial, the rabbits were weighed individually and thereafter on a weekly basis. body weight changes were determined by difference. a top loader weighing scale was used in determining the weight changes of rabbits. 2.9. feed conversion ratio the feed conversion ratio was determined as the ratio of feed consumed (g) to weight gain (g). agriculture and food sciences research, 2017, 4(1): 8-19 11 2.10. digestibility trial digestibility is an important measure of the nutritive value of feedstuffs. it is the difference in value between the feed eaten and materials voided by the animals expressed as percentage of feed eaten [43]. a direct method (in vivo) involving feeding experiment with rabbits was used in this study [44]. the digestibility trial was carried out at the ninth week and lasted for 15 days, using thirty six rabbits (i.e.3 rabbits) per treatment. no faeces were collected for the first 7 days, but faecal material and feed intake were recorded for the last 8 days. faecal samples collected from each day were oven – dried at 600c, preserved in bags and stored in a refrigerator. at the end, all faecal samples collected from each replicate were pooled together for the determination of proximate composition using the a.o. a.c [42] methods. percentage digestibility of diets was obtained as follows: percent digestibility = quantity in feed – quantity in faeces ____________________ × 100 quantity in feed 2.11. statistical analysis data obtained in this study were subjected to one – way analysis of variance (anova) using general linear model [45] for a completely randomized design (crd) of genstat release 10.3de (2011). significant means were separated using least significance difference (lsd) option [46] of the same statistical software. the model used was as follows: yij = µ + ti + eij where: yij: observed value µ: overall (population) mean value ti: random effect of the ith processing method of cphm eij: random residual error 3. results 3.1. proximate composition (determined analysis) of experimental diets, test ingredient (cocoa pod husk meal) and fibre fractions results of the proximate composition of the diets (growth phase) are presented in table 2. the proximate fractions (cp, cf, ee, ash and nfe) recorded significant differences (p < 0.05) between dietary treatments. the control diet (0 percent cphm) recorded the highest crude protein (cp) (18.30 percent) content while the least value was recorded in the raw cocoa pod husk meal (cphm) with 37.50 percent as 14.04 percent. the crude fibre content increases as the level of cphm inclusion increased across the diets, highest values were recorded in the 37.50 percent inclusion level with 23.53, 26.37 and 23.08 percent respectively for the raw, fermented and hot – water treated cphm. while the proximate composition and fibre fractions of the raw, fermented and hot water treated cphm are summarised in table 3. all parameters, except the total digestible nutrients (tdn) had significant differences (p < 0.05) between processing methods of cphm. the cp values were 9.37, 7.70 and 8.94 percent for the raw, fermented and hot – water treated cphm, respectively. the raw cphm recorded the highest cf content (61.80 percent), followed by the fermented cphm (57.42 percent) and the least value was recorded in the hot – water treated cphm (53.37 percent). the trend was however reversed for other fractions like ether extract (ee), ash and nitrogen free extract (nfe) contents as the highest values were recorded in the hot – water treated cphm, followed by the fermented cphm and least in the raw cphm. the ee values were 8.83, 9.52 and 11.68 percent while the values for ash content were 9.30, 10.80 and 10.85 percent and nfe values were 10.70, 14.56 and 15.16 percent respectively, for raw, fermented and hot – water groups. the acid detergent fibre (adf) and neutral detergent fibre (ndf) values were highest in the fermented cphm (73.09 and 79.45 percent, respectively), followed by the raw cphm (68.64 and 75.08 percent, respectively) and least in the hot – water treated cphm (64.60 and 71.09 percent, respectively). 3.2. growth performance characteristics of rabbits results of the pre – pubertal growth performance attributes of weaned rabbits fed cocoa pod husk meal based diets are summarised in table 4. the final weight, total weight gain and average daily feed intake (adf) of rabbits recorded significant differences (p < 0.05) between dietary treatments. the total weight gain recorded highest values in the respective control diets for the raw (1048.00g/rabbit), fermented (907.50g/rabbit) and hot – water treated (710.00g/rabbit) cphm. dietary treatments with the highest inclusion level (37.50 percent) of cphm recorded the least values of total weight gain, with the raw, fermented and hot – water treated forms of cphm having 557.50, 570.83 and 390.00g/rabbit, respectively. results further revealed that the fermented and raw cphm recorded better total weight gain across dietary treatments compared to the hot – water treated cphm (which recorded the least total weight gain) in this study. average daily gain (adg) values for the raw cphm were 16.63, 10.95, 13.84 and 8.85g/rabbit/day in the control, 12.50, 25.00 and 37.50 percent inclusion levels, respectively. the fermented cphm recorded adg values of 14.40, 10.42, 13.85 and 9.06g/rabbit/day in 0, 12.50, 25.00 and 37.50 percent levels, respectively. while the hot – water treated cphm recorded adg values of 11.27, 10.13, 6.24 and 6.19g/rabbit/day, respectively for 0, 12.50, 25.00 and 37.50 percent inclusion levels. the average daily feed intake (adf) of rabbits declined with increasing levels of raw cphm in the dietary treatments. the values were 66.94, 62.56, 59.27 and 52.98 g/rabbit/day in the control, 12.50, 25.00 and 37.50 percent inclusion levels, respectively. the fermented and hot – water treated cphm however recorded higher values of adf compared to the raw cphm. the hot – water treated cphm recorded the highest and worst values of feed conversion ratio (fcr), compared with the two other forms of cphm and the control diets. agriculture and food sciences research, 2017, 4(1): 8-19 12 3.3. apparent nutrient digestibility co efficients results of nutrient digestibility co – efficients of rabbits fed diets containing cocoa pod husk meal are presented in table 5. digestibility parameters like ash and nfe recorded significant effect (p < 0.05) of dietary treatments, while the values for dry matter, crude protein, crude fibre and ether extract were statistically similar. in the raw cphm, the highest inclusion level of 37.50 percent recorded the least digestibility value of 75.07 percent for crude protein (cp) compared to the control diet and other lower levels of inclusion. the trend was however reversed in the fermented and hot – water treated cphm as the highest digestibilty values for cp were recorded in the 37.50 percent inclusion level as 78.11 and 77.74 percent, respectively. the digestibility values for crude fibre (cf) were 77.92, 78.44, 75.05 and 74.50 percent for 0, 12.50, 25.00 and 37.50 percent, respectively in the raw cphm; while the fermented cphm recorded digestible cf values of 76.79, 71.03, 61.14 and 76.68 percent for 0, 12.50, 25.00 and 37.50 percent, respectively; the hot – water treated cphm had 77.42, 85.97, 80.80 and 82.19 percent, respectively for 0, 12.50, 25.00 and 37.50 percent inclusion levels. 4. discussion 4.1. determined (proximate) composition of experimental diets, cocoa pod husk meal and fibre fractions the proximate composition values of experimental diets in this study (table 2) are within the recommended ranges for growing rabbits. the crude protein content of diets (14.04 – 18.73 percent) falls within the recommended levels (16 – 18 percent) for both growing and breeding rabbits [47-51]. rabbits have been observed to adapt effectively to low and poor protein diets unlike poultry, because of coprophagy [52]. however, crude protein levels of 12, 15, 16 and 17 percent have been reported as optimum for rabbits in terms of maintenance, gestation, growth and lactation, respectively [10, 53]. the crude protein content in this study proved satisfactory, as the rabbits recorded effective growth and reproductive efficiency. the crude fibre content (13.38 – 26.37 percent) of the experimental diets was higher than the recommended level (10 – 14 percent) for rabbits [54] but within the range (22.96 – 32.06 percent) reported by ijaiya, et al. [15]. the high crude fibre levels were apparently due to the high crude fibre content of the test ingredient (cocoa pod husk meal) used in this study. according to champ and maurice [55] a crude fibre level in excess of 9 percent is needed for normal growth in rabbits and to reduce incidence of enteritis; while a high fibre level in excess of 20 percent may induce caecal impaction and limit energy intake [10]. however, rabbits fed higher levels of fibre in this study did not show any obvious adverse effect. furthermore, cheeke [56] reported that crude fibre levels of between 10 to 17 percent have been found to support weight gain; with the optimum gain of 41.30g/day/rabbit obtained at a fibre level of 14.80 percent. accordingly, satisfactory performance has been recorded in weaner rabbits fed high fibre (22.96 – 32.06 percent) and high energy (3,523.54 – 3,897.46 kcal/kg me) diets [15]. the energy content of the experimental diets ranges from 2,500.75 – 2,536.90 kcal/kg me. the energy content was within the recommended level (2,500 – 2,600 kcal/kg me) for growing and breeding rabbits [10, 48, 57]. the proximate composition and fibre fractions of the raw, fermented and hot – water treated cocoa pod husk meal (table 3) recorded significant effect (p < 0.05). the dry matter content in the raw cocoa pod husk meal (94.60 percent) was almost similar to the value (94.18 percent) reported by onifade, et al. [53] for the raw untreated form. the dry matter values for the fermented cocoa pod husk meal (84.40 percent) and hot – water treated cocoa pod husk meal (85.28 percent) are slightly lower than the value (88.96 percent) for processed forms of cocoa pod husk meal reported by alamowor, et al. [58]. the raw cocoa pod husk meal recorded highest values in dry matter, crude protein and crude fibre, followed by the hot – water treated cocoa pod husk meal. the fermented cocoa pod husk meal recorded the least values in dry matter and crude protein contents. the ether extract, ash, nitrogen free extract and fibre fractions were within the values reported by marcel, et al. [59] for cocoa pod husk meal. the ash content of the raw (9.30 percent), fermented (10.80 percent) and hot – water treated (10.85 percent) cocoa pod husk meal, respectively were slightly higher than the value of 9.07 percent reported by alamowor, et al. [58] and lower than the value of 11.14 percent reported by onifade, et al. [53] for unprocessed cocoa pod husk meal. the proximate fractions obtained in this study are comparable to those reported by hamzat and adeola [60] for dry matter and ash contents of cocoa pod husk, but fairly higher in crude protein, crude fibre, acid detergent fibre, neutral detergent fibre and total digestible nutrients. the differences in values could be attributed to the different processing methods adopted in the separate studies. nutritionally, the results revealed that cocoa pod husk meal (irrespective of the processing method) is low in nutrient contents (especially crude protein), but high in crude fibre. this is in agreement with the findings of oddoye, et al. [19] who reported that cocoa pod husk is not too nutritious, due to its low metabolizable energy, crude protein and high crude fibre contents. 4.2. growth performance characteristics the growth performance characteristics of rabbits (table 4) recorded significant differences across treatments in the final weight gain (fwg), average daily feed intake (adf) and total weight gain (twg). the average daily weight gain (adg) and feed conversion ratio (fcr) recorded statistically similar results across dietary treatments. the adf decreased significantly (p < 0.05) in the raw cocoa pod husk meal group from 66.94 g/rabbit/day in the control diet to 52.98 g/rabbit/day in the diet with 37.50 percent of raw cocoa pod husk meal, implying the rabbits consumed more feed in the control diet. the adf in the fermented cocoa pod husk meal decreased marginally from 66.47 to 62.75 g/rabbit/day across dietary treatments; while the hot – water treated cocoa pod husk meal recorded a progressive trend in adf ranging from 64.34 – 71.16 g/rabbit/day in dietary treatments. the values for adf obtained in this study are comparable to the range of 63.46 – 70.46 g/rabbit/day reported by isika, et al. [61] who evaluated the replacement value of processed cocoa bean meal for groundnut cake in rations for fryer rabbits. however, the adf values are far higher than the range of 24.08 – 37.16 g/rabbit/day reported by ahaotu, et al. [62] in dutch rabbits fed processed ripe banana peel meal. the differences in adf values could be attributed to age disparity, differences in feed materials and environmental influences. metabolic processes increase with age in animals, just as the feed intake to meet maintenance and performance functions [61]. the experimental rabbits in this study appeared to follow such assertion, especially in the hot – water treated cocoa pod husk meal group. the agriculture and food sciences research, 2017, 4(1): 8-19 13 feed intake in this group increased as the fibre level increased across the dietary treatments. this finding agrees with the report of onyema and iwuala [63] who stated that the consumption of high fibre diets resulted to increase in feed intake. however, the trend was reversed in the raw cocoa pod husk meal (cphm) group. the plausible reason for the observed depression in adf in the raw cphm could be the high antinutrient (theobromine) content in the raw form. theobromine has been reported to be a key anti – nutrient in cocoa products [64] its presence is reported to limit feed intake by animals and consequently, zootechnical parameters [65]. however, the adf was significantly (p < 0.05) highest in the hot – water treated cocoa pod husk meal group. this validates the report of adeyina, et al. [40] who stated that feed intake and weight gain were high in rabbits fed hot – water treated cocoa bean shell up to 200g/kg. this is so because the hot – water treatment and fermentation methods have been reported to reduce the theobromine content in cocoa by – products [40]. the values for adg fluctuated across dietary treatments with no definite pattern. in all the processed forms of cocoa pod husk meal, the highest inclusion level (37.50 percent) recorded the least adg, implying that cphm and its associated theobromine and unpalatable nature adversely affected weight gain. this agrees with the findings of marcel, et al. [59] who reported that the use of cocoa pod husk meal above certain permissible levels will depress the weight gain of animals. the feed conversion ratios obtained in this study were poor, since the values were highest in the hot – water treated cocoa pod husk meal group, compared to other processed forms. the poor feed efficiency may be attributed to the high levels of cphm in the rabbit diets that even fermentation and hot – water treatment could not address, and this agrees with devender [66]; day and dilworth [67]; marcel, et al. [59] who have recommended low levels of cphm and other cocoa by – products inclusion in animal diets. high intake of highly fibrous feeds by rabbits has been reported to cause reduced feed efficiency [68]. the mortality recorded may be unconnected to the diets, but due to attack by soldier ants. therefore, it could be stated that raw cphm depresses the performance of animals at levels exceeding 25 percent. 4.3. apparent nutrient digestibility co efficients the apparent nutrient digestibility co efficients in rabbits fed raw, fermented and hot – water treated cocoa pod husk meal baseddiets did not record any significant difference between dietary treatments in digestible dry matter, crude protein, crude fibre and ether extract; except the digestible ash and nitrogen free extract which were significantly affected (p < 0.05) (table 5). the digestibility co efficients obtained in this study were generally high in all parameters, implying that the rabbits were able to utilize nutrients in the diets for their growth. the digestibility of crude protein declined marginally across dietary treatments in the raw cocoa pod husk meal (cphm) group, while other processed methods recorded fluctuating trend without a definite pattern. the decline in crude protein digestibilty in the raw cphm may be associated with the theobromine content of the raw cocoa pod husk, which has been reported to limit its utilization in animals [59] and also due to the increasing level of fibre in the cocoa pod husk meal. the decline in protein digestibility confirms the result of negative influence of fibrous substances on nutrient digestibility in farm animals [69, 70]. reduced digestibility could be attributed to the reduction in mean retention time of feed in the alimentary canal [59]. it has also been reported that increasing the concentration of cocoa by – products in diets will increase the concentration of neutral detergent fibre, acid detergent fibre and acid detergent lignin that reduce the action of digestive enzymes in the intestine [71]. the digestible crude fibre values in this study were quite high. meffeja, et al. [72] reported that high fibre concentrations would lead to copious production of mucus which helps in protecting the lining of the digestive tract. the co efficients of nutrient digestibilty in this study were higher than the values reported by meffeja, et al. [72] for cocoa shell meal. this is because the food value of cocoa shell meal is lower than that of cocoa pod husk meal and because of the low digestibilty of the nutrients in cocoa shell meal; it has been recommended for animals at the finishing phase than those under rapid growth [72]. this justifies the high value of cocoa shell meal in diets meant for small ruminants and dairy cows. the digestibility of nutrients obtained in this study for dry matter (95.41 – 96.34 percent), crude fibre (61.14 – 85.97 percent), ether extract (71.51 – 83.57 percent) and ash (56.84 – 83.33 percent) are fairly higher than the ranges reported by adedire, et al. [73] for digestible dry matter (63.63 – 69.37 percent), crude fibre (55.61 – 62.88 percent), ether extract (62.14 – 64.15 percent) and ash (51.75 – 58.05 percent) who fed crop based residues fermented with rhizopus oligosporus to experimental rabbits. the crude protein digestibility in this study ranges from 72.18 – 78.54 percent and slightly lower than the range (80.40 – 88.01 percent) recorded by adedire, et al. [73]. the differences in nutrients digestibilty between the separate studies could be attributed to age disparity of the rabbits, different feeding materials used as test ingredients, processing methods and associated fibre levels. the trend of dry matter digestibility was statistically similar across dietary treatments and was proportional to crude fibre level in the diets especially with the degree of lignification [74] implying that the higher the fibre content of diets, the better the dry matter digestibility. this is in line with the report of aboushour and baraket [75] that dry matter intake and digestibility increased as dietary fibre increases. the result obtained revealed that digestible protein did not improve significantly with fermentation and hot – water treatment of cocoa pod husk meal compared with the raw group. this may be due to the insoluble fibre in the cocoa pod husk which was not effectively digested; as insoluble fibre can be further fermented in the caeca of rabbits to derive more protein and vitamins through coprophagy or caecotrophy [76]. the presentation of cocoa pod husk meal as raw, fermented and hot – water treated forms did not have any significant effect in the digestibility of dry matter, protein, fibre and ether extract. this suggests that the processing of cocoa pod husk meal in any of the three forms did not exert any stimulating effect that could have enhanced better digestibility. this observation is in tandem with the report of adedire, et al. [73] who opined that fermentation of plant materials may not enhance nutrient digestibility. agriculture and food sciences research, 2017, 4(1): 8-19 14 5. conclusion within the present experimental conditions, this study concludes that processed cocoa pod husk meal (cphm) could be included in diets meant for rabbits. the order of importance is the fermented cphm, followed by the hot – water treated cphm and lastly the raw cphm, respectively. the fermented cocoa pod husk meal – based diets performed best compared to other forms in terms of growth performance and nutrient utilization of rabbits. 6. recommendations based on the outcome of this study, it is therefore recommended as that: i. cocoa pod husk meal (cphm) should be fermented before it can be included in diets meant for rabbits. hence, fermentation is the best and cheapest method of detheobromination. ii. the level of inclusion of fermented cphm should not exceed 25.00 percent in rabbit diets. references [1] fao, "food & agriculture organization. livestock, a major threat to environment. rome, italy. retrieved from http://web.archive.org/fao.org/news. 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[76] o. o. oduguwa, o. o. sobamiwa, d. sanni, and a. y. akintunde, "effect of urea treatment on the utilization of cocoa pod husk in rabbit diets," tropical journal of animal science, vol. 1, pp. 23 – 28, 1999. view at google scholar | view at publisher [77] a. a. odunsi and o. g. longe, "the nutritive value of hot water or cocoa pod ash solution treated cocoa bean cake for broiler chicks," british poultry science, vol. 39, pp. 519 – 525, 1998. view at google scholar | view at publisher https://scholar.google.com/scholar?hl=en&q=effect%20of%20urea%20treatment%20on%20the%20utilization%20of%20cocoa%20pod%20husk%20in%20rabbit%20diets http://dx.doi.org/10.4314/tjas.v1i2.49575 https://scholar.google.com/scholar?hl=en&q=the%20nutritive%20value%20of%20hot%20water%20or%20cocoa%20pod%20ash%20solution%20treated%20cocoa%20bean%20cake%20for%20broiler%20chicks http://dx.doi.org/10.1080/00071669888700 agriculture and food sciences research, 2017, 4(1): 8-19 17 table-1. gross composition of experimental diets dietary treatments & levels of rcphm dietary treatments & levels of fcphm dietary treatments & levels of hcphm ingredients t1 (0%) t2 (12.50%) t3 (25.00%) t4 (37.50%) t5 (0%) t6 (12.50%) t7 (25.00%) t8 (37.50%) t9 (0%) t10 (12.50%) t11 (25.00%) t12 (37.50%) yellow maize 36.00 33.00 30.00 27.00 36.00 33.00 30.00 27.00 36.00 33.00 30.00 27.00 soybean m. 15.70 16.00 16.00 17.00 15.70 16.00 16.00 17.00 15.70 16.00 16.00 17.00 fish meal 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 pkc 4.00 6.00 7.00 8.00 4.00 6.00 7.00 8.00 4.00 6.00 7.00 8.00 wheat offal 10.00 6.25 5.50 1.00 10.00 6.25 5.50 1.00 10.00 6.25 5.50 1.00 rice husk 27.00 18.95 9.20 2.20 27.00 18.95 9.20 2.20 27.00 18.95 9.20 2.20 rcphm 0.00 12.50 25.00 37.50 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 fcphm 0.00 0.00 0.00 0.00 0.00 12.50 25.00 37.50 0.00 0.00 0.00 0.00 hcphm 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 12.50 25.00 37.50 bone meal 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 palm oil 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50 methionine 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 lysine 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 salt (nacl) 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 premix 0.20 0.20 0.20 0.20 0.20 0.20 0.20 0.20 0.20 0.20 0.20 0.20 total 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 calculated analysis : % cp 16.00 16.04 16.32 16.61 16.00 16.04 16.32 16.61 16.00 16.04 16.32 16.61 %cf 10.00 11.65 13.54 15.87 10.00 11.65 13.54 15.87 10.00 11.65 13.54 15.87 *theob.(g/kg) 0.00 2.83 5.67 8.50 0.00 2.83 5.67 8.50 0.00 2.83 5.67 8.50 me (kcal/kg) 2,500.75 2,505.80 2,524.85 2,536.90 2,500.75 2,505.80 2,524.85 2,536.90 2,500.75 2,505.80 2,524.85 2,536.90 note: gross composition of bio -super premix per kg: vitamin a -1,5000,000iu; vitamin d3 300,000iu; vitamin e 400mg; vitamin k3 100mg; vitamin b2 400mg; vitamin b12 2,000mg; nicotinamide 2,000mg; calcium d panto thenate 800mg; choline chloride 40,000mg; ferrous sulphate 2,000mg; manganese sulphate 5,000mg; copper sulphate 80mg; zinc oxide 3,000mg; cobalt sulphate 10mg; potassium iodide 120mg; dl-methionine 10,000mg and antioxidant 18,000mg. the premix was manufactured by bio pharmachemie company, hcm city, vietnam. cf crude fibre cp crude protein *theob.: theobromine content calculated based on standard method proposed by odunsi and longe [77]. me metabolizable energy agriculture and food sciences research, 2017, 4(1): 8-19 18 table-2. proximate composition (determined analysis) of experimental diets rcphm diets fcphm diets hcphm diets parameter t1 t2 t3 t4 t5 t6 t7 t8 t9 t10 t11 t12 (%) 0% 12.50% 25.00% 37.50% 0% 12.50% 25.00% 37.50% 0% 12.50% 25.00% 37.50% s.e.m dry matter 88.28c 89.03b 92.64a 87.10f 88.28c 88.70b 88.75b 88.32b 88.28c 87.32c 87.99d 88.61b 0.05 crude protein 18.30b 17.86d 18.08c 14.04f 18.30b 17.80d 15.89e 17.45c 18.30b 14.16f 15.56e 18.73a 0.009 crude fibre 14.13f 18.55cd 20.03e 23.53b 14.13f 14.67f 13.38f 26.37a 14.13f 22.66c 21.46d 23.08b 0.007 ee 10.53e 12.17b 11.57c 10.63e 10.53 11.36c 14.81a 10.98d 10.53e 9.91f 11.13c 10.95d 0.009 ash 10.21e 10.94a 8.36d 9.31d 10.21e 7.60f 8.48d 10.82b 10.21e 10.45c 10.38d 10.38d 0.006 nfe 46.83b 40.48d 41.96c 42.49c 46.83b 48.57a 47.44b 34.38e 46.83b 42.82c 41.47d 36.86e 0.09 note: a, b, c…f: means on the same row with different superscripts are significantly different (p < 0.05) all mean values are obtained from triplicate samples per treatment/diet rcphm: raw cocoa pod husk meal fcphm: fermented cocoa pod husk meal hcphm: hot -water treated cocoa pod husk meal ee: ether extract nfe: nitrogen free extract s.e.m: standard error of mean table-3. proximate composition and fibre fractions of raw and processed cocoa pod husk meal parameter test ingredient (%) rcphm fcphm hcphm s.e.m dry matter 94.60a 84.40b 85.28b 3.27 crude protein 9.37a 7.70b 8.94a 0.50 crude fibre 61.80a 57.42b 53.37c 2.44 ether extract 8.83b 9.52b 11.68a 0.86 ash 9.30b 10.80a 10.85a 0.51 nitrogen free extract 10.70b 14.56a 15.16a 1.40 adf 68.64b 73.09a 64.60c 2.46 ndf 75.08b 79.45a 71.09c 2.42 tdn 1.52 1.28 1.43 0.07 note: a,b, c means on the same row with different superscripts are significantly different (p < 0.05) all mean values are obtained from triplicate samples per treatment/diet rcphmraw cocoa pod husk meal fcphm fermented cocoa pod husk meal hcphm hot water treated cocoa pod husk meal cp: crude protein ndf: neutral detergent fibre cf: crude fibre tdn: total digestible nutrients ee: ether extract s.e.m: standard error of mean nfe: nitrogen free extract adf: acid detergent fibre agriculture and food sciences research, 2017, 4(1): 8-19 19 table-4. pre-pubertal growth performance characteristics of weaned rabbits fed cocoa pod husk mealbased diets rcphm diets fcphm diets hcphm diets performance trait t1 t2 t3 t4 t5 t6 t7 t8 t9 t10 t11 t12 0% 12.50% 25.00% 37.50% 0% 12.50% 25.00% 37.50% 0% 12.50% 25.00% 37.50% s.e.m initial wt. (g/rabbit) 602.00 610.00 608.00 605.00 605.00 610.00 602.50 612.50 600.00 612.00 600.00 610.00 1.32 final wt. (g/rabbit) 1650.00a 1300.00b 1480.00ab 1162.50b 1512.50ab 1266.67b 1475.00ab 1183.33b 1310.00b 1250.00b 993.33c 1000.00c 58.52 total feed intake (g/rbt.) 4217.48 3941.12 3734.17 3337.72 4187.72 4351.90 3956.66 3953.34 4053.66 4418.97 4466.96 4483.06 97.49 wkly. feed intake (g/rbt.) 468.61 437.70 414.91 370.86 465.30 483.54 439.63 439.26 450.41 490.99 496.33 498.12 10.83 adf (g/rabbit/day) 66.94b 62.56c 59.27d 52.98d 66.47b 69.08b 62.80c 62.75c 64.34bc 70.14a 70.90a 71.16a 3.87 total wt. gain (g/rbt.) 1048.00a 690.00c 872.00b 557.50d 907.50b 656.67c 872.50b 570.83d 710.00bc 638.00c 393.33e 390.00e 26.85 wkly. wt. gain (g/rbt.) 116.44 76.67 96.89 61.94 100.83 72.96 96.94 63.42 78.89 70.89 43.70 43.33 24.13 adg (g/rabbit/day) 16.63 10.95 13.84 8.85 14.40 10.42 13.85 9.06 11.27 10.13 6.24 6.19 0.93 fcr 4.03 5.71 4.28 5.99 4.62 6.63 4.53 6.93 5.71 6.92 11.36 11.50 0.72 mortality (%) 20.00* 00.00 0.00 00.00 00.00 0.00 00.00 20.00* 0.00 00.00 00.00 00.00 2.61 note: a,b,c,d,e means on the same row with different superscripts are significantly different (p < 0. 05) rcphm: raw cocoa pod husk meal fcr: feed conversion ratio fcphm: fermented cocoa pod husk meal s.e.m.: standard error of mean hcphm:hot -water treated cocoa pod husk meal rbt.: rabbit wkly.wt. gain: weekly weight gain * mortality due to soldier ants invasion adf: average daily feed intake adg: average daily gain table-5. nutrient digestibilty co efficients of rabbits fed diets containing cocoa pod husk meal (cphm) parameter rcphm fcphm hcphm (%) t1 t2 t3 t4 t5 t6 t7 t8 t9 t10 t11 t12 0% 12.50% 25.00% 37.50% 0% 12.50% 25.00% 37.50% 0% 12.50% 25.00% 37.50% s.e.m dry matter 96.32 96.43 95.41 96.43 96.35 95.58 96.42 96.33 96.34 96.43 96.27 96.31 0.09 crude protein 78.14 77.88 77.21 75.07 76.78 78.54 72.18 78.11 77.32 73.31 74.42 77.74 0.61 crude fibre 77.92 78.44 75.04 74.50 76.79 71.03 61.14 76.68 77.42 85.97 80.80 82.19 1.78 ether extract 71.51 83.57 78.39 76.48 76.26 82.39 79.74 77.23 81.01 74.77 82.03 81.74 1.05 ash 79.82a 63.25b 48.44c 64.23b 79.82a 66.05b 56.84c 72.64b 79.82a 63.35b 75.92ab 83.33a 3.12 nitrogen free extract 73.99a 65.49b 62.04b 63.92b 74.18a 73.96a 65.68b 55.12c 75.66a 68.97b 69.62b 67.42b 1.75 note: a,b,c means on the same row with different superscripts are significantly different (p < 0.05) rcphm: raw cocoa pod husk meal fcphm: fermented cocoa pod husk meal hcphm:hot -water treated cocoa pod husk meal s.e.m.: standard error of mean asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 1 agriculture and food sciences research vol. 5, no. 1, 1-5, 2018 issn (e) 2411-6653 / issn (p) 2518-0193 doi: 10.20448/journal.512.2018.51.1.5 nutrient composition of mushroom: pleurotus ostreatus (jacaum, ex. fr. kummer) grown on different agricultural wastes patience c. obinna-echem1  franklyn a. chukunda2 1department of food science and technology, rivers state university, nkpolu oroworukwo, port harcourt, nigeria 2department of forestry and environment, faculty of agriculture, rivers state university, nkpolu oroworukwo, port harcourt, nigeria ( corresponding author) abstract the study evaluated the nutrient compositions of pleurotus ostreatus (jacaum, ex. fr. kummer) grown using different agricultural waste: sawdust, sugarcane (sacharum officinarum) peels, plantain leaves and corn cobs. the proximate compositions of p. ostreatus fruit bodies varied significantly (p≤ 0.05). matured fruit bodies of p. ostreatus harvested from sawdust and sugarcane peel substrates had significantly (p≤ 0.05) the highest content of moisture (15.20%) and ash (5.23%) respectively. the corn cob substrates yielded p. ostreatus with significantly (p≤ 0.05) the highest amount of protein (9.38%) and lipids (8.5%); and lowest amount of carbohydrates (51.76%). crude fibre was found more in the fruit bodies of p. ostreatus harvested from sugarcane peel substrates (8.23%) followed by corn cobs (7.28% ). the mineral content varied significantly (p≤ 0.05) from 0.43 0.90, 0.12 0.30, 9.20 14.6, 7.30 -13.0 and 1.5 2.7 mg/100g for iron, sodium, potassium, phosphorus and calcium respectively. p. ostreatus from sugarcane peels had significantly (p≤ 0.05) the highest mineral content except for calcium. vitamin c content ranged from 1.3 mg/100g for corn hob to 1.8mg/100g for sawdust. the study revealed that sugarcane peel and corn hob in terms of the proximate and mineral composition are better substrate for the growth of p. ostreatus. it is therefore recommended that these agricultural wastes should be used in the cultivation of p. ostreatus. besides producing nutritious mushrooms, this will help to reduce the huge agricultural wastes within our environment that sometimes when not properly managed caused serious environmental pollution. keywords: mushroom, pleurotus ostreatus, substrates, proximate and mineral composition. citation | patience c. obinna-echem; franklyn a. chukunda (2018). nutrient composition of mushroom: pleurotus ostreatus (jacaum, ex. fr. kummer) grown on different agricultural wastes. agriculture and food sciences research, 5(1): 1-5. history: received: 6 november 2017 revised: 10 january 2018 accepted: 15 january 2018 published: 17 january 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ......................................................................................................................................................................................... 2 2. materials and methods ...................................................................................................................................................................... 2 3. results and discussion ...................................................................................................................................................................... 3 4. conclusion ............................................................................................................................................................................................ 4 references ................................................................................................................................................................................................. 4 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.51.1.5&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/0000-0002-1158-6268 https://orcid.org/orcid-search/quick-search?searchquery=franklyn%20a.%20chukunda agriculture and food sciences research, 2018, 5(1): 1-5 2 1. introduction mushroom is a macro-fungus with a distinctive fruiting body, which may be above ground (epigeous) or below ground (hypogeous) of sufficient size to be seen by the naked eye and to be picked up by hand [1]. all mushrooms belong to the kingdom mycetae, a group distinct from animals, plants and bacteria. most mushrooms belong to either ascomycetes or basidiomycetes. like plants, fungi have a distinct cellular structure but they lack the most important feature of plants, the ability to use energy from the direct sunlight through chlorophyll [2]. since mushrooms do not possess chlorophyll, they depend on other plant material the substrate for their food. they may simply be referred to as saprophytic, parasitic or symbiotic in nature [3]. saprophytic mushrooms depend and grow in plant and animal remains and waste such as animal droppings, dead animals, twigs, fallen leaves, dead wood and stumps organic matter, [3, 4] thus forming the first menu in the food chain or food web. they secrete acids and enzymes which degrade plant and animal remains, breaks down large and insoluble molecules such as carbohydrates, lipids and protein into smaller molecules that can be absorbed [5, 6]. parasitic mushroom depend on other organisms for their existence. they are either obligate or facultative. parasitic mushroom such as honey mushroom destroys hectares of forest; and act as agents of habitat restoration. honey mushroom (arinilla bulbosa) covered 37 acres in michigan forest and was estimated to be 1500 years old [5]. facultative mushroom are weak parasitic mushroom found in dying host plant. the deaths of these are caused by other organisms, or environmental stress. symbiotic mushrooms live in harmony with other organism and they derive mutual benefit from each other. they are referred to as symbiotic or mutualistic mushroom. the hypha/mycelium releases or delivers salts, water and produces antibodies to protect the host plant from pathogens and other fungi while the mushroom benefits carbohydrates and other nutrients [7-9]. commonly cultured mushrooms are saprophytic organisms that feed on dead plant materials. the part of the organism that we see and call a mushroom is really the fruiting body. unseen is the mycelium-tiny threads that grow throughout the substrate deriving nutrients by breaking down organic materials. some species of mushrooms can grow on a wide range of materials and others cannot. after the mycelium has grown throughout the substrate, and when its specific requirements are met, the mushroom will fruit [5]. pleurotus ostreatus (jacaum, ex. fr. kummer) is one among the 40 known species of the genus pleurotus, they are commonly referred to as “oyster mushroom”, grow widely in tropical and subtropical areas and easily artificially cultivated [10]. it is a facultative parasitic mushroom and commonly found in dying oak and cotton [11]. p. ostreatus has a broad, fan or oystershared cap spanning 5 to 25 cm natural specimens range from white to grey or tan to dark-brown; the margin is inrolled when young and smooth often somewhat lobed and wavy. p. ostreatus is popularly consumed all over the world due to their taste, flavor, high nutritional values and medicinal properties [10]. they provide many of the nutritional attribute commonly found in meat, beans, grains or other produce [12]. mushrooms are rich sources of protein [12, 13] ash, lipid, crude fibre, and carbohydrate, minerals (ca, mg, na, p, k, s, mn and fe). mushroom are good sources of vitamins like riboflavin, biotin and thiamine chang and buswell [14]. buigut [15] reported that good number of essential amino acids and water soluble vitamins are present in most edible mushrooms. the nutritional and several medicinal properties of mushroom of the genus pleurotus has been reviewed by some authors [10, 16]. some of the medicinal properties include antitumour a property associated to the presence of polysaccharides [17] antigenotoxic and bio-antimitagenic acidity [18] antihypertensive and anti-hyperglycaemic activities [19]. pleurotus tuber-regium is a common species in southern part of nigeria and it is very useful to cure headache, stomach ailments, colds and fever [20]. mushroom has the ability to biodegrade organic matter and bioaccumulate potential nutrient essential for human wellbeing. hence the cultivation of p. ostreatus using different agricultural wastes such as sawdust, sugarcane (sacharum officinarum) peels, plantain leaves and corn cobs as substrate will be relevant in the reduction of the huge amount of these wastes. this may provide a partial solution to the nation’s waste management problems and the pollution challenges around the environment. in addition to the nutritional benefit that would be derived from the consumption of the cultivated mushroom. this study was therefore aimed at using different agricultural wastes to cultivate p. ostreatus and determine the proximate and mineral compositions of p.ostreatus fruit bodies from the different agricultural wastes. 2. materials and methods 2.1. study site the study was carried out at the laboratory of forestry and environment and food science and technology, rivers state university, nkpolu oroworukwo, port harcourt. 2.2 source of spawn pleurotus ostreatus spawn used for the production was obtained from mushroom farm dilomat nigeria limited located in rivers state university, nkpolu oroworukwo, port harcourt. 2.3. collection and preparation of substrates the sawdust residues were collected from the local wood industry at elabuchi port harcourt. sugarcane (sacharum officinarum) peels and corn cobs were collected from sugarcane sellers’ dump and corn sellers at ogbunabali port harcourt, plantain leaves were also collected from plantain plantations at ogbunabali. the substrates were prepared as described by sales-campos [21]. briefly, the substrates were sun-dried, crushed and packed individually in black polypropylene bags and labelled. two hundred gram (200g) each of the substrate was humidified with 50% of water and the ph of 6.5 maintained using 5g of calcium carbonate (caco3) in relation to the dry sample weight of the substrates [22]. agriculture and food sciences research, 2018, 5(1): 1-5 3 2.4. pasteurization and inoculation the substrates were sterilized using autoclave at 1210c at 15 lbs pressure for 15 munities to kill microbes or contaminates associated with the substrates. after inoculation in aseptic conditions, incubation was performed inside a humid chamber for one month at the temperature of 28+ 20c and relative humidity of 85-90%. water was sprinkled regularly until primordial emergence [23, 24]. 2.5. proximate analysis the proximate compositions of the mushroom from the different substrates were analyzed using the standard analytical methods [25]. moisture was determined by drying at 80oc to a constant weight in an air oven (sanyo gallenkamp, weiss technik, west midlands, uk). ash was determined gravimetrically in a muffle furnace (sanyo gallenkamp, weiss technik, west midlands, uk) at 550°c for 6 h. determination of protein was by kjeldahl method. after distillation and titration, the nitrogen was corrected using a factor of 4.38 for mushrooms [1]. lipids were estimated by exhaustive extraction of 2g of sample with petroleum ether in a rapid soxhlet extraction apparatus (gerhardt, bonn, germany). crude fibre was obtained as the difference from the weight of ash subtracted from the increase of weight on the paper due to the insoluble material after the acid hydrolysis of the fat free samples and the filtration into ashless filter paper. carbohydrate was determined by difference. 2.6. mineral assay the atomic absorption method was employed for mineral element assay. mineral element of pleurotus ostreatus was determined by preparing solutions of their ashes. the quantitative measurement of each element: iron, magnesium, sodium, calcium phosphorus and potassium using gbc avanta ver 2.02 atomic absorption spectrophotometer (japan). phosphorus was determined on the ash solution using the molybdenum blue method [25]. 2.7. vitamin c determination the vitamin c content of the pleurotus ostreatus mushrooms was determined using standard analytical methods [25]. briefly, 10 ml of the prepared sample in a 50ml conical flask was titrated against 2, 6 – dichlorophenol solution until a fiant pink colour persisted for 15 seconds. 2.8. experimental design and analysis of data the experiment was conducted in a completely randomized design (crd); the treatment was replicated five times. data were statistically analyzed using duncan multiple range test (dmrt) at a probability of 5%. 3. results and discussion 3.1 proximate composition of pleurotus ostreatus grown on different agricultural wastes the proximate compositions of mature fruit bodies of p. ostreatus grown on different agricultural substrates varied significantly (p≤ 0.05) as shown in table 1. moisture is required for the normal biochemical activities in living things and also plays a role in the keeping quality of the harvested plant. the moisture content of the mature fruit bodies of p. ostreatus harvested from the different agricultural substrates ranged from 11.03 14.21 % for plantain leaves and sugarcane peels respectively. the moisture content in this study is lower than the report by roy, et al. [12]. the variation may be dependent on various factors, such as harvesting time, maturation period, substrate and environmental conditions of growth and storage [26]. giving the uniform condition of growth of p. ostreatus in this present study, the variation could be attributed to the substrate used. table-1. proximate composition (%) of pleurotus ostreatus grown on different agricultural wastes agricultural substrates moisture total ash protein total lipids crude fibre total carbohydrate plantain leaves 11.03±0.18d 2.85±0.15c 17.82± 3.05c 5.8 ± 0.05b 14.25±0.20d 59.28±0.60c sawdust 15.20±0.20a 5.23±1.20a 16.80±0.25d 5.3 ± 0.04c 16.20±0.25c 56.47±0.40d sugarcane peels 14.21±0.25b 3.10±0.11b 18.20±0.42b 5.6 ± 0.08c 18.23±0.21a 54.86±0.52b corn cobs 12.14±0.22c 3.08±0.20b 19.38± 0.58a 8.5 ± 0.06a 17.28±0.30b 51.76±0.37a note: mean values with the same superscript in the same column are not significantly (p≤0.05) different. n= 5±sd. the amount of protein, lipids and carbohydrate in the mature fruit bodies of the p. ostreatus grown on different agricultural substrates were in the range of 19.38 16.80%, 8.5 5.3% and 51.76 59.28% respectively. the result showed that the different agricultural wastes produced p. ostreatus with high protein content but corn hob substrate had significantly the highest amount. this suggests that p. ostreatus can be a valuable source of protein for use in other products like incorporation into complementary foods. the carbohydrate content is in agreement with the report that carbohydrates constitute the prevailing component of mushroom dry matter, usually about 50-60% [10]. according to kalac [27] the various carbohydrate compounds of mushrooms which include monosaccharide, their derivatives and oligosaccharides are important in the proper functioning of the alimentary tract. the lipid content varied from 5.3% for p. ostreatus harvested from sawdust to 8.5% for corn hob. the low lipid content confirms the report that mushrooms are generally low in oil and fat, although they may contain some essential fatty acids and can be recommended as good supplements for patients with cardiac problems [10, 28]. muhammad, et al. [29] reported that protein, and fat content of various pleurotus species was enhanced by different agro wastes used for the cultivation. matured fruit bodies of p. ostreatus harvested from sugarcane peel substrates had significantly (p≤ 0.05) the highest content of ash (5.23%). ash is an indication of mineral component. the high percentage of protein, carbohydrate, crude fibre moisture in the mushroom fruit bodies cultivated on different substrates conforms to the report of chukunda, et al. [30] and ukoima, et al. [31]. agriculture and food sciences research, 2018, 5(1): 1-5 4 table-2. mineral and vitamin c compositions of pleurotus ostreatus grown on different agricultural wastes agricultural substrates mineral (mg/100g dry weight basis) vitamin c iron (fe) sodium (na) potassium (k) phosphorus (p) calcium (ca) plantain leaves 0.43±0.01c 0.12±0.02b 9.20±0.02c 7.30±0.04c 2.7±0.02a 1.6±0.03 sawdust 0.65±0.02b 0.22±0.01a 12.6±0.04b 10.8±0.03b 1.5±0.03b 1.8±0.04 sugarcane peels 0.90±0.04a 0.30±0.03a 14.6±0.05a 13.0±0.05a 1.7±0.04b 1.5±0.01 corn cobs 0.85±0.03a 0.25±0.01a 12.3±0.04b 10.5±0.04b 1.2±0.04b 1.3±0.01 mean values with the same superscript in the same column are not significantly (p<0.05) different. 3.2. mineral and vitamin c composition of pleurotus ostreatus grown on different agricultural waste the results of mineral composition of pleurotus ostreatus grown in different substrates showed significant differences (p≤0.05) among the various substrates (table 2). sodium, potassium, phosphorus, calcium, and iron found in the fruit bodies of p. ostreatus were in the range of 0.12 0.30, 9.20 -14.6, 7.30 13.0, 1.2 2.7 and 0.43 0.85 mg/100g respectively. mature fruit bodies from sugarcane substrate had significantly (p≤0.05) the highest amount of minerals followed by those of the corn hobs except for calcium. fruit bodies from plantain leaves substrate was significantly (p≤0.05) the highest in calcium content. the rich minerals found could be attributed to the composition of the substrate and the ability of the mushroom to utilize these components from the substrates. the sodium content of the mature fruit bodies of p. ostreatus was lower than the report of roy, et al. 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licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 98-108, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.98.108 © 2019 by the authors; licensee asian online journal publishing group simulation of grain quantity, fan and solar collector sizes for an experimental forced convection grain dryer booker osodo1 daudi nyaanga2 jeremiah kiplagat3 ( corresponding author) 1lecturer, department of industrial and energy engineering, egerton university, kenya, 2associate professor, department of agricultural engineering, egerton university, kenya. 3director, institute of energy research, kenya power, nairobi, kenya. abstract forced convection grain dryers are more efficient and achieve greater drying rates than natural convection dryers. however, it is necessary to dry an appropriate grain layer thickness in such a dryer for the drying process to occur efficiently and at an appropriate rate. a well sized fan is also essential if the drying process is to proceed effectively. an oversize fan will be unnecessarily expensive to buy and operate due to high fan power, while an undersized one will not be able to supply adequate air flow. the solar collector must be properly sized if it is to heat the air to the required temperature. all these factors need to be addressed during the design of a grain dryer. lengthy and expensive trial and error processes can be avoided by applying simulation in the design process. this study developed an experimental grain dryer, addressing the above mentioned issues in the process. simulation of air flow within an initial model of the dryer was done and the results used to size the fan and drying cabinet. the solar collector was also sized. the experimental grain dryer developed consisted of a drying cabinet of dimensions 0.5 m x 0.5 m x 1.0 m and was equipped with a 0.039 kw centrifugal fan. the solar collector area was of dimensions 1.2 m x 1.8 m. keywords: simulation, forced convection dryer, dryer sizing, fan, drying cabinet, solar collector. citation | booker osodo; daudi nyaanga; jeremiah kiplagat (2019). simulation of grain quantity, fan and solar collector sizes for an experimental forced convection grain dryer. agriculture and food sciences research, 6(1): 98-108. history: received: 28 february 2019 revised: 27 march 2019 accepted: 1 may 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 99 2. materials and methods ................................................................................................................................................................. 100 3. results and discussion ................................................................................................................................................................. 103 4. conclusions ..................................................................................................................................................................................... 107 references ............................................................................................................................................................................................ 107 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.98.108&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.98.108&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/125 agriculture and food sciences research, 2019, 6(1): 98-108 99 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this article demonstrates that it is possible to use simulation in sizing the various components of a solar dryer. this will result in savings in time and resources which usually results from the normal design process which involves design, fabrication and testing before a prototype is eventually made. 1. introduction a large proportion of food product is often lost between harvesting and consumption. adebayo, et al. [1] stated that loss of crop occurs in the field (15%), during harvesting (13-20%), as well as during processing and storage (15-25%). post-harvest loss of crop may be attributed to different causes. pests, such as large grain borer account for 10-20% loss, while 5-10% of the losses may be attributed to poor storage facilities. diseases, on the other hand, contribute to 5% of post-harvest crop loss [2]. the problem of post-harvest food loss is particularly significant in developing countries, where food losses are estimated to be of the order of 40%, though this can rise to be as high as 80% under very adverse conditions [3]. for example, incidences of post-harvest product loss in kenya have been estimated at 30%, and can rise to be as high as 100% with the advent of afflotoxin [4]. postharvest loss of maize in kenya in 2007 was 21.1% [5]. one reason for loss of grain after harvesting is spoilage resulting from high moisture content. moist and partly moist crop is prone to fungus infection, which renders it unusable. high moisture content also encourages loss due to attacks by insects, pests and increased respiration. drying of grain is therefore necessary to avoid loss between harvesting and consumption tiwari [6];twidell and weir [7]. barawal and tiwari [8] reported that drying of crop helps to achieve better product quality, longer safe storage and reduction of post-harvest loss hence ensuring more food is available for the growing world population. grain drying may be carried out using different sources of energy. however, solar energy is preferred to other alternative sources of energy such as wind and shale since it is abundant, inexhaustible and non-polluting [9]. in forced convection solar drying, a fan is used to force the air through the grain in order to enhance the circulation of the heated air. such dryers produce greater drying rates and make it easier to control the drying process [10, 11]. the performance of a dryer may also be evaluated based on other criteria such as drying and dryer efficiency, uniformity of drying and quality of final product (extent of cracking and discoloration of grain) as well as total drying time [12, 13]. determination of an optimum design that will ensure best ventilation for any particular application is essential to ensure best performance [14]. although forced convection solar dryers achieve greater drying rates than natural convection dryers, their performance is often not optimal. one reason for this is inadequate distribution of air flow, resulting in inadequate drying air in some sections of the dryer and hence uneven drying of the grain. according to misha, et al. [15] uneven drying is the consequence of poor air flow distribution in the drying chamber. product closer to the air inlet is better dried than that further, due to reduced temperature and air velocity. sometimes, the fan is undersized, leading to insufficient air flow (and velocity), or oversized, leading to excessive energy consumption and in extreme cases grain being blown upward. also use of inappropriate grain layer thickness leads poor performance. if the grain layer is too thick, some sections do not dry well as they receive air which is saturated with moisture. if too thin, air exits while still having capacity to remove moisture, leading to low thermal efficiency. design of dryers that meet these criteria, without use of simulation, would require troublesome development stages, involving trial and error, continued testing and use of prototypes, a process which would be expensive and time consuming. simulation, which is the imitation or reproduction of the behavior of a system or process [16] is useful in the design process, as it saves on the time and resources that would otherwise be required to obtain optimal performance. in this study, the process of simulation was used to improve on the dryer design and performance. the objective of the study was to simulate the grain quantity, fan and solar collector sizes for an experimental forced convection grain dryer. 1.1. solar thermal collectors a solar thermal collector serves the purpose of trapping solar radiation which is then used for heating the working fluid. it usually consists of a black surface, the absorber, and a transparent cover. the absorber does not trap all the incident energy from the sun. it incurs losses due to reflection by the encapsulation (cover) or the absorber itself, convection as a result of exchange with the surrounding air, as well as radiation from the hot absorber surface. the efficiency of the collector depends on two factors: the extent to which solar radiation is converted to heat, and the extent of heat losses to the surroundings [17]. for a flat plate collector, solar collector area ac may be determined from equation 1 used by dabra, et al. [18] and aduewa, et al. [19]. 𝐴𝑐 = �̇�𝑎𝑐𝑝𝑎(𝑇𝑜−𝑇𝑎) 𝐼𝑐𝜂 (1) in the equation, �̇�𝑎 and cpa represented air mass flow rate and specific heat capacity respectively, while 𝐼𝑐 and 𝜂 stood for maximum insolation on collector surface and solar collector efficiency, also respectively. to and ta were used to represent optimum dryer temperature and inlet temperature at ambient. 1.2. pressure drop jia, et al. [20] explain that air flow through packed material may be described using the ergun equation 2. according to this equation, pressure drop , for fluid velocity depends on particle diameter ( , length of bed , fluid viscosity ( void space ( and fluid density ( the effect of cross sectional area (due to container diameter) is ignored in this equation. (2) although there are many channels through packed material, fluid will normally only flow through a few of them, a phenomenon called channeling. this leads to lack of distribution of fluid flow. another limitation is that of formation of hot spots, which leads to damage to the bed and packing materials [21]. the pressure drop in the agriculture and food sciences research, 2019, 6(1): 98-108 100 © 2019 by the authors; licensee asian online journal publishing group drying chamber limits the number of trays that may be used. due to high resistance to air flow through drying product, only a few drying shelves can be used without significantly affecting air movement [22]. 1.3. fan sizing according to wilcke and morey [23] different crops have different airflow requirements for drying, necessitating selection of a fan that will deliver airflows within the recommended range. greater airflows will require larger fans, leading to increased costs, while smaller ones may result in unacceptable crop quality. also, the fan must develop sufficient pressure to overcome resistance to airflow. typical air flow rates range from 0.25-0.51 m3/s.m2 of perforated screen area, these flow rates creating relatively low static pressures of 0.249-1.25 kpa in cross flow and mixed-flow dryers. the fan to be used for such a dryer must be of sufficient capacity to overcome this static pressure, being the resistive force the fan works against while trying to push the air through the grain column [24]. fan power ( may be obtained either from manufacturers’ charts or from equation 3 and 4 as suggested by wilcke and morey [23] as well as maier and bakker-arkema [24]. (3) (4) (v̇ is air volume flow rate while ps represents static pressure) the equations are similar, although for equation 4 an impeller efficiency of 60% has been assumed and incorporated. weiss and buchinger [25] stated that fan efficiency ranges between 30 % and 70 %, hence the assumption is reasonable to cater for most fans if cost of fan and that of power is the major consideration. in the current study, equation 4 was adopted to cater for a general situation where efficiency of fan does not have to be used every time. 2. materials and methods 2.1. research site the study was carried out in njoro, nakuru county, kenya. njoro is located 18 km south west of nakuru town. it lies at an altitude of 1800 m above sea level, and experiences temperature ranges between 17-22 ºc. nakuru county is a moderate to high solar energy potential area. the amount of available solar energy is season dependent, with the december-february season receiving the highest amount of insolation of 678 kwh/m2. the september-november season receives the least insolation of 602.6 kwh/m2. harvesting is normally carried out between august and december, depending on the type of grain [26-28]. 2.2. simulation to estimate grain layer thickness and number of trays simulation was carried out in order to determine the greatest grain layer thickness that would be penetrated by the drying air. this was determined by observing the velocity profile for simulated air flow up different layer thicknesses, with the expectation that air velocity would gradually increase up the layer. according to ergun’s equation 2 the governing equation in the simulation process, fluid velocity increases as pressure drop increases. the latter increases as fluid moves up the grain layer, hence fluid velocity is expected to vary in a similar manner. simulation was first carried out of hot air flowing through a single grain layer of 0.1 m thickness, at an air velocity of 1 m/s. a parametric sweep was then carried out in order to simulate air flow through various grain layer thicknesses ranging between 0.1 m – 0.3 m for air velocities ranging between 1 m/s 5 m/s. a parametric sweep enables simulation within the specified range of parameter values in one simulation process, without having to do it for discrete values. the purpose of the parametric sweep was to determine the maximum grain layer thickness that would allow the fan to overcome static resistance to airflow. once the maximum allowable layer thickness was determined, simulation of air flow up increasing number of grain layers was carried out in order to determine the number of layers the air was able to penetrate. in this case, variation of pressure up the drying cabinet with different grain layer numbers was observed. it was expected that pressure would decrease gradually up the drying cabinet. any behavior to the contrary would suggest air was not able to penetrate. the simulation process was carried out in two major stages: creation of the model and simulation of the model. before simulating air flow up the drying cabinet, a 2-d model of it had to be developed using the software solidworks. once created, the model was imported into the comsol multi-physics simulation software and the process of simulation carried out. the simulation process was also in two major stages: pre-processing and post-processing. figure 1 summarises the model creation process while figure 2 summarises the preprocessing stage of simulation. agriculture and food sciences research, 2019, 6(1): 98-108 101 © 2019 by the authors; licensee asian online journal publishing group figure-1. model creation. source: osodo [29]. figure-2. flow chart of preprocessing stage of simulation. source: osodo [29]. once simulation was complete, post-processing, summarized in figure 3, was carried out to extract the results in various forms as required. figure-3. post-processing stage of simulation. source: osodo [29]. 2.3. sizing of solar dryer 2.3.1. fan power determination having found the grain layer thickness that would allow penetration by the air, the simulated pressure drop for this layer thickness was taken as being equivalent to the static pressure to be overcome by the suction fan. this static pressure (𝑃𝑆) as well as the corresponding air flow rate (�̇�) was applied in equation 4 to determine the power of the appropriate fan. 2.3.2. drying cabinet and solar collector i. drying cabinet to determine the cross sectional area of the drying cabinet, a capacity of 18 kg per tray (a mass that an average family would dry for milling, and also that can be carried comfortably when loading) was assumed. the volume (vgr), of grain per tray was determined using equation 5 the grain density for maize being 0.76 g/cc. the cross agriculture and food sciences research, 2019, 6(1): 98-108 102 © 2019 by the authors; licensee asian online journal publishing group sectional area of the drying cabinet acb was then determined from equation 6 the value of 𝑡𝑔𝑟 (maximum grain layer thickness) having been determined as 0.1 m. 𝑉𝑔𝑟 = 𝑚𝑔𝑟 𝜌𝑔𝑟 (5) 𝐴𝑐𝑏 = 𝑉𝑔𝑟 𝑡𝑔𝑟 (6) (mgr and 𝜌𝑔𝑟 represent mass and density grain respectively) the height of the drying cabinet was sized to carry two trays, each holding a grain layer thickness equal to 𝑡𝑔𝑟. the void space between the trays, having a height equal to that of the grain layer, and a plenum chamber, as well as space above the second tray to accommodate the suction fan were also catered for. ii. solar collector the solar collector area (ac) was determined using equation 1. to find the air mass flow rate (�̇�𝑎) equation 7 and 8 were used. air velocity was measured at dryer exit, which had a diameter of 0.1 m, using a thermoanemometer, cabinet cross sectional area having been found as shown above. 𝑄 = 𝐴𝑣 (7) �̇� = 𝑄𝜌𝑎 (8) (q = flow rate in m3/s, a= cross sectional area in m2, v = air velocity in m/s, �̇� = mass flow rate in kg/s and 𝜌𝑎= density of air in kg/m3) ti was taken to be 23°c (the ambient temperature measured in research area during drying period) and to as 58°c (the maximum temperature to maintain grain quality). a value of 1200 w/m2 was used as insolation ic (estimated from measurements in the research area), while a solar collector efficiency (η) value of 83.28 % was used. this solar collector efficiency was similar to that reported by aduewa, et al. [19] at insolation of 1199.46 w/m2. plate-1. side view of experimental solar grain dryer. source: osodo [29]. plate-2. rear view of experimental solar grain dryer. source: osodo [29]. agriculture and food sciences research, 2019, 6(1): 98-108 103 © 2019 by the authors; licensee asian online journal publishing group 3. results and discussion 3.1. grain layer thickness in order to select the appropriate grain layer thickness, air flow up different layer thicknesses was simulated. the expectation was that air velocity would increase gradually up the grain layer. any variation from this expectation would imply an inappropriate layer thickness. figures 4 – 7 show velocity profiles for different layer thicknesses. figure-4. air velocity up single grain layer of height 0.1 m & inlet velocity 1 m/s. source: osodo [29]. figure-5. air velocity up single grain layer of height 0.2 m & inlet velocity 1 m/s. source: osodo [29]. figure-6. air velocity up single grain layer of height 0.25 m & inlet velocity 1 m/s. source: osodo [29]. agriculture and food sciences research, 2019, 6(1): 98-108 104 © 2019 by the authors; licensee asian online journal publishing group figure-7. air velocity up single grain layer of height 0.3 m & inlet velocity 1 m/s. source: osodo [29]. it is evident that for grain layer thickness of 0.1 m, air velocity increased gradually up the entire grain layer, leveling off at the top. this was according to expectations. this implied that 0.1 m would be an appropriate grain layer thickness for this dryer, and that the suction fan would be able to overcome the static pressure i.e. resistance to air flow, in this case. for other layer thicknesses, this was not the case. for example for grain layer thickness of 0.2 m at inlet velocities 1 m/s, velocity increased gradually up to a grain layer height of 0.04 m, before falling sharply. air velocity was again showed to be increasing sharply at the upper sections of the grain layer. the trend at the section at a height between 0.07 m0.18 m did not show, making it difficult to explain what happened. however, because the expectation was for the air velocity to rise steadily up the grain layer, it was concluded that this was not an appropriate grain layer thickness to use. for the velocity profile up 0.25 m grain layer thickness at 1 m/s, air velocity increased up the grain layer, but only up to a height of 0.03 m before decreasing, showing again that this was not an appropriate grain layer thickness to use. in the case of a 0.3 m grain layer thickness, once again air velocity increased up the grain layer, but the increase was not sustained. the velocity dropped way before the top of the grain layer, at a height of about 0.2 m. this showed that this was not an appropriate grain layer thickness to use. thus it was concluded that the maximum grain layer thickness should be 0.1 m. 3.2. number of layers and trays pressure profiles for simulated air flow up a drying cabinet with different numbers of grain layers are shown in figures 812. figure-8. variation of pressure for four (4) 0.1 m thickness grain layers. source: osodo [29]. agriculture and food sciences research, 2019, 6(1): 98-108 105 © 2019 by the authors; licensee asian online journal publishing group figure-9. variation of pressure for five (5) 0.1 m thickness grain layers. source: osodo [29]. figure-10. variation of pressure for eight (8) 0.1 m thickness grain layers. source: osodo [29]. figure-11. variation of pressure for eleven (11) 0.1 m thickness grain layers. source: osodo [29]. agriculture and food sciences research, 2019, 6(1): 98-108 106 © 2019 by the authors; licensee asian online journal publishing group figure-12. variation of pressure for sixteen (16) 0.1 m thickness grain layers. source: osodo [29]. it was found that beyond four (4) grain layers, the linear trend in pressure drop ceased. from five (5) grain layers and above, there were sections where there was little or no change in pressure, suggesting that there was little or no air flow. sections where the curve remained horizontal indicated no pressure drop. this trend intensified as number of grain layers increased, with the horizontal sections of the curve becoming longer, indicating no pressure drop for greater distances up the grain layers. this continued to the extent that for sixteen (16) grain layers and beyond, the graph was a horizontal line, showing that there was no pressure drop at all, hence suggesting that no air flow through the grain layers occurred. it was therefore concluded that the grain dryer should be loaded with at most four (4) trays, each with a maximum grain layer thickness of 0.1 m. 3.3. sizing of drying cabinet and solar collector figure13 shows variation of pressure for simulated air flow up a single 0.1 m grain layer. it indicates that there was a linear drop in pressure from the lower section of a grain layer upwards. figure-13. variation of pressure for a single layer of thickness 0.1 m. source: osodo [29]. as shown in figure 13 the total pressure drop for a single 0.1 m thick layer was 1.28 x 104 pa, this being the difference between the highest (6.9533 mpa) and the lowest pressure (6.9405 mpa). for two layers or trays, the total pressure drop would therefore be equal to twice this value. this was due to the assumption that pressure drop would be the same for each layer, since they were of equal thickness. equation 2 indicates that pressure drop depends on length of bed and void space which were constant. the other variables, namely particle size, fluid viscosity and density were also assumed to be constant. this yielded a total pressure drop of 2.56 x 104 pa for two (2) grain layers, which was found to be equivalent to a static pressure 𝑃𝑠 of 102.8 inches of water. using equation 5 – 8 the air volume flow rate 𝑉𝑠 was determined to be 1.984 cfm. by applying equation 4 the fan power 𝑃𝑓 was found to be 0.053 hp (0.039 kw). agriculture and food sciences research, 2019, 6(1): 98-108 107 © 2019 by the authors; licensee asian online journal publishing group 3.3.1. drying cabinet sizing although it was found that maximum number of trays should be four, the experimental dryer was designed to carry two trays. each drying tray with a capacity of 18 kg was sized to be of square cross section 0.5 m x 0.5 m. using equation 5 and 6. the lowest tray was to be placed 0.3 m from the bottom to allow for the plenum chamber, and the second one 0.2 m above (0.1 m each for grain layer and void space). leaving 0.3 m above the upper tray for fitting the fan, this resulted in a total drying cabinet height of 1 m. 3.3.2. solar collector sizing using an air velocity, v of 0.3 m/s (the lowest recommended for drying of grains) and drying cabinet cross section 0.25 m2, the volume flow rate q, through the collector was determined to be 0.075 𝑚3/s (mass flow rate �̇� = 0.092 kg/s), using equation 7 and 8. assuming a maximum temperature to of 58°c (to maintain grain quality), and an ambient temperature ti of 23°c (measured in research area during drying period) as well as an insolation ic of 1200 w/m2, the required solar collector area was determined to be 3.25 m2 after applying equation 1. a solar collector efficiency η of 83.28 %, as achieved by aduewa, et al. [19] at an insolation of 1199.46 w/m2 was used in the determination. 4. conclusions as a result of this study, an experimental grain dryer shown in figures 4-6 and plates 1 and 2 was sized and fabricated. simulation led to the conclusion that the grain dryer should be loaded with at most four (4) trays, each with a maximum grain layer thickness of 0.1. having decided to design a dryer with two drying trays, a drying cabinet of dimensions 0.5 m x 0.5 m x 1.0 m was adopted. it was designed to be equipped with a 0.039 kw centrifugal fan. the solar air heater was designed to have a collector area of 3.25 m2. references [1] a. b. adebayo, g. ndunguru, p. mamiro, b. alenkhe, n. mlingi, and m. bekunda, "post-harvest food losses in a maize-based farming system of semi-arid savannah area of tanzania," journal of stored products research, vol. 57, pp. 49-57, 2014. available at: https://doi.org/10.1016/j.jspr.2013.12.004. 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[29] b. o. osodo, "simulation and optimisation of a drying model for a forced convection grain dryer." unpublished phd thesis, kenyatta university, kenya, 2018. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 3, no. 1, 19-24, 2016 http://www.asianonlinejournals.com/index.php/aesr 19 comparison of some quality characteristics of milk from white fulani cow, ouda ewe, and kano brown doe reared under the same environment hauwa l. yusuf1 amin o. igwegbe2  paul y. idakwo3 1 department of animal science, faculty of agriculture, bayero university kano, kano, kano state, nigeria 2,3 department of food science and technology, faculty of engineering, university of maiduguri, maiduguri, borno state, nigeria ( corresponding author) abstract this study was carried out on three lactating animal species namely: ouda ewe (a), white fulani cow (b), and kano brown doe (c), at the middle of their second lactation period or parity. the purpose was to compare the gross chemical composition (ash, moisture, total solids, protein, fat, lactose, calcium, ph and titratable acidity) as well as the calorific value of the fresh milk from the three species reared under the same environment. the components were determined using standard analytical methods while the lactose contents were determined by difference. the ouda ewe milk recorded significantly higher (p 0.05) levels of ash (0.77 0.21%), total solids (19.31 0.17%), protein (4.38 0.04%), lactose (5.36 0.59%), calcium (182.33±0.22mg/100g, and calorific value (88.01±1.22 cal/100g); and significantly lower (p 0.05) level of moisture (84.04 1.64%) than the white fulani cow and kano brown doe milks. and the lowest levels of ph (6.78 0.06) and titratable acidity (0.22 0.03%) were recorded in the white fulani cow milk. the results were in agreement with those obtained elsewhere around the world and, it was recommended that efforts should be intensified in creating awareness in nigeria about the higher nutritional profile of milk from small ruminants and towards breeding of more lactogenic ewe. keywords: calorific value, titratable acidity, ouda ewe, kano brown doe, white fulani cow, lactogenic. contents 1. introduction .................................................................................................................................................................................20 2. materials and method ................................................................................................................................................................20 3. results and discussion ...............................................................................................................................................................21 4. conclusion ...................................................................................................................................................................................22 references ........................................................................................................................................................................................23 bibliography ....................................................................................................................................................................................24 citation | hauwa l. yusuf; amin o. igwegbe; paul y. idakwo (2016). comparison of some quality characteristics of milk from white fulani cow, ouda ewe, and kano brown doe reared under the same environment. agriculture and food sciences research, 3(1): 19-24. doi: 10.20448/journal.512/2016.3.1/512.1.19.24 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: contribution/acknowledgement: this work is licensed under a creative commons attribution 3.0 license all authors contributed to the conception and design of the study. the authors are very grateful to the managements of the department of animal science, faculty of agriculture, bayero university, kano; and the department of food science and technology, faculty of engineering, university of maiduguri, for providing the facilities and the enabling environment to carry out this study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 11 march 2016/ revised: 30 march 2016/ accepted: 5 april 2016/ published: 12 april 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://creativecommons.org/licenses/by/3.0/ http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.19.24 https://orcid.org/orcid-search/quick-search?searchquery=hauwa l. yusuf https://orcid.org/orcid-search/quick-search?searchquery=amin o. igwegbe https://orcid.org/orcid-search/quick-search?searchquery=paul y. idakwo http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.19.24 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.19.24 https://orcid.org/orcid-search/quick-search?searchquery=hauwa l. yusuf https://orcid.org/orcid-search/quick-search?searchquery=amin o. igwegbe https://orcid.org/orcid-search/quick-search?searchquery=paul y. idakwo http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.19.24 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.19.24 https://orcid.org/orcid-search/quick-search?searchquery=hauwa l. yusuf https://orcid.org/orcid-search/quick-search?searchquery=amin o. igwegbe https://orcid.org/orcid-search/quick-search?searchquery=paul y. idakwo http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.19.24 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.19.24 https://orcid.org/orcid-search/quick-search?searchquery=hauwa l. yusuf https://orcid.org/orcid-search/quick-search?searchquery=amin o. igwegbe https://orcid.org/orcid-search/quick-search?searchquery=paul y. idakwo http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.19.24 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.19.24 https://orcid.org/orcid-search/quick-search?searchquery=hauwa l. yusuf https://orcid.org/orcid-search/quick-search?searchquery=amin o. igwegbe https://orcid.org/orcid-search/quick-search?searchquery=paul y. idakwo http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.19.24 agriculture and food sciences research, 2016, 3(1): 19-24 20 1. introduction milk is an important source of all basic nutrients for mammals. milk from various ruminants is used for producing different dairy products including milk cream, butter, yoghurt, sour milk, etc. [1-3]. nigeria is the largest milk producer in west africa and the third largest producer of cow milk in africa, yet africa contributes just over two percent of the world's milk supply [4, 5]. milk accounts for twenty to twenty-five percent of the agricultural sector in sub-saharan africa, two percent of its calories, thirty-three percent of its calcium, and four percent of protein for its people. by value, livestock products make up to eleven percent of the food. the cattle had been the main source of milk in nigeria for a very long time [6] providing more than 90% of the total annual domestic milk output; with the white fulani cow or „bunaji‟ breed recognized as the principal milk producer [7]. unfortunately, the domestic output of about 407,000 metric tonnes of milk [8] from an estimated 14 million cattle can hardly satisfy the dairy demands of an ever increasing population of nigerians [9]. local sheep and goat breeds in nigeria have the potential to supply a significant portion of the milk deficit in the country, because their numbers far exceed that of cattle in both rural and urban communities [9, 10]. sheep milk can be used in every way that cow‟s milk is used: over cereal, cooking, baking, and desserts. the taste is often preferred over cow‟s milk bland taste. it has been observed that after drinking the sheep milk, many people often conclude that cow‟s milk is “tasteless” whereas sheep milk is sweet and creamy. sheep milk is also comparable to goat‟s milk, though the sheep milk does not have the faint bitterness or tanginess of goat‟s milk, which makes it a more preferable alternative to milk from other animal species [11-13]. results of various studies have shown that significant percentage of the world‟s population are lactose intolerant, having difficulty in digesting cow‟s milk (due to deficiency in lactase enzyme) causing symptoms such as gas, cramp and diarrhea. however, most people with lactose intolerance have been reported to be able to drink and enjoy sheep and goat milks without the symptoms because the milk from particularly the goat, is low in lactose content [14-21]. the fats in sheep and goat milks are both monounsaturated and polyunsaturated; they contain higher proportion of shortand medium-chain fatty acids, which have recognized health benefits. for example, short-chain fatty acids have little effects on cholesterol levels in people. they also make milk easier to digest [22]. sheep milk is also higher in calcium, as well as containing higher amounts of vitamins a, d, and e. it also offers more essential minerals such as zinc and magnesium [23]. on the other hand, the fat globules in goat milk are known to be smaller in size than those of the cow milk, and consequently, the fat remains suspended (little cream rose to the top) in goat milk as compared with the cow milk when the samples of the two milks were stored at low temperatures [10]. thus, goat milk is naturally homogenized and therefore it is easier to digest than cow milk because of smaller nature of the fat globules [24]. although the mineral contents of the goat and cow milks are generally similar, goat milk has been reported to contain 13% more calcium, 25% more vitamin b6, 47% more vitamin a, 134% more potassium, and three times more niacin than cow milk [25-27]. it is also four times higher in copper. goat milk also contains 27 percent more of the antioxidant selenium than cow milk [28]. furthermore, cow milk contains five times as much vitamin b12 as goat milk and ten times as much folic acid (12 g in cow milk versus 1 g in goat milk per eight ounces with a recommended daily allowance (rda) of 75-100 g for children [23, 28]. like cow milk, goat milk is low in essential fatty acids, because goats also have efa-destroying bacteria in their ruminant stomachs. despite that, goat milk is reported to contain more of the essential fatty acids linoleic and arachedonic acids, in addition to a higher proportion of short-chain and medium-chain fatty acids. these are easier for intestinal enzymes to digest [13, 16, 17, 29]. goat milk protein forms a softer curd  the term given to the protein clumps that are formed by the action of the intestinal acid on the protein [30] which makes the protein more easily and rapidly digestible. theoretically, this more rapid transit through the stomach could be an advantage to infants and children who regurgitate cow milk easily. goat milk may also have advantages when it comes to allergies. goat milk has been observed to contain only trace amounts of an allergenic casein protein, alpha-s1, found in cow milk [1, 30]. in nigeria, poor nutrition [31] and low reproductive performance have been highlighted as some of the major factors affecting milk production and quality from our indigenous ruminants. consumers always demand nutritionally rich milk and dairy products provided only from cow milk [8]. in addition, the current processes of milk collection from a large number of subsistence farmers is time consuming, inadequate, costly, and prone to adulteration. above all, milk collection is limited to cattle, with milk from other sources being uncommon. there is need to investigate the chemical characteristics of milk from some common milk producing ruminants such as the white fulani cow, the kano brown goat, and the ouda ewe. there is also the need to check on the possibility of including kano brown doe milk and ouda ewe milk as alternative sources of milk for regular consumption. the present study was therefore designed to compare the chemical composition and calorific values of the fresh milk from the three animal species. this information is necessary to developing optimal breeding conditions and marketing strategies for both the meat and milk production programmes using these animals which are in abundant supplies in the area. 2. materials and method 2.1. selection of the lactating animals five (5) each of the three healthy animal species ouda ewe (a), white fulani cow (b), and kano brown ewe (c), were randomly selected at the middle of their second lactation or parturition period. this is due to the fact that age and stage of lactation play important role in both the quality and quantity of milk from various lactating animals [12, 32]. the animals were confined under one good farm condition in the animal science farm, faculty of agriculture, bayero university, kano; and were fed with concentrated feeds consisting of grains, seeds and dry fodders. drinking water was also given to the animals on regular basis. agriculture and food sciences research, 2016, 3(1): 19-24 21 2.2. samples collection the five lactating animals from each group were milked manually (hand milking) twice a day (in the morning and evening) for five consecutive days, and their respective milk was pooled before sampling for the individual component analyses. samples were stored below the temperature of 5 0 c and the gross chemical analyses was conducted within twelve hours from the last day of sample collections. approximately 100ml of milk sample was collected from each of the pools in previously sterilized sample bottles, and then transferred to the animal science laboratory in iced coolers, for the analyses. 2.3. chemical analysis the fresh milk samples were analyzed for the following parameters: ash, total solids (ts), protein, moisture fat and lactose; in addition to the measurement of the ph and titratable acidity (ta), in accordance with the procedures outlined in atherton and newlander [33]; aoac [34] and nielson [35]. protein was determined through the quantification of the nitrogen content by the standard micro-kjeldahl methods [34] and nielson [35] and multiplying the total nitrogen obtained by a conversion factor of 6.38 to arrive at the protein content. the ash content was determined following the procedures described by al-wabel [27] and igwegbe, et al. [36] and while the calcium content in the ash was quantified using a buck 205 atomic absorption spectrophotometer (aas, buck scientific inc., usa). fat content was determined by gerber method [33, 34]. the moisture determination was carried out by step-up oven drying of five (5) ml of fresh milk at 115 0 c until a constant weight was obtained. the lactose content was obtained by subtracting the sum of protein, fat, ash and moisture from 100 [10]. the ph was measured with a ph meter (model wtw410d8120, welheim, german), while the titratable acidity was determined by titration of 9 ml of the fresh milk with 0.1nnaoh in the presence of phenolphthalein indicator [33]. all determinations were carried out in triplicates. 2.4. calculation of the calorific values of milk from the three species calorific (or energy) values of the milk samples (a, b, and c) were calculated from the results of the chemical analysis, using the following generalized equation [37]: cal /100g of milk = (% protein × 4) + (% fat × 9) + (% lactose × 4). 2.5. statistical analysis data were analyzed through analysis of variance (anova). test for significant differences between means were determined using duncan‟s multiple range test [38, 39] at 5% level of significance. 3. results and discussion milk can be obtained from all mammals. the first milk produced by mammals after the birth of an offspring looks different and has different composition. it is called colostrum or immature milk [14, 32]. after about 72 hours, a transition to white milk occurs and it is called mature milk. the results obtained from the chemical analyses of the fresh milk samples from the three animal sources are presented in table 1, along with their statistical parameters. the constituents varied in amount from one species of the animals to another, though the variations are not significant between some of the components. an interesting phenomenon that has been observed in nature is the relationship between the composition of milks among various mammalian species and the growth rate of the respective offspring. nature is, perhaps through evolution, conservative and designs the milk of a given species specifically for growth and development of the offspring of that particular species. this extraordinary wisdom and conservation of energy by nature are demonstrated by and can be appreciated through a close observation of the data presented in table 1. note the relatively higher percentages of ash (0.770.21%), protein (4.380.04%), fat (5.450.06%) and lactose (5.360.59%) in the ouda ewe milk (a) compared with the milks of white fulani cow (b) and kano brown doe (c), which are recorded as 0.680.04%, 3.700.01%, 3.540.03%, 4.880.03% and 0.670.02%, 2.640.10%, 4.750.07% and 4.490.21%, for ash, protein, fat and lactose, respectively. consequently, the calorific value of the sheep milk was observed in this study to be significantly higher than that of goat and cow milks  calculated to be 71.60±0.79, 70.27±1.05 and 88.01±1.22 cal / 100g of goat, cow and sheep milks, respectively. consequently, it has been reported that the growth rate (time in days for the newborn to double its birth weight) among the offspring of sheep is higher than that of the goat and cow due to the higher concentration of nutrients, particularly protein and ash, the nutrients required to develop muscle and skeleton, in the sheep milk [11, 19, 25, 40]. the result of this study also shows the sheep milk (a) to have significantly higher (p0.05) total solid (ts) contents than the cow (b) and goat (c) milks; and that the ts contents of the goat milk was slightly higher than that of the cow milk (table 1). also, the value of ash recorded in sample a (ouda ewe), was significantly higher (p0.05) than the values recorded for samples b (white fulani cow) and c (kano brown doe). the values were, however, in agreement with those obtained by asif and sumaira [12] for sheep, cow, and goat milks. moreover, the values are remarkably lower than those observed in a similar study in malawi by banda, et al. [41] who recorded 0.88% and 0.94% ash in goat and sheep milks, respectively. as indicated above, the total solid values recorded in this study was highest (p0.05) in ouda sheep (19.31%) followed by kano brown doe (13.30%), while the lowest percent total solid (12.35%) was obtained from the white fulani cow milk. the ts content in kano brown doe recorded in this study is slightly higher than the value obtained by igwegbe, et al. [10] while the value from the white fulani cow milk was significantly lower than that obtained from cow milk in a similar study  the values were 13.17% and 13.80% from goat and cow milks, respectively, from the north eastern part of nigeria. the higher levels of ts in sheep and goat milks may be attributed to the greater concentration of nutrients in their milks. the higher concentrations of ts in sheep and goat milks have also been observed in various studies by many other researchers [12, 13, 25, 40]. agriculture and food sciences research, 2016, 3(1): 19-24 22 table-1. comparison between the chemical composition of fresh milk of ouda ewe, white fulani cow and kano brown doe1 parameters (%) type of animal 2 a (ouda ewe) b (white fulani) c (kano brown doe) ash 0.77 ± 0.21a 0.68 ± 0.04b 0.67 ± 0.02b total solid 19.31 ± 0.17b 13.35 ± 0.13c 13.30 ± 0.14c moisture 84.04 ± 1.64a 87.20 ± 0.60b 87.45 ± 1.43c protein 4.38 ± 0.04c 3.35 ± 0.01d 2.64 ± 0.10e fat 5.45 ± 0.06a 3.54 ± 0.03b 4.75 ± 0.07c lactose3 5.36 ± 0.59c 5.23 ± 0.03d 4.49 ± 0.21e titratable acidity 0.35 ± 0.01b 0.22 ± 0.03c 0.40 ± 0.04b ph 6.81 ± 0.02a 6.78 ± 0.06a 6.81 ± 0.03a calcium (mg/100g) 182.33 ± 0.22a 115.56 ± 0.22b 125.07 ±1.06c calorific value(cal/100g) 88.01 ± 1.22b 70.60 ± 0.79c 71.27 ± 1.05d 1values are means of three determinations ± standard deviation (sd) 2in any row, means bearing similar superscripts are not significantly different (p>0.05) 3lactose was determined by difference with regards to protein, fat, lactose, moisture and mineral (particularly calcium) contents of the milk from the three species investigated in this study, significantly higher (p0.05) percentages of protein (4.38±0.04), fat (5.45±0.06), lactose (5.26±0.59) and calcium (182.33±0.22mg/100g) were recorded from ouda ewe (table 1) than the values recorded from the white fulani cow and kano brown doe milks. the mean percentage values of protein and lactose were significantly higher (p0.05) in white fulani cow milk (3.35±0.01 and 4.88±0.03) than in kano brown doe milk (2.64±0.01 and 4.49±0.21), respectively; while the latter has significantly higher percentage of fat (4.75±0.07) than the former. the higher protein content in sheep milk compared to the other livestock species may be due to the fact that more protein is required for wool development in the lamb. the moisture contents were observed to be inversely related with the percent concentration of other macro nutrients, especially protein and lactose, and thus, the kano brown doe milk recorded the highest mean percent level of moisture (87.45±1.43) than the white fulani cow and ouda sheep milks (87.20±0.60 and 84.04±1.64), respectively. also, the doe milk recorded significantly higher (p0.05) concentration of calcium (125.07±1.06mg/100g) than the cow milk (115.56±0.22mg/100g). these observations are in collaboration with the results of similar studies around the globe  including those of banda, et al. [41]; soliman [26]; asif and sumaira [12] and salman, et al. [37]. moreover, the levels of fat (7.0% and 6.7%) reported in sheep milk by george [42] and kolesnikov [21] respectively, are in complete disagreement with the percent fat (5.45±0.08) observed in sheep milk in the present study. in terms of the ph of the fresh milk, the mean ph of the doe and ewe milks were 6.81 and 6.78, respectively. although, the titratatble acidity (ta) expressed as percent lactic acid (la), was observed to be lowest in the cow milk (0.22±0.03), the ta of the fresh milks from the three animal species were within the range obtainable from good quality fresh milk suitable for the processing of any dairy products. the production of acid in milk is normally termed "souring" and the sour taste of such milk is due to lactic acid. the ta of fresh milk is a rough indication of the microbial activity in the milk and the manner in which the milk has been handled; and the hygienic condition of the environment, utensils, freedom from colostrum, prompt cooling of milk after milking and transportation of milk under refrigerated temperature, are the most important factors that determine the level of acidity and suitability of the milk for heat treatment and its use in preparation of dairy products such as yoghurt [10, 43]. generally, the females of all mammal species can by definition produce milk, but cow's milk dominates commercial production. in 2011, the food and agricultural organization (fao) estimated that 85% of milk worldwide was produced from cows [4, 5]. the report also indicated that aside from cattle, many kinds of livestock including goat, sheep, camel, donkey, horse, reindeer and yak, provided milk used by humans; and that goat, sheep, camel and donkey produced about 11%, 2%, 1.4% and 0.2%, respectively, of all milk produced globally in 2011. the main constituents of all milks are water, fat, protein, lactose and ash. the constituents other than the water are known as the total solids (ts) and the total solids minus the fat are known as solids-not-fat (snf) [33]. the composition of normal milk from all the animal species varies to a great extent, but the results from many studies on normal whole milk correspond closely to the figures obtained in the present study. many factors have been reported to affect the overall composition of milk positively or negatively. these factors include: type or species of the animal, the health condition of the animal (presence or absence of mastitis); stage of lactation; season of the year; type of feed, and to a lesser extent, some constituents  particularly fat, is affected by the time and milking frequency [7, 11, 12, 30, 32]. from the nutritional point of view, sheep milk is highly nutritious, containing more calcium and other minerals as well as vitamins (such as vitamin a, b, and e) than the goat and cow milks. both sheep and goat milks are more easily digested due to their smaller fat globules. and from the technological aspect, the sheep milk has higher solids content than goat or cow milk. as a result, more cheese can be produced from a liter of sheep milk than a liter of goat or cow milk. in fact, cheese yield of up to 18 to 25% from sheep milk compared with 9 to 10% from cow or goat milks, has been reported; and that freezing of the sheep milk did not affect the cheese-making qualities of the milk. goat is also termed as walking refrigerator for the storage of milk and can be milked a number of times in a day as compared with twice per day for sheep and cows [44]. moreover, because sheep usually produce far less milk than goats and cows, sheep milk was observed during the course of this study, to sell for a significantly higher price per liter than that of goat and cow milks. thus, a serious breeding programme is required to raise more milk-producing sheep particularly in the arid conditions common in the northern part of nigeria. 4. conclusion in conclusion, this study has ascertained that the milk of all mammals has similar chemical composition that varies somewhat in the proportion of the constituents from one species to another. the ouda ewe investigated in this agriculture and food sciences research, 2016, 3(1): 19-24 23 study appeared to produce the richest milk from compositional point of view, as the milk is characterized by higher total solids, fat, protein, calcium, and ash, than the milk from the white fulani cow and kano brown doe. this finding is in consistent with those of similar studies in various parts of the globe. efforts should be intensified in creating awareness among the nigerian populace concerning the higher nutritional benefits that can be derived from the sheep and goat milks. references [1] j. barlowska, m. szwajkowska, z. l. nczuk, and j. krol, "nutritional value and technological suitability of milk from various animal species used for dairy production," compreh. rev. food sci. food safety, vol. 10, pp. 291-302, 2011. 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[44] a. grandison, "causes of variation in milk composition and their effects on coagulation and cheese making," dairy ind. intl., vol. 51, pp. 21-24, 1986. http://www.attra.ncat.org/ http://www.sheep101.info/dairy.html agriculture and food sciences research, 2016, 3(1): 19-24 24 bibliography [1] m. borkova and j. snaselvo, "possibilities of different animal milk detection in milk dairy products ¾ a review," czech. j. food sci., vol. 23, pp. 41 – 50, 2005. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn(e) : 2411-6653 issn(p) : 2518-0193 vol. 3, no. 1, 37-44, 2016 http://www.asianonlinejournals.com/index.php/aesr 37 evaluation of porridge made from composite flour of orange-fleshed sweet potato and enset (bulla) flours abebe haile1  martha geribo2 esayas kinfe3 1,2,3hawassa university, awassa college of agriculture, school of nutrition, food science and technology, awassa, ethiopia ( corresponding author) abstract bulla is the water insoluble starchy product which is separated from kocho during the decortications process of enset. however, this extract lacks pro-vitamin a. this study enriched the bulla with orangefleshed sweet potato (ofsp) a plant with high levels of β-carotene that can be useful for combating vitamin a deficiency. the porridge was made of composite flours of bulla blended with 30, 35, 40 and 45% ofsp flours were investigated for proximate composition, β-carotene, functional property and sensory acceptability of products using standard methods. the data were analyzed using sas 1.9 software. the composite flours percentage moisture, crude (fiber, protein, fat), ash, carbohydrate, gross energy, β-carotene, ph, bulk density and water absorption capacity were found to be in the range of 29.54 to 40.25, (1.66 to 2.05, 2.11 to 2.55, 0.55 to 0.71), 1.76 to 2.11, 53.86 to 63.31, 227.49 to 268.11 kcal/100 g, 386.68 to 558.05 μg/100 g, 4.46 to 5.71, 0.58 to 0.80 and 1.70 to 4.87 ml/g, respectively. while, the composite flours porridge percentage moisture, crude fiber, ash, crude protein, crude fat, carbohydrate, gross energy, β-carotene, ph and viscosity content found to be in the range from 43.42 to 58.03, 1.32 to 1.79, 2.39 to 2.78, 2.01 to 2.31, 0.71 to 0.82, 35.21 to 48.63, 155 to 211.11 kcal/100 g, 201.46 to 301.50 μg/100 g, 5.71 to 5.82 and 515 to 728 cps, respectively. the porridge ob4pm2 (45% ofsp and 55% bulla) was most preferred due to its color, odor, flavor and overall acceptability with panelist scored value of 4.28, 4.33, 4.39 and 4.45, respectively. moreover, ob4pm2 porridge was found to be good source of ash, energy and β-carotene content. keywords: bulla, composite flour, orange-fleshed sweet potato, porridge, β-carotene, viscosity. contents 1. introduction ............................................................................................................................................................................... 38 2. materials and methods ............................................................................................................................................................. 38 3. results and discussions ............................................................................................................................................................ 40 4. conclusion .................................................................................................................................................................................. 43 references ...................................................................................................................................................................................... 43 citation | abebe haile; martha geribo; esayas kinfe (2016). evaluation of porridge made from composite flour of orange-fleshed sweet potato and enset (bulla) flours. agriculture and food sciences research, 3(1): 37-44. doi: 10.20448/journal.512/2016.3.1/512.1.37.44 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: contribution/acknowledgement: this work is licensed under a creative commons attribution 3.0 license all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 14 april 2016/ revised: 6 may 2016/ accepted: 18 may 2016/ published: 13 june 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.37.44 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=abebe%20haile https://orcid.org/orcid-search/quick-search?searchquery=martha%20geribo https://orcid.org/orcid-search/quick-search?searchquery=esayas%20kinfe http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.37.44 agriculture and food sciences research, 2016, 3(1): 37-44 38 1. introduction bulla is a processed starch made from enset (enset ventricosum). it is obtained when the scrapings and pulps were squeezed, separating of the liquid. collecting the pulpy white juice and left for one or two days to settle the resultant starch to concentrate into white flour. the starch that separates out from the liquid concentrates into a white powder. this product can be either fermented in pit or directly cooked without fermentation [1, 2]. bulla is a good source of calories. it can be prepared as pancake, porridge and made into soup. bulla fulfills both the nutritional requirement and income of the family there by providing household financial security that is by frequent sale of some amounts of fermented bulla from the storage pit. the price of bulla is higher than kocho even at local market [1]. bulla can be stored for a long period of time without spoiling. the shelf life of bulla depends on the age of the harvested enset plant and the way of packing system. proper packaging is needed to protect the spoilage. if there are pockets of air, or exposed to air, molds and other spoilage organisms can grow and soften the product. due to its moisture content there will be taste and color change happen to the product [3]. bulla was considered the best quality enset food. however, it is low in protein, fat content and has no adequate pro-vitamin a for the daily requirement of a normal person. according to the study of minaleshewa [2] and tilahun, et al. [4] protein and fat content of bulla were renged from 0.4-0.8 and 0.2-0.4g, respectively. the vitamin “a” content of enset products record was 0-0.2% [4]. as a result, it is a problem for those people who use enset product (bulla) as staple or co-staple food have a chance of being affected by vitamin a deficiency. orange-fleshed sweet potato (ipomoea batatas ) is an important tuber crop grown in the tropics, sub-tropics and warm temperate regions of the world for its edible roots crop. the roots contain significant amount of carbohydrate constitute mainly in the forms of starch, sugar and dietary fibers, which play an important role of providing low-cost energy in the diet of consumers. beside carbohydrate, it contains high levels of ß-carotene and appreciable amount of minerals. orange-fleshed sweet potato (ofsp) varieties are being promoted as food based intervention to combat vitamin a malnutrition [5]. the flesh color of the root varies from various shades of white, cream, yellow to darkorange depending upon the carotenoids content [6]. carotenoids have been linked with the enhancement of immune system and decreased risk of degenerative diseases such as cardiovascular problems. dietary fiber has the potential to reduce the incidence of a variety of diseases in man including colon cancer, diabetes, heart diseases and digestive disturbances [7]. the primary role of enriching the enset product (bulla) with orange-fleshed sweet potato was to upgrade the vitamin a content of the products. it is the intervention falls in the latter category of food based approaches which is particularly suited for rural areas where the enset products used as a staple food. a food based strategy has the advantage of bringing more nutrients, vitamin “a” and more sustainable approach. enriching the indigenous root crop enset product (bulla) by mixing with orange-fleshed sweet potato can be a good solution to improve the nutritional quality of household diets. vitamin a is a fat-soluble vitamin that was needed in small quantities for several metabolic activities in the body. when vitamin a intake is below required levels, a number of manifestations collectively known as vitamin a deficiency (vad) disorders occur. this vitamin deficiency has manifested as blindness, increased susceptibility to infection and closely associated with increased mortality and morbidity [8]. the causes of vad occurs when vitamin a intake or liver stores fail to meet daily metabolic requirements and the most common cause is a persistently low intake of vitamin a-rich foods. therefore, the objective of this study is enriching of enset product (bulla) flour with orange-fleshed sweet potato to prepare porridge of having high vitamin a content from the blended flours. moreover, to determine the physicochemical, β-carotene content and sensory acceptability of the products. 2. materials and methods 2.1. sample collection raw bulla sample was purchased from hawassa local market. raw orange-fleshed sweet potato of tulla variety (cip-420027) was collected from southern agricultural research center of primary quality sweet potato vine multiplication center (wondo genet farm). all the samples were properly packed and transported to the laboratory of food science and technology, hawassa university. the ofsp was stored at -18 0c (haier chest freezer model hcf 588) to protect spoilage. while bulla was stored at dry and ambient temperature until further processing. 2.2. bulla flour preparation bulla sample was prepared according to procedure outlined by minaleshewa [2]. the semi dried bulk bulla sample was spreaded on clean dry wood board and the long fibers were removed manually. the sample was then sieved through 500µm sieve (fritsch, test sieve, made in german) to remove the hard particles and made it uniform size and the contents were mixed. bulla flour was packed by polyethylene bag and stored at room temperature. 2.3. orange-fleshed sweet potato flour preparation orange-fleshed sweet potato flour was prepared according to the procedure outlined by owori, et al. [9]. the ofsp was taken out from deep freezer and kept on the laboratory table for four hours until its temperature was raised to room temperature. the ofsp was washed with potable water to remove any foreign bodies and then the outer skin was peeled by peeling machine (robot coupé h. biaugeaup model cl 30). the remaining unnecessary parts were removed manually and rinsed by clean potable water. the ofsp was chopped to minimize the size for the slicer machine. the chopped sweet potato was then sliced to a uniform size of 3 mm by the slicer machine (h. biaugeaup model w1 9901). the slices were soaked in 1% w/v sodium chloride solution for 30 minutes. the function of salt is to inhibit the growth of microorganisms on the slices while drying. the slices were dried on the flat racks of solar dryer. the dried sample was milled by electronic grinder and sieved by 500µm sieve (fritsch, test sieve, made in german). the flour obtained was double packed by dark polyethylene bags stored in dry and dark place to protect oxidation of ßcarotene. agriculture and food sciences research, 2016, 3(1): 37-44 39 2.4. preparation of ofsp added bulla composite flour the formulation of ofsp with bulla for porridge making were done based on the recommended dietary intake (rdi) of adult person to maintain a healthy life. it also allows creating unique and delicious recipes based on the acceptability of the users, see table 1 below. table-1. proportion of ofsp powder used to formulate the composite flour bulla composition /kg ob4pm1 ob3pm1 ob2pm1 ob1pm1 b1pm1 ofsp 45% 40% 35% 30% bulla 55% 60% 65% 70% 100% total 1kg 1kg 1kg 1kg 1kg note: * pm1→ is the code of treatments before cooking table-2. proportion of ingredients used to prepare porridge bulla composite flour porridge /kg ob4pm2 ob3pm2 ob2pm2 ob1pm2 b1pm2 ofsp 45% 40% 35% 30% bulla 55% 60% 65% 70 100% water (ml/kg) 1500 1500 1500 1500 1500 oil (ml/kg) 20 20 20 20 20 note: *pm1→ treatments before cooking, *pm2 → treatments after cooking. b1pm1=100% of bulla flour, ob1pm1=30% ofsp + 70% of bulla, ob2pm1=35% ofsp + 65% of bulla, ob3pm1=40% ofsp + 60% of bulla and ob4pm1=45% ofsp + 55% of bulla composite flour; b1pm2=100% bulla porridge, ob1pm2=30 % ofsp + 70% bulla, ob2pm2=35% ofsp + 65% of bulla, ob3pm2=40% ofsp + 60% bulla and ob4pm2=45% ofsp + 55% bulla composite flour porridge. 2.5. porridge preparation bulla based porridge was prepared using the methods outlined by yewelsew, et al. [10]. one kilogram of composite flours was mixed with 1500 ml/kg of water, kneaded well in the pot, and boiled until all particles became soluble while boiling 20ml oil was added. then the composite flour porridge was cooked at the temperature of 100 0c for 10 minutes. 2.6. chemical analysis 2.6.1. determination of ß-carotene contents ß-carotene content in the food was determined as the procedure outlined by rodriguez, et al. [11] based on spectro-photometric test. 2.6.2. determination of proximate composition the proximate composition of flour and composite flour porridge were determined by using the standard method of association of official analytical chemists (aoac) [12]. the total carbohydrate amount in the sample was determined by difference: total c𝐚𝐫𝐛𝐨𝐡𝐲𝐝𝐫𝐚𝐭𝐞 % = 𝟏𝟎𝟎 − (𝐏 + 𝐅 + 𝐀 + 𝐌) whereas: p = the mass percentage of crude protein, f = the mass percentage of crude fat a = the mass percentage of ash, m = the mass percentage of moisture 2.6.3. determination of gross energy the energy value was determined by calculation using the relationship from fat, carbohydrate and protein contents of the atwater’s conversion factors; (4kcal/g) for protein, (9kcal/g) for fat and (4 kcal/g) for carbohydrates and expressed in calories. gross energy (kcal/g) = (4 × protein) + (4 × carbohydrate) + (9 × fat) 2.7. determination of physical and functional properties 2.7.1. ph value the ph of the samples was determined from 1/10 dilution of sample by glass electrode attached to digital ph meter as their combination ratio before cooking and after cooking of each composite flours samples were done according to the method described by yirmaga [13]. the ph meter (mp 511 lab. ph meter, china) was calibrated using ph 4.0 and 7.0 buffers prior to determination of the ph of the samples. 2.7.2. bulk density the bulk density of ofsp added bulla composite flours were determined by using method [14]. agriculture and food sciences research, 2016, 3(1): 37-44 40 2.7.3. water absorption capacity water absorption capacity ofsp added bulla composite flours were determined using the method [15]. 2.8. viscosity the viscosities of the composite flours porridge were measured using the method of [16]. 2.9. sensory evaluation sixty-two panelists who were the students of the school of nutrition, food science and technology, hawassa university did the consumer acceptability of composite flour porridge. the panelists ranked the color, odor, flavor, and overall acceptability of composite flours porridge using a five point hedonic scale (5 extreme like, 4 like very much, 3 like, 2 dislike and 1 dislike very much). the composite flours porridge samples were presented in triplicate. samples were being ordered on the table without providing any information to the panelists. a bottle of tape water with white plastic cups was given to rinse their mouth after each test. they use their observation and sense organ on making decisions, and they interpret the nature of a sample. the results were recorded and analyzed to determine the significance of variations of sensory attributes of the products average scores. 2.10. ethical approval the study protocol was approved by the ethical committee of hawassa university college of medicine and health science for sensory evaluations. 2.11. statistical analysis the data were analyzing using the sas 9.1 software. analysis of variance (anova) was used for the analysis. fisher`s least significance difference (lsd) test was used to determine the significance of mean differences of the results. the level of statistical significance was set at (p<0.05). 3. results and discussions 3.1. proximate composition, physicochemical and functional properties table-3. proximate composition (%) of ofsp added bulla composite flours and porridge. treatment moisture content crud fiber ash crude protein composite flours b1pm1 48.25±0.01d 0.39±0.01i 1.26± 0.01i 1.06±0.01h ob1pm1 40.38± 0.00g 1.66±0.01e 2.11±0.01g 1.76± 0.01g ob2pm1 33.72± 0.01h 1.81±0.00c 2.24± 0.01f 1.88±0.01e ob3pm1 31.62±0.01i 1.92±0.01b 2.38±0.01e 2.01±0.01d ob4pm1 29.54±0.01j 2.05±0.01a 2.55±0.00c 2.11±0.01c porridge b1pm2 60.30±0.01a 0.12± 0.01j 2.00±0.01h 1.90±0.00f ob1pm2 58.03±0.01b 1.32± 0.01h 2.39±0.01e 2.01±0.00d ob2pm2 50.11±0.01c 1.49± 0.01g 2.51±0.01d 2.12±0.01c ob3pm2 46.75±0.01e 1.62± 0.01f 2.62±0.01b 2.21±0.01b ob4pm2 43.42±0.00f 1.79± 0.01d 2.78±0.01a 2.31±0.01a note: *pm1→ treatments before cooking, *pm2 → treatments after cooking. b1pm1 = 100% of bulla flour, ob1pm1 = 30% ofsp + 70% of bulla, ob2pm1 = 35% ofsp + 65% of bulla, ob3pm1=40% ofsp + 60% of bulla and ob4pm1=45% ofsp + 55% of bulla composite flour; b1pm2 = 100% bulla porridge, ob1pm2 = 30 % ofsp + 70% bulla, ob2pm2 = 35% ofsp + 65% of bulla, ob3pm2 = 40% ofsp + 60% bulla and ob4pm2 = 45% ofsp + 55% bulla composite flour porridge. means ± sd, values within the same column with different superscript letters are significantly different from each other (p<0.05). table 3 shows that the moisture content of ofsp and bulla composite flours and porridges were ranged from 31.62-48.25% and 43.42-60.30 %, respectively. the highest moisture content observed in b1pm2 (60.30%) followed by ob1pm2 (58.03%) and the least was for the ob4pm1 (29.54%). moisture content of ofsp and bulla composite flour and porridge were significantly different (p<0.05) to each other. according to kalekristos [3] the moisture content of bulla bought from addis ababa supper market was found in the range of 44 to 57%, that was contain more moisture than the present study. there were significant differences in moisture content between the treatments, before and after cooking of the porridge. the highest moisture content of composite flours and porridges scores were (ob1pm1) 40.38% and ob1pm2 (58.03%), respectively. the difference might be due to addition of more water while cooking the porridge. as the amount of ofsp flour increases in the composite flours, the moisture content of flour and porridge were decreased gradually. crud fiber contents of bulla 0.39%, ofsp added bulla composite flour ranged from 1.66 to 2.05%, while bulla porridge ranged from 0.12 to 1.79%. there were significant difference (p<0.05) among the treatments. higher crude fiber (2.05%) content was obtained in bulla based composite flour ob4pm1, while the lowest (0.12%) was obtained for porridge b1pm2. according to the previous reported literature, bulla was the decanted fluid of the decortications processing of enset plant and it is very poor in fiber content of recorded values were in the range from 0.6 to 0.8% [3]. as the amount of ofsp flour increases in the composite flours, the fiber content increased gradually. this might be due to the crude fiber content of ofsp more than bulla flour. however, the fiber content of ofsp added bulla composite flours were not fulfilling the rda of fiber for healthy adult. according to fao food standards program agriculture and food sciences research, 2016, 3(1): 37-44 41 [17] of the rda of fiber for healthy person set at 25 to 38 g/day which was significantly different from the present study. hence, bulla composite flours porridge needs additional fiber source of food. ash actually a much more general term that can refer to a number of substances in the food. the ash is residue that remains after heating and removing water or it refers to any inorganic material present in food [18]. the ash content of ofsp added bulla composite flour and porridge were ranged from 1.26-2.55% and 2.39 2.78%, respectively. according to minaleshewa [2] and yirmaga [13] the ash content of bulla was found 0.2 and 2.39 %, respectively. the highest ash content was observed for ob4pm2 (2.78%) whereas the least for b1pm1 (1.26%). the ash content of composite flours and porridges were significantly different (p<0.05) to each other. it might be due to the leaching of some minerals from cooking utensils. as the amount of ofsp flour increased in the composite flours, the crude ash content increased. the increment of ash content show that the increasing the density of inorganic material such as minerals present in food. the protein value for different bulla based composite flour and porridge were presented in table3 shows that the highest protein record was for ob4pm2 (2.31%) and the least was observed for b1pm1 (1.06%). there were significant different (p<0.05) observed among protein content except for ob4pm1 and ob3pm1. the crude protein content of bulla was recorded 1.6g/100 g while the protein content of ofsp added bulla composite flour was in the range of 1.76 to 2.11%. these, value were greater than the crude protein content of bulla reported values of 0.6 and 0.4-0.8 g/100g by tilahun, et al. [4] and minaleshewa [2] respectively. the highest protein content of bulla composite porridge score was (2.31%) observed in ob4pm2. when the percentage of ofsp increases, the protein content of composite flour increased gradually. this might be due to relatively higher protein content of ofsp compared to bulla. however, protein contents of the composite flours were not enough for daily requirement of healthy person which was in the range of 0.80 to 1.52 g/kg/d [17, 18]. table-4. proximate composition (%) of crude fat, carbohydrate, energy and β-carotene of composite flours and porridges note: *pm1→ treatments before cooking, *pm2 → treatments after cooking (porridge). b1pm1=100% of bulla flour, ob1pm1=30% ofsp + 70% of bulla, ob2pm1=35% ofsp + 65% of bulla, ob3pm1=40% ofsp + 60% of bulla and ob4pm1=45% ofsp + 55% of bulla composite flour; b1pm2=100% bulla porridge, ob1pm2=30 % ofsp + 70% bulla, ob2pm2=35% ofsp + 65% of bulla, ob3pm2=40% ofsp + 60% bulla and ob4pm2=45% ofsp + 55% bulla composite flour porridge. means ± sd, values within the same column with different superscript letters are significantly different from each other (p<0.05). as it is indicated in table 4 the fat content of ofsp added bulla composite flour and porridge were ranged from 0.26 0.70 % and 0.220.82%, respectively. the present studied samples were slightly different from minaleshewa [2] fat content observed value ranged from 0.2 to 0.4 g/100 g. the fat content of the ofsp added bulla composite flours and porridges were significantly different (p<0.05) to each other. the highest and lowest fat content values were observed for ob4pm2 (0.82%) and b1pm2 (0.22%), respectively. however, fat content of bulla composites were not enough to satisfying the daily requirements of adult person. the results indicating that both bulla and ofsp are poor sources of crude fat. table 4 show that the carbohydrate content of ofsp added bulla composite flour ranged from 48.78 to 63.31%, while that of porridge ranged from 35.00 to 48.63%. the highest and lowest carbohydrate content values were observed for composite flour ob4pm1 (63.31%) and porridge b1pm2 (35%), respectively. according to minaleshewa [2] the carbohydrate content of bulla recorded to be 43.6 to 55.4g/100g, which was less than from the present study finding. there were significant difference (p<0.05) observed between carbohydrate content of the composite flours and porridges. due to the dilution of more water for porridge processing, the sample contained high amount of water. on the other hand as the percentage of ofsp is increased the carbohydrate content of both the composite flours and porridges were significantly increased. the carbohydrate content of the bulla composite flour products were somehow enough for satisfying daily requirements of adult person. the findings indicating both bulla and ofsp were good sources of carbohydrate. the rda of carbohydrate for healthy person was found to be 130 g/day [18]. the gross energy content of ofsp added bulla composite flours and porridges were ranged from 201.07 268.11 and 149.58-211.11kcal/100gm, respectively. the highest gross energy content was found for ob4pm1 (268.11 kcal/ 100gm), while the least was for b1pm2 (149.58kcal/ 100gm). the gross energy content were significantly different from each other (p<0.05). according to the study of almaz [19] and minaleshewa [2] the energy content of bulla was scored 1860 and 1410 -1950 kj/kg, respectively which were different from present study. the highest energy content of composite flour and porridge were scored for ob4pm1 (268.11 kcal/100g) and ob4pm2 (211.11 kcal/100g), respectively. there were significant difference (p<0.05) between the energy content of composite flour and porridge. when the percentage of ofsp increases in the composite flour, the energy content of composite flour and porridge were treatment crude fat total carbohydrate gross energy, kcal/100gm β-carotene, μg/100gm composite flours b1pm1 0.26±0.00h 48. 78±0.02e 201.07±0.00f nil ob1pm1 0.55±0.01g 53.86±0.01d 227.49±0.07d 386.68±0.04d ob2pm1 0.59±0.01f 60.09±0.01c 253.19±0.04c 451.13±0.02c ob3pm1 0.64±0.00e 61.73±0.02b 260.33±0.61b 515.58±0.03b ob4pm1 0.71±0.01d 63.31±0.00a 268.11±0.05 a 558.05±0.00a porridges b1pm2 0.22±0.01i 35.00±0.00h 149.58±0.05 j nil ob1pm2 0.71±0.01d 35.21±0.00h 155.27±0.00i 201.46±0.34h ob2pm2 0.74±0.01c 42.71±0.02g 185.98±0.05 h 231.54±0.03g ob3pm2 0.78±0.00b 45.71±0.02f 198.72±0.06g 265.71±0.01f ob4pm2 0.82±0.01a 48.63±0.01e 211.11±0.07e 301.50±0.02e agriculture and food sciences research, 2016, 3(1): 37-44 42 enhanced gradually. this might be due to the relatively higher energy content of ofsp compared to bulla. the results indicate that the composites are good source of energy. as mentioned on the previously reported literatures, bulla was poor in β-carotene content. therefore, it can be enhanced by the application of β-carotene rich food items to improve the β-carotene of the diet. the effect of addition of ofsp on β-carotene content of enset (bulla) based composite flour and porridge is shown in table 4. the β-carotene content of solar drayed ofsp powder was scored 1280 μg/100g, while the β-carotene content of ofsp added bulla composite flour values were ranged from 386.68 to 558.05 μg/100g. similarly, the β-carotene content of the ofsp added bulla composite porridge values were ranged from 201.46 to 301.50 μg/100g. the highest β-carotene content observed for the composite flour ob4pm1 (558.05μg/100gm) followed by ob3pm1 (515.58μg/100gm) and the least was observed for porridge ob1pm2 (201.46μg/100gm). the β-carotene content of composite flours and porridge were found to be significantly different (p<0.05) to each other. according to the study of stella [6] and aurélie [20] the β-carotene content of raw ofsp was observed in the ranged values from 1,255 to 2,400 and 1,596 to 2,382 μg /100g of edible portion, respectively which is different from present study. the βcarotene content of raw ofsp flour comparing with the composite four was significantly different (p<0.05). this might be due to the degradation of β-carotene in the drying process and mixing with food having none β-carotene. the β-carotene content of composite flour is significantly higher than bulla based porridge (p<0.05). this might be due to the effect of cooking temperature on β-carotene content in the preparation of the porridge. on the other hand when the percentage of ofsp increases, the β-carotene content of composite flour and porridge enhanced gradually. this is due to the higher β-carotene content of ofsp compared to bulla. the results indicate that the composites were good source of β-carotene even though it is less than the one indicated by stella [6] and aurélie [20]. according to kósambo [21] the average β-carotene loss on the finished products of porridge made from ofsp flour was 65%. in the present study the loss of β-carotene content during cooking process of ofsp and bulla composite flour porridge was 256.55 μg/100g (45.97%). the finding show that the β-carotene degradation decreased. this might be due to short cooking time, temperature and genetic variation of the crop. a person who eats 45% ofsp and 55% bulla composite porridge he/she gain 301.50μg that was (33.5%) of the daily requirement of adult. the daily requirement of β-carotene of healthy person range between 250 to 900µg/day [17, 18]. 3.2. physical and functional properties composite flours or/and porridge table-5. physical and functional properties, and ph of bulla based composite flours and porridge treatment ph /100mg bulk density, g/ml wac, g/ml3 viscosity, cps composite flours b1pm1 4.0±0.01c 0.92±0.01a 0.76±0.00e ob1pm1 4.46±0.01c 0.80±0.01b 1.70±0.10d ob2pm1 5.27±0.01b 0.69±0.02c 3.07±0.12c ob3pm1 5.64±1.15b 0.61±0.01d 4.00±0.00 b ob4pm1 5.71±0.03a 0.58±0.01e 4.87±0.15a porridge b1pm2 5.13±0.01c 515±1.00e ob1pm2 5.71±0.00a 525.67±0.58d ob2pm2 5.73±0.01a 553.00±1.00c ob3pm2 5.77±0.01a 624.67±0.58b ob4pm2 5.82±0.02a 728.00±1.73a note: *pm1→ treatments before cooking, *pm2 → treatments after cooking. b1pm1=100% of bulla flour, ob1pm1=30% ofsp + 70% of bulla, ob2pm1=35% ofsp + 65% of bulla, ob3pm1=40% ofsp + 60% of bulla and ob4pm1=45% ofsp + 55% of bulla composite flour; b1pm2=100% bulla porridge, ob1pm2=30 % ofsp + 70% bulla, ob2pm2=35% ofsp + 65% of bulla, ob3pm2=40% ofsp + 60% bulla and ob4pm2=45% ofsp + 55% bulla composite flour porridge. means ± sd, values within the same column with different superscript letters are significantly different from each other (p<0.05). wac-water absorption capacity, cps-centi pose second. the ph value of bulla was 4.00/100mg; ofsp added bulla composite flour and porridge were found in the range from 4.46-5.82/100mg as presented in table 5. the highest ph value observed for bulla based porridge ob4pm2 (5.82/100mg), while the least value was seen for composite flour b1pm1 (4.00/100mg). this might be due to cooking temperature increases the ph value of the samples by denaturing the amino acids in the food. as the percentage of ofsp increases, the ph value of composite flour and porridge were enhanced. the composite flour ph values were significantly different (p<0.05) to each other. while the ph values for porridges were not significantly different (p>0.05) to each other. according to the study of kalekristos [3] bulla bought from addis ababa supper market ph value was 4.9/100mg which was less acidic compared to present study. as mentioned in previous sections, the bulk density of a powder depends on how closely individual particles packed together and it play important roles during mixing in dough formation [14]. the bulk densities for bulla was 0.92 g/ml, ofsp added bulla composite flour were ranged from 0.58-0.91g/ml. the highest bulk densities was observed for b1pm1 (0.92g/ml) while the least was for ob4pm1 (0.58g/ml). the bulk densities of ofsp added bulla composite flours were significantly different from each other (p<0.05). as the amount of ofsp powder increases in the composite, the bulk density decreased gradually. this is due to particle size of ofsp is less than bulla. the water absorption capacity of flour indicates how much water to add during food preparation and interrelated to gelatin properties. hence, the low water absorption capacity recorded by the flours could explain the less gel formation capacity [16]. the finding of the water absorption capacity of bulla in this study was 0.76 ml/g, while the agriculture and food sciences research, 2016, 3(1): 37-44 43 water absorption capacity of ofsp added bulla composite flours were ranged from 1.70 to 4.87 ml/g as shown in table 5. as the amount of ofsp flour increases in the composite, the water absorption capacity increased gradually. this might be due to the water absorption capacity of ofsp powder was higher compared to bulla flour. the highest water absorption capacity of ofsp added bulla composite was for ob4pm1 (4.87 g/ml3) and the least was for b1pm1 (0.76 g/ml3). there were significantly different from each other (p<0.05). the value of viscosity of bulla porridge was 515.00 c.p.s, ofsp added bulla composite flours porridge were ranged from 525.67-728.00 c.p.s. (see above table 5). the highest viscosity value was observed for ob4pm2 (728.00 c.p.s) followed by ob3pm2 (624.67) and the least for b1pm2 (515.67 c.p.s.). there were significantly difference (p<0.05) between each treatment. as the amount of ofsp flour increases in the composite flours, the viscosity of the porridge was also increased. this might be due to the water absorption capacity of the ofsp flour. the higher the viscosity is a desirable quality attribute of flour for cooking purpose. liquids make up of small molecules have a low viscosity and liquids with long chain molecules have a much higher viscosity [22]. 3.3. sensory evaluation of bulla based composite flours porridge table-5. organoleptic evaluation score of ofsp added bulla composite flours porridge note: *pm1→ treatments before cooking, *pm2 → treatments after cooking. b1pm1= 100% of bulla flour, ob1pm1=30% ofsp + 70% of bulla, ob2pm1=35% ofsp + 65% of bulla, ob3pm1=40% ofsp + 60% of bulla and ob4pm1=45% ofsp + 55% of bulla composite flour; b1pm2=100% bulla porridge, ob1pm2=30 % ofsp + 70% bulla, ob2pm2= 35% ofsp + 65% of bulla, ob3pm2=40% ofsp + 60% bulla and ob4pm2=45% ofsp + 55% bulla composite flour porridge. means ± sd, values within the same column with different superscript letters are significantly different from each other (p<0.05). the porridges were prepared by blending of ofsp and bulla in different combination were analyzed for various sensorial attributes for their acceptance by using 5 point hedonic scale. when you are offering a new product or changing ingredients of food items, it is important to know what likely the consumer reaction is to be to the characteristics of your product. at the present study all panelists were instructed to make their own individual assessments, according to the evaluation criteria provided for each samples on the basis of, color, odor, flavor and overall acceptability. the organoleptic evaluation of ofsp and bulla composite flours porridge score values ranged from 1.05-4.28, 1.00-4.33, 1.3-4.39 and 1.25-4.45 for attributes of color, odor, flavor and overall acceptability, respectively as indicated in above table 6. there were significant difference (p<0.05) was observed among the treatments. the highest preferences of bulla composite porridge by color, odor, flavor and overall acceptability were scored 4.28, 4.33, 4.39 and 4.45, respectively. it was observed that the bulla based porridge sample ob4pm2 , prepared by addition of 45% ofsp flour was liked most by sensory panelist as compared to the other combinations. the least preferred bulla composite porridge ob1pm2 was scored 1.05, 1.00, 1.30 and 1.25 for color, odor, flavor and overall acceptability, respectively. the evaluation results of the panelist’s preferences were increases as the ratio of ofsp increases in the composite flours. 4. conclusion as orange-fleshed sweet potato (ofsp) percentage increased in the composite flours and porridge; gross energy, β-carotene content, ph, water absorption capacity and viscosity were enhanced. however, an increased amount of ofsp in bulla flour and its corresponding products do not made them to be good source of crude fiber, protein and fat. the composite flours porridge ob4pm2 was preferred by panelists in terms of the sensory attributes of color, odor, flavor and overall acceptability. references [1] n. almaz and n. anke, "the significance of enset culture and biodiversity for rural household food and livelihood security in south western ethiopia. wageningen university and research center," agriculture and human values, pp. 62-65, 2004. [2] a. minaleshewa, "studies on commercially available enset (enseteventricosum welw.cheesman) food products (kocho and bulla) for major, minor and trace elements," thesis of in partial fulfillment of requirements for the m.sc. graduate project (chemistry.774) addis ababa university, 2007. [3] y. kalekristos, "studies on of influence of baking time and temperature on the quality of kocho biscuit enriched with faba bean and wheat," in partial fulfillment of the requirements for the m.sc. thesis of institute of technology department of chemical engineering addis ababa university, ethiopia, 2010. [4] a. tilahun, s. ann, and a. jens, "advancing human nutrition without degrading land resources through modeling cropping systems in the ethiopian high lands. the united nations university," food and nutrition bulletin, vol. 25, pp. 344-351, 2004. [5] t. assefa, t. engida, a. teshome, and t. tesfaye, "summary of progress on orange-fleshed sweet potato research and development in ethiopia," in proceedings of the 13th istrc symposium, arushac 2007 tanzania, 2007, pp. 728 – 730. [6] o. stella, promotion of high quality sweet potato flour in the food industry, evidence from rural women in nigeria university. ibadan, nigeria, 2004. [7] l. jan, u. rafael, a. mariaisabel, and h. julie, "orange-flesh sweet potato promising partnerships for assuring the integration of nutritional concerns into agricultural research and extension," pp. 1-4, 2000. [8] ethiopian national guidelines for control and prevention of micronutrient deficiencies, federal ministry of health family health department. ethiopia, 2004. parameters porridge color odor flavor overall acceptability b1pm2 1.05±0.12e 1.00±00 e 1.30±1.00 e 1.25±0.30 e ob1pm2 2.63±1.08d 2.62±1.09d 2.60±1.06d 2.58±0.99d ob2pm2 2.80±0.78c 2.84±0.85c 2.83±0.80c 2.89±0.77c ob3pm2 3.36±0.92b 3.36±0.95b 3.37±1.05b 3.53±0.94b ob4pm2 4.28±0.87a 4.33±0.82a 4.39±0.83a 4.45±0.76a agriculture and food sciences research, 2016, 3(1): 37-44 44 [9] c. owori, b. lemaga, r. o. m. mwanga, a. namutebi, and r. kapinga, "sweet potato recipe book, sweet potato processed product from eastern and central africa kampala uganda. [accessed january, 2014]," 2007. [10] a. yewelsew, b. j. stoecker, m. j. hinds, and g. e. gates, "nutritive value and sensory acceptability of corn and kocho based foods supplemented with legumes for infant feeding in southern ethiopia," african journal of food agriculture nutrition and development, vol. 6, pp. 3-13, 2006. [11] a. rodriguez, a. rodriguez, and k. mieko, "assessment of provitamin a determination by open column chromatography/visible absorption spectrophotometery," journal of chromatographic science, vol. 26, pp. 624-629, 1988. [12] association of official analytical chemists (aoac), standard official methods of analysis of the association of analytical chemists, s.w. williams (ed), 14th ed. washington, d c.: aoac, 1984. [13] m. yirmaga, "improving the indigenous processing of kocho, an ethiopian traditional fermented food. nutrition & food department of food science and post harvest technology. institute of technology, haramaya university, ethiopia," journal of nutrition and food science, vol. 3, p. 1000182, 2013. [14] a. k. oladele and j. o. aina, "chemical composition and functional properties of flour produced from two varieties of tigernut (cyperuse sculentus). department of food technology, university of ibadan, ibadan, nigeria," african journal of biotechnology, vol. 6, pp. 2473-2476, 2007. [15] s. k. sathe and d. k. salunkhe, "functional properties of great northern bean (phaseolus vulgaris) proteins. emulsion, foaming, viscosity and gelatin properties," journal of food science, vol. 4, pp. 71-75, 1981. [16] a. v. ikujenlola, "chemical and functional property of complementary food prepared from malted and un melted achaea soybean and defatted sesame seed," journal of food engineering and applied science, vol. 3, pp. 471-473, 2008. [17] fao food standards program, "codex alimentarius commission. codex committee on vegetable proteins draft codex standard for wheat protein products including wheat gluten (advanced to step 8 of the procedure) 24th session geneva, switzerland," 2001. [18] food and nutrition board, "dietary references of energy, carbohydrate, fiber, fat, fatty acid, protein and amino acid (macronutrient). us department of agriculture, national agricultural library and national academy of sciences," institute of medicine, pp. 48, 107, 265, 339, 422 & 589, 2005. [19] n. almaz, diversity and conservation of enset (enset ventricosum welw. cheesman) and its relation to household food and livelihood security in south western ethiopia: wageningen university, 2001. [20] b. aurélie, "investigating of carotenoid loss after drying and storage of orange-fleshed sweet potato," a thesis submitted in partial fulfillment of the requirements of the university of greenwich phd thesis, 2010. [21] l. kósambo, "effect of storage and processing on all trans β-carotene content in fresh sweet potato (ipomoea batatasl) roots and its products," kenya industrial research and development institute conference, annual report (july 2003 – june 2004) kenya industrial research project, nairobi2004. [22] p. a. okorie, functional and pasting properties of lesser known nigerian yams as a function of blanching time and particle size. department of food science and technology. abakaliki: ebonyi state university, 2011. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 23 agriculture and food sciences research vol. 5, no. 1, 23-29, 2018 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2018.51.23.29 physico-chemical and sensory quality of tigernut (cyperus esculentus) –coconut (cocos nucifera) milk drink patience c. obinna-echem1  charles n. torporo2 ( corresponding author) 1,2rivers state university, department of food science and technology, nkpolu-oroworukwo, port harcourt, rivers state, nigeria abstract physico-chemical and sensory qualities of blends of milk drink from tigernut (cyperus esculentus) and coconut (cocos nucifera) were evaluated. twelve blends (sample a – k) were formulated using different ratios of tigernut to coconut milk as follows: 100:0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90 and 0:100. the ph, total titratable acidity, specific gravity and viscosity ranged between 5.86-6.37, 0.10-0.15%, 0.99-1.08, and 0.25-0.69 centipoise (cp) respectively. viscosity decreased with increase in coconut milk. the solid non-fat, protein, crude fibre and carbohydrate content decreased with increase in coconut milk substitution and the values ranged from 8.45 13.70%, 2.39 3.93%, 0.46 – 0.87% and 4.10 8.65% respectively. the moisture content ranged between 83.80% for sample a and 89.50% for sample i. the fat (2.09 – 3.64%) and ash content (0.13 – 3.00%) varied significantly among blends. the colour, flavor, mouth feel, taste and overall acceptability had increase in likeness with increase in coconut milk. the result indicated that the tigernut-coconut drink is of good quality and the 10% tignernut substitution based on the sensory properties was more acceptable. this forms a baseline data for further studies. keywords: tigernut-coconut milk drink, physico-chemical, proximate, sensory qualities. citation | patience c. obinna-echem; charles n. torporo (2018). physico-chemical and sensory quality of tigernut (cyperus esculentus) –coconut (cocos nucifera) milk drink. agriculture and food sciences research, 5(1): 23-29. history: received: 6 november 2017 revised: 30 january 2018 accepted: 2 february 2018 published: 6 february 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: the authors appreciate the laboratory assistance of mr friday owuno of the department of food science and technology, river state university, port harcourt, nigeria. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 24 2. materials and methods ................................................................................................................................................................... 24 3. results and discussion ................................................................................................................................................................... 25 4. conclusion ......................................................................................................................................................................................... 28 references .............................................................................................................................................................................................. 28 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.51.23.29&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=patience%20c.%20obinna-echem https://orcid.org/orcid-search/quick-search?searchquery=charles%20n.%20torporo agriculture and food sciences research, 2018, 5(1): 23-29 24 1. introduction milk, a lacteal secretion from mammary gland is a rich source of protein and essential component such as vitamins and minerals which promote growth, bone and teeth development [1]. it serves as an excellent source of all nutrients except ascorbate and has been recognized as an important food for infant and growing children [2]. in developing countries, the cost of dairy milk and their product are expensive for the commoners to purchase. the dramatic decrease in the consumption of milk and milk products stimulated in part the processing of milk from different seeds and nuts [3]. in view of the scarce milk supply in various countries and the ever increasing gap between the requirement and population, efforts have been made over the years to develop alterative milklike products from vegetable sources [4]. among the sources of vegetable milk, soybean has received very high research attention and more research is still being designed to improve the quality of soymilk [5]. however, hardly any attention has been given to the use of locally available underutilized tubers like tigernut in milk production or in combination with other milk to produce a palatable ready–to–serve bottled beverage, like “horchata de chufas” as done in south europe especially in spain [6]. tigernut (cyperus esculentus) is an underutilized tuber of the family cyperaceae, which produces rhizomes from the base of the tuber that is somewhat spherical [7]. it is a tuber that grow freely and is consumed widely in nigeria and other parts of west africa, east africa, parts of europe particularly spain as well as in the arabian peninsula [8]. in nigeria, tigernut is known as “aya” in huasa, “ofio” in yoruba and “akiausa” in igbo. three varieties (black, brown and yellow) are cultivated in the country and among these, only two varieties (yellow and brown) are readily available in the market. the yellow variety is preferred to all other varieties because of its inherent properties like its bigger size and attractive colour. the yellow variety also yields more protein and possess less anti-nutritional factors especially polyphenols [9]. tigernut produces high quality oil of up to about 25.5% content and 8% of protein [10]. the protein in tigernut is of high biological value considering the many essential amino acids it contained [11]. these amino acids are higher than those proposed in the standard by the fao/who [12] and satisfy amino acid need of adults [13]. tigernut has been reported to be an excellent source of some useful minerals (iron, calcium phosphorus, potassium, sodium, magnesium, zinc and traces of copper) and vitamin e that are essential for body growth and develpment [14]. it is said to be aphrodisiac, carminative, diuretic, emmanogogue stimulant and tonic and also plays a role in the treatment of flatulence, indigestion, diarrhea, dysentery and excessive thirst [10]. tigernut is used in the production of refreshing purely natural vegetable milk. the milk originated from spain where it is known as chufa de horchata while it is commonly called kunnu aya in northern nigeria. it is a very nutritive energy drink both for the young and the old. the qualities of tigernut in this context stimulate its inclusion in the preparation of beverage so as to provide protein energy-rich drink at affordable price in place of animal protein/fat which is scarce and expensive [15]. coconut (cocos nucifera) is a large palm, growing up to 30 m (98 ft) tall, with pinnate leave 4-6 m (13-20 ft) long and pinnate 60-90 cm long; old leaves break away cleanly, leaving the trunk smooth, they are generally classified into two types: tall and dwarf [16]. coconut is one of the most important palm extensively grown and used nut in the world. it is an important commercial crop in many tropical countries, contributing significantly to their economies [17]. coconuts grows extensively in nigeria and are eaten as snack usually for the pleasure of it, they may be excellent sources of raw materials for the development of dairylike products [18]. coconut milk is being used by confectionaries, bakeries, biscuit and ice cream industries worldwide to enhance flavour and taste of various products [19]. this study is therefore aimed at the preparation of a refreshing and affordable milk drink from tigernut and coconut, and to evaluate the physico-chemical and sensory properties of the milk drink. 2. materials and methods 2.1. tigernut tubers and coconut dried tigernut tubers and matured coconut used in this study were procured from rumuwoji market, mile 1, port harcourt, rivers state, nigeria. 2.2. extraction of tigernut milk tigernut milk was prepared by the method described by adedokun, et al. [3]. briefly, 1 kg of dried tigernuts was manually sorted to remove foreign matters. the sorted nuts were washed with tap water and soaked in 6 l of warm distilled water at room temperature for 48 h to hydrate. thereafter the nuts were washed and wet milled into slurry using (qbl-18l40, taipei city, taiwan). the milk was extracted by sieving using a cheese cloth. the chaff was discarded and the milk was packaged in sterile plastic bottles and refridgerated till required for use. 2.3. extraction of coconut milk coconut milk was also prepared as described by belewu and belewu [20]. the coconuts were shelled and the meat separated from the shell with a kitchen knife. the brown layer was also removed using a kitchen knife. the meat was thoroughly washed and grated using (qbl-18l40, taipei city, taiwan). warm water was added to the grated coconut meat and allowed to stand at room temperature for 1 h. thereafter, the milk was extract using a cheese cloth. the milky-white opaque emulsion was packaged in sterile plastic bottles and refrigerated till when needed for use. 2.4. tigernut/coconut milk blends the extracted tigernut milk (tm) and coconut milk (cm) were mixed at different proportions as shown in table 1. a homogeneous mixture was achieved with the use of a food q-link blender and grinder (qbl-18l40, taipei city, taiwan) operated at full speed for 5 minute. the resultant blend was pasteurized at 700c for 15 minute in a water-bath and cooled immediately to room temperature (28±20c) for further analysis. agriculture and food sciences research, 2018, 5(1): 23-29 25 table-1. ratios for the blend of tigernut-coconut milk sample id a b c d e f g h i j k blend (tm:cm) 100:0 90:10 80:20 70:30 60:40 50:50 40:60 30:70 20:80 10:90 0:100 2.5. physical analyses the physical properties of the products determined were ph, total titrable acidity, total solids, specific gravity and viscosity. the ph of 10 ml of the milk blends was determined with a ph meter (ts 652, germany). thereafter, the 10 ml of the sample was titrated against 0.1 mol l-1 naoh with phenolpthaline as indicator to determine the amount of acid in the sample as total titratable acidty (tta). the result was expressed as % lactic acid. total solid total solid was determined by difference after analyzing for moisture using an automatic moisture analyser (and ml-50, japan). specific gravity of the samples was determined using the aoac [21] method. the density of the milk was measured against the density of water. viscosity of the samples was determined with the aid of a rotary digital viscometer (ndj-85, china) at 20oc. the rate of flow of 250 ml of the milk blend was compared with the rate of flow of the same amount of water. 2.6. proximate analysis the moisture, protein and ash content of the tigernut-coconut milk blends were determined using standard analytical methods [21]. moisture was determined by using an automatic moisture analyser (and ml-50, japan). ash was determined gravimetrically in a murffle furnace (sanyo gallenkamp, weiss technik, west midlands, uk) at 500°c for 24 h. determination of protein was by kjeldahl method. after distillation and titration, the nitrogen was corrected using a factor of 5.7. fat was determined by rose-gottlieb method [22]. carbohydrate was obtained by difference. total solid was obtained by difference (%ts = 100 moisture content). the total solid non-fat was obtained as follows: (%) tsn= ts – fat [23]. where, ts =total solid, tsn=total solid non-fat 2.7. sensory analysis a panel of 20 panelists consisting of staff and students chosen from the university community were used for evaluation of the sensory attributes of the tigernut-coconut milk. they were asked to assess the samples based on the following attributes: colour, mouth feel, taste, flavour and overall acceptability. panelists rating were based on a 9-point hedonic scale with the degree of likeness of the product attribute express as: 1 dislike extremely, 2 dislike very much, 3 dislike moderately, 4 dislike slightly, 5 neither like nor dislike, 6 like slightly, 7 like moderately, 8 like very much and 9 like extremely. 2.8. statistical analysis results were analyzed statistically by the analysis of variance and difference between means separated by least significant difference (lsd) procedure. 3. results and discussion 3.1. physical properties the ph, titratable acidity (% lactic acid), specific gravity and viscosity of different blends of tigernut-cocount milk drink were shown in table 2. table-2. the ph, titratable acidity (% lactic acid), specific gravity and viscosity of different blends of tigernut-cocount milk drink sample identity blend tm:cm ph total titratable (%lactic acid) specific gravity viscosity (cp) a 100:0 5.99±0.02 0.15±0.07 1.10±0.1 0.69±0.06a b 90:10 6.05±0.05 0.13±0.07 1.02±0.01 0.61±0.01a c 80:20 6.18±1.18 0.12±0.01 0.99±0.04 0.40±0.01b d 70:30 5.95±0.04 0.12±0.01 1.03±0.01 0.33±0.03bc e 60:40 5.95±0.06 0.13±0.01 1.02±0.01 0.35±0.01bc f 50:50 5.95±0.07 0.11±0.01 1.02±0.01 0.33±0.03bc g 40:60 5.86±0.2 0.17±0.01 1.03±0.02 0.38±0.04b h 30:70 6.12±0.07 0.10±0.01 1.00±0.01 0.35±0.01bc i 20:80 6.20±0.01 0.11±0.01 0.99±0.01 0.49±0.02ab j 10:90 6.10±0.06 0.10±0.01 1.00±0.01 0.25±0.01c k 0:100 6.37±0.04 0.10±0.02 1.08±0.01 0.25±0.01c note: the ph, total titratble acidiy and specific gravity did not vary significantly (p ≥ 0.05) values of viscosity with the same superscript do not vary significantly (p ≥ 0.05). n=3±sd there was no significant difference (p ≥ 0.05) in the ph, titratable acidity and specific gravity of the blends. the ph of the milk blends from 5.86 6.37 for sample g and sample k (the blend with 60% coconut milk and 100% coconut milk) respectively. this range of ph is an indication of low acidity. ph indicates the level of acidity (h+) and alkalinity (oh-) in the milk and is used to determine the wholesomeness of the milk. the level of titratable acidity in all the treatment ranged between 0.1 0.17% lactic acid. this compared favorably with soymilk 0.10 0.30% and tigernut milk 0.13 0.20% [24] and cow milk 0.17 0.19% as reported by bowen and lawrence [25]. the level of acidity of the milk drink is also an indication of a good quality, as lactic acid is usually produced from the activities of microorganism where the milk sugar is converted to lactic acid. levels of lactic acid more than what has been obtained in this study is not required since the product is not a fermented product. the viscosity varied from 0.25 0.69 centipoise (cp) for sample k and a respectively. the viscosity seemed to decrease with increase in coconut milk. this may be attributed to the carbohydrate content of tigernut. the specific gravity ranged between 0.99 1.08 for sample c and k respectively. this level of specific gravity is an indication of good quality. higher specific gravity would mean that there is more water than milk solid. the agriculture and food sciences research, 2018, 5(1): 23-29 26 amount of water would depend on the quantity used for extraction and result indicated that the quantity used was acceptable for a good quality milk drink. 3.2. proximate composition of tigernut-coconut milk drink table 3 shows the proximate composition of milk drink produced from different blends of tigernut and coconut milk. the total solid content varied between 16.20±0.04 to10.50±0.07% for sample a and i respectively. the total solid non-fat ranged from 8.45±0.10 13.70±0.04% for sample k and sample a respectively. the variation could be attributed to the solid content of the coconut milk as the trend was that of decrease with increase in the substitution of coconut milk. it also indicated consistency in the milk extraction method. the solid non-fat content followed a similar trend ranging from 13.70±0.04 6.86±0.07%. except for sample i, the solid non-fat content of the tigernut and coconut milk drink were either greater than or comparable with cow’s milk the crude protein obtain ranged between 3.93 2.93% for sample a and k respectively. this observation indicated that tigernut milk had more protein than the coconut milk as the decrease in protein was with increase in coconut milk. the result obtain was lower than 6.64% as reported by ogbonna, et al. [26]. however, the crude protein found in the milk blend compared favourably with 3.20% and 3.4% for coconut natural milk blend and cow milk respectively [27] as reported by adedokun, et al. [3]. the fat content of the milk blend varied from 2.09 to 3.75% for sample c and sample i respectively. this result compared favourably with 2.36 and 3.12 % for coconut-natural milk and tigernut-natural milk blend [3]. crude fibre varied from 0.46 for sample k to 0.87 for sample a. tigernut contributed significantly to the crude fibre content of the milk blend. the high crude fibre content of blends with more tigernut milk makes for its recommendation for result people with problems of digestion, flatulence, and diarrhea. ash content is a measure of mineral elements in a food. the ash content of the samples varied significantly and the unsubstituted coconut milk (sample k). the result obtained compare unfavorably with the findings of adedokun, et al. [3] in which 1.08% was reported from natural tigernut blend. sample a and k had significantly the highest (8.65%) and lowest (4.10%) carbohydrate content. the trend was a decrease in carbohydrate content with the increase in coconut milk substitution, confirming the high carbohydrate content of tigernut. this could be of health importance as it can be used by diabetic patient due to its high content of arginine, which liberates the hormone insulin [28]. agriculture and food sciences research, 2018, 5(1): 23-29 27 table-3. proximate composition of different blends of tigernut-cocount milk drink sample identity blend tm:cm parameter (%) moisture total solid solid non-fat ash crude fiber fat protein carbohydrate a 100:0 83.80±0.04a 16.20±0.04 13.70±0.04 0.26±0.02ab 0.87±0.03a 2.50±0.01c 3.93±0.01a 8.65± 0.07a b 90:10 84.25±0.04ab 15.75±0.04 13.22±0.04 0.23±0.01b 0.85±0.03a 2.53±0.03c 3.38±0.01b 8.28±0.02a c 80:20 86.65±0.02b 13.35±0.02 11.26±0.02 0.13±0.03c 0.67±0.03b 2.09±0.01d 3.79±0.01a 6.45±0.08b d 70:30 85.75±0.30b 14.25±0.03 11.56±0.03 0.30± 0.01a 0.67±0.02b 2.69±0.03c 3.61±0.07ab 6.85±0.07b e 60:40 86.10±0.60b 13.90±0.60 10.91±0.05 0.15±0.02c 0.62±0.02b 2.99±0.01c 3.72±0.02ab 5.62±0.02b f 50:50 86.90±0.07bc 13.10±0.07 9.94±0.06 0.16±0.01c 0.81±0.02a 3.16±0.08bc 3.61±0.02ab 4.99±0.07bc g 40:60 88.30±0.90cd 11.70±0.09 8.68±0.09 0.25±0.01b 0.52±0.02c 3.02±0.01bc 3.46±0.05b 4.76±0.04c h 30:70 88.30±0.40cd 11.70±0.40 8.43±0.30 0.16±0.01c 0.51±0.05c 3.27±0.05bc 3.08±0.01bc 4.61±0.03c i 20:80 89.50±0.70d 10.50±0.07 6.86±0.07 0.18±0.01c 0.51±0.02c 3.64±0.05a 2.87±0.02c 4.88±0.16cd j 10:90 88.20±0.04cd 11.80±0.04 9.06±0.04 0.29±0.01a 0.49±0.07c 2.74±0.03c 2.63±0.04cd 4.49± 0.07b k 0:100 88.70±0.10cd 11.30±0.10 8.45±0.10 0.17±0.01c 0.46±0.02c 2.85±0.07c 2.39± 0.07d 4.10±0.07bc note: values with the same superscript in the same column do not vary significantly (p ≥ 0.05). n=3±sd agriculture and food sciences research, 2018, 5(1): 23-29 28 3.3. sensory properties the sensory properties (colour, flavor, mouth feel, taste and overall acceptability) of the various blends of tigernut and coconut drink are shown in figure 1. the general trend was that of increase in likeness with increase in coconut milk substitution. this may be related to the viscosity of the drink which followed a similar trend. the flavor and overall acceptability of the 100% coconut milk (sample k) was liked very much, while the colour, taste and mouth were liked moderately. the attributes for sample a, b, c and d (the 100, 90, 80 and 70% tigernut substitution) were slightly disliked. sample g, i and j with 30, 20 and 10% tignernut substitution had values of slight and moderate likeness for all the sensory properties. the addition of 10% tigernut would be recommended based on the sensory properties. figure-1. sensory properties of different blends of tigernut-cocount milk drink note: blends (tm:cm): a(100:0), b(90:10), c(80:20), d(70:30), e(60:40), f(50:50), g(40:60), h(30:70), i(20:80), j(10:90), k(0:100) 4. conclusion the physico-chemical properties of tigernut-coconut drink gave an indication of a good quality milk drink. although the solid non-fat, protein, crude fibre and carbohydrate content decreased with increase in coconut milk substitution, the 10% substitution of tignernut had moderate likeness for all the sensory attributes. the 10% tignernut substitution based on the sensory properties would be recommended. this forms a baseline data for further studies. references [1] b. j. akinyele, m. o. fawole, and f. a. akinyosoye, "microorganisms associated with fresh cow milk, wara, and nono; two local milk products hawked by fulani women in ilorin, kwara state, nigeria," nigerian food journal, vol. 17, pp. 11-17, 1999. 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licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 9-14, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.9.14 © 2019 by the authors; licensee asian online journal publishing group influences of storage conditions, cultivation and culinary practices, on the antioxidant capacity of red bulbs of some onion varieties grown in burkina faso virginie marie dakéné1 abdoulaye sérémé2 moumouni koala3 marius k. somda4 hagrétou sawadogo lingani5 alfred s. traoré6 ( corresponding author) 1national laboratory of public health; 09 bp 24 ouagadougou 09; burkina faso 2substances department / irsat / cnrst, 03 bp 7047 ouagadougou 03; burkina faso 3department of medicine and traditional pharmacopoeia / irss / cnrst, 03 bp 7047 ouagadougou 03; burkina faso 5food technology department / irsat / cnrst, 03 bp 7047 ouagadougou 03; burkina faso 4,6department of biochemistry-microbiology; university ouaga i pr joseph ki-zerbo; 03 bp 7021 burkina faso abstract red onion bulb is a vegetable containing micronutrients with antioxidant properties. however, certain factors can have an impact on the content of these compounds. the present research evaluate the effects of parameters such as cultural practices, storage conditions and culinary practices, on the antioxidants content of red onion bulb of some varieties grown in burkina faso. the identification of the factors was carried out through field surveys, and their effects on the antioxidants content were assessed and appreciated through especially specific related bibliographic and laboratory test data. the results showed that84.78% of farmers use chemical fertilizers and all of them use chemical pesticides on onion crop. they don’t use premises or equipment suitable for storage. about dishes,7.61% of cooks, peel, cut and wash onion bulb before steam cooking, boiling water cooking or oil frying them at high temperature during 30 minutes to 3 hours. some cooks use to braise onion bulb scales as ingredients for different dishes such as barbecue. only 02,72% of cooks grind the onion bulbs with a pestle, before using the crushed as such on barbecue, or slow cooking it in different sauces. research results show that these culinary practices contribute declining significantly the final intake of antioxidant for the consumer's body. keywords: crops, cultivation, storage, culinary practices, onion. contents 1. introduction ...................................................................................................................................................................................... 10 2. material and methods ..................................................................................................................................................................... 10 3. results and discussion ................................................................................................................................................................... 11 4. conclusion ......................................................................................................................................................................................... 13 references .............................................................................................................................................................................................. 13 citation | virginie marie dakéné; abdoulaye sérémé; moumouni koala; marius k. somda; hagrétou sawadogo – lingani; alfred s. traoré (2019). influences of storage conditions, cultivation and culinary practices, on the antioxidant capacity of red bulbs of some onion varieties grown in burkina faso. agriculture and food sciences research, 6(1): 9-14. history: received: 22 october 2018 revised: 30 november 2018 accepted: 10 january 2019 published: 7 march 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this research project was supported by a grant from the west africa agricultural productivity program/national center of specialization fruits and vegetables (waapp / ncs-fv). competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.9.14&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.9.14&domain=pdf&date_stamp=2017-01-14 http://asianonlinejournals.com/index.php/aesr/article/view/1720 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2019, 6(1): 9-14 10 © 2019 by the authors; licensee asian online journal publishing group 1. introduction onion is a plant native from central asia [1]. it is now grown worldwide and is one of the basic ingredients in many dishes [2]; [3]. onion is rich in trace elements including manganese, selenium, phosphorus, zinc, vitamins (b, c, e, β-carotene), polyphenols [4]. some of these compounds have antioxidant properties [5] which gives onion therapeutic virtues. indeed, many studies attribute to onion bulb the properties to fight against certain diseases related to oxidative stress such as cancer, cardiovascular disease, diabetes, age-related diseases [2]; [6]; [7]; [8]; [9]. all of these diseases are increasingly important causes of disability and premature death in both developing countries such as burkina faso and developed countries. in 2001, they were responsible for about 60% of the 56.5 million deaths worldwide, and 46% of the global burden of disease. this morbidity rate will rise to 57% in 2020, and these oxidative stress-related diseases will account for nearly 3/4 of all deaths worldwide [10]. in developing countries, deaths from these diseases dominate mortality statistics [11]. burkina faso is not left out of this situation. the epidemiological profile of the country is marked, among other things, by the gradual increase in the prevalence of diseases related to oxidative stress [12]. although there are no national survey data, these diseases are currently a real public health problem nowadays [13]. one of the main causes of these diseases is the action of free radicals, chemical molecules naturally produced in the body during different metabolic reactions. when they are in excess in the body, they are responsible for oxidation of biological molecules such as proteins, lipids, carbohydrates and even dna [5]; [14]; [15]; [16]. antioxidants are chemical molecules that help the body fight free radicals and repair the damage they cause [17]; [18]; [19]. they play a very important role in the prevention of diseases related to oxidative stress. synthetic antioxidants exist but are very difficult to access because of their cost. in addition, in high doses, they become pro-oxidant agents potentially dangerous for health favier [5]; pelli and lyly [20]; defraigne and pincemail [21]; edeas [22]. pincemail, et al. [23] and also many studies pointed out that a high level of antioxidants in the body obtained thanks to a diet rich in fruits and vegetables, makes it possible to reduce without danger the relative risk mortality due to oxidative stress. many foods are used as a source of antioxidants, but do not cover the need of this natural substance. it is necessary to look for other more abundant and easily accessible sources of antioxidants and the way to optimize the body intake, hence the present investigation on the onion. burkina faso is one of the major producers of onions in west african (4th rank after nigeria, niger and senegal). this production is lower than domestic demand [24]. it could be used as a natural source of antioxidants and would therefore enable vulnerable populations such as children, pregnant women, old people, and aids patients to prevent diseases linked to oxidative stress by strengthening the immunity of their bodies against other opportunistic diseases. however, some factors such as cultivation techniques, environmental conditions, could have an impact on the biosynthesis of antioxidant compounds in plants [25]; [26]; [27]; [28]; [29]. the aim of this work is to evaluate the effect of storage conditions, cultivation and culinary practices, on the antioxidant content of red bulb of some onion varieties grown in burkina faso, and thus to propose the ways for optimizing antioxidant intake in the consumer’s body. . 2. material and methods 2.1. study areas data on cropping practices, storage conditions and culinary practices were collected through field surveys carried out in nine (09) regions of burkina faso, namely mouhoun, waterfalls, high-basins, north, central, westcentral, central-plateau, east-central and east. figure 1. figure-1. map of burkina faso and location (in yellow) of the study areas. source: pafasp (agro sylvo pastoral support program) [24]. agriculture and food sciences research, 2019, 6(1): 9-14 11 © 2019 by the authors; licensee asian online journal publishing group 2.2. methodology for field data collection 2.2.1. sampling the nine (09) regions selected are those where the production activity of onion bulb is very promising. among these regions are the three largest regions producing onions, namely, the regions of mouhoun, north and westcentral. sampling was random and involved 30 producers and 20 households in each locality. 2.2.2. field data collection tools field data collection was conducted from surveys by interviews of producers and consumers in the nine (09) regions. the technique adopted was the semi-structured interview where the respondent must answer exhaustively with his own terms and his own frame of reference to the questions asked to him by the investigator [30]; [31]. a survey form was designed and administered in local languages (mooré, dioula, karaboro, goulmatchéma). information about culinary practices was collected only from women as they are those who deal with culinary issues within households. the survey data were processed, analyzed and commented in relation to specific bibliographic data and laboratory test results. 3. results and discussion a total of 276 producers and 184 households were surveyed. 3.1. cultural practices and influence on the antioxidant content of onion bulb the data collected on the cultural practices applied to onion bulb among 276 people surveyed are presented in table 1. these practices concern soil fertilization and the use of chemical pesticides for the treatment of plants. table-1. cultural practices applied to onion bulb by 276 respondents. operation number of respondents practicing it percentage use of chemical fertilizers 234 84,78 % use of organic manure 42 15,22 % use of chemical pesticides 276 100 % source : field work data. data show that the majority of respondents (84.78%) use chemical fertilizers, which are generally npk (nitrogen, phosphorus, potassium) and urea in onion bulb production, and only 15.22% use organic manure for this purpose. all the producers surveyed (100%) use chemical pesticides for the treatment of onion crops. the use of chemical fertilizers and pesticides may have a negative impact on the antioxidant content of onion bulbs grown. indeed, previous work has shown that the application of chemical fertilizers including large amounts of nitrogen allow an increase in yield at the expense of the antioxidant content of plants [32]; [33]; [34]. this phenomenon called "dilution effect" has also been highlighted by farrell [35] and davis, et al. [36]. as for the effect of plant chemical pesticides studies have shown that more the plant is left on its own to deal with aggressors and diseases, more it synthesizes antioxidants to defend itself [37]; [38]; [39]. thus, the application of pesticides could prevent plants from being in a defense situation and thus synthesize antioxidants. organic production methods are characterized by a culture without chemical fertilizers or chemical pesticides. organic production of onions is difficult because of the inaccessibility of biological fertilizers and pesticides due to their high cost. while a lot of research has shown that such production methods could therefore make it possible to ensure that bulbs have better levels of phenolic compounds and a high antioxidant capacity. ren, et al. [40] have shown that the antioxidant capacity of organic onions is 20 to 50% higher than those of conventional onions. these data were confirmed by benbrook [41] who pointed out that 85% of the cases studied showed that organic farming practices can increase the antioxidant content of plants by more than 30% compared with plants derived from conventional agriculture. the same source states that, soils rich in humus with high biological activity can positively modify the concentration and the combination of antioxidants in harvested fruits and vegetables, compared to soils that are regularly fumigated and extensively chemical fertilized, and which have a generally lower bioactivity [41]. it could be argued that the ability of organic farming practices to increase the antioxidant content of fruits and vegetables would come from the use of organic fertilizers and pesticides, which are very rich in organic and probably other agro-ecological parameters could explain the increase of antioxidant content in organic farming according to many authors, dakéné, et al. [42]; sereme, et al. [43]; dabire, et al. [44]. 3.2. storage conditions and it influence on the antioxidant content of the onion bulb onion bulb storage facilities used by producers in surveyed areas are presented in table 2. data shows that only 8.33% have a sufficiently ventilated store with shelves for onion bulbs storing photo 1. some (15.22%) have a room (shed or straw hut) photo 2 for the bulbs storage, and 56.52% of the respondents use their dwelling house photo 3 for the storage. it is noticed that 19, 93% of the respondents have no means of storing for the bulbs after the harvest. table-2. storage conditions of the onion bulb among the respondents (n = 276). storage medium number of respondents owning it percentage store ventilated and provided with shelves 23 8,33% case or shed made of straw 42 15,22 % living house 156 56,52 % no way 55 19,93 % note: storage temperature: ambient temperature, uncontrolled. relative humidity (rh), oxygen and carbon dioxide content (co2): uncontrolled. source: field work data. agriculture and food sciences research, 2019, 6(1): 9-14 12 © 2019 by the authors; licensee asian online journal publishing group photo-1. store ventilated and. equipped shelves. photo-2. straw box. photo-3. living house. (field work photos). in addition, the storage is done in the ambient air and the shelves are often overloaded. the temperature, relative humidity, oxygen content and carbon dioxide (co2) of the storage room are not controlled. these storage conditions, if they make it possible to minimize losses due to rotting of the bulbs, could have a significant impact on the nutritional quality of the product, particularly its antioxidant content. indeed, agblor and waterer [45] have shown that to maintain the nutritional quality of onion bulbs during storage, the optimal temperature should be 0 °c and the relative humidity between 65 and 70%. the storage could also be done at room temperature as is the case for the producers surveyed, but provided that it is between 20 and 30 °c and that the bulbs are properly dried [46]. the oxygen in the air should be as low as possible to limit bulb respiration as renard and chervin [47] have shown. in this study, among the producers surveyed, no storage room is equipped with a device for monitoring and adjusting these parameters in order to ensure that they are suitable for the proper storage and preservation of the nutritional quality of the bulb. it is therefore possible that such an environment is unfavorable to maintaining the antioxidant content of stored bulb. 3.3. culinary practices and influence on the antioxidant content of onion bulb in order to identify the culinary practices and their influence on the antioxidant concentration of the onion bulb, a household survey was carried out especially among women those are in charge of the family's cooking issues. one hundred and eighty-four (184) households were surveyed for this purpose. the information sought was related to onion bulb culinary practices as an ingredient of different dishes. 3.3.1. modes of preparation of the onion before consumption or cooking data on the methods of processing of onion bulb before consumption are presented in table 3. table-3. modes of preparation of the onion bulb before consumption or cooking in the households surveyed (n = 184). operation number of households surveyed practicing it percentage peeling and cutting 23 12,50 % peeling, cutting, washing 106 57,61 % peeling, washing, cutting 55 29,89 % source : field work data. from these data, it appears that 57.61% of respondents peel, cut the onion scales into small pieces and wash them before eating without any processing, or after cooking. 29.89% peel, but wash before cutting the onion scales, and 12.50% only peel before cutting them. peeling, cutting and washing are a set of practices that could contribute to a decrease in the antioxidant content of bulbs. indeed, previous work has shown that antioxidant micronutrients are generally more concentrated in the epidermal parts of fruits and vegetables [41]; [48]; [49]. peeling, although essential in some cases, such as the red bulb of onion, could therefore contribute to reducing these compounds in ready-to-eat products. the losses during the cutting would be explained by the fact that during this operation, there is an alteration of the cellular integrity. this would then bring into contact the soluble compounds generally accumulated within the vacuoles, and the degradation enzymes initially present in the cell walls and the cytoplasm [50]. washing, meanwhile, would lead to the loss of water-soluble compounds by diffusion in water, as shown by arroqui, et al. [51]. however, the losses could be at different levels: 57.61% of the households surveyed practice washing after cutting, which could cause more losses of water-soluble compounds because the diffusion of these is facilitated and therefore higher. 29.89% of the households surveyed practice washing before cutting, which would cause fewer losses. thus, a best practice will be to perform: (i) peeling consisting simply of the removal of dry outer scales; (ii) washing of peeled bulbs before cutting; (iii) cutting while avoiding the reduction of onion bulb into too small pieces. by taking into account these improvements, the loss of anti-oxidants during cooking operations could be significantly reduced. 3.3.2. cooking methods used and cooking time of the bulbous onion in the households surveyed the different cooking methods of the onion bulb used by the respondents are presented in table 4. agriculture and food sciences research, 2019, 6(1): 9-14 13 © 2019 by the authors; licensee asian online journal publishing group table-4. modes, time and cooking temperature of the onion bulb among the respondents (n =184). onion bulb cooking method among respondents number of households surveyed practicing it percentage cooking with oil and then with water 184 100 % cooking with water 117 63,59 % oil cooking 49 26,63 % steam cooking 20 10,87 % braising 13 07,07 % muffled cooking 5 02,72 % note: duration of cooking: between 30 minutes to 3 hours. cooking temperature: uncontrolled. source: field work data. the data show that frying followed by cooking with water is the one most practiced by the households surveyed (100% of the respondents).for 63.59% cooking is done with water and 26.63% of the households surveyed are oil frying. steaming and braising are used respectively by 10.87% and 07.07% of respondents. as for the braising, it is used only by 02.72% of the households surveyed. it is noted that heat processing is the common factor in all these practices. heat causes loss of antioxidant compounds such as carotenoids and vitamin c. this loss is explained by the sensitivity of these compounds to heat and oxidation as shown [47]. cooking time is also a factor that could contribute to the decline the antioxidant content of onion bulbs. indeed, renard and chervin [47] have shown that the importance of losses during cooking depends on the time-temperature pair: the higher the cooking temperature and the longer the duration, the more the destruction of the heat-sensitive compounds is important. this destruction can reach 95% of the content with regard to vitamin c. in the case of the consumers surveyed, the possible losses of antioxidant compounds could be explained by maladjustment of the time-temperature pair: the cooking times (30 min to 3h) are sufficiently long to cause losses. indeed, eduvigis, et al. [52] showed that for a heat processing up to 100 o c, the cooking time should be 11-17 minutes to maintain a good antioxidant capacity. the time-temperature pair must therefore be taken into consideration when cooking the onion bulb. for a high cooking temperature, the time should be short and vice versa. 4. conclusion this study shows that: (i) certain cultural practices, namely the use of chemical fertilizers and chemical pesticides, (ii) room storage conditions such as temperature, relative humidity, oxygen and carbon dioxide content, (iii) culinary practices such as peeling, cutting, washing, cooking time and cooking temperature, are a combination of factors that could contribute to a decrease in the antioxidant content of onion bulbs. therefore, the use of an organic farming, the conservation of the onion bulb under ideal conditions of storage (temperature of 0 °c, relative humidity between 65 and 70%, limitation of the respiration of the bulb by reducing the degree of oxygenation in the storage room and increasing the carbon dioxide content); the use of appropriate cooking process (peeling only external dry scales, washing peeled bulb before cutting, cutting onion bulb into big pieces, cooking time from 11 to 17 minutes for a temperature under 100oc, cooking with water retention or steaming), could on the one hand maintained the antioxidant content of the bulb, and on the other hand, minimize losses in antioxidant compounds during the processing. references [1] m. pitrat and c. foury, stories of vegetables: origins at the dawn of the 21st century vol. 410. paris, france: inra, 2003. 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[52] r. eduvigis, c. sanchez-moreno, d. a. m. begona, and c. pilar, "characterisati on ofonion (allium cepa l.) by-products as food ingredients with antioxidant and antibro wningproperties," food chemistry, vol. 108, pp. 907–916, 2007. available at: https://doi.org/10.1016/j.foodchem.2007.11.058. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research vol. 4, no. 1, 1-7, 2017 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2017.41.1.7 1 comparative studies on proximate and some mineral composition of selected local rice varieties and imported rice brands in ghana faustina dufie wireko-manu1  cleopatra amamoo2 ( corresponding author) 1department of food science and technology, kwame nkrumah university of science and technology, kumasi, ghana 2bayport savings and loans plc, 71 osu badu street, airport west, dzorwulu, accra, ghana abstract purpose: the objective of this study was to determine and compare the proximate and mineral (fe, zn, mn, mg, se) composition of locally produced polished and unpolished rice and two imported rice brands. design/methodology/approach: the proximate and mineral (fe, zn, mn, mg, se) composition of five local rice varieties (maawuwoe coop brown rice, maawuwoe coop white rice, shigafa brown rice, shigafa white rice and long grain parboiled rice) and two imported brands of rice (imported 1 and 2) were evaluated using standard methods after which they were compared to establish which had the higher percentage of nutrients. findings: the percentage protein of the imported rice brands (7.0-8.4%) were significantly higher (p<0.05) than those of the local rice varieties (5.69–6.89%). the local varieties had comparable carbohydrate (74.20-79.41%) but significantly higher fat (1.1-2.57%) and fibre (0.64-1.95%) contents than the imported brands (76.94-78.21% carbohydrate, 0.49-0.88% fat and 0.5% fibre). maawuwoe coop brown rice had significantly higher fe and zn contents of 96 mg/100g and 2.9 mg/100g respectively and shigafa brown rice recorded the highest mn content of 1.81 mg/100g. imported brand 1 however had the highest mg content (96 mg/100g). for the local varieties, the brown had higher fibre contents (1.95-1.65%) than that observed in the white. originality/value: this study is meant to provide knowledge on the nutritional quality of local rice to inform consumer and subsequently enhanced patronage in ghana. keywords: local brown rice, local white rice, long grain, nutrients, quality. citation | faustina dufie wireko-manu; cleopatra amamoo (2017). comparative studies on proximate and some mineral composition of selected local rice varieties and imported rice brands in ghana. agriculture and food sciences research, 4(1): 1-7. history: received: 2 august 2016 revised: 16 march 2017 accepted: 13 april 2017 published: 5 may 2017 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ......................................................................................................................................................................................... 2 2. materials and methods ...................................................................................................................................................................... 2 3. results and discussion ...................................................................................................................................................................... 4 4. conclusion ............................................................................................................................................................................................ 6 references ................................................................................................................................................................................................. 6 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2017.41.1.7 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=faustina%20dufie%20wireko-manu https://orcid.org/orcid-search/quick-search?searchquery=cleopatra%20amamoo agriculture and food sciences research, 2017, 4(1): 1-7 2 1. introduction rice is currently the second most important food staple after maize in ghana and its consumption keeps increasing as a result of population growth, urbanization and change in consumer habits [1]. from 2000 to 2003 the estimated consumption levels recorded commercial rice imports of approximately 61%, food aid accounted for 2%, and domestic rice production accounted for approximately 37% [2]. despite the increase in production, the level of consumer demand far exceeds the supply. this has resulted in the continuous increase in the importation of the rice into the economy which is registered the highest demand deficit of 215,000 metric tonnes among selected crops in ghana [3]. in the same year, computations by the ghana statistical service indicated a custom value of imported rice of gh¢ 108,523,136 compared to a significantly low amount of gh¢ 1,186 of exported rice in ghana. rice is an important food for urban dwellers and is taking over from traditional staples, mainly root crop. rice costs are rising, according to the mofa, and imported rice is particularly expensive. in january 2012 an 84-kg bag of locally produced paddy rice cost ghs66.20 ($34.10), but by december it had jumped by 24% to reach ghs81.86 ($44.60). this is still low compared to import prices, which rose from ghs97.60 ($50.10) for a 50-kg bag in january 2012, to ghs117.85 ($60.60) in december 2012, a 21% increase. comparing the costs, local rice prices sat at ghs0.97 ($0.49) per kg in december 2012, compared to ghs2.36 ($1.20) per kg for imported rice. according to the ministry, rice imports cost the country an estimated $450m annually [4]. many urban consumers are said to prefer imported rice to local rice even though it is more expensive because it looks cleaner, more polished and more prestigious to eat and to offer to visitors. usually quality and price of the commodity is of utmost importance when purchased. the difference in consumer preference has been attributed to several factors including poor post harvest handling, non-availability, and a general perception of poor quality physical characteristics, presence of foreign matter, nutritional quality and cooking behaviours [5]. consumers who lived in urban area and had high standard of living, high income and education, tend to purchase rice of high quality based on their nutritional content [6]. consumers in ghana prefer to purchase imported rice brand rather than the local ones [7]. however local rice is said to taste better. the need to make local rice more competitive is recognized; particularly in light of nutritional composition. different studies have shown that the local rice varieties (emo kokoo, mr. more, mui, abodwese, long grain, beta, tema, amavi, emo fitaa, abibifoo mo) exhibit nutritional superiority especially in minerals such as potassium, calcium and protein contents [5]. even though ghanaians are aware of the locally produced rice in ghana, most consumers are more familiar with imported rice [8]. lack of knowledge on the nutritional quality of local rice, and the misconception that imported foods and drinks are of better quality among others might have contributed to low patronage of local rice. the situation has created low profitability. the local rice farmers are not earning enough resulting in high loss rate and unemployment in the sector [5]. the objective of this study was to determine and compare the proximate and mineral (fe, zn, mn, mg, se) composition of locally produced white and brown rice and two imported rice brands. 2. materials and methods 2.1. material source five local rice varieties namely maawuwoe coop brown rice (mcbr), maawuwoe coop white rice (mcwr), shigafa brown rice (sbr), shigafa white rice (swr) and long grain parboiled rice (lgpr) as well as two imported brands of rice (ib 1 and ib 2) were used for the study. the local varieties were purchased from central market in kumasi with the exception of maawuwoe coop brown and white rice which were obtained from the csir-crop research institute, fumesua, kumasi, ghana. the two imported rice brands were obtained from a retail outlet in kumasi based on their high consumer patronage, as revealed by a preliminary consumer survey of local and imported aromatic rice varieties sold on the market [5]. table 1 shows the list and the sources of the selected rice varieties. table-1. list of selected rice varieties and their country of origin grouping name of rice origin of rice imported rice imported brand 1 thailand imported brand 2 thailand local varieties maawuwoe coop brown rice ghana maawuwoe coop white rice ghana shigafa brown rice ghana shigafa white rice ghana long grain parboiled rice ghana source: these are rice samples collected by authors of this study from market and csir-crops research institute in ghana 2.2. sample preparation the rice samples were ground with a blender into fine powder and stored in labelled polythene bags under room temperature until required for analysis. 2.3. chemical analysis 2.3.1. proximate analysis the methods for analyses were the standard procedures of aoac [9]. all proximate components were analyzed in duplicate and reported as mean on % dry weight basis. agriculture and food sciences research, 2017, 4(1): 1-7 3 2.3.1.1. determination of % moisture content two grams of each sample was weighed in a crucible and dried to constant weight at 105 °c for 6 hours in a drying oven. crucible plus sample was cooled in a desiccator and re-weighed. the moisture content was calculated as percentage moisture lost. 2.3.1.2. determination of % crude lipid and crude fibre content two grams of the rice samples were weighed in a porous thimble of a soxhlet apparatus. the thimble was placed in an extraction chamber, which was suspended above a pre weighed receiving flask containing petroleum ether. the flask was heated on a heating mantle for 8 hr to extract the crude lipid. after the extraction, the thimble was removed from the soxhlet apparatus and the solvent distilled off. the flask containing the crude lipid was heated in the oven at 100 ºc for 30 min to evaporate the solvent, then cooled in a desiccator, and reweighed. the difference in weight was expressed as percentage crude lipid content. crude fibre was estimated by acid-base digestion with 1.25% h2so4 (prepared by diluting 7.2 ml of 94% conc. acid of specific gravity 1.835 g/ml per 1000 ml distilled water) and 1.25% naoh (12.5 g per 1000 ml distilled water) solutions. the residue after crude lipid extraction was put into a 600 ml beaker and 200 ml of boiling 1.25% sulphuric acid added. the contents were boiled for 30 minutes, cooled, filtered through a filter paper and the residue was washed with boiling water until washings were no longer acidic. the washed residue was returned to the digestion flask and further digested by boiling in 200 ml of 1.25% sodium hydroxide for 30 minutes. the digest was filtered through a porous crucible to obtain the residue. this was washed with boiling water and finally with 15 ml of 95% ethanol. the washed residue was dried in an oven at 110 ºc to constant weight and cooled in a dessicator. the residue was scraped into a pre–weighed porcelain crucible, weighed, ashed at 550 ºc for 30 minutes, cooled in a dessicator and reweighed. crude fibre content was expressed as percentage loss in weight on ignition. 2.3.1.3. determination of % nitrogen content and estimation of crude protein 2.3.1.3.1. digestion two grams of each air dried sample was weighed into 500 ml long-necked kjeldahl flask and 10 ml distilled water was added to moisten the sample. one spatula full of kjeldahl catalyst (mixture of 1 part selenium + 10 parts cuso4 + 100 parts na2so4) was added followed by 20 ml conc. h2so4. the solution was digested until clear and colourless. the flask was allowed to cool; the fluid decanted into a 100 ml volumetric flask and made up to the mark with distilled water. 2.3.1.3.2. distillation an aliquot of 10 ml fluid from the digested sample by means of a pipette was transferred into kjeldahl distillation flask. ninety (90) ml of distilled water was added to make it up to 100 ml in the distillation flask. twenty (20) ml of 40 % sodium hydroxide was added to the content of the distillation flask. distillate was collected over 10 ml of 4 % boric acid and 3 drops of methyl red in a 200 ml conical flask. the presence of nitrogen gave a light blue colour. 2.3.1.3.3. titration the collected distillate (about 100 ml) was titrated with 0.1 n hcl till the blue colour changed to grey and then suddenly flashed to pink. a blank determination was carried out without a sample. crude protein was estimated by multiplying the value obtained for percentage nitrogen content by a factor of 6.25. 2.3.1.4. % ash determination ash crucibles were preheated in an oven, cooled in a desiccator and weighed. two grams of each sample was weighed into the crucibles and then placed in a muffle furnace at 600 °c for 4 hours. crucibles plus ashed samples were cooled in a desiccator and weighed. 2.3.1.5. % carbohydrate determination carbohydrate was determined as the difference between 100 and the sum of the percentages of water, protein, total lipid (fat), and ash. 2.3.2. mineral analysis samples were ashed in a porcelain crucible at 550 ºc in a muffle furnace for four hours, removed from the furnace and cooled. the ash was moistened with 2 ml of distilled water and slowly and carefully the ash was dissolved in 5 ml of 8 n hcl. the crucible was covered and placed on steam water bath for 20 minutes, cooled and transferred into 100 ml volumetric flask by filtering using whatman no. 42 filter paper catching the filtrate in the 100 ml volumetric flask and made up to standard volume. the crucible was washed well with water, the washing going through the filter. the filter paper was washed several times with distilled water and made up to the 100 ml mark and shaken vigorously to assure complete mixing. selenium, iron, zinc, magnesium, and manganese were analysed by icp-aas (induced coupled plasma – atomic absorption spectrometry). all determinations were carried out in duplicate and reported as mean mineral content in mg/100g dry matter (dm). concentrations for each element were determined from previously calibrated standard curves. 2.4. statistical analysis data were reported as mean ± standard deviation for at least duplicate analyses of the same extract. anova was used to analyse the means ofz samples and the least significant difference was used to further determine exactly where the differences occurred, at 5% significance level. agriculture and food sciences research, 2017, 4(1): 1-7 4 3. results and discussion 3.1. proximate composition results of proximate composition for the local and imported rice types are shown in table 2. moisture content (11.46-15.17%) of the varieties (both local and imported) were significantly different (p<0.05). these values closely correspond to the results of 11.65% to 13.43% reported by eshun [10] and 12% reported by abbey, et al. [11]. maawuwoe coop brown rice (mcbr) had higher moisture content of 15.17% which may be attributed to its drying temperature used [12]. the relatively higher moisture content of mcbr may have effect on its storage. long grain parboiled rice (lgpr) and shigafa white rice (swr) had the least percentage moisture of 12.07% and 11.46% respectively which may be due to the parboiling process [12] and may have the potential to store relatively longer. the significant difference observed in the moisture content of the rice varieties may be attributed to temperature and duration of drying as well as difference in storage conditions at source. the crude fat values obtained ranged between 0.49 2.57% with the imported brands having lower values compared to local ones even though the values are well in line with the range of 0.4 to 0.8% reported by laureys [13]. the variety mcwr had the highest fat content of 2.57% with import brand 1 (ib 1) recording the lowest fat content of 0.49% respectively (table 2). statistical analysis revealed a significant difference (p < 0.05) in the crude fat values for the rice varieties. rice bran, being an excellent fat source, which is lost during milling contributes to the low fat content in the well-polished imported rice brands. thus the observed difference may be attributed to the degree of milling. milling and polishing of rice removes the outer layer of the grain where most of the fats are concentrated [14]. the values obtained for percentage ash content in this study ranged between 0.5 1.5%. maawuwoe coop white rice had the highest ash content of 1.50% followed by lgpr which had 1% while the other rice varieties had the least ash contents of 0.5%. values obtained in this study corresponds closely with the values obtained by ebuehi and oyewole [15] who reported ash values of 0.53% and 0.80% for raw ‘aroso’ and ‘ofada’ rice varieties respectively in nigeria. kwarteng, et al. [6] also reported 0.48% to 0.6%. the ash content of a food sample gives an idea of the mineral elements present in the food sample [16]. minerals are more concentrated in the bran and thus get lost during milling and polishing when the bran is removed from the grain [17]. the ash content of 0.5% observed in the other varieties may be due to the degree of milling/polishing which is influenced by variety. as stated earlier, rice bran contains much more minerals than the actual endosperm and the tendency of rice bran to stick to the grains during milling and polishing (which is a varietal trait) influences ash content [6]. table-2. proximate composition of some local and imported rice types in ghana type sample moisture (%) ash (%) fat (%) protein (%) fiber (%) carbohydrate (%) local mcbr 15.17±0.26bfhj 0.5±0.00 1.85±0.20bdfg 5.69±0.00b 1.65±0.21bd 75.14±0.06a mcwr 14.19±0.05bfgj 1.5±0.04 2.57±0.18bdfh 6.79±1.55b 0.75±0.07b 74.2±2.70adfh lgpr 12.07±0.26adehi 1.0±0.007 1.1±0.17ae 6.89±0.16b 0.64±0.05a 78.30±0.58ag sbr 13.83±0.05bcfg 0.5±0.00 1.35±0.17bde 6.45±0.16b 1.95±0.07bd 75.92±0.04ae swr 11.46±0.028ad 0.5±0.00 1.46±0.00bde 6.45±0.16b 0.72±0.07bc 79.41±0.26bc imported brand ib 1 12.78±0.65a 0.5±0.00 0.88±0.18a 8.4±0.44a 0.50±0.00a 76.94±0.38a ib 2 13.30±0.18ac 0.5±0.00 0.49±0.002bc 7.0±0.00a 0.50±0.00a 78.21±0.18ac note: values are means ± standard deviations of duplicate determinations. values in the same column having the same superscript letters are not significantly different at 5% significant level. key: sample sample code maawuwoe coop brown rice mcbr maawuwoe coop white rice mcwr long grain parboiled rice lgpr shigafa brown rice sbr sbr shigafa white rice swr swr imported brand 1 ib 1 imported brand 2 ib 2 carbohydrate values in the varieties ranged from 74.20% for mcwr to 79.41% in swr. virtually all the rice varieties have appreciably high carbohydrate content (table 2). the variation in values for carbohydrate among the varieties was statistically significant (p<0.05). these values also correspond closely to that reported by abbey, et al. [11] and laureys [13]. ebuehi and oyewole [15] had 78.3% and 81.1% for raw ‘ofada’ and ‘aroso’ rice varieties respectively. the observed high carbohydrate content of both local and imported rice varieties affirm that rice is a carbohydrate food source. generally, the local varieties had significantly higher crude fibre than the imported brands. the range of values for crude fibre observed in this study was between 0.5 1.95% for the two imported rice types and shigafa brown rice respectively. these values agree with crude fibre values of other varieties which range from 0.59% for idsa 85 to 0.89% for jasmine 85 and 0.86% for sikamo and 0.87% for nerica-2 [10]. statistical analysis revealed a significant difference (p < 0.05) in the crude fibre values for the rice varieties. maawuwoe coop brown rice does not have any of its parts, including the germ, the bran, and the endosperm, removed (has not been refined) hence its high fibre content of 1.95%. the high crude fibre content in shigafa brown rice may also be due to the fact that is a whole grain which contains all three components: bran, germ, and endosperm [18]. milling of rice during the production of polished rice generally decreases the fibre contents, hence the relatively low fibre in the imported brands [19]. the bran is particularly rich in dietary fibre and contains significant quantities of starch [20] which also contributes to the carbohydrate content of brown rice. the observed difference may be due to the fact that the imported rice brands are more polished than the local varieties. percentage carbohydrate could also be influenced by other environmental factors under which rice is grown (soil type, crop management practices, rainfall, solar radiation, and growth temperature) [19]. http://en.wikipedia.org/wiki/dietary_fiber http://en.wikipedia.org/wiki/starch agriculture and food sciences research, 2017, 4(1): 1-7 5 the protein contents of the rice varieties ranged from 5.64% 8.40% (table 2). the findings of this study showed that the imported rice brands had higher protein contents of 7% and 8.4% respectively. these values compare closely to those reported by abbey, et al. [11] and laureys [13] as 7.0% for polished rice. maawuwoe coop brown rice had the least protein value of 5.64% but was even higher than what was reported earlier by kwarteng, et al. [6] a range of 5.10-5.30%. the variations observed may be due to varietal and environmental influences. [21, 22] reported that deposition of protein in rice grains depends on plethora of interrelated metabolic pathways involved in uptake of n, fe and zn from soil, their transport to source tissues such as culms and leaves and mobilization and/or remobilization to developing grains. each of these processes is governed by several genes and influenced by environmental factors such as soil type, drought, fertilizers application, genotype, environment interaction [23, 24]. the significant difference in percentage of crude protein between the imported varieties and the local varieties may be as a result of genotype and environmental differences [25]. most of the protein is found in the endosperm layer with additional protein found in the bran layer which constitutes about 7-9% of the whole grain. these contributes to the percentage of crude protein in the local varieties [26]. 3.2. mineral composition the results of the mineral (iron, zinc, magnesium, manganese and selenium) analysis of the local and imported rice types are shown in table 3. the values of iron obtained agree to those reported by ebuehi and oyewole [15] and diako, et al. [5]. the sbr recorded the highest iron content of 96 mg/100 g and ib 1 had the lowest iron content of 11.1 mg/100 g. significant differences (p < 0.05) were observed in iron contents among the local rice varieties and between the local and imported varieties. this may be due to the fact that the brown variety is a whole grain which has its bran particularly rich in dietary minerals such as iron. the difference in iron content among the rice varieties may also be influenced by nitrogen application and soil quality [27]. the magnesium values obtained in the rice varieties ranged from 66.7 mg/100 g in sbr to 96 mg/100 g in ib 2 with a significant difference (p < 0.05). the values of magnesium obtained for the rice varieties correspond with the 67.0 mg/100 g and 82.0 mg/100 g respectively for raw ΄ofada΄ and raw ΄aroso΄ rice varieties reported by ebuehi and oyewole [15]. the local intra varietal differences could be due to genetic factors or the mineral content of the soil on which they were grown [17]. in general, the more rice bran is removed from the grain during polishing, the more vitamins and minerals are lost [19]. the manganese values obtained from the study ranged from 1.81 mg/100 g in sbr to 1.09 mg/100 g in ib 1. the values of manganese obtained for the rice varieties are similar to those reported by tinsley and emeritus [28]. imported brand 1 (ib 1) recorded the lowest manganese content which may be due to the fact that minerals are highest in the bran which is lost during processing [27]. the difference in the manganese contents among the rice varieties might be as a result of fertilizer application, rate of parboiling and the amounts of soil nutrients all of which affect the mineral contents of rice. the zinc values obtained in table 3 ranged from 1.05 mg/100 g in lgpr to 2.9 mg/100 g in mcbr. these values agreed with earlier findings by trinidad, et al. [29] who found a range of 1.40 to 1.79 mg/100 g and 1.97 mg/100g reported by tinsley and emeritus [28]. the bran and the germ in brown rice contain many vitamins and minerals contributing to mcbr having the highest zinc content. the zinc content in the parboiled rice may be due to prior cooking which preserves some of the nutrient content, as micronutrients are transferred from the aleurone and germ into the starchy endosperm [30]. selenium contents were <0.1 mg/100 g for all the rice varieties which did not agree with the 0.15 mg/100 g reported by tinsley and emeritus [28] soils are frequently low in available se [31, 32]. se availability in soils depends upon soil ph, redox potential, calcium carbonate level, cation exchange capacity, and organic carbon, iron (fe) and aluminium (al) levels [33]. the low selenium contents in both the local and imported varieties may be due to the effect of non-application of selenium enriched fertilizer on the soil where the rice varieties were cultivated. selenium content and assimilation of rice could be promoted by selenium foliar application on rice. table-3. mean mineral composition of some local and imported rice types in ghana type minerals (mg/100g) sample iron zinc magnesium manganese selenium local mcbr 96±0.03bdfg 2.9±0.14bdfh 62.4±1.40 bdfhjk 1.16±0.01 bdfhjk <0.10 mcwr 48±0.01bdfh 1.94±0.01a 76.0±0.00 bdfhjl 1.59±0.01 bdfhjl <0.10 lgpr 73±0.01bdf 1.05±0.007bdfg 72.0±1.40 bdfhi 1.67±0.01 bdfhi <0.10 sbr 20.7±0.01bde 1.72±0.03bde 66.72±0.00 bdfg 1.81±0.03 bdfg <0.10 swr 11.47±0.42be 1.3±0.00bc 48.0±1.40 bde 1.29±0.00 bde <0.10 imported brand ib 1 89.0±0.00a 1.91±0.00a 96.0±1.40 a 1.09±0.00 a <0.10 ib 2 11.1±0.04bc 1.92±0.03a 79.0±2.80 bc 1.47±0.03 bc <0.10 note: values are means ± standard deviations of duplicate determinations. means with the same alphabet as superscript within columns are not significantly different at 5% significant level in general, the levels of minerals from some of the varieties like shigafa brown rice and maawuwoe white rice varieties are high in comparison to the other local varieties and imported brands. this may be due to genetic factors or the mineral content of the soil as well as polishing and processing [17]. the rate of fertilizer application and the native fertility of paddy fields have been shown to affect the mineral levels of rice [34]. an analysis of nutrient content of rice suggests that the nutritional component of rice is one of the most important indicators of quality; rice is predominantly a starchy food though it also contributes useful quantities of proteins and vitamins, mineral and fiber. the chemical composition (nutrients) of rice grain varies considerably depending on factors like plant variety (breeds), environmental condition (i.e. location and season in which grown), fertilizer treatment, degree of milling, http://en.wikipedia.org/wiki/dietary_mineral agriculture and food sciences research, 2017, 4(1): 1-7 6 and condition of storage [35]. as with all natural foods, the precise nutritional composition of rice varies depends on the variety, soil conditions, environmental conditions and types of fertilizers. 4. conclusion the local rice compared well with imported rice in terms of carbohydrate (74.20-79.41%) but had significantly higher fat (1.1-2.57%) and fibre (0.64-1.95%) while the imported rice had higher protein content (7.0-8.4%). the local rice maawuwoe coop brown rice had significantly high fe and zn contents, 96 mg/100g and 2.9 mg/100g respectively; while shigafa brown rice recorded the highest for mn, 1.81 mg/100g. the imported brand 1, however had the highest mg content (96 mg/100g). the results show that local rice is a nutritious staple crop, containing most of the essential nutrients. efforts to improve local rice consumption in ghana should introduce other desirable attributes such as taste into breeding programmes to make local rice attractive to consumers. education and advertisement of nutritional characteristics of local rice are important steps to improve the patronage of local rice in ghana. references [1] mofa, "agriculture in ghana: facts and figures, statistics," research and information directorate (srid), vol. 1, 2009. 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[35] p. roy, t. ijiri, h. okadome, d. nei, t. orikasa, n. nakamura, and t. shiina, "effect of processing conditions on overall energy consumption and quality of rice (oryza sativa l.)," journal of food engineering, vol. 89, pp. 43–348, 2008. view at google scholar | view at publisher asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://scholar.google.com/scholar?hl=en&q=effect%20of%20processing%20conditions%20on%20overall%20energy%20consumption%20and%20quality%20of%20rice%20(oryza%20sativa%20l.) http://dx.doi.org/10.1016/j.jfoodeng.2008.05.015 http://dx.doi.org/10.1016/j.jfoodeng.2008.05.015 agriculture and food sciences research issn(e) : 2411-6653 issn(p) : 2518-0193 vol. 3, no. 2, 75-79, 2016 http://www.asianonlinejournals.com/index.php/aesr 75 fruit growth characteristics of rose hips (rosa sp) ümit dölek1  mehmet güneş2 ( corresponding author) 1gökhöyük vocational and technical anatolian high school,05100, amasya, turkey 2department of horticulture, agricultural faculty, gaziosmanpaşa university, 60240-tokat, turkey abstract this study was carried out to determine the fruit growth characteristics of two rose hip (rosa) species selected from wild population in tokat region of the northern anatolia, turkey. shrubs were propagated by cuttings and planted in 2000 at the research station of the horticultural department of agricultural faculty of gaziosmanpaşa university, tokat, turkey. fruit weight, fruit diameter, fruit length, fruit flesh percentage and seed weight of genotypes belonging rosa dumalis and rosa jundzillii were investigated in the study. fruit samples were picked up weekly from full flowering to optimal harvest time for measurement and weighing. as a result, fruit development characteristics of species measured exhibited single sigmoidal curve except seed weights. in general, developing of measured properties were fast until last 3-4 weeks, then became stable or slightly decreased. results obtained in 2011 were higher than the results obtained in 2010 in both genotypes. keywords: rosa, fruit growth, sigmoid, curve, fruit diameter, fruit flesh contents 1. introduction ............................................................................................................................................................................... 76 2. material and methods ............................................................................................................................................................... 76 3. results and discussion .............................................................................................................................................................. 76 4. conclusion .................................................................................................................................................................................. 79 references ...................................................................................................................................................................................... 79 citation | ümit dölek; mehmet güneş (2017). fruit growth characteristics of rose hips (rosa sp). agriculture and food sciences research, 3(2): 75-79. doi: 10.20448/journal.512/2016.3.2/512.2.75.79 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: contribution/acknowledgement: this work is licensed under a creative commons attribution 3.0 license all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 2 november 2016/ revised: 25 november 2016/ accepted: 3 december 2016/ published: 7 december 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.2/512.2.75.79 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=%c3%9cmit%20d%c3%b6lek https://orcid.org/orcid-search/quick-search?searchquery=mehmet%20g%c3%bcne%c5%9f http://search.crossref.org/?q=10.20448/journal.512/2016.3.2/512.2.75.79 agriculture and food sciences research, 2016, 3(2): 75-79 76 1. introduction growth, most commonly, means an irreversible increase in size [1]. growth is occurred by dividing and increasing of cells. therefore, increasing and dividing of cells are two important processes for fruit development during growing period. fruit development begins from full blooming and continues to harvest time. the developing young fruit uses nutrient from plant at the first stage of growth period. then, fruit partly produces own energy requirement by photosynthesis. but this production is not enough for optimal fruit development. that’s why, young fruit continues to feed from plant at the all stages of development and maturation processes [2]. fruit growth is influenced by some genetic factors such as cultivar/variety, rootstock, age, biennial bearing, the position of the flower on spur, cultural factors such as pruning, defending against pests, thinning, irrigation and nutrition, exogenous hormone applications and environmental factors such as temperature, light, soil, moisture, wind and gases [3]. the length of developing period may be short or long depending on fruit species. the length of this period varies from ten to twenty-week for many deciduous fruit species. for example, this period is sixty-week for valencia orange cv. while it is only three-week for strawberries. during the developing period fruit volume increases several times when compared with its initial weight and volume [4, 5]. some parameters, such as fruit fresh/wet weight, fruit volume increasing, fruit diameter etc. are used for determination of fruit development. selected properties are measured from full blooming to harvest time for a certain time intervals. developing rate varies during fruit growing period depending on some factors. fruit development is expressed by growth curves. growth curve shapes can change related to species/cultivars, ecological conditions, and cultural practices. fruit species are divided two main groups related to growth curve shapes as single and double sigmoid. apple, pear, orange, banana, strawberry, avocado, mango, etc. show single sigmoidal curve. peach, apricot, cherries, olive, currant, grape, fig, raspberry, blackberry etc. show double sigmoidal curve. some researches [3, 6, 7] were conducted on development of mentioned fruit species but there is no research on rose hip fruit development. understanding of fruit development helps us to use of fertilizing management, pruning, growth regulators, fruit thinning and shedding fruit weight, fruit diameter, fruit length and fruit flesh ratio, and size prediction [5, 8]. it is desirable that the growth progress of plant know under orchard conditions in real time for many practical reasons. while on the one hand growers may better fine-tune management practices, on the other hand, the ability to forecast average fruit weight at harvest is of great value for markets [9]. in light of this information, the aim of this study is to determine the fruit growth by measurement of some pomological characteristics of two rose hip species. as far as we know that this is the first study conducted on fruit growth characteristics of rose hips. 2. material and methods this study was carried out on two selected rose hip genotypes belonging to rosa dumalis (accession number: ar-11) and rosa jundzillii (accession number: yl-04), grown in the research station of the horticultural department of agricultural faculty of gaziosmanpaşa university, tokat, turkey, during two years (2010 and 2011). accessions were selected from wild rose hip population in the tokat region of the northern anatolia [10]. shrubs were propagated by hardwood cuttings and planted on their own roots at a spacing of 3 x 3 m in the research and treatment station in 2000. the soil texture of area is silty clay, and ph is slightly alkaline. the rose hip bushes were pruned in january or february (dormant period) of each year with removal of dry, old or low shoots. the plants were fertilized with mineral nutrients and manure together with drip irrigation at regular intervals during the vegetation period. mean fruit weight (g), fruit diameter and length (mm), fruit flesh percentage and seed weight (g) of studied species were measured. fruits were randomly sampled from each shrub. the fruit and seed weight and flesh ratio, and diameter and length values of the fruits were examined in order to form growth curves. physical characters were measured weekly in 2010 and 2011 from full bloom to optimal harvest time. 30 fruits with three replicates per species were measured or weighed for determination of growth curves of each studied physical character. 3. results and discussion growth curves of the species and the years associated with studied characteristics were presented in figure 1a-d, 2a-d, 3a-d, 4a-d and 5a-d. a linear fast growth was recorded in fruit weights of species in both years from full bloom to last 3-4 weeks. growth during last 3-4 weeks was stable or decreased slightly. fruit weights of both species were higher in 2011 than 2010 (figure 1a,b,c,d). figure-1a. fruit weight developing of r.dumalis in 2010-2011 figure-1b. fruit weight developing of r. jundzillii in 2010-2011 source: unpublished data source: unpublished data agriculture and food sciences research, 2016, 3(2): 75-79 77 figure-1c. fruit weight developing of r.dumalis and r. jundzillii in 2010 figure-1d. fruit weight developing of r.dumalis and r. jundzillii in 2011 source: unpublished data source: unpublished data velocity of fruit flesh were similar in both species and years (figure 2a,b). fruit flesh percentage of rosa dumalis was higher than rosa jundzillii in both years (figure 2c,d). figure-2a. fruit flesh ratio developing of r. dumalis in 20102011 figure-2b. fruit flesh ratio developing of r. jundzillii in 20102011 source: unpublished data source: unpublished data figure-2c. fruit flesh ratio developing of r.dumalis and r. jundzillii in 2010 figure-2d. fruit flesh ratio developing of r.dumalis and r. jundzillii in 2011 source: unpublished data source: unpublished data fruit diameter growth of r. dumalis were fast in the beginning weeks in both years (figure 3a). fruit diameter of r. jundzillii in 2011 were faster and more regular than 2010 (figure 3b). fruit diameter of r. dumalis was higher than r. jundzillii in 2010 (figure 3c) but were similar and closer in 2011 (figure 3d). figure-3a. fruit diameter developing of r. dumalis in 2010-2011 figure-3b. fruit diameter developing of r. jundzillii in 2010-2011 source: unpublished data source: unpublished data agriculture and food sciences research, 2016, 3(2): 75-79 78 figure-3c. fruit diameter developing of r.dumalis and r. jundzillii in 2010 figure-3d. fruit diameter developing of r.dumalis and r. jundzillii in 2011 source: unpublished data source: unpublished data fruit length developments of r. dumalis and r. jundzillii were higher and more regular in 2011 than 2010 (figure 4a and figure 4b). fruit size development of r. dumalis showed a faster improvement (figure 4c) in 2010 when species compared. however, a closer or similar development occurred in 2011 (figure 4d). figure-4a. fruit length developing of r. dumalis in 2010-2011 figure-4b. fruit length developing of r. jundzillii in 2010-2011 source: unpublished data source: unpublished data figure-4c. fruit length developing of r.dumalis and r. jundzillii in 2010 figure-4d. fruit length developing of r.dumalis and r. jundzillii in 2011 source: unpublished data source: unpublished data seed weight of genotypes exhibited an irregular development depending on year and species (figure 5a,b,c,d). figure-5a. seed weight developing of r. dumalis in 2010-2011 figure-5b. seed weight developing of r. jundzillii in 2010-2011 source: unpublished data source: unpublished data agriculture and food sciences research, 2016, 3(2): 75-79 79 figure-5c. seed weight developing of r.dumalis and r. jundzillii in 2010 figure-5d. seed weight developing of r.dumalis and r. jundzillii in 2011 source: unpublished data source: unpublished data measured properties of r. jundzillii in the second year were higher than the result of the first year as determined for r. dumalis. it was concluded that the fruit load per bush and ecological conditions affected the fruit weight and other parameters. as a matter of fact, fruit growth is influenced by some genetic, cultural and environmental factors [3]. the fruit growth of persimmon indicated that fresh weight measurements exhibit the general sigmoid curve pattern for a typical botanical berry, with only slight deviation schroeder [8]. filiz [6] investigated the weight, volume and size developing of peach from full bloom to harvest. in the growth rate of measured characteristics occurred rapid increase during the initial weeks, but towards to the last weeks this rate continued to decrease. öz [7] reported that fruit diameter and length of kiwifruit increased rapidly in first 4-5 weeks but developing rate decreased gradually after that time. growth investigations of various fruits based on diameter, weight or volume indicate that each botanical fruit type exhibits a characteristic mode of development, depending somewhat upon its basic morphology. thus the stone fruits, as typified by the peach or cherry, follow a general pattern of dimensional growth which consists of three rather distinct periods from flower to mature fruit. rapid growth of all tissues characterizes the first period. very little grown can be detected during the second or “rest” period during which the endocarp hardens. the third period is marked by a very rapid increase in size as fruit maturity is approached. the apple and pear, pomes, exhibit a different type of dimensional growth, which is nearly constant in rate during the entire period of fruit development. citrus fruits, such as lemon or orange, each of which is morphologically a specialized berry, the hesperidium, exhibits a continuous increase in volume throughout the season, provided water is available. shrinkage of citrus fruits may occur during the growing season when a water deficit occurs within the plant. the fig, morphologically a fleshy receptacle which contains many true botanical fruits, achenes, follows the growth pattern of the stone fruits with two periods of size increase based on diameter measurements. the olive, a drupe, however, has three periods of increase in volume. fresh fruit weight and fruit length increases in the pericarp of the date, a berry, are represented by simple "s" curves which reach a maximum in late summer, sometime before full fruit maturity, and fall off as maturity is attained [8]. while the rose hip is similar in some of the trends exhibited by these fruits, it is distinct in others. 4. conclusion it can be stated that fruit growth characteristics of species measured exhibited the single sigmoidal curves in two years except seed weights. seed weight growth curves formed the irregular patterns depending on year and species. developing rate of fruit weight, fruit diameter, fruit length and fruit flesh ratio were fast up to last 3-4 weeks. results of two species obtained in 2011 were higher than the results of 2010, in general. this means that fruit development characteristics can be affected by environmental factors as well as genetic and maintenance conditions. references [1] f. b. salisbury and c. w. ross, plant physiology, 4th ed. california, usa: wadsworth publishing company belmont, 1992. [2] i. karaçalı, storaging and marketing of horticultural crops, 4th ed. i̇zmir, turkey: ege university agricultural faculty publications no: 494, 2004. [3] e. atay, l. pırlak, and a. n. atay, "determination of fruit growth in some apple varieties," journal of agricultural sciences, vol. 16, pp. 1–8, 2010. [4] a. n. miller, c. s. walsh, and c. d. cohen, "measurement of indole-3-acetic acid in peach fruits (prunus persica l. batsch cv redhaven) during development," plant physiology, vol. 84, pp. 491–494, 1987. [5] m. n. westwood, temperate zone pomology: physiology and culture, 3rd ed. oregon, usa: tımber press, 1995. [6] a. filiz, "evaluation of various indicators of maturity for harvest of the some peach (prunus persica l.) cultivars in tokat province," phd thesis gaziosmanpaşa university, natural and applied science institute, tokat, turkey, 2004. [7] a. t. öz, "effects of controlled (ca) and normal atmosphere (na) storage on ethylene biosynthesis in hayward (actinidia deliciosa) kiwifruits harvested at different periods," phd thesis uludağ university, natural and applied science institute, bursa, turkey, 2006. [8] c. a. schroeder, "fruit growth in the oriental persimmon," california avocado society, 1960 yearbook, vol. 44, pp. 130-133, 1960. [9] g. costa, m. noferini, f. bucchi, and l. corelli-grappadelli, "methods for early forecasting apple size at harvest," acta horticulturae, vol. 363, pp. 651–659, 2004. [10] m. güneş, "a research on improvement of rose hips (rosa spp.) by selection and propagation with cuttings, wild grown in tokat province," phd thesis yuzuncu yil university, natural and applied science institute, van, turkey, 1997. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 30 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 30-40, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.30.40 © 2019 by the authors; licensee asian online journal publishing group the mysterious domination of food/drinking water contaminants and adulterants in bangladesh a. k. mohiuddin department of pharmacy, world university of bangladesh, dhaka, bangladesh. abstract food adulteration and contamination is nothing new in this age. it is the consequence of modern civilization, people moving from places with altered social value and ethics, industrialization and rapid progression of economic activities. it is highly prominent in urban areas of many developed or underdeveloped countries and so is with in dhaka city. commercialism and business mind drive people toward such unethical activities knowingly or unknowingly. most of the cases it is done by uneducated or illiterate people, having least idea about what evil they are doing to mankind. people who are health conscious mostly avoid these but many of them have to go with this because of the busy life schedule or carelessness. by definition, safe food or drink means pollutants or adulterants presence within the limit of the standard such as pathogenic micro-organisms, natural toxins and potentially harmful chemicals that may cause health hazards beyond a certain limit, either deliberately added or naturally present in them. again, the economic development of the country doesn't reveal basic literacy and awareness of general people. necessary steps should be taken by the authority and mass people should change their mind set up and have to avoid those who create harm. purpose of the study: brief review of chemical induced food and drinking contamination, their consequences and control. healthcare providers/policy makers have a major role play to concerned field. findings: both general people and the old system, are responsible for this unlivable condition of bangladesh. population is not the sole for this instance. a sense of poor rules and regulation is always found everywhere. negligence is becoming a wide spread disease contaminating illiterate to well educated, all kind of people. keywords: carcinogen, chemical intoxication, food poisoning, health hazard, pathogen, adulterants. citation | a. k. mohiuddin (2019). the mysterious domination of food/drinking water contaminants and adulterants in bangladesh. agriculture and food sciences research, 6(1): 30-40. history: received: 9 january 2019 revised: 18 february 2019 accepted: 21 march 2019 published: 4 june 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 31 2. materials and methods ................................................................................................................................................................... 31 3. results and discussion ................................................................................................................................................................... 32 4. recommendations ............................................................................................................................................................................ 38 5. conclusion ......................................................................................................................................................................................... 39 references .............................................................................................................................................................................................. 39 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.30.40&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.30.40&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/1828 https://orcid.org/0000-0003-1596-9757 agriculture and food sciences research, 2019, 6(1): 30-40 31 © 2019 by the authors; licensee asian online journal publishing group 1. introduction contamination is the presence of an unwanted substance where it should not be or at concentrations above recommended. pollution is contamination that results in adverse biological effects to resident communities. food contaminants and adulterants gave a new dimension in city life, together rest of the country. rural people are less exposed to adulteration than urban people because of busy life, arrangement of food/spice item are mostly obtained from nearby grocery stores, chain shops and nearby open markets. recent media reports on the malpractice in the food sector has revealed horrifying scenario leading to massive public outcry. although there is no official statistics on food borne illness, it is perceived to be a major problem in urban areas of bangladesh. contaminated food, filthy and unhygienic environment of urban cities result in health hazard which may even cause deaths. the healthcare providers/policy makers’ role in environmental health is related primarily to being alert to the conditions prevailing in the community and of working with others to adequately control any of the attendant hazards. 2. materials and methods comprehensive literature search followed by consulting healthcare professionals, hospital, clinic associates, newspaper journalists, ngo workers about adulteration and food contamination. a few high officials were asked help for necessary books, journal, and newsletters. a few local magazine and newspapers also observed to get the necessary concern. projections were based on various types of adulteration in food served/sold to general people, contaminated drinking water supply from a variety of sources. 2.1. gross outcomes of chemical contamination food remains a significant vehicle of disease organisms. foodborne disease, more commonly but often incorrectly called “food poisoning,” is grossly underreported. in most instances the illness produced by contaminated food is mild and of short duration, but more severe outbreaks (such as hepatitis a, most commonly seen in public restaurants) can occur. epidemics of food-borne disease are dramatic and sudden, and most people become sick within 6 to 24 hours after consuming the contaminated foodstuffs. the epidemic pattern of food-borne disease presents differently from the gastrointestinal symptoms (e.g., nausea, vomiting, and diarrhea) induced by intestinal enteroviruses. the safety laws and regulations of bangladesh are as given in table 1. who published – chemical exposure to toxic level is suspected to be involved in causing – • carcinoma • cardiovascular disease • kidney, liver dysfunction hormonal imbalance • premature birth • suppression of immune system • impaired development of nervous system • mental health problems and • learning disabilities/cognitive dysfunction table-1. food safety laws and regulations and food standards in bangladesh. agricultural products market act, 1950 (revised in 1985) fish protection & conservation act, 1950 (latest amendment in 1995) the food grain supply (prevention of prejudicial activity) ordinance, 1956 the bangladesh pure food ordinance, 1959 (bangladesh ordinance no. lxviii of 1959) agricultural pest ordinance 1962 agricultural produce markets regulation act, 1964 (revised in 1985) the cantonments pure food act 1966 destructive insects and pests rules, 1966 (plant quarantine) amended up to 1989 the bangladesh pure food rules 1967 the special powers act, 1974 the animals slaughter (restriction) and meat control (amendment) ordinance, 1983 marine fisheries ordinance, 1983 and marine fisheries rules, 1983 fish and fish products (inspection and quality control) ordinance, 1983 the pesticide ordinance, 1971 and the pesticide rules, 1985 bangladesh standards and testing institution ordinance, 1985 (xxxvii of 1985) the radiation protection act, 1987 the iodine deficiency disorder prevention act, 1989 the essential commodity act, 1990 national food policy 1996 national agriculture policy 1996 fish and fish products (inspection and quality control) rules, 1997 national food and nutrition policy 1997 national fisheries policy 1998 national policy for safe water and sanitation 1998 national health policy 2000 bangladesh standards and testing institution (amendment) act, 2003 the bangladesh pure food (amendment) act, 2005 product labeling policy 2006 national livestock policy 2007 fish feed and animal feed act 2010 export and import policy 2009-2012 the bangladesh food safety act 2013 bsti ordinance and many others source: nasreen and ahmed [1]. agriculture and food sciences research, 2019, 6(1): 30-40 32 © 2019 by the authors; licensee asian online journal publishing group 2.2. laboratories for food analysis • public health laboratory (iph) • bsti (ministry of industries) • food testing laboratory (ministry of food & disaster management) • food testing laboratory (dhaka city corporation) • institute of food radiation biology, bangladesh atomic energy commission • institute of food science technology, bcsir • institute of nutrition & food science, university of dhaka 3. results and discussion food adulteration is the most notorious enemy of mankind. civilization has its own drawback that even causing destruction of itself. very few people raised voice on this but crippled by the facts of commercialism. the scope of this article is limited to chemical food contaminants and adulterants. a few discussions based on real life experience and recent studies or reports from various journals and news articles are summarized here. 3.1. food and supply water contamination dhaka city, among huge amount of solid wastes per day from industrial discharge, fertilizers, fossil fuels, sewage sludge and municipality wastes and they are the major sources of heavy metals in soils and subsequent uptake by crops, vegetables and other food items causing serious health hazards to human beings [2-4]. a significant transfer of arsenic, cadmium, chromium, copper, lead, mercury, nickel, zinc, molybdenum and vanadium like heavy metals took place from soils to vegetables (spinach, tomato and cauliflower) grown in industrially polluted soils of konabari at gazipur and keraniganj in dhaka [5]. in the absence of effluent treatment plants (etp), the factory wastes are drained out at will into the farmlands, and ultimately contaminate the farm produce. in our country we have allowed things like pollution and food contamination to run riot. till now, no agency, either under the health ministry or the ministry of science and technology or the ministry of industries, has conducted any examination of the pesticideresidue levels or toxic chemicals in the foodstuff being marketed [6]. apart from these, the slum areas are both populated and are in greatest risk of notorious pathogen found both in food sample and supplied water table 2. according to dhaka water supply and sewerage authority (dwasa), it can currently supply 75% of water demand, out of which 85% is from groundwater sources (deep tube wells). the presence of toxic metal lead in elephant road, dhaka university, jatrabari, and demra area and toxic penta chloro phenol (pcp) and existing pathogenic bacterial load in the wasa supplied drinking water from different areas of dhaka city were found to be unsuitable for human consumption table 3. 3.2. arsenic issue of drinking water twenty million people in bangladesh are still drinking water contaminated with arsenic, two decades after the potentially deadly toxin was discovered in the supply [7]. the bangladesh government is failing to adequately respond to naturally occurring arsenic in drinking water across large areas of rural bangladesh, table-2. detection of foodborne pathogens in food and household water samples collected at point of use from four slums of dhaka city, bangladesh, december 2015 to may 2016 [8]. presence of organisms in food/water overall n = 56 a. organisms present in food n (%) 95% ci yeast and mould (>100 cfu/mg) 48.0 (85.7) 0.74–0.93 coliforms (>100 cfu/mg) 41.0 (73.2) 0.59–0.84 b. cereus (>100 cfu/mg) 27 .0 (48.2) 0.35–0.62 e. coli (>100 cfu/mg) 17.0 (30.4) 0.19–0.44 staphylococcus (>100 cfu/mg) 8.0 (14.3) 0.08–0.27 v. cholera 2.0 (3.5) 0.01–0.14 b. organisms present in water overall n = 16 n (%) 95% ci total coliforms 16.0 (100) – fecal coliforms 16.0 (100) – total aerobic bacterial count 16.0 (100) – yeast 16.0 (100) – mould 16.0 (100) – staphylococcus 16.0 (100) – e. coli 10.0 (62.5) 0.35–0.86 fecal streptococci 9.0 (56.3) 0.29–0.79 pseudomonas 7.0 (43.8) 0.21–0.71 note: total coliforms and fecal coliforms count (cfu/g). human rights watch said in a report. approximately 20 years after initially coming to international attention, an estimated 20 million people in bangladesh – mostly rural poor – still drink water contaminated over the national standard [11]. bangladesh’s health system largely ignores the impact of exposure to arsenic on people’s health. an estimated 43,000 people die each year from arsenic-related illness in bangladesh, according to one study [12]. the government identifies people with arsenic-related illnesses primarily via skin lesions, although the vast majority of those with arsenic-related illnesses don’t develop them. those exposed are at significant risk of cancer, cardiovascular disease, and lung disease as a result, but many receive no health care at all [13]. agriculture and food sciences research, 2019, 6(1): 30-40 33 © 2019 by the authors; licensee asian online journal publishing group table-3. lead, cadmium, chromium and arsenic content in first 14 water samples [9]. sample no. sampling area pb content (mg/l) cd content (mg/l) cr content (mg/l) as content (µg/l) total bacterial count c.f.u./100ml 1 dhaka university 0.52 0.05 bdl 0.78 4.0 × 105 2 bangshal bdl 0.03 bdl 0.43 2.1 × 104 3 dmch bdl 0.04 bdl 0.25 1.0 × 104 4 basabo bdl bdl bdl 5.12 4.2 × 106 5 komlapur bdl bdl bdl 0.21 6 badda bdl 0.04 bdl 1.29 1.0 × 105 7 sobujbagh bdl 0.04 bdl 0.42 5.2 × 106 8 shagun bagichaa bdl 0.06 bdl bdl 5.0 × 103 9 demra 0.46 0.07 bdl 0.44 10 jatrabari 0.51 0.07 bdl 0.15 1.5 × 104 11 mohammadpur bdl 0.07 bdl 0.53 5.0 × 103 12 panthapath bdl 0.07 bdl 0.29 3.0 × 104 13 elephant road 0.53 0.08 bdl 0.10 2.5 × 104 14 shampur bdl 0.08 bdl 0.56 3.5 × 104 source: sabrina, et al. [10]. 3.3. food adulterants important food hazards include microbial hazards, pesticide residues, misuse of additives, chemical contaminants, including biological toxins and adulteration. although microbiological contamination and chemical hazards have received most attention, it is recognized that food adulteration and food fraud should not be neglected considering their role in public health [14]. food adulteration includes various forms of practices, such as mixing, substituting, concealing the quality of food by mis-labelling, putting up decomposed or expired food, and adding toxic substances [15]. about the proportion of adulterated food items in the market varied between 70% to 90%. more than 76 percent food items in the market were found adulterated in a random survey by public health laboratory of dhaka city corporation in 2004 [16, 17]. according to the international centre for diarrheal disease & research, bangladesh (icddr, b), there is approximately 150 food items in the country. a study by the institute of public health (iph) revealed that more than 50% of the food samples they tested were adulterated. textile dyes, which are highly injurious to health, are being randomly used to color many types of food. textile dyes, which are highly injurious to health, are being randomly used to color many types of food [18]. clay powder is mixed with the mixture of turmeric powder and cold toxic yellow dye to make it yellow. water and salt are well mixed with these species to increase the weight. mangoes, watermelon, litchi, watermelon, pineapple, papaya and bananas are artificially ripened using a carcinogenic chemical called ethylene oxide table 4. in bananas, another chemical called calcium carbide is used that becomes a spray acetyl-gas to generate heat. dalda (hydrogenated vegetable oil/ fat popular in south asia) used in cooking is an example of a worst case false. "our stomach temperature is 37 degrees celsius and the melting point is 54 degrees celsius dalda. so, there is no way that dalda can be absorbed by the body [19]. most sweetened condensed milk products sold in the market actually contains palm oil which is used in substitute for cow’s milk and therefore most sweetened condensed milks out there do not contain milk at all [20]. fish is considered to be an essential protein for people of all ages. many fish sellers spray fish with formalin in an indiscriminate manner, it makes the fish or fruits stiff and keeps them looking fresh for longer table 5. undoubtedly human health is now under the possession of formalin, in our country about 400 tons formalin is being imported which are goes to human stomach, creates deadly mistreats on long term exposure table 6, even though for laboratory or research purposes 100 tons of formalin is quite enough, 80% of the imported formalin being added to food only for business purposes. three-fourths of the marketed vegetables, fruits, and fish contain pesticides and formalin residues. the consumption of such foods might lead to serious diseases [21]. ironically even food color is being adulterated. substandard food color is finding its way into many types of food. this includes the reddish jelapi, and the saffron beguni, peaju or alur chop. candy, chips, ice cream, chewing gum and even biryani may contain large amounts of poor-quality food color. cooking oil that is so commonly used to deep fry items should only be used once but many food vendors and restaurants recycle burnt oil. once the oil is used for cooking, it becomes oxidised. the more the oil is used, the more pre-oxide is created which is really harmful for the body. this gets more poisonous with continued usage. in 2017, police seized around 3,000 eggs suspected to be artificial during a raid in patiya upazila, chittagong. they also arrested two men – an egg shop owner and an egg supplier – suspected to be involved in the trade and marketing of fake eggs [24]. doubt also pointed in rice also. the term ‘plastic rice’ was coined by the online media around 2010, when china reported its use for adulteration of a premium rice called wuchang, which is known for its aroma [25]. the institute of public health (iph) in dhaka and the world health organization (who) in their joint study on food adulteration in 1994 tested 52 street vendors and found that all of their food samples were contaminated with different types of pathogenic microorganisms. they also conducted another study in 2003 in dhaka city and found that 96% of sweetmeats, 24% of biscuits, 54% of breads and 59% of ice creams table 7 were extensively adulterated [26]. agriculture and food sciences research, 2019, 6(1): 30-40 34 © 2019 by the authors; licensee asian online journal publishing group table-4. adulterants used in different food items of vegetable origin as reported in lay press reports [22]. food category and food item adulterant edible oil argemone oil, mineral oil and rancid oil given commonly** soybean oil mustard oil palm oil, chemical*, color*, burnt mobil† from rail locomotives, burnt oil from electric transformer chemicals food grain and grain products lentils, mugdal, chola, mosurdal, dabli, mashkolai, buter dal (lentil types) toxic coloring agents*, imported low-quality inedible lentils mixed with textile dye* and have fungal growth; less expensive mashkolai dal powdered with champa color* and sold as mugdal rice urea added to make it whiter dhekichata chal (husked rice), ata (course flour) red toxic color* mixed with rice and ata to sell as husked rice, red atta muri (puffed rice) urea fertiliser to make it whiter and puffier wheat, corn animal feed packaged as human food semai (vermicelli) dalda made with rotten potato, cow intestine, low-quality palm oil vegetables and tubers vegetables organophosphorus compounds and other pesticides tomato calcium carbide for artificial ripening potol (pointed gourd), peas textile dye* eggplant pesticide* green peas chemically colored* dabli potato red toxic color* spices mixed spices (powder) brick dust, saw dust, chaler kura (dust from outer layer of rice) turmeric powder brick dust, buter dal, kheshari dal (lentils), artificial powder, color chili powder powder with color coriander powder chaler kura (dust from outer layer of rice), toxic color* zeera (cumin) powder brick dust, toxic color*, powder pepper papaya seed salt no iodine bakery products cake‡ textile dye, chemicals*, inedible date expired ata/maida, fertiliser urea, substandard inedible dalda, rotten egg biscuit‡ ammonium bicarbonate, sodium cyclamate, fertiliser urea, toxic coloring agents*, palm oil, burnt oil, outdated inedible ata/maida bread‡ rotten egg, outdated ata/maida fruit and fruit products (lead arsenite, calcium carbide, ethephon, formalin, injection of dye in general) mango, banana, pineapple calcium carbide for artificial ripening cherry koromcha (carissa carandas, christ’s thorn) with chemical color orange and lychee juice water, flavor, textile dye*, sweet pumpkin, and color imported juices substandard, date expired with new sticker snacks noodles‡ dhekichata chal, lal atta (coarse wheat flour), red potato chanachur fried in burnt mobil†, no potato, imported powder, and color peyaju, beguni toxic dye* chocolate, sugar, and honey chocolate powder, sugar, color*, chemical sugar soda used instead of sugar in food honey sugar syrup others pickle inedible ingredients jorda (smokeless tobacco) wood dust, chemical mineral water and drinking-water‡ tap-water, arsenic contaminated, contaminated with bacteria, no mineral note: *chemical nature/composition not mentioned/specified; **argemone oil leads to epidemic dropsy, loss of vision, heart diseases, tumor, mineral oil cause liver damage and carcinogenic; rancid oil leads destroys vitamin a and e †polychlorinated biphenyl (pcb) used as coolant in automobiles and transformers; ‡prepared in unhygienic condition table-5. underlying reasons behind adulteration issue in fish supply chain of bangladesh [23]. reasons details a cheap method to prevent post harvest loss to prevent this post-harvest loss one of the cheap methods is to use formalin and other toxic chemical which helps to keep this fish fresh for a long time. lack of technical knowledge lack of proper technical knowledge contributes in use of waste material in fish culture, improper handling after post-harvest and use of formalin and other toxic elements. lack of ice box, unavailability of ice, high price of ice, lack of cold store to keep unsold fish. ice/icebox is expensive. main problem in the market of bangladesh is there is lack of cold storage in the market place. as a result, there is no proper way to preserve unsold fish. lack of awareness most of the traders do not aware of the danger of using these harmful chemicals. on the other hand, customers are also not properly aware of this issue. lack of government initiative co-ordination between authorities is a major issue, does not appear to be any cohesive view regarding procedures and penalties for the same offence by officials from same organisation. lack of policy framework food laws and regulations are mostly outdated and fragmented. even the new safe food act 2013 is not free from this as it is basically modeled on the pure food ordinance of 1959. inadequate penalties considering the extent of harmfulness of food adulteration, penalties mentioned in law is insufficient. for an example, the penalty for food adulteration is maximum term of six months of imprisonment or up to a maximum fine of bdt 1000 which is equivalent to eur 10.77. considering the gravity of the offences this punishment is not hard enough. source: fatema and moslah [23]. agriculture and food sciences research, 2019, 6(1): 30-40 35 © 2019 by the authors; licensee asian online journal publishing group table-6. effect of formalin treated food consumption on health [23]. exposure routes effect on human carcinogenicity formalin has the potential effect to cause cancer, repeated and prolonged exposure increases the risk of cancers of the lung, nasopharynx, oropharynx and nasal passage reproductive health it has a harmful effect on reproduction system by inducing oxidative stress. skin (dermal) prolonged and repeated contact with formalin could cause numbness (lack of feeling) and a hardening or tanning of the skin eye contact formalin solution splashed in the eye can cause injuries from transient discomfort to severe such as loss of vision source: fatema and moslah [23]. table-7. adulterants used in different food items of animal origin as reported in lay press reports [22]. food category and food item hen egg** white eggs of farm hens colored red with textile dye* to sell as local hen eggs. tortoise eggs sold as hen eggs fish inject formalin through the gills or dip fishes in water treated with chemicals, such as chloro-fluoro carbon (cfc); ddt† powder to prevent rotting; add red color* to give fresh look; sell rotten fish dry fish ddt† mutton buffalo, sheep and beef meat sold as mutton beef buffalo meat sold as beef halim‡ left over bones, intestine sweetmeats and dairy products butter cow’s intestine, dalda mixed with color*, powder* ghee, dalda (hydrogenated vegetable oil) ‡ banaspati, toxic chemical*, potato smash, cow’s fat, intestine sweetened curd‡ textile dye* sweetmeats‡ textile dye named ‘thousand power color’ and toxic chemicals*; rotten eggs; dalda made with cow’s intestine, saccharin, soybean oil and vegetable oil instead of milk fat; paste of ground rice and sulphuric acid mixed with milk to make posset jilapi (coil-like juicy sweet) fried with mobil halua rotten carrot and lau (bottle gourd), chemical* ice-cream‡ unsold foul-smelling ice-cream refined and re-packaged, almost no milk, palm oil for soap manufacturing, textile dye*, low-quality milk powder, sodium cyclamate imported milk powder adulterated, low-quality, date expired, without bsti approval fast food and restaurant food† jelly, sauce toxic coloring agents*, chemicals*, spirit chicken** dead chicken; cooked and raw meat refrigerated together shrimp sold rotten fish fried and raw fish refrigerated together note: *chemical nature/composition not mentioned/specified; ** blri also showed that broiler meat and egg showed presence of antibiotic residue of ciprofloxacin, sulfonamide, oxytetracycline and enrofloxacin in high level. †dichloro-diphenyl trichloroethane; ‡prepared in unhygienic condition; polychlorinated biphenyl (pcb) used as coolant in automobiles and transformers consumption of adulterated food items may cause asthma, sore throat, larynx constriction, bronchitis, skin infections, allergic reactions, diarrhea, hematuria, circulatory failure, numbness, dizziness, kidney failure, stomach cancer, liver cancer, nervous disorders and many other diseases table 8. after consumption of adulterated food items, thousands of people are becoming sick. children are the worst victims. about three million people suffered from diarrhea during 2005-2009 and about 15% of children died in 2011 as reported by the directorate general of health services [27]. the long-term effects are also very severe especially the incidence of renal failure, liver damage and cancer which are increasing alarmingly in bangladesh. heavy metals, such as lead, chromium and arsenic accumulate in the body that might cause kidney and liver damage and develop abnormality among children. indiscriminate and irrational use of antibiotics in poultry without following withdrawal period may result in unexpected residues in animal food and could cause serious health hazards to consumers. research reports on antibiotic residues in broiler meat and liver from different farms and local markets for the presence of residues of ciprofloxacin, enrofloxacin, oxytetracycline, doxycycline and amoxicilin antibiotics revealed significant level of exposure of antibiotic residues. there’s a greater chance of declining immunological responses and can detrimentally affect intestinal microbiota in susceptible individual. according to prof. muniruddin ahmed (clinical pharmacy and pharmacology, dhaka university) cooking cannot destroy antibiotic residues, which made them resistant to antibiotic treatment [28]. milk in rural areas is usually adulterated with dirty water, which can cause hepatitis. people have now come to know about a new milk adulteration technique that uses a thickening agent, sorbitol, and detergent. icddr, b recent studies shows nearly 75% samples from primary-level producers were contaminated with coliform and more than 50% with fecal coliform bacteria. at the collection points, samples were found contaminated with a high number of coliform bacteria and fecal contamination of more than 90% while more than 40 % of the samples had a high e coli count [29]. vegetable and fruit samples collected from around savar, dhamrai and tongi show the presence of textile dyes, which, in the short-term, will cause diarrhea, food poisoning and gastrointestinal problems, but in the long-term toxic materials will accumulate in the body with serious health implications [30]. in bangladesh, people allowed things like pollution and food contamination to run riot. till now, no agency, either under the health ministry or the ministry of science and technology or the ministry of industries, has conducted agriculture and food sciences research, 2019, 6(1): 30-40 36 © 2019 by the authors; licensee asian online journal publishing group any examination of the pesticideresidue levels or toxic chemicals in the foodstuff being marketed. these merchants and traders are the enemy not only of the nation and their own children but of the entire mankind. the holy prophet (pbuh) has disowned those who indulge in this immoral business. he said ‘’ the adulterator is not one of us’’ [31]. table-8. toxic elements in noxious addition of food/additives with possible outcomes [17]; [22]; [29]; [16, 32-42]. contaminants food/additives possible outcome coloring agents chrome, tartzine and erythrosine spices, sauces, juices, lentils and oils cancer in kidney, liver, skin, prostate and lungs rye flour (ibid) barley, bread and wheat flour convulsion and miscarriage hormone (ibid) cauliflower infertility of women coal tar and industrial dyes sweets, sauce, pastry cream, powders spices carcinogenic burnt oil crispy snack food poisoning, reflux, heartburn agenomato or monosodium glutamate (ibid) chinese restaurant food items nervous system disorder and depression flour chalk powder gi problems soap ghee/butter gi problems calcium carbide/ethylene dioxide ripening of fruits cancer in kidney, liver, skin prostate and lung urea (ibid) for whiten rice and puffed rice damage of kidney & nervous system, respiratory problem brick dust chili powder respiratory problem sulfuric acid and palm oil condensed milk cardiac function problem saw dust, used and exhausted tea leaves loose tea respiratory problem sodium cyclamate sweetmeat cancer, fetal abnormality metanil yellow aniline dyes turmeric powder carcinogenic melamine milk products kidney malfunction oleomargarine or lard butter asthma and weakened kidney function yellow and sudan red colors (ibid) chili powder tumors in liver and bladder and finally for cancer ddt dried fish (shutki) cancer especially breast cancer, liver cancer and pancreatic cancer, reproductive damage (weaken semen, early menopause, exposure of teratogen and birth defects) and some neurological damage reported. bottle and jar water bottle and jar water bottle and jar water formalin preservation of fish, meat, fruit and milk throat cancer, blood cancer, childhood asthma and skin-diseases. poisonous coloring agents like auramine, rhodamine b, malachite green, yellow g, allura red, and sudan red applied on food items for coloring, brightness and freshness damage liver and kidney and cause stomach cancer, asthma and bladder cancer source: mohiuddin [42]. 3.4. energy drinks vs carbonated drinks the government has decided to ban the production, marketing and import of energy drinks under the guise of carbonated beverages with immediate effect [43]. the bangladesh standard and testing institution (bsti) at a council meeting on also resolved to take legal action against the companies that have been advertising their products as energy drinks after having them listed as carbonated beverages with bsti. the move came after the bangladesh food safety authority (bfsa) wrote to bsti to say the production of energy drinks in the name of carbonated beverages should not be allowed, as the ingredients of the two are significantly different. there is no government fixed standard for energy drinks in the country, industries minister told parliament once. authority received complaints about the use of various highly addictive substances, including caffeine and opium, in energy drinks [44]. in 2012, a department of narcotics control test conducted on energy drinks from several local and foreign companies had found excessive amount of artificial caffeine, viagra (sildenafil citrate), beer and alcohol as ingredients [45]. the manufacture and advertising of energy drinks under a license for carbonated beverage is fraud, punishable under several laws including the bsti act and the food safety act. initially, the government will write to the companies to instruct them to discontinue their production and marketing of energy drinks. punitive actions will follow if they do not comply, said the official. the government will also amend the import policy in order to blacklist the foreign energy drinks with harmful ingredients [46]. 3.5. safety issues of bottled water so-called mineral water supplied to houses and offices in jars are not tested. in most cases, the water is filled in the jars right away from the tap and sometimes in the empty bottles of some of the well-known mineral bottle brands [47]. the bangladesh agricultural research council (barc) has of late made a sensational revelation regarding the quality of water different companies supply for drinking in and around dhaka. according to the findings of the government study, 'coliform bacteria' (pathogens from feces of humans and animals) has been found in 97% of so-called filtered water supplied in jars to households, shops and offices in the capital city of bangladesh. a team of barc researchers determined the 'horrifying' data while studying the level of minerals in jars and bottled water marketed in dhaka city. the researchers sampled 250 jars from across the city's farmgate, karwan bazar, elephant road, new market, chawk bazar, sadarghat, keraniganj, jatrabari, motijheel, basabo, malibagh, rampura, mohakhali, gulshan, banani, uttara, airport, dhanmondi, mohammadpur, mirpur, gabtoli, and on the city's outskirts at aminbazar, savar and ashulia. the level of 'coliform bacteria' in the samples collected from agriculture and food sciences research, 2019, 6(1): 30-40 37 © 2019 by the authors; licensee asian online journal publishing group elephant road, chawk bazar, basabo, gulshan and banani areas was found significantly high in the research [21]; [48]. a few months ago, a mobile court busted six fake mineral water plants in bosila area of dhaka’s mohammadpur and sentenced six staffers to different terms of jail. the team also seized 2000 jars of water and destroyed those later [49]. 3.6. food adulteration during ramadan the crime of those selling adulterated and unhygienic food items is very serious and strict action needs to be taken as per law against such guilty persons. according to media reports some 600 field-level sanitary inspectors are working across the country to ensure food safety for all during the holy month of ramadan. this is welcome news but if previous records are anything to go by people should not get their hopes up too high. even more unfortunate is the fact that this nefarious practice increases exponentially during the month of ramadan. it was found that coloring agents are used in spices, sauces, juices, lentils and oils. formalin and carbide used in fish, fruit, meat and milk [50]. the shopkeepers and the merchants many of them with a pious fade try to earn a large amount of profit by this unethical practice, and so they play with the life and health of the people. they mix dangerous things in the daily eatables. 3.7. sub-standard vermicelli (shemai) and cow fattening ahead of eid adulterated vermicelli flooded different markets in the capital ahead of eid-ul-fitr, one of the biggest religious festivals of the muslims, posing a serious threat to public health. a section of unscrupulous businessmen is busy in manufacturing sub-standard vermicelli in the port city ahead of eid-ul-fitr as the food item has a high demand in the day. according to local sources, some of the factories are using unrefined palm oil and animal fat to produce laccha vermicelli while hazardous chemicals and toxic color were also used to make the food items. bangla vermicelli is produced using flour and water and laccha is produced with oil, flour sugar and water. most cases, the vermicelli was being dried in unhygienic condition in the rooftop of the factories [51]. seeking anonymity, a seasonal vermicelli maker said a section of businessmen in the city bought the adulterated vermicelli and sell them tagging labels of renowned companies. bsti personnel left a comment as bangla vermicelli is used to make by seasonal factory for a certain time, it is not mandatory to take license from bsti, however, the factory should take health and hygienic certificate issued by the sanitary inspector [52, 53]. with eid-ul-azha around the corner, the poribesh bachao andolon (poba) has recently urged the government to monitor how cattle is being reared and fattened in the farms in the country [54]. knowingly causing such damage to public health in order to hike up the price of cows is a crime, and must be dealt with swiftly. the use of antibiotics, growth hormones, harmful chemicals, and steroids is prohibited by the animal feed act 2010. violators may be faced with a year in prison, a tk 50,000 fine, or both. a large number of farmers involved in bull fattening just before 3 or 4 months of eid-ulazha (muslim festival), when they sell the animals with profitable price. visiting different villages in bera, santhia and ataikula upazilas of pabna, and shahjadpur and baghabari areas in sirajganj, these correspondents found that almost every household was using steroids, antibiotics and other chemicals for months in blatant violation of law. everyone -from cattle farm owners to landless farmers -wanted to take full advantage of this. these cattlefattening drugs are also widely used in chuadanga, jhenidah, nilphamari, barisal, faridpur, manikganj and some other districts. consumption of meat of these animals poses serious health risks for humans, according to experts [55]. the changes to the cow caused by these injections are not merely cosmetic – severe health damage is done to humans by the consumption of this meat. while most traders would still claim that the fattening supplements were not harmful, there is reason to believe these chemicals may cause cancer, kidney disease, and infertility in women [56]. 3.8. penalty imposed on famous eateries isn't it surprising that like many occasional drives against various crimes, the fight against unsafe food, too, has assumed the character of a seasonal activity? while this should have been a continuous and unrelenting activity round the year across the country, sporadic and infrequent moves here and there, leave no permanent impact on the sellers and producers of spurious food items of all varieties. newspaper reports say that the drive against food adulteration is currently on in the capital, launched last week. the bangladesh food safety authority (bfsa), the state watchdog to regulate the sector is reportedly monitoring the capital's food markets under the supervision of an executive magistrate. one has reasons to question the rationale behind the drive in the capital alone, that too with just one magistrate. the effectiveness of the drive is bound to cause nothing more than a ripple with mobile courts punishing a few sweetmeat shops, restaurants and if at all, some kitchen market sellers. the fact that such drives, sporadic and half-hearted, failed to bring any discipline in the country's food market cannot be disputed. however, famous eateries/food chains are not devoid of these cases of adulteration and substandard food staffs. penalty imposed on these famous shops/food chain should at least give an idea to general people that paying high price is not always an indicator of good quality. even a few of them were penalise more than once or twice for the same reason but substandard food serves never ended table 9. for a better view references regards are attached in this table with date published. government officials monitoring food markets in capital dhaka fined businesses millions of bdt only after finding that their food items contained ingredients harmful to human health. inadequate monitoring of food markets may have exposed people’s health to serious hazards from consumption of substandard and adulterated foods. in the absence of corrective measures, punishing the offenders-at times by way of hefty monetary penalties-is not an appropriate method of dealing with the problem. a properly institutionalized mechanism with sufficient manpower and regular monitoring round the year can only bring things to some semblance of order. in this regard, it is important that the bsfa and other agencies such as the bsti and the city corporations which also run such drives maintain a well-coordinated plan of action. it is also important that punitive actions should be backed by actions to improve the quality and standard of food of all varieties. to monitor the situation, inspection and sudden raids are welcome, but it must not be forgotten that inspection is just one of the many ways to rein in food adulteration. agriculture and food sciences research, 2019, 6(1): 30-40 38 © 2019 by the authors; licensee asian online journal publishing group table-9. eateries/chain shops raid list for adulteration/substandard food serving. eateries/super shops possible reasons/issues reference date published khushbu biriyani, gulshan textile dyes for coloring biriyani daily sun & daily bangladesh 29.05,18 & 28.05.18 ‘kosturi restaurant, gulshan unhygienic environment inside the restaurant. unb news 28.05.18 swapno’s banani date-expired food daily sun 29.05.18 kfc, american burger, dawat-e mejban, united catering, dynamic food court, dhanmondi rotten and unhealthy foods daily sun 29.05.18 meenabazar’s shantinagar soda water without the bsti approval and other issues. daily sun 29.05,18 agora and nandan super shop selling different local and foreign products having no bsti approval. daily sun 29.05,18 boomers café, baily road keeping food in unhygienic environment daily sun 29.05,18 sbarro cafe keeping sauce without a bsti approval. daily sun 29.05,18 kutumbari restaurant, chittagong preserving date-expired fish and meat. dhakatribune 28.06.16 bonoful sweets, chittagong harmful ammonia for making biscuits crispy. dhakatribune 28.06.16 sizzle, chittagong applying color in making cakes which is unfit for human consumption dhakatribune 28.06.16 flavor sweet and bakers and fulkoli food products, chittagong date-expired food colour in manufacturing cake and sweetmeat. dhakatribune 28.06.16 fakhruddin biryani, baily road using expired and rotten ingredients for making kebab and halim the news today 06.02.18 ‘mr bakers, turag area preserving date-expired and stale cake and making biscuits and toasts without bsti license. the daily sun 16.06.17 arabians sweets and bakery for not writing manufacturing and expiry dates on their packets. the daily sun 16.06.17 ‘pusty dairy and bakery, malibagh for not taking bsti license for producing curd. the daily sun 16.06.17 ‘lucky vermicelli factory’ at matuail in jatrabari making vermicelli in an unhygienic environment. the daily sun 16.06.17 yammi yammi’ and ‘ujjal food products’ at pallabi, mirpur making food items in an unhygienic manner the daily sun 16.06.17 new food hotel and restaurant, shibganj sweets, tuhin food hotel, new bidyut hotel, tripti hotel and cafe raj hotel, memory biriani house, amir ali fish shop in rajshahi city selling stale and adulterated foods the daily star 17.09.18 maloncho restaurant at the new elephant road poor hygiene dhakatribune 25.07.15 food corner, kings fast food, penji fast food, kepray fast food, capital fast food, al amin food, saikat fast food, welcome fast food and al jaber fast of newmarket, dhaka selling unhygienic food. united news of bangladesh unb beta 20.05.18 kfc’ & boomers baily road serving expired and chemical-mixed food items. dhakatribune 25.07.15 pizza hut, baily road using chemicals in two of its popular sauces without the authority’s approval. dhakatribune 25.07.15 chandrima restaurant and mini chinese, kasturi chhayanir and thai chinese restaurant and new star kebab preserving and selling unhygienic food items the independent & daily prothom alo 19.06.17 & 18.06.17 dawar-e-mejban, dhanmondi using harmful chemicals to color foods. newage 25.05.18 ma-moni hotel and restaurant, suprema restaurant, kosturi restaurant, rangpur city preserving foodstuffs in unhygienic and dirty environment bss news and bangladesh post 26.09.18 agora, shwapno and meena bazar chittagong for selling rotten meat and fish bdnews24.com 11.06.16 meena bazar, agora, swapna dhaka for selling low-grade packed products. bdnews24.com 09.04.12 agora, meena bazar and fast food chain coopers, shantinagar for keeping expired and stale food items (rotten fish, meat and expired food items) the daily star and the independent 15.05.16 & 11.05.16 agora in prabartok, the grocer, and khulshi mart in khulshi, meena bazar in sholoshahar and shwapno in gol pahar chittagong selling products at hiked up prices and also selling rotten and stale items the daily star 11.06.16 platinum suites, king’s confectionary, kobe restaurant, pizza inn selling expired, stale and rotten foods. bdnews24.com 02.06.18 disney dine restaurant kabab jangson limited nababi bhoj solution lounge, shamoli selling adulterated food items in unhygienic conditions dhakatribune 18.05.18 mir al-amin hotel and sharif hotel producing food items in an unhygienic condition during ramadan daily sun 19.06.17 alauddin sweetmeat, royal restaurant lalbagh, star hotel and kabab, voot restaurant, ambala sweetmeat, iftari bazar and dominous pizza in dhanmondi using illegal food colorings, poisonous chemicals and used cooking oil in ramadan the daily star 29.05.12 source: mohiuddin [42]. 4. recommendations while contamination of food may be due to negligence, deliberate adulteration by toxic chemicals or radioactive materials for long shelf life of products and increasing the volume in size and weight-among the many crooked methods-is so rampant that it is difficult to find anyone who does not encounter an unpleasant moment of agriculture and food sciences research, 2019, 6(1): 30-40 39 © 2019 by the authors; licensee asian online journal publishing group food-related illness at least once a year. taking care of the situation thus calls for a whole package of initiatives. in advanced countries this involves producing, handling, storing and preparing foods in such a way as to prevent infection and contamination in the entire chain. however, in situations prevailing in this country, it is not merely about maintaining a clean chain but putting in strong deterrents so that criminality in the business could be stopped. sources of harmful stuffs must be plugged, if necessary, by way of ban on imports or local production. strong advocacy on the detrimental effects of consumption should be routinely done. at the same time, training on safe and scientific methods of preservation of food products should also be a high priority in an attempt to curb adulteration. adulteration and contaminant control are a never ending, on the other hand a continuous process. it will increase with time as the civilization go ahead. pharmacists should be aware of the local occupations, companies, and factories and to be cognizant of the initial symptoms of disease. again, pharmacists should become acquainted with the local community and to adapt the principles of health and medical care to the particular situations encountered. the pharmacist’s continuing education requirements should include watching the local pattern of society and its diseases, and changing the emphasis toward evolving disease patterns and their control. government and regulatory authorities are to play strong role in controlling food contaminants and adulteration. 5. conclusion with constant change to the physical, biological, cultural, social, and economic environment, both healthcare providers and citizens should cultivate an informed awareness of these changes, and health providers should adapt their methods of health education, disease prevention, and disease control to the changes in each community. this is especially true food daily consumed, which require concerted community action for their control, but providers may play a much more fundamental and personal role in controlling food-borne diseases; often, the first indication of an outbreak of food-borne disease is time-limited, with an unusually large number of people seeking relief from health hazards. the necessary role in environmental health is related primarily to being alert to the conditions prevailing in the community and of working with others to adequately control any of the attendant hazards. government related authorities, ngos and other private organizations (e.g. pharmaceutical companies) should take initiatives further to ameliorate food and drinking water situation which is worst among all other previous times. general people should be aware of these facts of mischiefs and take necessary steps on their own. a yearround campaign regarding these issues in public places, electronic media and even in rural areas can bring a change as brought by diarrhea, vitamin a campaigns back in 70’s and 80’s. abbreviations bdl (below detection limit); bsti (bangladesh standards and testing institution); bcsir (bangladesh council of scientific and industrial research); icddr, b (international centre for diarrheal disease research, bangladesh); etp (effluent treatment plants); iph (institute of public health); ddt (dichloro-diphenyltrichloroethane); bfsa (the bangladesh food safety authority); barc (bangladesh agricultural research council). references [1] s. nasreen and t. ahmed, "food adulteration and consumer awareness in dhaka city, 1995-2011," journal of health, population, and 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[38] r. shafkat, "letters to the editor urea in puffed rice. the daily star august 27, 2013 richard p. the politics of arsenic-free water," dhakatribune, 2017. [39] k. rajib, "cover unsafe food unhealthy life," daily sun, 2015. [40] editorial, "editorial. look for the fliesthe future ahead!," bangladesh journal of medicine, vol. 25, pp. 40-41, 2014. [41] m. mahboob, "food adulteration: the bangladesh paradox," law journal bangladesh, vol. 2, 2015. [42] a. mohiuddin, "chemical contaminants and pollutants in the measurable life of dhaka city," european journal of sustainable development research, vol. 3, p. em0083, 2019. available at: https://doi.org/10.29333/ejosdr/5727. [43] business, "business, staff reporter. manufactures seek help to dispel doubts about beverages. the independent," 2018. [44] staff, "staff correspondent. no standard fixed for energy drinks amu tells js," the daily star, 2015. [45] s. i. ashif, "the dangers lurking in your energy drinks," dhakatribune, 2017. 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[56] tribune editorial, "put a stop to unethical cow-fattening practices. dhakatribune," 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 172 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 2, 172-181, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.62.172.181 © 2019 by the authors; licensee asian online journal publishing group camel milk composition, udder health and effect of different storage times and temperatures on raw milk quality using camel milking machine “stimulactor” kaskous s. department of research and development, siliconform, schelmengriesstrasse 1, d-86842 türkheim, germany. abstract camel milk is considered one of the most valuable food sources for people in arid and semi-arid areas and is in increasing demand in many european countries and north america. the objective of this study was to investigate the camel milk composition, udder health and effect of different storage times and temperatures on raw milk quality using camel milking machine “stimulactor” (st-c). this work was carried out in the department of research and development of siliconform türkheim, germany. five one-humped dromedary lactating camels were used in this experiment. milk samples were collected and fat, protein, lactose, somatic cell count (scc) and bacterial count (bc) were determined. in none of the tested milk samples pathogenic bacteria could be shown, i.e. all quarters were healthy during the study period. the mean contents of fat, protein, lactose, scc and bc of milk samples were 2.92±0.07%, 2.28±0.01%, 3.91±0.02%, 126.43±7.21 x 103 cells/ml and 23.88±0.57 x 103 bacteria/ml, respectively. after 24 h at room temperature or 48 h at 4 degree a refrigerator, storing raw milk samples had no significant changes in the milk composition and –quality. in conclusion, a good safe raw camel milk with normal composition was obtained if hygienic measures were taken into consideration in the farm and by using the camel milking machine for milk removal. furthermore, we could store the raw camel milk for 24 h at room temperature or for 48 h in the refrigerated temperature without any hygienic quality problems, and all milk components did not significantly change. keywords: dromedary camel, milk composition, stimulactor, udder health, milk quality, milk storage. citation | kaskous s. (2019). camel milk composition, udder health and effect of different storage times and temperatures on raw milk quality using camel milking machine “stimulactor”. agriculture and food sciences research, 6(2): 172-181. history: received: 2 april 2019 revised: 13 may 2019 accepted: 18 june 2019 published: 29 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 173 2. material and methods ................................................................................................................................................................... 173 3. results and discussion ................................................................................................................................................................. 174 4. conclusion ....................................................................................................................................................................................... 179 references ............................................................................................................................................................................................ 179 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.172.181&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.172.181&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://asianonlinejournals.com/index.php/aesr/article/view/878 agriculture and food sciences research, 2019, 6(2): 172-181 173 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study gives us very important information about raw camel milk composition under german condition and how to produce a good quality of raw camel milk. for that, hygienic measures and using the camel milking machine “stimulactor” for milk removal are important factors. in addition, this study shows, how you can store the raw camel milk without any significant change in the raw camel milk quality. 1. introduction camel milk and its products have nutritional and medical properties that make them valuable foods. in many regions in asia and africa, camel milk are used to treat some diseases and to combat health problems such as gastrointestinal disorders, sugar diseases, food allergy, psoriasis, hepatitis c and b, autism and tuberculosis [1-3]. traditionally, camel milk is consumed raw by the pastoralists without any heat treatments in order to exploit the healthy milk properly [4-8]. however, camel milk is an excellent culture medium for the growth of microorganisms [5, 9, 10] and non-heat treated milk and raw milk products are considered a major factors responsible for illnesses caused by food borne pathogens [10, 11]. the milk of a healthy udder is practically sterile [12] and it contains a very low concentration of microorganisms, usually less than 10 x 102 colony forming units of total bacteria per ml milk [10]. however, the milk is contaminated only with the passage of the teat canal with bacteria [9]. entering bacteria into the milk may come from the dirty udder-and teat-surfaces, the stall, the feed, the milker, the air, the water, the milking equipment and finally during storage and transport. among the microbe population, some categories are more dangerous such as pathogenic bacteria, because they may transmit diseases to humans or cause defects in the final product. the rate of multiplication of microbes in the milk depends mainly on the initial number of bacteria, the storage temperature and storage duration. our challenge is how to provide safe raw camel milk to consumer at any time or to the dairy industry for processing. sometimes, camel herds are located so far from the cities and the camel milk must be stored for many hours before reaching the consumer. the aim of this study was to investigate the composition of camel milk and total bacteria under german conditions, in which green grass or hay are available all year round. nevertheless, it is highly humid and cold in the winter. several studies dealing with the chemical composition of camel milk were conducted around the world but not in germany. furthermore, changes in the milk composition and total bacteria count in the raw camel milk was investigated after different storage time and temperature. many people around the world want to drink camel milk fresh for healthy reason. therefore, the best storage time and temperature was applied in this study after using machine milking technique “stimulactor”, which contributed to the stability of milk quality and composition. 2. material and methods 2.1. location this work was carried out in the department of research and development of siliconform company located in türkheim, germany. 2.2. animals and management in this study, five one-humped dromedary lactating camels in the late stage of lactation were used and were available in the department of research and development. the camels were kept outdoor most of the time. however, at night and in the cold winter they were kept in the barn using the loose housing system. camels fed primarily on pasture and they were also provided with hay in addition to vitamins and minerals supplement. meanwhile, drinking water was available at all time. 2.3. the used milking system st-c was used in this work, which developed in the siliconform company, türkheim, germany [13] (www.siliconform.com). this milking system has the following technical characteristics: it is based on a quarter individual milking system, which means that milking cups work independently from each other, without a claw. however, the system provides periodic air inlet into the teat cups. furthermore, the working vacuum level is set to 36 kpa and sequential pulsation (25% each quarter) was adopted. the pulsation rate was 90 cycles/min with a 65:35 pulsation ratio during the milking time. in addition, the system was equipped with a silicone liners and it had a very special pre-stimulation program and an excellent cleaning and sanitary process. 2.4. the milking routine the camels were milked once a day at 11:00 am with a unit milking machine st-c, and without the presence of calves during the milking process. the milking routine started with a pre-milking preparation, in which the teats were cleaned with a wet udder tissue and afterwards they were dried using another clean tissue. afterwards, the teat cups of the milking unit were individually or in pairs attached manually to the teats. subsequently, the milking process started on the control display and the pre-stimulation began. the pre-stimulation was programmed so that the teats were intensively stimulated with a normal pulse rate (90 cycles/min) and a reduction in the suction phase (b-phase) of 10% over a period of 90 sec. simultaneously, intensive movement of the teat cups were regulated as an additional stimulation which is operated by an actuator. it was an arm on which four milk tubes are situated. during the pre-stimulation and the milking time, this arm was moving up and down. this movement was transferred to the teat cups and made the teats erecting. after stimulation time, the main milk phase began and the milk flow was observed on the display. when the milk flow has decreased to a certain level, the milking process was automatically stopped by detaching the milking unit. after milking process is completed, the milking unit was cleaned and the milk volume was immediately measured using a graduated cylinder. agriculture and food sciences research, 2019, 6(2): 172-181 174 © 2019 by the authors; licensee asian online journal publishing group 2.5. milk sample collection milk samples were collected weekly during a period of 8 weeks and qualitative examinations were done. three groups of milk samples were collected, namely: 2.5.1. group 1 (gr.1) milk sampling used to detect the presence of pathogen bacteria in the udder: milk samples were taken from each quarter after the udder being washed, dried, disinfected, the first stripping was removed, then the sample tube was opened and about 5 to 10 ml of milk was taken from each quarter in each tube. afterward, the tubes were immediately closed, maintained in a cool box at 8 c° using ice cubes and transported to the laboratory for the pathogenic bacteria determination. 2.5.2. group 2 (gr.2) milk sampling to determine the milk composition and milk quality: after machine milking, milk samples were taken in duplicate, ca. 25 ml in each in special tubes with preservatives for the determination of fat, protein, lactose, scc and other tubes with other preservatives for determination of bc. 2.5.3. group 3 (gr.3) milk sampling to determine the effect of storage times and temperatures on raw milk parameters: milk samples were taken and divided as shown in the table 1: table-1. storage of raw milk in different times and temperatures. studied time room temperature (24 c° ) refrigerator temperature (4 c° ) immediately after milking (zero time, 0) x x after 2 hours of milking x x after 6 hours of milking x x after 24 hours of milking x x after 48 hours of milking x x source: kaskous, s. after the storage times and temperatures shown in the table, the milk samples were taken and analyzed for fat, protein, lactose, scc and bc. 2.6. milk sample analysis milk samples gr.1 in the individual quarters were examined using methods met-egd-001-005. the investigation was applied in the animal health service bavaria e.v. however, the determination of the milk parameters in milk sampling gr. 2 und gr. 3, they were performed on: fat and protein, routine procedure of ir spectroscopy was used. nevertheless, lactose content was determined using the routine methods mpr-met 01. scc of the milk was determined by fluorescence optical counting (asu l01.01-1). while, bc was determined by flow cytometric enumeration of microorganisms (asu l01.01-7). all of the milk samples were analyzed in milchprüfring bavaria e.v. 2.7. data analysis the data were subjected to statistical analysis of variance using sas program package [14] and the least square means were compared using f-test for the influence of the week or the time and temperature on milk quality and composition. the results were shown as lsm±se. 3. results and discussion 3.1. examination findings for quarter milk samples in none of the tested milk samples pathogenic bacteria could be shown, i.e. all quarters were healthy during the study period. it is known that the milk of a healthy udder is sterile and it is only contaminated with bacteria when the milk passes through the teat canal to the outside [9]. moreover, johnson, et al. [12] found the same results and reported that a high number of non-lactating dromedary glands are either sterile or harbor only a low number of mainly non-mastitis pathogens. the main reason that most udders remain sterile is the implementation of hygienic measures in the farm. however, the mammary gland of dromedaries is protected by a variety of defense mechanisms like innate or specific immunity as well as physiological particularities [12]. furthermore, dromedary milk itself possesses powerful antibacterial and antiviral proteins [3, 15, 16]. but, in many african and asian countries not all camels are healthy (apparently healthy), though their milk is sterile and has no pathogenic bacteria. it is shown in many studies that camel milk is a source for many bacteria which may lead to health hazard for men when it is taken raw [10, 17-20] because raw camel milk is produced and handled under poor hygienic conditions with high health risk to the consumers [21]. however, in this study there was no contamination to the camel’s milk either from the animal itself, the stable, the farmer or the feed, because the hygienic procedures at the research station were ideal. furthermore, we used sanitized milking machine (st-c) in this study, which kept the udder clean and healthy. consequently, the milk remains clean without pathogenic bacteria. therefore, we recommend the use of the milking machine in camels. since the contamination with pathogenic bacteria during hand milking is very high as compared to machine milking [22]. 3.2. camel milk parameters 3.2.1. fat content the mean content of fat in camel milk during the study period was 2.92±0.07% and it ranged between 2.70±0.18 and 3.30±0.18 %. the figure 1 shows significant (p<0.05) and continuous increase in fat content in the milk with progress in the milking season until the eighth week of the study. these results are identical with some agriculture and food sciences research, 2019, 6(2): 172-181 175 © 2019 by the authors; licensee asian online journal publishing group results in the international studies [23-27]. however, fat content of camel milk varies greatly from 2 to 5% percent in the literature depending on many factors such as weather conditions, stage of lactation, feed, presence of water, country, and milking method [28-30]. our results showed that the percentage of fat in the camel milk was somewhat low. perhaps these low levels of fat contents were due to the camel milk samples that were taken in the summer. this results were confirmed by haddadin, et al. [31] as they had noticed the lowest percentage of fat in august (2.5%), and a higher percentage in the middle of winter (3.9%). other factors may affect the low fat content in the camel’s milk may be due to the fact that animals were milked once daily only. this fact was shown in a study that total solids, fat content, and milk ph decreased by increasing milking interval, showing the greatest value at 8-hr intervals and the lowest at 24-hr intervals [32]. similar results were obtained by alshaikh and salah [33] that the organic component of camel milk such as fat, protein, solids-not-fat (snf) and lactose decreased with increasing milking intervals and concurrently the secretion rates of milk was decreased. moreover, the results indicated that the fat content in the milk increased continuously with advanced stage of lactation. this increase was known in all lactating ruminants, so that most of milk components (total solids, ts, fat, protein and ash) increased at the end of lactation. in addition, always the presence of feed and water also may affect the fat content of camel milk significantly, so that milk fat concentration became higher by increasing the degree of dehydration in the camel [30]. however, alwan, et al. [27] reported that the rearing conditions were observed to significantly affect some milk components. in our study, the feed was enough and the water was free for the animals during the investigation period and the ambient temperature was between 22 and 27 co. therefore the fat content in the raw camel milk was quite normal figure 1. week 1 2 3 4 5 6 7 8 f at % 2.4 2.6 2.8 3.0 3.2 3.4 3.6 a a ab ab ab bc bc c figure-1. lsm ± se of fat content of raw camel milk during the trial period. means without common letters differ significantly (p<0.05) source: kaskous, s. 3.2.2. protein content the mean content of protein in camel milk was 2.28±0.01% and it ranged between 2.21±0.03 and 2.37±0.03 %. the figure 2 shows significant (p<0.05) continuous increase in protein content of the milk with progress in the stage of lactation until the eighth week of the study. the protein content in this study was similar to protein content with other studies as in ellouze and kamoun [34] with 2.29%; raghvendar, et al. [35] with 2.30%, but more than the results of omer and eltinay [36] with 2.06%. however, the protein content in this study was quite low, compared to the majority of authors as mal, et al. [37]; mal, et al. [38] with 3.73% and 3.89%, respectively; bakheit, et al. [39] 3.4% and the average is 3.1±0.5% [40]. in general, the protein percentage of raw camel milk ranges between 2.15 and 4.90% [41] or between 2.30% and 3.95% [1] or between 3 and 3.90% [42]. on the other hand, this protein content in the milk under german conditions may be normal, since water is free and green forage or hay are available throughout the year. under these conditions the protein synthesis in the udder was going well. on the contrary, bekele, et al. [30] reported that, protein content of camel milk is not significantly altered in the presence of water and feed. in general, camel milk contained less protein, fat and lactose than bovine milk [43]. week 1 2 3 4 5 6 7 8 p ro te in % 2.15 2.20 2.25 2.30 2.35 2.40 2.45 a ab ab ab ab b ab ab figure-2. lsm ± se of protein content of camel milk during the trial period. means without common letters differ significantly (p<0.05) source: kaskous, s. agriculture and food sciences research, 2019, 6(2): 172-181 176 © 2019 by the authors; licensee asian online journal publishing group 3.2.3. lactose content the mean content of lactose in camel milk was 3.91±0.02% and it ranged between 3.81±0.04 and 4.04±0.04%. the figure 3 shows significant (p<0.05) and continuous decrease in lactose content in the milk with the progress in the lactation stage until the eighth week of the study. it shows that the lactose content of camel milk is slightly lower than cow’s milk with 4.9%. similar results were also reported by elamin and wilcox [44]. the continuous reduction of the lactose content in the milk in this study was normal, since all the camels reached the end of lactation, then. this process is not only limited to camels, but also with all the lactating ruminants. however, many researchers reported that lactose content of camel milk varied between too extreme values 2.4 and 5.8 % as compared to 4.4 and 5.8 percent in cow’s milk [41] and the average was 4.4±0.7% [40]. it is known that the lactose content in the milk in true ruminants remains constant, since the lactose content of the milk plays an important role for the osmotic pressure of the mammary gland. therefore the lactose content of the milk remains unchanged. but the climate and feeding situation of camels are extremely different, therefore slight changes in the lactose content of camel milk was found in different parts of the world [31, 45, 46] and the nature of vegetation eaten by the camels in the desert areas could play a significant factor in the variation of lactose content in camel milk [42]. week 1 2 3 4 5 6 7 8 l a c to s e % 3.75 3.80 3.85 3.90 3.95 4.00 4.05 4.10 a ab ab ab b b b ab figure-3. lsm ± se of lactose content of camel milk during the trial period. means without common letters differ significantly (p<0.05) source: kaskous, s. 3.2.4. scc content the mean number of scc in raw camel’s milk was 126.43 x 103 ±7.21 cells/ml and it ranged between 83.00 x 103 and 178.00 x 103 cells/ml. similar results have been obtained by saleh and faye [47] and the mean value of scc was 125 x 103 cells/ml camel milk. the figure 4 shows the significant (p<0.05) differences in scc content in the milk during the study period. however, the concentration of scc in the raw camel’s milk was normal under german conditions and within the physiological level. hence, no mastitis could be detected either in clinical or subclinical analysis in the camels, as was written above. it is known that scc of the milk has been an indicator of the udder health. according to abdurahman [48] scc in raw camel’s milk were composed of macrophages, polymorphonuclear neutrophil granulocytes (pmn) and lymphocytes. in addition to those cells, a large number of cell fragments can be found in camel milk. these particles may have an influence on the diagnostic of udder health by determining the scc, as they could be counted with the somatic cells of the immune system. the cell count of a healthy cow udder has not been more than 100 x 103 cells/ml. however, it has not been known value for the cell number in the healthy camel milk. therefore, a limit between healthy and sick udder based on the cell count in the milk in camels has not yet been established, because there has been a problem knowing the basal levels of cells and their physiological variations in the camel, until now. furthermore, investigations have clearly shown that, quarters infected with bacteria had higher mean values for scc than non-infected quarter [47] and in inflamed quarter’s polymorph nuclear granulocytes dominated the sample and in non-inflamed quarters the dominant cells were epithelial cells and cell fragments [49]. however, an increase in the number of scc, particularly polymorphonuclear neutrophil granulocytes in camel milk is a strong indication of inflammation. this has been the same reaction as in the cow when the udder is inflammated. interestingly enough, hamed, et al. [50] found that lymphocytes were the predominant cell type in camel’s milk and macrophage in cow’s milk, when the cell counts are less than 100.000 cells/ml milk. furthermore, in raw camel’s milk pmn numbers were higher immediately after parturition and declined gradually with advanced lactation, while macrophages number increased through lactation. in cow’s milk, pmn were the dominant cell type at the beginning of the lactation and tended to maintain at high levels as lactation progressed. however, limits must be laid down for assessing the state of udder health. nagy, et al. [25] reported that the mean of scc in camel milk tank was 394 x 103 cells/ml and it ranged between 113.702 x 103 and 927.423 x 10 3 cells/ml. however, most of the milk samples (75%) have higher cell numbers (between 300 to 500 x 103 cells/ml) compared to our results. the study acknowledges that, the collected camel milk samples for the scc determination were obtained from both the entire herd (tank milk) over 2 year period and not from a short period as in this study. perhaps such reason showed the high cell count difference in the camel milk in that study, moreover, the animals were on a farm in the united arab emirates (uae). another studies, explored that the cell count in the camel milk was high as in woubit, et al. [51] with a range from 3 x 105 to 1.5 x 107 leukocytes/ml of camels milk and in guliye, et al. [52] with a range from 1.01 x 105 to 11.78 x 106 cells/ml camel milk. surely enough under the asian and african conditions the cell number in the camel milk was higher than the ones in europe, and a good hygienic conditions were difficult to achieve due to their high temperature and lack of agriculture and food sciences research, 2019, 6(2): 172-181 177 © 2019 by the authors; licensee asian online journal publishing group laws and regulations as compared in europe. therefore, abdurahman [49] has concluded that early problem recognition with improved hygienic measures, will result in reduced milk losses due to mastitis hence, increase the availability of milk for human consumption and sale. week 1 2 3 4 5 6 7 8 s c c x 1 0 0 0 m l m ilk 50 100 150 200 a ab ab b ab ab ab ab figure-4. lsm±se of somatic cell count of camel milk during the trial period. means without common letters differ significantly (p<0.05) source: kaskous, s. 3.2.5. bc content the mean number of bc in raw camel milk was 23.88 x 103±0.57 bc/ml and it ranged between 21.00 x 103 and 26.00 x 103 bc/ml. figure 5 shows the differences (p>0.05) in bc content in the milk during the study period. it was found that the non-pathogenic bacteria number in the camel milk was normal during the course of the experiment, and the udders were healthy, too. nevertheless, the milk was contaminated with non-pathogen bacteria after milking process. however, in the literature, it has been little information available on the total bacterial number in the raw camel’s milk, and the authors did not directly measure the total bacteria count. bacteriological examinations (blood agar) were carried out as standard methods as mentioned in some studies like abdurahman, et al. [53]; woubit, et al. [51]; yam, et al. [54] and abera, et al. [6]. however, ayadi, et al. [55] examined the total flora in the camel milk (32,097 x 103±396 ufc/ml), and the camels were healthy by the california mastitis test (cmt) during the investigation period. abdurahman, et al. [53] reported that 43.5% of the quarter camel milk samples yielded pathogenic bacteria in sudan. streptococcus agalactiae, streptococcus aureus, coagulase-negative staphylococci, and escherichia coli were isolated from these milk samples. other studies of younan, et al. [56] in kenya showed that 12 % of camel's milk samples were contaminated with s. agalactiae and 11 % with s. aureus. while aljumaah, et al. [57] found that 33 % of the tested quarters had subclinical mastitis based on california mastitis test. but, abera, et al. [58] have shown that clinical and subclinical mastitis was prevalent to 8.3% and 20.7% on the studied camel herd in ethiopia, respectively. it was concluded from many investigations that improvement in the hygienic measures must be applied in the farms in order to keep udders healthy and milking machine must be applied too to make a good milk quality and the udder remains healthy. it is known that by the application of milking machine in the camel the somatic cell counts in the milk and the microbiological contamination can be significantly reduced as compared to the hand milking [22]. finally, two points must be emphasized, the milking machine availability and the hygienic measures taken in the farm which, would improve the udder health of the camel, hence the resulting milk quality. in order to confirm this point, aljumaah, et al. [57] reported that poor hygiene of milking process had a higher prevalence of subclinical mastitis and intramammary infections. therefore the poor hygiene during milking is identified as a risk factor for occurrence of mastitis in camel [59, 60]. week 1 2 3 4 5 6 7 8 b ac te ria l c ou nt x 1 00 0 m l m ilk 18 20 22 24 26 28 30 a a a a a a a a figure-5. lsm±se of bacterial count of camel milk during the trial period. means without common letters differ significantly (p<0.05) source: kaskous, s. agriculture and food sciences research, 2019, 6(2): 172-181 178 © 2019 by the authors; licensee asian online journal publishing group according to most authors the composition of camel milk varies greatly and this variation could be attributed to many factors such as rearing conditions [27, 61] season [31] management systems [62] geographical location, feeding regime, breeds or camel types [63] milking frequency [32, 64] stage of lactation and parity [25, 41, 57]. 3.3. effect of different of storage time and temperature on raw milk quality statistical analysis has shown that the storage time and temperature have influenced on the camel milk parameters. table 2 clearly shows the significant (p<0.001) effect of the storage time on the parameters of fat, protein, lactose, scc and bc in the camel milk. the temperature of the milk during storage also had an effect on the camel milk parameters, too. however, the table reveal that, the temperature had a significant (p<0.001) effect on protein, lactose, scc and bc, but not on fat content of camel milk. additionally, the interaction between storage time and temperature has shown a great effect on protein, lactose, scc and bc table 2. table-2. statistical analysis of the effect of time and temperature on raw camel milk quality. effect nr df den df fat protein lactose scc*** bc**** f value pr>f f value pr>f f value pr>f f value pr>f f value pr>f t* 4 36 8.63 p<0.001 36.70 <0.001 69.08 <0.001 42.77 <0.001 739.32 <0.001 tm* 1 36 0.71 0.4064 103.74 <0.001 102.86 <0.001 114.31 <0.001 1346.59 <0.001 txtm 4 36 0.15 0.9618 77.84 <0.001 62.15 <0.001 44.62 <0.001 652.12 <0.001 note: t*: time, tm**: temperature, scc***: somatic cell count, bc****: bacterial count. table 3 shows the chemical characteristics of camel milk stored at room temperature (24±1.7 c°). fat, protein, scc and bc contents do not change significantly at room temperature for up to 6 hours and afterwards they had changed significantly (p<0.05) in 24 and 48 hours of storage. which confirm that, fat content and scc in the raw milk were decreased significantly while protein content and bc increased significantly when the raw milk was stored for more than 24 hours at room temperature. also, lactose content in the milk was decreased significantly (p<0.05) after 2 hours of storage at room temperature. the results of this study showed that milk quality changed after being stored of 24 hours at room temperature and were completely denatured after storing the milk sample 48 hours at room temperature. similar results have been found at +25 c° in algeria in dairy camel milk samples [65]. contrary to our results, omer and eltinay [36] found that the mean values of fat were 2.4, 2.7 and 3.1% for the 1st, 2nd and 3rd days of camel milk stored at room temperature, respectively. the same authors found similar changes to our study in the lactose content of camel milk during storage time at the room temperature, and the mean value of lactose content at room temperature decreased from 4.4% (day 1) to 3.34% (day 2) to 3.28% (day 3) in camel milk collection. it seemed that room temperature storage (24±1.7 c°) contributed to decrease in milk quality parameters. however, storing the camel milk at room temperature led to significant changes in the total number of microbes in this milk. these changes have led to several physical and chemical alterations in this milk. therefore, changes in the milk components have occurred. in practice, camel milk mostly stored for 24 hours at room temperature in many countries [66]. after 24 hours storage time, lactic acid bacteria may grow up leading to decrease in the milk ph and at that then camel milk will not be used for human consumption, which we confirmed in this study. furthermore omer and eltinay [36] reported that ph value in raw camel milk decreased as in our study during the storage time (3 days) at room temperature, and the mean value of ph was 6.6 at the day of collection which was reduced to 5.6 (2nd day) and to 5.4 (3rd day). another study from arabha, et al. [67] explored that scc in the camel milk samples= without adding preservative decreased significantly (p<0.0001) during 4 days of storage, however, such decrease was not noticed in samples with preservative. the results on milk samples stored at +4 c° during 0, 2, 6, 24 and 48 hours showed that lactose content, scc and bc were not changed table 4. however, it was also found that milk fat and protein contents were lower after being stored at + 4 c° during 24 and 48 hours (p<0.05). generally, it can be said that it is possible to store the raw camel milk at room temperature for 24 hours and at +4 c° for 48 hours without noticeable change. some studies have shown that when the cow milk was stored at 7 c°, total bacteria count was increased [65, 68]. however, this increase in bacterial count was reduced when the temperature becomes lower than +7 c°. it was reduced further when the storage temperature of the milk samples was decreases to +4 c°. many farmers from kazakhstan have sent their camel milk to moscow, but when this milk arrived, it was no longer tenable or acceptable. that is why the cold chain (not more than 8 degrees) must be applied, starting from the date of collecting the milk from the camel udder until it is reached the consumer. the average generation time of microorganisms was around 20 to 30 minutes under optimal growth conditions (temperature: 25 to 35 c° and ph 6.65). table-3. raw camel milk quality after being stored at room temperature 24 c° during 0, 2, 6, 24 and 48 hours (t) after milking. t* (hour) fat (%) protein (%) lactose (%) ph scc** x 103/ml bc*** x 103/ml 0 2.70a 2.26a 4.04a 6.43a 178.00a 22.00a 2 2.70a 2.26a 4.03ab 6.43a 173.00a 42.20a 6 2.65ab 2.26a 4.00b 6.40b 178.00a 364.20a 24 2.62b 2.28a 3.99b 6.37c 140.81b 24620.00b 48 2.61b 2.57b 3.65c 5.57d 104.02c 63100.00c se**** 0.12 0.05 0.03 0.01 7.03 742.00 note: t*: time, scc**: somatic cell count, bc***: bacterial count, se****: standard error. least square means in the same column with different superscripts a, b, c, d are statistically significant different at p<0.05. agriculture and food sciences research, 2019, 6(2): 172-181 179 © 2019 by the authors; licensee asian online journal publishing group table-4. raw camel milk quality after being stored in the refrigerator at +4 c° during 0, 2, 6, 24 and 48 hours (t) after milking. t* (hour) fat (%) protein (%) lactose (%) ph scc** x 103/ml bc*** x 103/ml 0 2.70a 2.26a 4.04 6.43 178.00 22.00 2 2.67a 2.26a 4.04 6.43 177.20 35.40 6 2.64ab 2.28a 4.02 6.43 178.20 49.00 24 2.62b 2.22b 4.04 6.42 179.00 221.00 48 2.58b 2.20b 4.03 6.42 178.40 2099.80 se**** 0.12 0.05 0.03 0.01 7.03 742.00 note: t*: time, scc**: somatic cell count, bc***: bacterial count, se****: standard error. least square means in the same column with different superscripts a, b are statistically significant different at p<0.05. 4. conclusion • healthy and safe camel milk with normal composition can be obtained, since the hygienic measures in the farm are maintained and a milking machine could be used for the milk removal. • a healthy clean udder is a sterile udder and may produces milk free of pathogens germs. • it is possible to store the raw camel milk for 24 hours at room temperature and for 48 hours at the refrigerated temperature without any significant change in the milk quality. • the rapid cooling of raw camel milk after milking process may keep the milk quality in good condition, which may help the farmer to market the raw camel milk safely. however, the hygienic measures in the tropic and subtropical countries are not always available and most camel owners milk their camels with hand. • using milking machines “stimulactor” for camel’s guarantees to supply the market with healthy and high quality camel milk. references [1] a. k. yadav, r. kumar, l. priyadarshini, and j. singh, "composition and medicinal properties of camel milk: a review," asian journal of dairy and food research, vol. 34, pp. 83-91, 2015. available at: https://doi.org/10.5958/0976-0563.2015.00018.4. 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[68] b. faye and g. loiseau, "sources of contamination in dairy sectors and examples of quality approaches," in proceedings of the international workshop, cirad-fao, 11-13 december 2000, montpellier, france, 2000, p. 5. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn(e) : 2411-6653 issn(p) : 2518-0193 vol. 3, no. 2, 53-58, 2016 http://www.asianonlinejournals.com/index.php/aesr 53 study on biochemical compounds, antioxidant activity and organoleptic taste of some spice tea iftekhar ahmad1  tomal toru das2 md. yasin3 mohammad afzal hossain4 1,2,3,4department of food engineering and tea technology, shahjalal university of science and technology, sylhet-3114, bangladesh ( corresponding author) abstract many kinds of food additives are used in the food industry, among them spices are renowned. spice tea is good for health but little research has been done about this. so a study was conducted to determine the biochemical compounds, antioxidant activity and consumer acceptability of spice tea. three spices (cinnamon, cardamom and ginger) were used to develop individual spice tea. the results showed that protein content of cinnamon, cardamom and ginger tea are 14.41%, 18.59% and 17.5% respectively. caffeine, tannin, carbohydrate, lipid, moisture and ash content were found higher in different spice tea sample. antioxidant activity of cinnamon tea, cardamom tea, ginger tea and general black tea were found 94.82%, 90.33%, 86.33% and 87.80% respectively, which are higher than black tea (cntrl). during the tasting of cup quality, comparatively better result was found for cinnamon tea and ginger tea than black tea based on infusion, liquor color, briskness, strength and creaming down. keywords: spice tea, antioxidant activity, caffeine, tea, biochemical compounds. contents 1. introduction ............................................................................................................................................................................... 54 2. materials and methods ............................................................................................................................................................. 54 3. results and discussion .............................................................................................................................................................. 56 4. conclusion .................................................................................................................................................................................. 57 references ...................................................................................................................................................................................... 57 citation | iftekhar ahmad; tomal toru das; md. yasin; mohammad afzal hossain (2016). study on biochemical compounds, antioxidant activity and organoleptic taste of some spice tea. agriculture and food sciences research, 3(2): 53-58. doi: 10.20448/journal.512/2016.3.2/512.2.53.58 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: this work is licensed under a creative commons attribution 3.0 license contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 6 january 2016/ revised: 19 february 2016/ accepted: 23 august 2016/ published: 10 october 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.2/512.2.53.58 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=iftekhar%20ahmad https://orcid.org/orcid-search/quick-search?searchquery=tomal%20toru%20das https://orcid.org/orcid-search/quick-search?searchquery=md.%20yasin https://orcid.org/orcid-search/quick-search?searchquery=mohammad%20afzal%20hossain http://search.crossref.org/?q=10.20448/journal.512/2016.3.2/512.2.53.58 agriculture and food sciences research, 2016, 3(2): 53-58 54 1. introduction tea (camellia sinensis (l.) o. kuntze) was first used in china as a medicinal drink, and later became a popular beverage. nowadays, tea is one of the most popular beverages in the world and it is the most consumed beverage next to water. tea is an important crop in bangladesh. the total tea growing areas of bangladesh is divided into three fairly divergent ecological zonesnamely surma valley in greater sylhet, halda valley in chittagong and karatoa valley in panchagarh districts [1].at present, bangladesh producing 63 million kg of made tea per year and from there 93% of tea estates are situated in the greater sylhet zone. herbal teas are often consumed for their medicinal effects and especially for their sedative, relaxative, and stimulative properties. herbal teas are mostly popular because of their fragrance, antioxidant properties and therapeutic applications [2]. spices are mainly used for flavoring and they also have certain medicinal properties. a spice is a dried seed, fruit, root, bark or vegetative substance used in nutritionally insignificant quantities as a food additive for flavor, color, or as a preservative that kills harmful bacteria or prevents their growth. in bangladesh; cinnamon, cardamom and ginger are used as spice and they have many medicinal effects. so, addition of these spices with tea can play an important role in public health and can be used for medicinal purposes. cinnamon contains unique healthy and healing property due to the presence of active components.cardamom tea helps treat indigestion, prevents stomach pain, and relieves flatulence. drinking a cup of cardamom tea is helpful for women who experience mood swings during their menstrual period [3]. ginger is an energizer and a stimulator. drinking ginger tea both stimulates and soothes the digestive system. arthritic people have found ginger tea helpful since it has anti-inflammatory properties. it is good to fight against colds and flu [4]. spicing of the tea is one way of value addition but limited research has been done on the biochemical effect, antioxidant activity and organoleptic taste of spice tea for consumer acceptability. for this reason, it is very important to conduct this research and develop a spice tea to meet the following objectives. ➢ to evaluate the biochemical compounds (polyphenol, caffeine etc.) of some spice tea ➢ to determine the antioxidant activity of some spice tea. ➢ and organoleptic taste of some spice tea. 2. materials and methods this experiment was undertaken at the laboratory of department of food engineering and tea technology, shahjalal university of science and technology (sust), sylhet. before starting the experiment, samples (cinnamon, cardamom, ginger and black tea) were collected from different places of sylhet. preparation of spice tea: three types of spice (cinnamon, cardamom and ginger) are added with black tea as powder form and black tea without spice as control sample were used for this study. collected spices were sorted and heated in an oven at 105 ºc for 60 minutes to remove the moisture. then an electrical grinder was used to powder form the spices. in each sample, 50g of spice was added to 200g of black tea to make the spice tea [5]. 2.1. estimation of total carbohydrate about 1g of each tea sample was ground well (in mortar and pestle) with 100% ethanol. two ml of the alcoholic extract was diluted to 10 ml with distilled water and one ml of dilutent taken in a test tube and incubated under ice cold conditions; 4 ml of 0.2% ice cold acidified anthrone reagent was added to it. contents were then incubated in a boiling water bath for 8 minutes and cooled down to room temperature under running water. absorbance of the green color developed was read at 630 nm against the reagent blank in a uv-visible spectrophotometer and percent total carbohydrate (as dextrose equivalents) was expressed from standard calibration curve of dextrose [6]. 2.2. determination of protein content protein was determined using micro-kjeldahl. 2gm of sample was weighed and inserted in a 250ml kjeldahl digestion flask and taking care to see that no portion of the sample clings to the neck of the flask. then 2gm of digestion mixture and 25 ml of conc. sulphuric acid was also added to the digestion flask. the flask was kept in digestion chamber at 300°c to digest the sample. heating is continued for 2hrs until the color of the digest pale blue. the digest was cooled and transferred to a 250 ml volumetric flask. the digestion flask was rinsed 3 times with distilled water and transferred to the volumetric flask. the volume was made 250 ml by adding distilled water. 5ml of the digested sample was pipetted to a distillation flask and 60 ml of 40% naoh was also added to the flask. then it was transferred to the distillation chamber. on the other hand, 10ml of 2% boric acid, 10ml of distilled water were pipetted into a conical flask with 4 drops of distillation indicator that was worked as receiver. the conical flask was also set in receiving chamber. after that, the distillation flask was heated at 300°c for 1hr. the color of the solution in receiver turned pink to bluish green. the distillation process was turned off when the volume of receiver was made 60-70 ml.after distillation, titration was carried out with 0.1 n standardized hcl until the blue color was disappeared. for accuracy to determine the end point, the titration was further continued until a faint pink tinge appeared and subtracted from the burette reading 0.02 ml. blank was also carried out that contained no sample from digestion to titration. the calculation for protein is given below: nitrogen%= {𝑆𝑎𝑚𝑝𝑙𝑒 𝑡𝑖𝑡𝑟𝑒 − 𝐵𝑙𝑎𝑛𝑘 𝑡𝑖𝑡𝑟𝑒} × 𝑁𝑜𝑟𝑚𝑎𝑙𝑖𝑡𝑦 𝑜𝑓 𝐻𝐶𝑙 × 14 × 𝑣𝑜𝑙𝑢𝑚𝑒 𝑚𝑎𝑑𝑒 𝑢𝑝 𝑜𝑓 𝑡ℎ𝑒 𝑑𝑖𝑔𝑒𝑠𝑡 × 100 𝐴𝑙𝑖𝑞𝑢𝑜𝑡 𝑜𝑓 𝑡ℎ𝑒 𝑑𝑖𝑔𝑒𝑠𝑡 𝑡𝑎𝑘𝑒𝑛 × 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑎𝑚𝑝𝑙𝑒 𝑡𝑎𝑘𝑒𝑛 × 1000 protein % = (nitrogen % × 6.25) 2.3. quantification of lipids 1g of tea sample was taken in a separating funnel and 25 ml of chloroform and methanol mixture (2:1) was added into it. five ml of 0.9% sodium chloride was added and the mixture was shaken well. the mixture was allowed to stand still for 30 minutes and there was a separation of layers. the chloroform layer was carefully transferred to a pre-weighed china dish. entire extraction procedure was repeated twice and extractants were pooled together. pooled chloroform extract was evaporated to dryness on a boiling water bath. china dish with lipids was agriculture and food sciences research, 2016, 3(2): 53-58 55 dried in an oven at 105 oc and then weighed. from the difference in weights the percentage of lipids present in the leaf material was calculated gravimetrically and expressed as percentage according to ravichandran and parthiban [7]. lipid (%) = (weight of dried dish – weight of empty dish) × 100 2.4. moisture content determination the moisture content was measured according to the official method 44-o1 of aacc [8]. the moisture content of the sample is calculated using the following formula. dry matter (%) = {(𝑊𝑡. 𝑜𝑓 𝑑𝑟𝑖𝑒𝑑 𝑠𝑎𝑚𝑝𝑙𝑒(𝑔)) × 100}/𝑊𝑡. 𝑜𝑓 𝑓𝑟𝑒𝑠ℎ 𝑠𝑎𝑚𝑝𝑙𝑒(g) moisture (%) = 100 – dry matter. 2.5. determination of total ash total ash was estimated by directly incineration of sample taken in a crucible according to aacc [8] method 08-01. ash was calculated as 𝐴𝑠ℎ(%) = wt. of ash wt. of sample × 100 2.6. estimation of polyphenols about 1g of each tea sample was ground well (in mortar and pestle) with 100% ethanol and one ml of the alcoholic extract was diluted to 50 ml with distilled water. then two ml of diluted extract was added with 4 ml of 1:1 folin-ciocalteu’s reagent and water mixture, and 2 ml of 35% sodium carbonate. the contents were further made up to 10 ml with distilled water and the mixture was shaken thoroughly and allowed to stand still for 30 minutes. absorbance of the blue color developed was read at 700 nm against the reagent blank using uv-visible spectrophotometer. quantum of polyphenols present in tea was computed using the standard calibration curve derived from known concentrations (10 to 50 ppm) of gallic acid and the results were expressed as percent gallic acid equivalents [9]. 2.7. estimation of theaflavin (tf), thearubigin (tr), highly polymerized substances (hps) and total liquor colour (tlc) two grams of each tea sample were weighed and transferred in a 250 ml conical flask. 100 ml of boiled water was added to the sample and the contents were infused over the boiling water bath for 10 minutes with intermittent shaking. it was then filtered through cotton wool and the analysis was carried out. solvent extraction of tea extract was carried out in separating funnels with adequate shaking at every stage. contents of tf, tr, hps and tlc were calculated from the absorbance values where, tf (%) = (4.313 x c x 2 x 100) / (sample weight x dmc); tr (%) = (13.643 x (b+d-c) x 2 x 100) / (sample weight x dmc); hps (%) = (13.643 x e x 2 x 100) / (sample weight x dmc); tlc (%) = (10 x a x 2 x 100) / (sample weight x dmc). multiplication factors of tf and tr were derived from molar extinction coefficients of pure compounds and dilution factor [10]. in the case of tlc, value 10 is the dilution factor [11]. fig-1. estimation of tf, tr, hps, and tlc in black tea by spectrophotometer [10, 11] 2.8. estimation of caffeine at first 50 ml distilled water was heated at 40 ºc and then 100mg tea sample was added to the hot water and stirrer for 30 min with magnetic stirrer. then it was filtered and cooled to room temperature. 50 mi of chloroform was poured into the infusion and stirrer for 10 min with magnetic stirrer without additional heat. then the mixture was transferred to a separating funnel and allows standing still for 30 minute for the separation of caffeine. organic phase (chloroform) was separated from the water phase. organic solution was poured into quartz of uv cell and absorbance was taken in 260nm. quantum of caffeine present in tea leaves was computed using the standard calibration curve of caffeine and the results were expressed as percent caffeine equivalents [12]. 2.9. determination of total tannins content folin-ciocalteu phenol reagent was used to determine the total tannins content as reported by amorim, et al. [13]. at first, 0.2 ml of the sample extract was added with 8.3 ml of distilled water and 0.5 ml of folin-ciocalteu phenol reagent was added and kept at room temperature for 5 minutes. then 1 ml of 35% sodium carbonate was agriculture and food sciences research, 2016, 3(2): 53-58 56 added. the mixture was shaken well, kept at room temperature for 20 minutes and absorbance was measured at 725 nm. blank was prepared with water instead of the sample. a set of standard solution of tannic acid was read against a blank. total tannin content was determined as mg of tannic acid equivalent per gram using the equation obtained from a standard tannic acid calibration curve. 2.10. determination of antioxidant activity the scavenging effects of spice tea samples for 1,1-diphenyl-2-picrylhydrazyl (dpph) radical were measured according to the method by chan, et al. [14]. briefly, 2.0 ml aliquot of test sample (in methanol) was added to 2.0 ml of 0.16 mm dpph methanolic solution. the mixture was vortexed for 1 min and then left to stand at room temperature for 30 min in the dark because dpph is very much light sensitive, and then absorbance was measured at 517 nm in uv spectrophotometer. the ability to scavenge the dpph radical was calculated using the following equation: scavenging effect (%) = [1 (a sample – a sample blank)/ a control] × 100 where, ‘a control’ is the absorbance of the control (dpph solution without sample); ‘a sample’ is the absorbance of the test sample (dpph solution plus test sample) and ‘a sample blank’ is the absorbance of the sample only (sample without dpph solution). 2.11. organoleptic taste the liquor was prepared by pouring boiling water in a mug of a capacity of 142ml (about 0.25 pint) in which 2.5 gm (equal in weight to a 25 paisa coin) was contained. after 5 minutes of brewing the liquor was poured into a bowl and infused leaf was shaken from the mug into the inverted lid, which was placed on top of the mug. then the dry leaf, the infused leaf and the liquor was evaluated.in order to taste the liquor some of the liquor was taken into the mouth with a sucking noise. the liquor was swilled around the tongue and brought into contact with the plate and gums. in this way the thickness of the liquor was assessed by judging its viscosity, its bitterness by the taste on the back of the tongue, and its astringency and pungency by the sensation apprehended on parts of the cheek and the gums. all these factors together make up the briskness; strength and body of the liquor. tea aroma and flavor were assessed by drawing liquor to the back of the mouth up to the olfactory nerve in the nose. a mouthful of liquor is thus felt, tasted and smelled and after tasting spit out into a spittoon [15-17]. 2.12. statistical analysis the experimental data were statistically analyzed by ibm spss statistics version 20 statistical software. the results are expressed as mean and data were statistically analyzed by one-way anova, with the level of significance set at p<0.05. the mean values adjusted by duncan’s multiple range test (dmrt). 3. results and discussion 3.1. total carbohydrate, protein, lipid, moisture and total ash content tests were conducted to determine these values in different tea sample including a control sample. the results obtained from the tests are given below tea sample carbohydrate % protein % lipid % moisture % ash % cinnamon 66.25d±0.88 16.41a±1.09 4.80c±0.16 7.45a±0.24 4.71a±0.23 cardamom 58.85a±0.71 18.59b±1.09 3.89a±0.08 7.51a±0.18 6.38c±0.19 ginger 62.73c±0.81 17.5ab±1.09 4.28b±0.19 8.38c±0.17 5.69b±0.20 general(cntrl) 60.75b±0.89 20.78c±1.09 4.41b±0.20 7.92b±0.22 5.75b±0.12 note: {values are expressed as mean±sd of three observations, different letter(s) are significantly different by dmrt (p>0.05)} from the table it is clear that, the amount of carbohydrate and lipid is higher in cinnamon tea, protein is higher in general tea. moisture is higher in ginger and ash in cardamom tea. graham [18] and ahmad, et al. [17] found same kind of result in case of black tea [17, 18]. 3.2. estimation of polyphenol, caffeine, total tannins content polyphenol, caffeine and total tannins value are given below tea sample polyphenol (ppm) caffeine (ppm) total tannins (ppm) cinnamon 74.62c±1.54 52.72c±1.02 45.00c±1.02 cardamom 58.98a±1.18 37.72a±0.98 33.87a±1.25 ginger 68.46b±0.77 43.04b±1.26 32.37a±1.06 general (cntrl) 77.69d±0.77 38.64a±0.98 37.25b±0.95 note: {values are expressed as mean±sd of three observations, different letter(s) are significantly different by dmrt (p>0.05)} analyzing this table we can say that caffeine and tannin are found good in spice teas but in case of polyphenol it was found high in normal black tea.this kind of result also found by mohammad shameem, et al. [19] in case of black tea [19]. 3.3. estimation of tf, tr, hps and tlc theaflavin (tf), thearubigin (tr), highly polymerized substances (hps), total liquor color (tlc) was analyzed to know the difference between spice tea and general tea. agriculture and food sciences research, 2016, 3(2): 53-58 57 tea sample tf % tr % hps % tlc % cinnamon 0.5179 5.2777 9.4941 2.6258 cardamom 0.4878 4.8533 8.9838 2.3679 ginger 0.5359 5.8675 9.3077 2.4997 general (cntrl) 0.5621 5.9414 9.7196 2.9095 from the table, it is clear that, lowest values of tf, tr, hps and tlc contents are in cardamom tea and highest values of tf, tr, hps and tlc contents are in general tea (cntrl). 3.4. determination of antioxidant activity the method is based on the reduction of alcoholic dpph solution in the presence of a hydrogen-donating antioxidant due to the formation of non-radical form dpph-h by the reaction. tea sample scavenging effect (%) 500 (mg/ml) 50 (mg/ml) 5 (mg/ml) cinnamon 94.82d±0.57 60.81c±0.31 53.74c±0.40 cardamom 90.33c±0.59 58.16b±0.54 52.95bc±0.28 ginger 86.33a±0.69 57.79b±0.55 52.43b±0.55 general(cntrl) 87.80b±0.41 53.42a±0.55 43.54a±0.55 note: {values are expressed as mean±sd of three observations, different letter(s) are significantly different by dmrt (p>0.05)} from the table, it is clear that the antioxidant activity of cinnamon tea, cardamom tea and ginger tea is higher than the general tea. so, spice tea shows more scavenging activity than general tea. 3.5. organoleptic taste different types of spice tea were evaluated in regarding to the cup quality of made tea. the tea was remarked as ‘excellent’, ‘above average’, ‘average’ and ‘below average’ based on the score obtained from organoleptic taste. the tea obtained above 34 marks was remarked as “excellent”; above 32 to below 34 was remarked as “above average”; above 30 to below 32 was remarked as “average”; and below 30 was remarked as “below average” [15-17]. cup quality of cinnamon tea, cardamom tea and ginger tea was found as “average” grade but general tea was found as “below average”. spice tea was found having comparatively showed the better characteristics. ginger tea got the highest total mark. cinnamon tea and ginger tea was good in liquor color and strength. tea sample quality attributes total score 50 remarks infusion 10 liquor color10 briskness 10 strength 10 creaming down10 cinnamon tea 6.19 6.69 6.59 6.48 5.5 31.45 a cardamom tea 6.22 6.13 6.31 6.31 5.81 30.78 a ginger tea 6.69 6.25 6 6.38 6.44 31.76 a general tea (cntrl) 6.28 6.38 5.72 6 5.59 29.97 ba 4. conclusion tea is one of the most popular beverages and plays a vital role as a pharmaceutical agent. there are different brands of black and green tea which are commercially available in the market, having variation in their composition and quality. but addition of spices with black tea gives a better quality in its composition and also gives a better taste than general black tea. spice tea works as herbal and medicinal drinks. from the present study, we find that the antioxidant activity of different spice tea is higher than general black tea. as we know, some spices are used as herbs and works as medicine, so mixing spice with tea gives natural health benefits. from the result of the study show that these health benefits are found increased in tea because of addition of spices with black tea.spices have a great positive impact at consumer level in different ways and when it added with tea then popularity of spice tea increases. addition of spices with black tea has a good impact on the panelist rating as shown by the three best rated spice tea than general tea. ginger spiced tea and cinnamon spiced tea had the highest overall mean liking. therefore, this study recommends the best spice tea as cinnamon tea and ginger tea since they exhibited high antioxidant activity and the highest overall mean liking. from the obtaining results might be useful to define the formulation of spice tea, enhancing their health effect. references [1] m. s. a. mamun, "development of tea science and tea industry in bangladesh and advances of plant extracts in tea pest management," international journal of sustainable agricultural technology, vol. 7, pp. 40-46, 2011. [2] s. kundu, r. ghosh, p. choudhary, and a. prakash, "health benefits of various indian culinary herbs and comparative statistical analysis for organoleptic properties of indian teas by using analysis of variance anova," international journal of pharmacy and pharmaceutical sciences, 2014. [3] chandini and ravikumar, "review on herbal teas," journal of pharmaceutical sciences and research, vol. 6, pp. 236-238, 2014. [4] m.-k. hassan, t. behrouz, j. beman-ali, n. azadeh, and r. m. mohammad, "the effect of ginger powder supplementation on insulin resistance and glycemic indices in patients with type 2 diabetes: a randomized, double – blind, placebo – controlled trial," complementary therapies in medicine, vol. 22, pp. 9-16, 2014. [5] s. vaskar, production of different flavoured tea. the green tea book.kuthari agricultural management center. india: tamil nadu, 2011. [6] j. e. hedge and b. t. hofretier, in: carbohyd. chem. 17 (eds.) whistler, r. l. and bemiller, j.n. new york: academic press, 1962. agriculture and food sciences research, 2016, 3(2): 53-58 58 [7] r. ravichandran and r. parthiban, "lipid occurrence, distribution and degradation to flavor volatiles during tea processing," food chemistry, vol. 68, pp. 7-13, 2000. [8] aacc, approved methods of the american association of cereal chemists, 10th ed.: american association of cereal chemists, st. paul, mn, 2000. [9] m. n. d. choudhury and m. r. gowsamy, "a rapid method for determination of total phenolic matter in tea camellia sinensis l," two bud, vol. 30, pp. 59-61, 1983. [10] e. a. roberts and r. f. smith, "the phenolic substance of manufactured tea ix. the spectrophotometric evaluation of tea liquors," journal of the science of food and agriculture, vol. 14, pp. 689-700, 1963. [11] s. n. s. thanaraj and r. seshardi, "influence of polyphenol oxidase activity and polyphenol content in tea shoot on quality of black tea," journal of the science of food and agriculture, vol. 51, pp. 57-69, 1990. [12] a. b. m. a. maidon, a. o. mansoer, and h. sulistyarti, "study of various solvents for caffeine determination using uv spectrophotometer," journal of applied sciences research, vol. 8, pp. 2439-2442, 2012. [13] e. l. c. amorim, j. e. nascimento, j. m. monteiro, t. j. s. peixotosobrinho, t. a. s. araújo, and u. p. albuquerque, "a simple and accurate procedure for the determination of tannin and flavonoid levels and some applications in ethnobotany and ethnopharmacology," functional ecosystems and communities, vol. 2, pp. 88-94, 2008. [14] e. w. c. chan, y. y. lim, and y. l. chew, "antioxidant activity of camellia sinensisleaves and tea from a lowland plantation in malaysia," food chemistry, vol. 102, pp. 1214-1222, 2007. [15] btb, statistics on tea. nasirabad, chittagong: bangladesh tea board, 2011. [16] a. f. m. b. alam and s. k. l. haque, "comparative study on the productivity and cup quality of some promising test clones again standard clone bt1," tea journal of bangladesh, vol. 37, 2001. [17] i. ahmad, a. z. m. s. mazumder, f. b. sumi, m. a. hossain, and m. m. hoque, "physico-chemical characteristics and assesment of cup quality of tea collected from various tea estates of different countries," journal of applied science and technology, vol. 9, 2013. [18] h. n. graham, "green tea composition, consumption, and polyphenol chemistry," preventive medicine, vol. 21, pp. 334-350, 1992. [19] a. m. mohammad shameem, m. md, a. mainuddin, and y. md, "physiological and biochemical changes in tea leaves and made tea due to red spider mite infestation," asian journal of plant science, vol. 15, pp. 16-25, 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 30 agriculture and food sciences research vol. 5, no. 1, 30-46, 2018 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2018.51.30.46 impact of climatic and non-climatic factors on sustainable livelihood security in gujarat state of india: a statistical exploration ajay kumar singh1  jofri issac2 ( corresponding author): 1assistant professor (economics), department of humanities and social sciences, dit university, dehradun, uttarakhand-248009 india. 2research fellow, institute of rural management anand (irma), gujarat, india. abstract the present study estimates the district-wise sustainable livelihood security index (slsi) in gujarat using composite z-score technique during 2000-2011. it considers slsi as an integrated index of ecological security index (esi), economic efficiency index (eei) and social equity index (sei). accordingly, it applies linear and non-linear regression models to measure the impact of different climatic factors (i.e., maximum temperature, minimum temperature and precipitation) in winter, spring, summer and autumn seasons on slsi, esi, eei and sei. it shows that slsi, esi, eei and sei are varied across districts due to high diversity in socio-economic and ecological factors, and climatic change in gujarat. empirical results based on linear and non-linear regression models imply that climatic factors in different weather seasons have a negative and significant impact on slsi, esi, eei and sei. so, sustainable livelihood security (sls) would be in an alarming position due to climate change, urbanization, population growth and industrialization in gujarat. thus, policy makers need to adopt an effective and conducive policy to mitigate the adverse effects of those factors which have a negative impact on sustainable livelihood security. hence, this study provides several practical and viable policy suggestions to increase the sls in gujarat and other indian states. keywords: slsi, esi, eei, sei, gujarat. citation | ajay kumar singh; jofri issac (2018). impact of climatic and non-climatic factors on sustainable livelihood security in gujarat state of india: a statistical exploration. agriculture and food sciences research, 5(1): 30-46. history: received: 19 march 2018 revised: 6 april 2018 accepted: 9 april 2018 published: 12 april 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 31 2. socio-economic and geographical structure of gujarat ........................................................................................................ 32 3. theoretical outline of sustainable livelihood security index and associated index ...................................................... 32 4. material and research methodology ........................................................................................................................................... 32 5. origination of empirical models .................................................................................................................................................. 35 6. descriptive and empirical findings ............................................................................................................................................. 37 7. conclusion and policy suggestions .............................................................................................................................................. 41 references .............................................................................................................................................................................................. 42 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.51.30.46&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/0000-0003-0429-0925 https://orcid.org/0000-0003-0763-9336 agriculture and food sciences research, 2018, 5(1): 30-46 31 1. introduction over and above the economic and policy factors, and climate change will affect the sustainable livelihood security (sls) of poor people due to their inadequate adaptation and lower coping capabilities in developing economies. sls is a key focus area for researchers and policy makers in developing economies [1]. also, it became a central research area for international development agencies in the 1990s [2]. sls is a multi-dimensional, multiinteracting and complex phenomenon of the society [3] which varies across individual, household, region, states and country. thus identification of sls is very debatable and complex issue for researchers, international development organizations and global policy makers [4-8]. livelihood is defined as the capability and assets which maintain and it make a better living standard of the peoples [2, 3, 9]. livelihoods also include food system outcomes which contribute to food security and social welfare [10]. the capability of an individual comprises selfesteem, security, happiness, stress, vulnerability, power and exclusion [3, 11]. capability approach adopted by many research organizations to assess the situation of livelihood security of peoples [12]. livelihood assets are categorized into natural, physical, economic or financial, human and social capital [2, 3, 13-15]. natural capital comprises soil, forest area, irrigated area, genetic resource, hydrological cycle and water resource. cash, credit, debit, savings, basic infrastructure, physical assets, production equipment and technology are the indicators of economic or financial capital [14, 16]. human capital comprises skills, knowledge, ability to labour, good health and physical capability [16]. social capital includes the social network, social claims, social relationship, affiliations and association [16]. livelihood security is a situation in which peoples are capable to overcome all stress which makes them socially and environmentally vulnerable and weak. while, sls is a specific situation in which peoples are capable to mitigate the negative impact of all activities such as natural disaster, climate change, economic crisis and social conflicts [9, 10, 14]. sustainable livelihood is a necessary condition to achieve sustainable development that includes ecological security [9]. the idea of sustainable livelihood is introduced by the brundtland commission on environment and development and united nations conference on environment and development in 1992 [16]. it provides a holistic approach for poverty eradication [14]. sls includes five indicators (i.e., diversification index, food security index, input self-sufficiency, benefit-cost ratio and women‘s participation in agriculture) to estimate the sls in india [12]. while, böhringer and jochem [17] observed three components (i.e. economic development, environmental development and social development) of sls. sls has significant interconnection with different indicators, thus, assessment of sls draw the attention of policy makers, government representatives, local stakeholders and international development organizations to take precautionary action. it may be useful to adopt effective policies to maintain, regulate, utilize and conservation of natural resource (i.e., land, water and air) to mention sls. earlier studies have estimated the sustainable livelihood security index (slsi) as a proxy for sls in most developing economies [3-8]. however, existing researchers could not assess the influence of various socioeconomic and climatic factors on sls using empirical models in developing economies. 1.1. sustainable livelihood security in india in india, sls is negatively influenced due to overwhelming urbanization, high population growth, extensive industrialization, reduction in arable land and forest area, degradation in groundwater availability, and climate change [18]. india could not create sustainable development path due to chronic poverty, income inequality, food insecurity, ineffective and inappropriate government policies, unfair food distribution policies, and extensive utilization of natural resource in production activities. poverty remains a major obstacle to achieve sustainable development, socio-economic development and human development in india [13, 19]. sustainable development meets the requirements of present population without compromising the ability of future generations to meet their own needs [20-22]. it is an integration of economic, human, social, institutional, technological and environmental development. however, most economies are not giving any attention to protecting the quality and quantity of natural factors which have a positive association with sls. hence, it is vital to maintain and protect utilization of natural resource and other environmental factors to meet the livelihood needs of coming generation [12]. in india, agricultural production activities play a significant role to provide sls, employment opportunities, food security and poverty alleviation, as around 54% indian workforce are engaged in agricultural [23-25]. in india, agricultural sector feeds vast population which stands second on the world. thus, climate change and overexploitation of the natural resource would bring several threats for sls, poverty eradication policies, farmer's income, job opportunities, health situation and regional-development disparities across indian states [18, 23, 25]. india has a large pool of marginal and small farmers who are financially weak, therefore year to year variability in temperature, rainfall, precipitation, soil moisture, solar radiation would bring negative implications on the livelihood of a population [18, 23]. hence, agriculture is one of the significant factors which are positively associated livelihood security. application of fertilizer in agriculture has created several barriers to maintain environmental sustainability, human health, food quality and loss of bio-diversity [18]. 1.2. critical research questions and research objectives existing studies did not estimate sustainable livelihood index (slsi) for long-time data series in developing economies. also, there is insufficient literature available which applied robust and rigorous empirical model to analyze the influence of certain climatic and socio-economic variables on constructed slsi. limited studies estimate slsi for long-time data series at the district level in india [5, 8, 26]. nevertheless, these studies did not undertake an empirical investigation to assess the association of socio-economic and climatic factors with slsi in india using robust and rigorous empirical model. hence, it is essential to identify that how variability in socioeconomic variables (e.g., per capita income, urbanization, industrialization, population density, population growth, and forest area and literacy rate) and fluctuation in climatic factors (maximum and minimum temperature and precipitation) would affect the sls in india. due to existing research gap, the present study addresses the following research questions: • what is the association of sls with socio-economic and climatic factors in gujarat? agriculture and food sciences research, 2018, 5(1): 30-46 32 • what is the relation of sls with its components in gujarat? • how sls gets affected due to changing in climatic factors in gujarat? • which climatic and socio-economic factors have a significant influence on sls in gujarat? • how variability in socioeconomic and climatic factors would affect sls in gujarat in near future? • how sls varies across districts in gujarat? • how and why sls did get fluctuated during 2000-2011. pertinent to aforesaid research questions, the main aim of present study is to generate district-wise slsi using a composite z-index technique in gujarat. it investigates the association of slsi, esi, eei and sei with socioeconomic factors using correlation coefficient analysis technique. thereupon, it measures the influence of climatic factors on constructed slsi, esi, eei and sei using linear and non-linear regression models. 2. socio-economic and geographical structure of gujarat gujarat is the industrial hub of india, and its industrial sector contributes a large share in india’s gross domestic product (gdp) [27]. despite that, health and education status of gujarat is relatively at a lower position as compared to other states like kerala and tamil nadu. only 45% of the pregnant women in gujarat reach the hospital at the time of delivery.1 the maternal mortality rate is significantly higher in gujarat as compared to kerala and tamil nadu. so, primary health and education sector are in lower position in gujarat. in gujarat, total public expenditure on health sector has declined from 4.25% in 1990-95 to 0.77% in 2005-2010.2 it infers that health facilities in gujarat are not in a better position as compared to other states of india.3 sex ratio is a good indicator of human development and social development in an economy. the sex ratio has also declined from 920 in 2001 to 918 in gujarat in 2011 (census (goi), 2011). gujarat has around 18,539 villages, while the state has only 7,245 primary schools.4 thus gujarat could not create an appropriate platform for primary education. so, gujarat is in a poor position in social indicators, while economic growth of the state has increased due to heavy entrepreneurial activities of the people [27]. these, heavy entrepreneurial activities do have an insignificant impact on sls of peoples especially in the rural area in gujarat. so, high diversity in sls across districts is a very critical issue for policy makers in gujarat [5, 26]. 3. theoretical outline of sustainable livelihood security index and associated index numerous studies have estimated the sustainable livelihood security index (slsi) based on primary and secondary in different economies [4, 7, 17, 28-30]. existing studies estimate the slsi at the micro level (i.e., individual/household) using primary data and macro level (i.e., block/district/region/state/country) using secondary data. many indices have been developed by the scientific research community to measure the situation of human livelihood in term of slsi [4, 11] livelihood security index (lsi), household’s livelihood security index, livelihood vulnerability index (lvi). besides, scientific research community has also developed several indexes to assess the sustainable livelihood and environmental security of a region such as living planet index (lpi), ecological footprint (ef), city development index (cdi), human development index (hdi), environmental performance index (epi), environmental vulnerability index (evi), environmental sustainability index (esi), index of sustainable economic welfare (isew), genuine progress index (gpi), well-being index (wi), genuine savings index (gsi), and environmental adjusted domestic product (edp) [17, 20, 28-33]. these indexes integrate various components like socio-economic, demographics, livelihoods, social networks, health, food and water security and natural disaster to estimate slsi [7]. akter and rahman [6] estimated household's livelihood security index using economic, food, nutrition, health, education, empowerment, water and sanitation indicators of human livelihood. ajaero [3] estimates the livelihood asset index using asset indices analytical technique, and estimate the effects of livelihood asset indices on household's livelihood status in nigeria. kamaruddin and samsudin [7] used sustainable livelihood analysis framework to examine the capacity and preparedness of the rural poor in receiving entrepreneurial project channeled in malaysia. ponnusamy and gupta [4] developed a sustainable livelihood index (sli) using seven components such as environmental conservation, permanent asset creation, food security, nutritional security, input recycling, employment generation and annual income from different enterprises. singh and hiremath [5] estimated the slsi, which includes composite indices of ecological security index (esi), economic efficiency index (eei) and social equity index (sei) in india. sajjad, et al. [8] generates block-wise slsi with its three composite indices, i.e. esi, eei and sei for agricultural sustainability in bihar (india). ghabru, et al. [26] also estimated district-wise slsi using aforesaid indicators in gujarat. 4. material and research methodology brief outline of study area: gujarat is one of the leading industrialized states in india. it contributes more than 7.5% to india's gdp and 18% to india's fixed capital. it occupies 10% of india's factories and its manufacturing sector contributes 28% of its state gross domestic product. it has achieved strong annual gsdp of 10% during 2005-13, which is more than the national average during the same time period. however, gujarat is more environmentally unsustainable state of the country as compared to other indian states. there are several problems to sustain the livelihood security of the peoples. therefore, the present study focusses on gujarat. this study is based on secondary data, which includes district-wise panel data during 2000-2011. for this, eighteen districts with various economic zones of gujarat are considered in this study. 1 http://peoplesdemocracy.in/2014/0504_pd/education-health-gujarat. 2 http://peoplesdemocracy.in/2014/0504_pd/education-health-gujarat. 3 http://peoplesdemocracy.in/2014/0504_pd/education-health-gujarat. 4 http://peoplesdemocracy.in/2014/0504_pd/education-health-gujarat. http://peoplesdemocracy.in/2014/0504_pd/education-health-gujarat http://peoplesdemocracy.in/2014/0504_pd/education-health-gujarat http://peoplesdemocracy.in/2014/0504_pd/education-health-gujarat http://peoplesdemocracy.in/2014/0504_pd/education-health-gujarat agriculture and food sciences research, 2018, 5(1): 30-46 33 data sources and description: economic, food security, agriculture, education, women empowerment, ecological, climatic and geographical related variables at district level are taken from various sources like centre monitoring indian economy (cmie) pvt. limited; ministry of agriculture (gog); census (goi); department of animal husbandry dairying and fisheries (gog); planning commission (goi); national sample survey organization, department of statistics (goi); ministry of statistics and programme implementation (goi); indian meteorological department (goi). the spss, stata, minitab statistical software is used to construct slsi, esi, eei, and sei, and to run the proposed regression models. as some values are unavailable in time series, so interpolation and extrapolation techniques are applied to estimate the missing values to complete the time series [25, 34, 35]. 4.1. technique to measure the sustainable livelihood security index (slsi) (i) selection of variables: according to literature review, the selected variables have a scientific connection with sls [36]. also, there should not be high correlation among the variables [17]. as per the literature review, livelihood security is positively and negatively associated with different indicators, therefore, it essential to segregate undertaken variables in three categories i.e. ecological security, economic efficiency and social equity [8]. (ii) normalization or standardization-index of variables: normalization index is a technique in which a variable can be converted into one scale i.e. 0-1 [8, 15, 17, 18, 25, 26, 34, 36-41]. it makes individual variable as comparable across districts. if a variable is positively associated with sls, then standardization-index is estimated as ghabru, et al. [26]: siidt = {[xidt – min(xidt)]/[max(xidt) – min(xidt)]) (1) here, si is standardization-index or normalization-index for ith variables; d is cross-sectional districts; and t is time. min(xidt) and max(xidt) are the minimum and maximum values respectively in an individual indicator across districts [25, 34, 39, 40]. the value of estimated si lies between 0-1 for each indicator [15, 25, 37, 40-42]. if the value of an indicator (e.g., population density, population growth rate, urbanization, fertilizer consumption/hectare land, infant mortality rate) is negatively associated with sls [8] then si is estimated using below formula [26]: siidt = {[xidt – max(xidt)]/[min(xidt)max(xidt)]) (2) (iii) weight of arbitrary variables: appropriate weight to each indicator make index more scientific and rational [15, 17, 18, 25, 34, 36-41]. it provides right interrelationship between variables. weight of each indicator is examined as: 𝑊𝑖 = {𝐾/√(𝑉𝑎𝑟 (𝑆𝐼)) (3) here, wi is estimated weight (0<w>1 and ∑ 𝑊𝑖 = 1𝑚 𝑖=1 ) which is allotted to ith indicator [15, 25, 37, 39-41] var(si) is statistical variation across standardization-index for all indicators [25, 34, 37, 39, 41]. in equation (3), weight shows the reputation of an individual indicator. while, k is measured as: here, 𝐾 = 1 {∑ ( 1 √𝑉𝑎𝑟 (𝑆𝐼) )𝑚 𝑖=1 } (4) (iv) final slsi: slsi is a linear sum of all standardization-index that is multiplied by estimated weight of a specific indicator [17, 18, 25, 36, 37]. mathematically, it can be specified as: (slsi)dt=w1*(si_1)dt+w2*(si_2)dt+w3*(si_3)dt+…+wn*(si_n)dt (5) here, slsi is estimated sustainable livelihood security index; w1, w2, w3, …wn are the weightages for associated variables; si_1, si_2, si_3…si_n are the standardization-indexes for corresponding indicators [25, 37, 39, 40]. 4.2. mathematical functional relationship of slsi with its associated indicators as per literature review, sls is an integration of ecological security, economic efficiency and social equity [2, 4-8, 11, 12, 17, 26, 30]. hence, sls may be considered as a function of ecological security, economic efficiency and social equity [26]. in the present study, slsi is defined as a relative index which covers most variables and provides the relative position of a specific district in sustainable livelihood security as compared to other districts. hence, slsi is a function of ecological security index (esi), economic efficiency index (eei) and social equity index (sei). mathematically, above-mentioned relationship may be expressed as: (slsi)dt = f{(esi)dt, (eei)dt, (sei)dt) (6) here, slsi is sustainable livelihood security index; esi, eei and esi are the ecological security index, economic efficiency index and social equity index respectively; d is cross-sectional districts; and t is time period (2000-2011). these indicators can be indicated as: (esi)dt = f{(pd)dt, (pgr)dt, (fagca)dt, (ur)dt, (ci)dt, (hht)dt) (7) here, esi -ecological security index; pdpopulation density; pgrpopulation growth rate; fagcaratio of forest area with gross cropped area; ururbanization rate; cicropping intensity; and hhthouseholds having toilets. (eei)dt = f{(pcamp)dt, (fgy)dt, (giagca)dt, (fcphl)dt, (pcddp)dt, (aophl)dt) (8) here, eeieconomic efficiency index; pcampper capita availability of milk production; fgyfood-grain yield; giagcaratio of gross irrigated area with gross cropped area; fcphlfertilizer consumption/hectare land; pcddpper capita district domestic product; and aophlagriculture output/hectare land. (sei)dt = f{(lr)dt, (flr)dt, (hhea)dt, (phasw)dt, (imr)dt, (br)dt) (9) here, seisocial equity index; lrliteracy rate; flrfemale literacy rate; hheahouseholds having electricity accessibility; phaswpopulation having accessibility to safe water; imrinfant mortality rate; and br birth rate. esi, eei and sei would be linear sum of standardization-index or normalization-index which is multiplied by assigned weight of associated variable that is estimated as: (esi)dt =w1*(pd_si)dt +w2*(pgr_si)dt +w3*(fagca_si)dt +w4*(ur_si)dt +w5*(ci_si)dt +w6*(hht_si)dt (10) agriculture and food sciences research, 2018, 5(1): 30-46 34 (eei)dt =w1*(pcamp_si)dt +w2*(fgy_si)dt +w3*(giagca_si)dt +w4*(fcphl_si)dt +w5*(pcddp_si)dt +w6*(aophl_si)dt (11) (sei)dt =w1*(lr_si)dt +w2*(flr_si)dt +w3*(hhea_si)dt +w4*(phasw_si)dt +w5*(imr_si)dt + w5*(br_si)dt (12) here, w1, w2, w3, w4, w5 and w6 are the weights for related variables; sis is the standardization-index or normalization-index for corresponding variables in equation (10), (11) and (12). final, slsi would be the sum of esi, eei and sei, which is assessed as: (slsi)dt = (esi)dt + (eei)dt + (sei)dt (13) 4.3. justification on variables (a) ecological security related indicators: ecological security is helpful to develop natural resource based economy in long-term [26]. therefore, following variables are considered under ecological security related indicators 1. population density: high population density is caused to increase the additional climate vulnerability and high pressure on agricultural [8, 15, 18, 26, 34, 36, 39, 40, 43-45]. it also increases additional pressure on ecological variables. ecological security is also negatively associated with extensive population density. so, high population density would be negatively associated with sls. 2. population growth rate: high population growth requires more natural resource to sustain basic livelihood (e.g., food, land, transport, water, air) of peoples [8, 11, 13, 15, 18, 26, 39, 40, 44, 45]. it also increases ghgs emission in the atmosphere, thus it is significant cause to increase climate change. subsequently, high population growth is negatively associated with environmental factors and sls. 3. ratio of forest area with gross cropped area: forest area is an important factor to absorb co2 emission from human and economic activities and it mitigates the negative impact of climate change in agricultural and other production activities [5, 8, 13, 15, 18, 25, 26, 35, 36, 39, 40, 43, 45]. forest area also provides the livelihood opportunity in term of hunting, wood, timber, other [1]. also, forest area maintains the balance in ecological variables, therefore it is a better adaptable technique to mitigate the adverse effects of climate change in production activities, and thus it is helpful to increase sls. hence, the ratio of forest area with a gross sown area is used as a proxy as a crucial indicator of ecological security in this study. 4. urbanization rate: the association between urbanization rate and sls is complex. as urbanization is caused to increase carbon emission, therefore it is a significant contributor to increase climate change [15, 36, 40, 4345]. hence, sls is negatively correlated with urbanization rate in those economies which are unable to maintain the quality and quantity of natural resource due to higher urbanization [25]. 5. cropping intensity (ratio of a gross cropped area with net sown area): cropping intensity is a crucial indicator to increase the crop production [8, 15, 18, 25, 26, 36, 39, 40, 43, 45]. thus, it useful to improve the farmer’s income and their livelihood security. 6. households having toilets: sanitation facilities are an essential component to maintain the ecological security, thus households having toilets is considered as a proxy for sanitation facility in this study. table-1. description of ecological security related indicators and sources of data ecological security related indicators indicators unit symbol source population density number pd gujarat social development infrastructure board society, general administrative department (planning), government of gujarat population growth rate % pgr ratio of forest area with gross cropped area number fagca cmie pvt. limited urbanization % ur http://censusindia.gov.in/ cropping intensity (ratio of gross cropped area with net sown area) number ci cmie pvt. limited households having toilets % hht http://censusindia.gov.in/ source: author's compilation based on review of literature. (b) economic efficiency related indicators: economic efficiency provides the appropriate way to use of the natural and human resource through the application of technological advancement [26]. hence, following variables are compiled to estimate the eei: 1. per capita availability of milk production: milk is useful to meets the nutritional security to peoples [26, 43]. so, it is useful to increase sls. 2. food-grain yield: food-grain yield is useful to increase per capita availability of food-grain, thus food security and subsequently sls [18, 26, 39, 40]. 3. ratio of a gross irrigated area with gross cropped area: as irrigated area has a high yielding capacity than non-irrigated area [23, 25, 26, 34, 39, 40]. irrigated area plays a significant role to increase agricultural production and farmer’s income. irrigated area is supportive to increase the sls [9, 15, 18, 25, 36]. 4. fertilizer consumption/ha cropped area: suggested consumption of fertilizer in cultivation is useful to increase the crop productivity and agricultural sustainability [23, 26, 39, 40]. however, extensive application of fertilizer in cultivation deteriorates agricultural production activities. in addition, abundant fertilizer application in cultivation is caused to increase co2 emission in the atmosphere, thus it is caused to increase high probability for climate change [25]. hence, sls may decline due to extensive fertilizer application in cultivation. 5. per capita district domestic product: per capita income is crucial indicator to increase the food security. food security is a prime component of sls, thus it is helpful to maintain the livelihood security [39, 40, 43, 44]. hence, per capita district domestic product is included to capture the influence of this factor in sls. agriculture and food sciences research, 2018, 5(1): 30-46 35 6. agricultural output/hectare land: agricultural output/hectare land is useful to improve the farmer’s income [8]. income of agricultural labour and landless labour also improve as agricultural output/hectare land increases. subsequently, sls would be improve as agricultural output/hectare land increases. table-2. description of economic efficiency related indicators and sources of data economic efficiency related indicators indicators unit symbol source per capita availability of milk production kg/ annum pcamp livestock census, department of animal husbandry, dairying & fisheries, ministry of agriculture, goi food-grain yield tonne/ ha fgy land use statistics, ministry of agriculture, government of india, ministry of agriculture, goi ratio of gross irrigated area with gross cropped area number giagca agriculture and co-operation department, government of gujarat fertilizer consumption/ hectare land tonne/ ha fcphl cmie pvt. limited per capita district domestic product rs pcddp using sdp data from states of india, cmie agriculture output/ hectare land rs aophl cmie pvt. limited source: author's compilation based on review of literature. (c) social equity related indicators: social equity provides an equitable distribution of available resource, thereby present and future population can get the equal economic and social benefits from development [26]. therefore, following variables are used to create social equity index (sei): 1. literacy rate: literate population have more understanding to use available resources to achieve more benefits for livelihood security than the illiterate person [18, 25, 26, 36, 40, 43, 44]. so, it is a significant driver to increase sls for long-term. 2. female literacy rate: female literacy is the critical indicator of women empowerment which ensure the contribution of women in national development [5, 8, 26]. it is also useful to increase social development, thus female literacy rate is useful to increase sls. 3. population having electricity: availability of electricity may be useful to increase the social development [8, 26]. consequently, sls is positively and significantly associated with availability of electricity. 4. population having accessibility to safe water: safe water is useful to maintain the health status of peoples. so, sls would be improved as availability of safe drinking water increases. in this study accessibility of safe water is used as a proxy for health indicator. 5. infant mortality rate (rural + urban): as sls is positively associated with income, health and assets [10]. subsequently, it is also a critical indicator of social development [26, 36]. therefore, infant mortality rate is undertaken to capture the impact of health indicator on sls. 6. birth rate (rural + urban): birth rate is a critical indicator which is highly associated with sls. birth rate measures the effective position of the health sector, thus birth rate is included in this study. ghabru, et al. [26] used a number of primary health centres to create social equity index in gujarat (india). table-3. social equity related indicators and sources of data social equity related indicators indicators unit symbol source literacy rate % lr cmie pvt. limited female literacy rate % flr households having electricity accessibility % hhea http://censusindia.gov.in/ population having accessibility to safe water % phasw infant mortality rate number imr health and family welfare department, government of gujarat birth rate (rural + urban) number br source: author's compilation based on review of literature. (d) climatic factors average precipitation, maximum temperature and minimum temperature in winter, spring, summer and autumn seasons are used as climatic factors in this study [18, 25, 36, 38, 40, 43]. 5. origination of empirical models this study assesses the impact of climatic factors on sls and its associated indicators. so, slsi, esi, eei and sei are used as independent variables and regresses with climatic factors using linear and non-linear regression models. for this, the models are adopted from earlier studies which also uses estimated index as dependent variables to assess the impact of explanatory variables on it. demeke, et al. [46] used estimated food security index (fsi) as a dependent variable to assess the impact of socio-economic and climatic factors on it in ethiopia. kumar and sharma [47] also assess the impact of climatic and non-climatic factors on estimated fsi in india. ye, et al. [48] investigate the influence of climatic factors on estimated fsi in different climate change scenarios in china. belloumi [49] empirically investigate the impact of socio-economic and climatic factors on estimated fsi in esa countries. kumar, et al. [44] examine the impact of socio-economic variables on estimated global food security index across economies. tripathi [36] assessed the influence of socio-economic variables on estimated vulnerability index in uttar pradesh (india). singh [40]; sharma and singh [39] also assessed the influence of climatic factors on estimated fsi in india. also, singh, et al. [41] used several indexes like intellectual property awareness index, science & technological development index and socio-economic development index for empirically investigation across economies. in the present study, slsi, esi, eei and sei are considered as independent agriculture and food sciences research, 2018, 5(1): 30-46 36 variables and recursively regressed with climatic factors using linear and non-linear regression models. for this, following empirical models are used: (slsi)dt = β0 +βt (ttf) +β1 (amaxtwise)dt +β2 (amaxtspse)dt +β3 (amaxtsuse)dt +β4 (amaxtause)dt +β5 (amintwise)dt +β6 (amintspse)dt +β7 (amintsuse)dt +β8 (amintause)dt +β9 (apcpwise)dt +β10 (apcpspse)dt +β11 (apcpsuse)dt + β12 (apcpause)dt + udt (14) here, β0 is the constant coefficient; βt is the regression coefficient of time trend factor; β1… β12 are the regression coefficients of associated climatic factors; udt is the error term; d is the cross-sectional districts and t is time period in equation (14). the detail description of climatic factors is given in table: 4. table-4. description of climatic factors description of climatic factors indicators unit symbol sources average maximum tempereture in winter season 0c amaxtwise indian meteorological department (goi); indian institute of tropical meteorology (iitm); and geographic information system (gis) software average maximum tempereture in spring season 0c amaxtspse average maximum tempereture in summer season 0c amaxtsuse average maximum tempereture in autumn season 0c amaxtause average minimum tempereture in winter season 0c amintwise average minimum tempereture in spring season 0c amintspse average minimum tempereture in summer season 0c amintsuse average minimum tempereture in autumn season 0c amintause average precipitation in winter season mm apcpwise average precipitation in spring season mm apcpspse average precipitation in summer season mm apcpsuse average precipitation in autumn season mm apcpause source: author's compilation based on review of literature. (esi)dt = θ0 +θt (ttf) +θ1 (amaxtwise)dt +θ2 (amaxtspse)dt +θ3 (amaxtsuse)dt +θ4 (amaxtause)dt +θ5 (amintwise)dt +θ6 (amintspse)dt +θ7 (amintsuse)dt +θ8 (amintause)dt +θ9 (apcpwise)dt +θ10 (apcpspse)dt +θ11 (apcpsuse)dt +θ12 (apcpause)dt +ϕdt (15) here, θ0 is constant coefficient; θt is the regression coefficient of time trend factor; θ1… θ12 are the regression coefficients of associated climatic factors; and ϕdt is the error term in equation (15). (eei)dt= λ0 +λt (ttf) +λ1 (amaxtwise)dt +λ2 (amaxtspse)dt +λ3 (amaxtsuse)dt +λ4 (amaxtause)dt +λ5 (amintwise)dt +λ6 (amintspse)dt +λ7 (amintsuse)dt +λ8 (amintause)dt +λ9 (apcpwise)dt +λ10 (apcpspse)dt +λ11 (apcpsuse)dt +λ12 (apcpause)dt + €dt (16) here, λ0 is constant coefficient; λt is the regression coefficient of time trend factor; λ1… λ12 are the regression coefficients of related climatic factors; and €dt is error term in equation (16). (sei)dt= ξ0 +ξt (ttf) +ξ1 (amaxtwise)dt +ξ2 (amaxtspse)dt +ξ3 (amaxtsuse)dt +ξ4 (amaxtause)dt +ξ5 (amintwise)dt +ξ6 (amintspse)dt +ξ7 (amintsuse)dt +ξ8 (amintause)dt +ξ9 (apcpwise)dt +ξ10 (apcpspse)dt +ξ11 (apcpsuse)dt +ξ12 (apcpause)dt + τdt (17) here, ξ0 is the constant coefficient; ξ1… ξ12 are the regression coefficients of associated climatic factors; and τdt is the error term in equation (17). for non-linear regression models, the square term of all climatic factors are included in the above-mentioned equation (14), (15), (16) and (17) which are given as: (slsi)dt= α0 +αt (ttf) +α1 (amaxtwise)dt +α2 (amaxtwise^2)dt +α3 (amaxtspse)dt +α4 (amaxtspse^2)dt +α5 (amaxtsuse)dt +α6 (amaxtsuse^2)dt +α7 (amaxtause)dt +α8 (amaxtause^2)dt +α9 (amintwise)dt +α10 (amintwise^2)dt +α11 (amintspse)dt +α12 (amintspse^2)dt +α13 (amintsuse)dt +α14 (amintsuse^2)dt +α15 (amintause)dt +α16 (amintause^2)dt +α17 (apcpwise)dt + α18(apcpwise^2)dt +α19 (apcpspse)dt +α20 (apcpspse^2)dt +α21 (apcpsuse)dt +α22 (apcpsuse^2)dt +α23 (apcpause)dt +α24 (apcpause^2)dt +ɷdt (18) here, α0 is the constant coefficient; αt is the regression coefficient of time trend factors; α1 … α24 are the regression coefficients of corresponding variables; and ɷdt is the error term in equation (18). (esi)dt= €0 +€t (ttf) +€1 (amaxtwise)dt +€2 (amaxtwise^2)dt +€3 (amaxtspse)dt +€4 (amaxtspse^2)dt +€5 (amaxtsuse)dt +€6 (amaxtsuse^2)dt +€7 (amaxtause)dt +€8 (amaxtause^2)dt +€9 (amintwise)dt +€10 (amintwise^2)dt +€11 (amintspse)dt +€12 (amintspse^2)dt +€13 (amintsuse)dt +€14 (amintsuse^2)dt +€15 (amintause)dt +€16 (amintause^2)dt +€17 (apcpwise)dt + €18(apcpwise^2)dt +€19 (apcpspse)dt +€20 (apcpspse^2)dt +€21 (apcpsuse)dt +€22 (apcpsuse^2)dt +€23 (apcpause)dt +€24 (apcpause^2)dt + εdt (19) here, €0 is the constant coefficient; €t is the regression coefficient of time trend factors; €1 … €24 are the regression coefficients of corresponding variables; and εdt is the error term in equation (19). (eei)dt= ¥0 +¥t (ttf) +¥1 (amaxtwise)dt +¥2 (amaxtwise^2)dt +¥3 (amaxtspse)dt +¥4 (amaxtspse^2)dt +¥5 (amaxtsuse)dt +¥6 (amaxtsuse^2)dt +¥7 (amaxtause)dt +¥8 (amaxtause^2)dt +¥9 (amintwise)dt +¥10 (amintwise^2)dt +¥11 (amintspse)dt +¥12 (amintspse^2)dt +¥13 (amintsuse)dt +¥14 (amintsuse^2)dt +¥15 (amintause)dt +¥16 (amintause^2)dt +¥17 (apcpwise)dt +¥18(apcpwise^2)dt +¥19 (apcpspse)dt +¥20 (apcpspse^2)dt +¥21 (apcpsuse)dt +¥22 (apcpsuse^2)dt +¥23 (apcpause)dt +¥24 (apcpause^2)dt +µdt (20) here, ¥0 is the constant coefficient; ¥t is the regression coefficient of time trend factors; ¥1 … ¥24 are the regression coefficients of corresponding variables; and µdt is the error term in equation (20). (sei)dt= δ0 +δt (ttf) +δ1 (amaxtwise)dt +δ2 (amaxtwise^2)dt +δ3 (amaxtspse)dt +δ4 (amaxtspse^2)dt +δ5 (amaxtsuse)dt +δ6 (amaxtsuse^2)dt +δ7 (amaxtause)dt +δ8 (amaxtause^2)dt +δ9 (amintwise)dt +δ10 (amintwise^2)dt +δ11 (amintspse)dt +δ12 (amintspse^2)dt +δ13 (amintsuse)dt +δ14 (amintsuse^2)dt +δ15 (amintause)dt + δ16 (amintause^2)dt +δ17 (apcpwise)dt +δ18(apcpwise^2)dt +δ19 (apcpspse)dt +δ20 (apcpspse^2)dt +δ21 (apcpsuse)dt +δ22 (apcpsuse^2)dt +δ23 (apcpause)dt +δ24 (apcpause^2)dt +ψdt (21) here, δ0 is the constant coefficient; δt is the regression coefficient of time trend factors; δ1 … δ24 are the regression coefficients of corresponding variables; and µdt is the error term in equation (21). 5.1. selection of appropriate empirical model as the sample size of this study includes 18 districts of gujarat, which is compiled as district-wise panel data. these districts have high variation in climatic and socio-economic factors. thus, it is essential to select an agriculture and food sciences research, 2018, 5(1): 30-46 37 appropriate, consistent and viable empirical model to provide a viable statistical inference on empirical results. firstly, im-pesaran-shin test is applied to identify the existence of panel unit root in the individual time series. variance inflation factor (vif) is estimated to recognize the multicollinearity in explanatory variables [36]. ramsay reset test is considered to identify that whether functional forms of linear and linear regression models is appropriate or not. thereafter, random effect model is used to estimate the regression coefficients of explanatory variables in all model under the assumption that variation across districts is random and independent variables do have an insignificant impact on predictors [23, 25, 34, 47]. the consistency of this model is checked through breusch-pagan lagrange multiplier test [25, 34, 47]. fixed effect model is useful to capture the variation across districts and to make results unbiased. therefore, fixed effect model is also applied, while the appropriateness of this model is recognized through hausman specification test [23, 25, 34, 47]. pesaran's test is also used to identify the cross-sectional independence in panel data [23, 25, 34, 47]. the presence of group-wise heteroskedasticity is recognized using modified wald test [25, 34, 47]. autocorrelation is a major problem in panel data, thus the presence of autocorrelation is identified through wooldridge test [23, 25, 34, 47]. finally, prais winsten models with panels corrected standard errors estimations (pcses) and feasible generalize least square (fgls) estimations are used to estimate the regression coefficients of explanatory variables for proposed regression models. it gives preference to pcses model because this model provides better results than fgls model [23, 25, 34, 47, 50]. 6. descriptive and empirical findings 6.1. brief explanation on descriptive results figure: 1 indicates the relative position of across districts in estimated slsi during 2000-2011. the mean values of slsi for time periods 2000-2005 and 2006-2011, implies that bharuch, mahesana, amreli, junagadh, sabarkantha and rajkot have slsi’s more than 1.5, thus these districts have a relatively better position in sls as compared to others districts in gujarat. kachchh, valsad, surendranagar, dangs, panchmahals and surat have a slsi’s value less than 1.40, thus districts are in relatively lower position in sls. jamnagar, bhavnagar, kheda, vadodara, ahmedabad and banaskantha have slsi's value between 1.501.40, thus these districts in middle position in estimated slsi. since the slsi’s value lies between 1.88-1.17, therefore, sls significantly varies across districts in gujarat. the variation exists due to high variation in population density, population growth rate, ratio of forest area with gross cropped area, urbanization rate, cropping intensity (ratio of gross cropped area with net sown area), households having toilets, per capita availability of milk production, food-grain yield, ratio of gross irrigated area with gross cropped area, fertilizer consumption/hectare land, per capita district domestic product, agriculture output/hectare land, literacy rate, female literacy rate, households having accessibility to electricity, population having accessibility to safe water, infant mortality rate and birth rate (rural + urban) across districts in gujarat. also, estimated correlation coefficients of slsi with socio-economic variables imply that slsi is negatively associated with population density (r= -0.360), population growth rate (r= -0.56), urbanization rate (r= 0.144) and infant mortality rate (r= -0.247) (refer to table: c1 in appendix: c). cropping intensity (r= 0.127), households having toilets ( r= 0.161), food-grain yield (r= 0.308), ratio of gross irrigated area with gross copped area (r= 0.136), fertilizer consumption/hectare land (r= 0.045), per capita district domestic product (r= 0.388), agricultural output/hectare land (r= 0.267), literacy rate (r= 0.312), household having electricity accessibility (r= 0.271), population having accessibility of safe water (r= 0.403) and birth rate (r= 0.152) have a positive association with sustainability livelihood security in gujarat. figure-1. estimated slsi across districts in gujarat source: author's estimation. agriculture and food sciences research, 2018, 5(1): 30-46 38 figure-2. estimated esi across districts in gujarat source: author's estimation. figure: 2 provide the relative position of across districts in estimated esi during 2000-2011. the estimates demonstrate that bharuch, junagadh, dangs, amreli, panchmahal, valsad, vadodara, sabarkantha, banaskantha, mahesana, jamnagar and kachchh have an esi’s value more than 0.50. thus, these districts have a better position in ecological security as compared to other districts of gujarat. other districts have an esi’s value less than 0.50. therefore, these districts are in a relatively poor position in the estimated value of esi. a value of esi lie between 0.66-0.25, thus it shows that these districts have a significant variation in ecological security due to high variation in population density, population growth rate, ratio of forest area with gross cropped area, urbanization rate, cropping intensity (ratio of gross cropped area with net sown area) and households having toilets across districts in gujarat. estimates based on karl-person correlation coefficients infer that ecological security is negatively correlated with population density (r= 0.592), population growth rate (r= 0.491) and urbanization rate (r= 0.620). while, ecological security is positively associated with forest area (r= 0.260), cropping intensity (r=0.082), per capita availability of milk production (r= 0.288) and per capita district domestic product (r= 0.186) (refer to table: c1 in appendix: c). figure-3. estimated eei across districts in gujarat source: author's estimation. variation in economic efficiency across districts is measured through economic efficiency index, which is given in figure: 3. the estimated value of eei is less than 0.5 across districts, thus these districts are inefficient to maintain economic efficiency. however, the values of eei are lies between 0.48-0.18 during 2006-2011. therefore, all districts have a high and significant variation in economic efficiency due to high diversity in per capita availability of milk production, food-grain yield, ratio of a gross irrigated area with gross cropped area, fertilizer consumption/hectare land, per capita district domestic product and agriculture output/hectare land across districts in gujarat. estimated correlation coefficients also indicate that economic efficiency is negatively associated with population density (r= -0.483), population growth rate (r= -0.362), urbanization rate (r= -0.287) and infant mortality rate (r= -0.232) (refer to table: c1 in appendix: c). while, economic efficiency is positively associated with per capita availability of milk production (r= 0.387), food-grain yield (r= 0.262), ratio of gross irrigated area with gross cropped area (r= 0.499), per capita district domestic product (r= 0.499) and agricultural output per hectare land (r= 0.362). the variation in social equity across districts is measured through estimation of social equity index (sei) during 2000-2011, which is given in figure: 4. the mean value of sei during 2000-2005 and 2006-2011 shows that these districts have a significant variation in social equity. the estimated values of sei are observed more than 0.50 during 2006-2001 for mahesana, ahmedabad, bharuch, surat, rajkot, sabarkantha, amreli, kheda, vadodara, agriculture and food sciences research, 2018, 5(1): 30-46 39 bhavnagar, junagadh, valsad, jamnagar, kachchh, and panchmahals. also, the value of sei lies between 0.79-0.28 for across districts in gujarat. the variation in sei exists due to high diversity in literacy rate, female literacy rate, households having electricity accessibility, the population having accessibility to safe water, infant mortality rate and birth rate (rural + urban) across districts in gujarat. as estimated correlation coefficients of sei with socioeconomic variables demonstrates that social equity is positively associated with literacy rate (r= 0.767), female literacy rate (r= 0.700), household having electricity accessibility (r=0.651), population having accessibility of safe water (r= 0.743), infant mortality rate (r= 0.196) and birth rate (r= 0.085), food-grain yield (r= 0.174), ratio of gross irrigated area with gross cropped area (r= 0.183) and fertilizer consumption per hectare land (r= 0.211) (refer to table: c1 in appendix: c). while social equity is negatively associated with population growth rate (r= 0.171). figure-4. estimated sei across districts in gujarat source: author's estimation. 6.2. validity and consistency of estimated slsi the present study creates district-wise slsi and its associated indexes i.e. esi, eei and sei using composite z-score technique. therefore, it is essential to check the validity and consistency of estimated index to make the unanimity among the researchers and policy makers [25, 34, 36, 44, 47]. it also measures the accuracy and viability of estimated index. in fact, the estimated index can be used for further empirical investigation after validation. the validity of an estimated index can be checked through estimating correlation coefficients among it and its associated index [25, 36, 44, 47]. if an estimated index has a positive or negative but statistically significant association with its associated index, then it may be considered as ‘good’ [25, 36, 44, 47]. in order to check the validity and consistency of estimated slsi, esi, eei and sei, the karl pearson correlation coefficients are estimated among these indicators [25, 36, 44, 47]. the correlation coefficients among the slsi, esi, eei and sei are given in table: 5. estimates imply that slsi is positively associated with esi, eei and sei. further, it infers that sls would be improved with increase in ecological security, economic efficiency and social equity in gujarat. while esi is negatively associated with sei, thus social equity may be declined as increase in ecological security. economic efficiency and social equity is also negatively correlated with each other. so, it is a great concern to increase the economic efficiency and social equity in gujarat. table-5. pearson correlation coefficients among slsi and its associated indexes parameters slsi esi eei sei slsi 1 0.529** 0.650** 0.576** esi 0.529** 1 0.259** -0.188** eei 0.650** 0.259** 1 -0.002 sei 0.576** -0.188** -0.002 1 note: slsisustainable livelihood security index; esiecological security index; eei economic efficiency index; seisocial equity index. source: author's estimation; *and** shows that correlation coefficients are statistically significant at 0.05 and 0.01 significance level. 6.3. description of empirical results 6.3.1. association of slsi with climatic factors the association of slsi with climatic factors are given in equation (22) and (23). as f-value under ramsey reset is statistically insignificant, thus linear regression model produces better results than non-linear regression model. however, results based on non-linear regression model also provide the relationship between slsi and climatic factors. regression coefficients based on linear regression model imply that sls is negatively associated with maximum and minimum temperature, and precipitation during winter, spring, summer and autumn seasons [note *, ** and *** shows that regression coefficients are statistically significant at 1%, 5% and 10% significance level in equation (22)… (29)]. empirical results also indicate that maximum temperature during spring, summer and autumn seasons show a negative influence on sls. minimum temperature during winter and autumn seasons also has a negative association with sls. besides, regression coefficients of precipitation during agriculture and food sciences research, 2018, 5(1): 30-46 40 spring, summer and autumn seasons appeared negative, thus these factors have a negative influence on sls in gujarat. the regression coefficient of time trend factor with slsi under linear and non-linear regression model is found positive. hence, the technological change would be helpful to increase sls in gujarat. estimates based on non-linear regression model indicate that climatic factors have a non-linear relationship with sls. (slsi)dt= -10.1103 +0.0048 (ttf) +0.0971* (amaxtwise)dt -0.0515** (amaxtspse)dt -0.0582 (amaxtsuse)dt -0.0042 (amaxtause)dt -0.0902* (amintwise)dt +0.0240 (amintspse)dt +0.1998* (amintsuse)dt -0.0742* (amintause)dt +0.7360* (apcpwise)dt -0.0114 (apcpspse)dt -0.0052*** (apcpsuse)dt -0.0258 (apcpause)dt +udt [n= 216; wald chi2 =58*; r-squared =0.1985; mean vif= 5.77; aic= -120.4305; bic= -73.1766; f-value (ramsey reset)= 0.35] (22) (slsi)dt = -3.9216 +0.0055 (ttf) +0.7385** (amaxtwise)dt -0.0096*** (amaxtwise^2)dt -0.6937*** (amaxtspse)dt +0.0084 (amaxtspse^2)dt +0.3273 (amaxtsuse)dt -0.0067 (amaxtsuse^2)dt -0.8773*** (amaxtause)dt +0.0131*** (amaxtause^2)dt 0.7264* (amintwise)dt +0.0202* (amintwise^2)dt +1.4593** (amintspse)dt -0.0280** (amintspse^2)dt -0.6018 (amintsuse)dt +0.0168 (amintsuse^2)dt -0.1416 (amintause)dt +0.0012 (amintause^2)dt -0.2353 (apcpwise)dt +1.6931 (apcpwise^2)dt +0.0140 (apcpspse)dt -0.0029*** (apcpspse^2)dt -0.0126 (apcpsuse)dt +0.0002 (apcpsuse^2)dt -0.0882 (apcpause)dt +0.0117 (apcpause^2)dt + ɷdt [n= 216; wald chi2 = 84.05*; r-squared =0.2889; mean vif =2149.81; aic= -122.2791; bic= 34.5219; f-value (ramsey reset)= 2.23***] (23) 6.3.2. association of esi with climatic factors the empirical relationship between estimated esi with climatic factors is given in equation (24) and (25). the regression coefficients of explanatory variables are estimated through linear and non-linear regression models. as the f-value under ramsay reset test is found statistically insignificant for both the models, thus these models produce better and viable results. estimates indicate that maximum temperature during spring and summer seasons have a negative influence on ecological security. also, regression coefficients of minimum temperature during winter and summer seasons with esi is found negative. therefore, ecological security is negatively impacted due to change in minimum temperature in aforesaid seasons. precipitation during spring and summer seasons also brings negative implications on ecological security. as regression coefficients of the original term of climatic factors and square term of climatic factors with esi have an opposite sign. so, climatic factors have a nonlinear relationship with ecological security. it also found that climatic factors have u-shape and hilly-shape relationship with ecological security. (esi)dt= 3.4819 -0.0012 (ttf) +0.0042 (amaxtwise)dt -0.0125 (amaxtspse)dt -0.0104 (amaxtsuse)dt +0.0015 (amaxtause)dt -0.0118 (amintwise)dt +0.0213 (amintspse)dt -0.0122 (amintsuse)dt +0.0015 (amintause)dt +0.1776* (apcpwise)dt -0.0104*** (apcpspse)dt -0.0006 (apcpsuse)dt +0.0556* (apcpause)dt + ϕdt [n= 216; wald chi2 = 48.53*; r-squared =0.1022; mean vif = 5.77; aic= -330.0421; bic= -377.296; f-value (ramsey reset)= 0.21] (24) (esi)dt = 28.9022** -0.0026 (ttf) +0.5457* (amaxtwise)dt -0.0083* (amaxtwise^2)dt -0.4087** (amaxtspse)dt 0.0053*** (amaxtspse^2)dt -0.3795 (amaxtsuse)dt +0.0055 (amaxtsuse^2)dt -0.3999*** (amaxtause)dt +0.0061*** (amaxtause^2)dt 0.2754* (amintwise)dt +0.0083* (amintwise^2)dt +0.2342 (amintspse)dt -0.0044 (amintspse^2)dt -1.0333** (amintsuse)dt +0.0212** (amintsuse^2)dt +0.0589 (amintause)dt -0.0018 (amintause^2)dt +0.2123 (apcpwise)dt -0.1447 (apcpwise^2)dt 0.0171*** (apcpspse)dt +0.0009 (apcpspse^2)dt -0.0095** (apcpsuse)dt +0.0003*** (apcpsuse^2)dt +0.1152** (apcpause)dt 0.0400 (apcpause^2)dt +εdt [n= 216; wald chi2 = 95.81*; r-squared = 0.1973; mean vif = 2149.81; aic= -377.4848; bic= -289.7276; f-value (ramsey reset)= 1.70] (25) 6.3.3. association of eei with climatic factors the empirical association between economic efficiency index (eei) and climatic factors are estimated through linear and non-linear regression models. regression coefficients of climatic factors with eei are given in equation (26) and (27). f-value under ramsay reset test is found statistically insignificant for the linear regression model, therefore this model produces better results than non-linear regression model. the regression coefficient of time trend factor with eei is observed positive and statistically significant. it implies that adoption of technology in production activities would be helpful to increase the economic efficiency of resources. thus, it is suggested to apply advanced technology in production activities in those districts and rural areas which are using lower technologies in production activities. regression coefficients of maximum temperature during spring, summer and autumn seasons are found negative, thus it shows that maximum temperature during aforesaid periods would be caused to decrease economic efficiency. minimum temperature during winter, spring and autumn seasons also have a negative influence on economic efficiency. economic efficiency is also negatively influenced due to variability in precipitation during summer and autumn seasons. the estimates can be consistent with those studies which estimate climate change impact on the individual indicator of economic efficiency i.e. per capita availability of milk production, food-grain yield [25] ratio of gross irrigated are with a gross cropped area, per capita district domestic product, agricultural output/ha land [50]. estimates based on non-linear regression model provide crucial evidence that climatic factors have a non-linear relationship with economic efficiency in gujarat. (eei)dt = -13.0881*** +0.0074*** (ttf) +0.0355** (amaxtwise)dt -0.0372* (amaxtspse)dt -0.0171 (amaxtsuse)dt -0.0122 (amaxtause)dt -0.0136 (amintwise)dt -0.0219 (amintspse)dt +0.0466 (amintsuse)dt -0.0274*** (amintause)dt +0.3377* (apcpwise)dt +0.0028 (apcpspse)dt -0.0062* (apcpsuse)dt -0.0122 (apcpause)dt + €dt [n= 216; wald chi2 = 88.63*; r-squared = 0.2454; mean vif= 5.77; aic= -418.3421; bic= -371.0882; f-value (ramsey reset)= 0.21] (26) (eei)dt = 9.8363 +0.0052 (ttf) +0.1606 (amaxtwise)dt -0.0022 (amaxtwise^2)dt +0.3257*** (amaxtspse)dt -0.0047*** (amaxtspse^2)dt -0.0047*** (amaxtsuse)dt +0.0137** (amaxtsuse^2)dt +0.0878 (amaxtause)dt -0.0016 (amaxtause^2)dt 0.0619 (amintwise)dt +0.0018 (amintwise^2)dt +0.2005 (amintspse)dt -0.0044 (amintspse^2)dt -1.4442* (amintsuse)dt +0.0297* (amintsuse^2)dt +0.0190 (amintause)dt -0.0008 (amintause^2)dt -0.0256 (apcpwise)dt +0.6172 (apcpwise^2)dt +0.0262** (apcpspse)dt -0.0026** (apcpspse^2)dt +0.0015 (apcpsuse)dt -0.0003*** (apcpsuse^2)dt -0.0352 (apcpause)dt +0.0126 (apcpause^2)dt +µdt [n= 216; wald chi2 =193.19*; r-squared =0.3375; mean vif =2149.81; aic= -422.4792; bic= -334.722; f-value (ramsey reset)= 2.970**] (27) agriculture and food sciences research, 2018, 5(1): 30-46 41 6.3.4. association of sei with climatic factors the association of climatic factors with social equity index (sei) is estimated through linear and non-linear regression models. the regression coefficient of climatic factors with sei is presented in equation (28) and (29). regression coefficients of maximum temperature during spring and summer seasons has a negative and statistically significant influence on sei. it means that social equity is negatively impacted due to variability in climatic factors. the influence of minimum temperature during winter and autumn seasons on sei is also found negative. it infers that minimum temperature during the aforesaid periods have a negative impact on social equity. regression coefficients of precipitation during spring and autumn seasons with sei are appeared negative. thus, social equity also negatively influenced due to change in precipitation during spring and autumn seasons. estimates based on non-linear regression model indicates that climatic factors have a non-linear relationship. also, climatic factors have a u-shaped and hilly-shaped relation with social equity. since, the social equity index includes several factors i.e. literacy rate, female literacy rate, households having electricity accessibility, population having accessibility to safe water, infant mortality rate and birth rate. (sei)dt = -0.5576 -0.0013 (ttf) +0.0571* (amaxtwise)dt -0.0017 (amaxtspse)dt -0.0306*** (amaxtsuse)dt +0.0066 (amaxtause)dt -0.0647* (amintwise)dt +0.0245 (amintspse)dt +0.1652* (amintsuse)dt -0.0482* (amintause)dt +0.2211 (apcpwise)dt -0.0037 (apcpspse)dt +0.0016 (apcpsuse)dt -0.0691** (apcpause)dt +τdt [n= 216; wald chi2 = 110.73*; rsquared =0.3201; mean vif = 5.77; aic= -326.1622; bic= -278.9083; f-value (ramsey reset)= 5.12] (28) (sei)dt = -42.6509* +0.0029 (ttf) +0.0302 (amaxtwise)dt +0.0009 (amaxtwise^2)dt -0.6089** (amaxtspse)dt +0.0077** (amaxtspse^2)dt +1.5811* (amaxtsuse)dt -0.0259* (amaxtsuse^2)dt -0.5662 (amaxtause)dt +0.0086 (amaxtause^2)dt -0.3879* (amintwise)dt +0.0101** (amintwise^2)dt +1.0255*** (amintspse)dt -0.0192* (amintspse^2)dt +1.8689* (amintsuse)dt 0.0341* (amintsuse^2)dt -0.2185** (amintause)dt +0.0039 (amintause^2)dt -0.4217 (apcpwise)dt +1.2210*** (apcpwise^2)dt +0.0049 (apcpspse)dt -0.0012 (apcpspse^2)dt -0.0046 (apcpsuse)dt +0.0001 (apcpsuse^2)dt -0.1684* (apcpause)dt +0.0393 (apcpause^2)dt + ψdt [n= 216; wald chi2 = 367.92*; r-squared =0.4392; mean vif = 2149.81; aic= -343.7822; bic= 256.0249; f-value (ramsey reset)= 2.23***] (29) 7. conclusion and policy suggestions the main aim of this study is to estimate the district-wise sustainable livelihood security index (slsi) in gujarat using composite z-score technique during 2000-2011. for this, the study creates ecological sustainability index (esi), economic efficiency index (eei) and social equity index (sei). slsi is measured as an integrated index of esi, eei and sei for aforesaid period. eighteen different variables are compiled as a single number to estimate the district-wise slsi. accordingly, it used linear and non-linear regression models to examine the influence of climatic factors (i.e., maximum temperature, minimum temperature and precipitation) in winter, spring, summer and autumn seasons on slsi, esi, eei and sei. descriptive results indicate that there is existence high variation in sustainable livelihood security, ecological security, economic efficiency and social equity across districts in gujarat. it is also observed that variation in slsi, esi, eei and sei exists due to the significant diversity of socio-economic and health related factors (i.e. population density, population growth rate, ratio of forest area with gross cropped area, urbanization rate, cropping intensity (ratio of gross cropped area with net sown area), households having toilets, per capita availability of milk production, food-grain yield, ratio of gross irrigated area with gross cropped area, fertilizer consumption/hectare land, per capita district domestic product, agriculture output/hectare land, literacy rate, female literacy rate, households having electricity accessibility, population having accessibility to safe water, infant mortality rate and birth rate) across districts in gujarat. moreover, the estimated correlation coefficients imply that slsi is positively and significantly associated with esi, eei and sei. so, it shows that sls would be improved as the increase in ecological security, economic efficiency and social equity in gujarat. empirical results based on linear and nonlinear regression models show that sustainable livelihood security, ecological security, economic efficiency and social equity are negatively associated with climatic factors during winter, spring, summer and autumn seasons. estimates also demonstrate that sustainable livelihood security, ecological security, economic efficiency and social equity have a non-linear relationship with climatic factors. here, it can be concluded that sls would be in alarming position due to climate change in gujarat. as slsi is negatively associated with population density, population growth rate, urbanization rate and infant mortality rate. thus national policy maker need to adopt a policy which must reduce the negative implications of these factors on sls in gujarat. cropping intensity, households having toilets, food-grain yield, ratio of gross irrigated area with gross copped area, fertilizer consumption/hectare land, per capita district domestic product, agricultural output/hectare land, literacy rate, household having electricity accessibility, population having accessibility of safe water and birth rate have a positive association with sls in gujarat. thus, these factors may be considered for further policy formulation. furthermore, it is essential to control population density, population growth rate and urbanization rate in order to maintain ecological security. for this, protection of forest area, and to increase cropping intensity, per capita availability of milk production and per capita district domestic product would be helpful to sustain ecological security. it is also suggested to maintain the pace of population density, population growth and urbanization, infant mortality rate to increase economic efficiency in gujarat. for this, increase in per capita availability of milk production, food-grain yield, ratio of gross irrigated area with gross cropped area, per capita district domestic product and agricultural output/hectare land would be imperative to improve economic efficiency in this state. social equity would be improved as increase in literacy rate, female literacy rate, household having electricity accessibility, population having accessibility of safe water and birth rate, food-grain yield, ratio of the gross irrigated area with gross cropped area and fertilizer consumption/hectare land. empirical results also suggested that there is indispensable to implement the effective management policies to preserve the natural resource to maintain sls in gujarat [1]. forest area play a significant role to maintain the ecological security, consequently forest area may be useful to conserve the other ecosystem services [31, 35, 44]. it is also desirable to apply labour-intensive and small-scale farming to contribute more to productivity 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1.551 1.398 junagadh 1.650 1.677 1.692 1.679 1.853 1.714 1.633 1.648 1.617 1.705 1.770 1.607 kachchh 1.528 1.416 1.354 1.388 1.396 1.476 1.466 1.346 1.365 1.360 1.430 1.344 kheda 1.500 1.551 1.447 1.515 1.453 1.387 1.375 1.358 1.380 1.516 1.607 1.533 mahesana 1.836 2.084 1.969 1.953 1.920 1.857 1.754 1.764 1.742 1.817 1.896 1.795 panchmahals 1.322 1.256 1.308 1.263 1.198 1.155 1.208 1.190 1.213 1.245 1.352 1.327 rajkot 1.463 1.367 1.411 1.415 1.692 1.718 1.724 1.580 1.594 1.503 1.631 1.531 sabarkantha 1.638 1.579 1.518 1.491 1.643 1.585 1.457 1.514 1.594 1.652 1.744 1.627 surat 1.478 1.255 1.272 1.212 1.233 1.294 1.238 1.116 1.232 1.131 1.159 1.134 surendranagar 1.211 1.179 1.183 1.159 1.298 1.375 1.346 1.271 1.315 1.314 1.446 1.412 vadodara 1.396 1.293 1.306 1.375 1.397 1.348 1.398 1.293 1.477 1.508 1.578 1.487 valsad 1.579 1.471 1.382 1.394 1.349 1.305 1.465 1.324 1.359 1.378 1.300 1.481 source: author’s estimation. table-a2. estimated district-wise ecological security index (esi) during 2000-2011 district 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 ahmedabad 0.435 0.404 0.297 0.311 0.301 0.277 0.282 0.296 0.296 0.360 0.363 0.350 amreli 0.631 0.591 0.493 0.510 0.527 0.508 0.570 0.529 0.561 0.581 0.589 0.581 banaskantha 0.623 0.622 0.563 0.621 0.612 0.485 0.445 0.498 0.481 0.588 0.569 0.528 bharuch 0.660 0.517 0.588 0.579 0.586 0.565 0.748 0.571 0.698 0.644 0.656 0.638 bhavnagar 0.529 0.512 0.441 0.480 0.475 0.428 0.449 0.457 0.470 0.534 0.536 0.516 dangs 0.554 0.499 0.436 0.452 0.467 0.641 0.462 0.663 0.600 0.645 0.675 0.674 jamnagar 0.519 0.481 0.438 0.449 0.496 0.449 0.482 0.477 0.503 0.538 0.540 0.526 junagadh 0.641 0.606 0.577 0.569 0.646 0.560 0.527 0.605 0.582 0.690 0.680 0.643 kachchh 0.644 0.574 0.452 0.486 0.464 0.444 0.548 0.454 0.488 0.520 0.530 0.514 kheda 0.531 0.494 0.393 0.431 0.478 0.407 0.373 0.434 0.428 0.567 0.574 0.554 mahesana 0.504 0.649 0.578 0.606 0.601 0.507 0.488 0.525 0.516 0.522 0.528 0.523 https://scholar.google.com/scholar?hl=en&q=application%20of%20livelihood%20vulnerability%20index%20in%20assessing%20smallholder%20maize%20farming%20households'%20vulnerability%20to%20climate%20change%20in%20brong-ahafo%20region%20of%20ghana http://dx.doi.org/10.1016/j.kjss.2017.06.009 https://scholar.google.com/scholar?hl=en&q=an%20overview%20and%20evaluation%20of%20composite%20indices%20of%20development https://scholar.google.com/scholar?hl=en&q=assessment%20of%20social%20vulnerability%20to%20climate%20change%20in%20the%20eastern%20coast%20of%20india https://scholar.google.com/scholar?hl=en&q=assessment%20of%20social%20vulnerability%20to%20climate%20change%20in%20the%20eastern%20coast%20of%20india http://dx.doi.org/10.1007/s10584-015-1379-1 https://scholar.google.com/scholar?hl=en&q=carbon%20emission%20and%20global%20food%20security:%20a%20cross%20country%20analysis https://scholar.google.com/scholar?hl=en&q=implications%20of%20climatic%20and%20non-climatic%20variables%20on%20food%20security%20in%20developing%20economies:%20a%20conceptual%20review http://dx.doi.org/10.15406/mojfpt.2018.06.00138 https://scholar.google.com/scholar?hl=en&q=using%20panel%20data%20to%20estimate%20the%20effect%20of%20rainfall%20shock%20on%20smallholders%20food%20security%20and%20vulnerability%20in%20rural%20ethiopia http://dx.doi.org/10.1007/s10584-010-9994-3 https://scholar.google.com/scholar?hl=en&q=climate%20change%20impact%20on%20china%20food%20security%20in%202050 https://scholar.google.com/scholar?hl=en&q=assessing%20the%20impacts%20of%20climate%20change%20on%20land%20productivity%20in%20indian%20crop%20agriculture:%20an%20evidence%20from%20panel%20data%20analysis agriculture and food sciences research, 2018, 5(1): 30-46 44 panchmahals 0.587 0.586 0.622 0.517 0.524 0.495 0.543 0.503 0.486 0.602 0.614 0.614 rajkot 0.540 0.500 0.432 0.426 0.478 0.424 0.437 0.439 0.442 0.466 0.464 0.449 sabarkantha 0.600 0.571 0.493 0.452 0.530 0.470 0.472 0.512 0.515 0.585 0.579 0.553 surat 0.481 0.444 0.345 0.314 0.294 0.248 0.240 0.224 0.266 0.269 0.264 0.250 surendranagar 0.415 0.379 0.325 0.339 0.360 0.356 0.444 0.399 0.441 0.473 0.485 0.481 vadodara 0.543 0.507 0.442 0.451 0.452 0.415 0.461 0.428 0.570 0.616 0.611 0.577 valsad 0.583 0.525 0.494 0.483 0.498 0.461 0.576 0.477 0.494 0.577 0.595 0.600 source: author’s estimation. table-a3. estimated district-wise economic efficiency index (eei) during 2000-2011 district 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 ahmedabad 0.327 0.294 0.259 0.304 0.380 0.389 0.344 0.306 0.321 0.324 0.372 0.319 amreli 0.267 0.278 0.418 0.425 0.504 0.403 0.438 0.460 0.487 0.364 0.478 0.432 banaskantha 0.378 0.396 0.367 0.426 0.479 0.451 0.413 0.404 0.399 0.360 0.466 0.386 bharuch 0.332 0.356 0.458 0.508 0.453 0.525 0.501 0.457 0.460 0.417 0.509 0.501 bhavnagar 0.289 0.211 0.309 0.372 0.348 0.349 0.373 0.364 0.394 0.346 0.367 0.391 dangs 0.558 0.568 0.566 0.601 0.455 0.436 0.423 0.386 0.412 0.403 0.399 0.379 jamnagar 0.256 0.210 0.275 0.286 0.465 0.367 0.408 0.445 0.452 0.438 0.527 0.368 junagadh 0.347 0.413 0.459 0.442 0.539 0.468 0.451 0.442 0.465 0.450 0.486 0.379 kachchh 0.368 0.370 0.383 0.406 0.436 0.416 0.338 0.356 0.370 0.339 0.408 0.301 kheda 0.371 0.406 0.408 0.447 0.329 0.338 0.351 0.301 0.341 0.326 0.374 0.319 mahesana 0.567 0.620 0.580 0.526 0.494 0.464 0.436 0.440 0.446 0.517 0.584 0.477 panchmahals 0.274 0.243 0.266 0.311 0.231 0.201 0.180 0.167 0.221 0.134 0.203 0.152 rajkot 0.262 0.196 0.339 0.352 0.574 0.516 0.501 0.445 0.500 0.394 0.507 0.408 sabarkantha 0.354 0.279 0.300 0.324 0.392 0.400 0.388 0.337 0.391 0.356 0.438 0.363 surat 0.356 0.140 0.272 0.234 0.267 0.265 0.226 0.195 0.293 0.185 0.208 0.154 surendranagar 0.341 0.296 0.388 0.362 0.479 0.527 0.435 0.430 0.437 0.386 0.486 0.483 vadodara 0.285 0.239 0.299 0.331 0.346 0.338 0.298 0.269 0.314 0.278 0.349 0.283 valsad 0.391 0.327 0.276 0.297 0.231 0.255 0.242 0.245 0.286 0.225 0.158 0.297 source: author’s estimation. table-a4. estimated district-wise social equity index (sei) during 2000-2011 district 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 ahmedabad 0.753 0.822 0.817 0.765 0.766 0.770 0.876 0.772 0.720 0.702 0.728 0.693 amreli 0.653 0.648 0.677 0.673 0.676 0.691 0.677 0.666 0.655 0.653 0.635 0.661 banaskantha 0.447 0.472 0.461 0.440 0.444 0.458 0.437 0.488 0.476 0.478 0.468 0.492 bharuch 0.696 0.726 0.725 0.722 0.727 0.739 0.770 0.735 0.728 0.739 0.747 0.746 bhavnagar 0.589 0.600 0.576 0.600 0.605 0.698 0.647 0.609 0.596 0.589 0.598 0.602 dangs 0.269 0.284 0.271 0.258 0.260 0.338 0.278 0.272 0.277 0.287 0.281 0.303 jamnagar 0.592 0.643 0.644 0.637 0.637 0.629 0.616 0.545 0.508 0.499 0.485 0.504 junagadh 0.662 0.658 0.656 0.667 0.667 0.686 0.655 0.601 0.569 0.565 0.604 0.585 kachchh 0.516 0.473 0.519 0.495 0.497 0.616 0.580 0.536 0.507 0.501 0.492 0.528 kheda 0.598 0.651 0.647 0.637 0.646 0.642 0.651 0.624 0.610 0.623 0.659 0.660 mahesana 0.765 0.815 0.812 0.821 0.825 0.886 0.830 0.799 0.779 0.779 0.785 0.794 panchmahals 0.461 0.427 0.420 0.435 0.443 0.459 0.486 0.520 0.506 0.509 0.535 0.561 rajkot 0.661 0.671 0.640 0.636 0.640 0.778 0.786 0.696 0.652 0.644 0.660 0.674 sabarkantha 0.684 0.728 0.725 0.715 0.720 0.715 0.597 0.665 0.687 0.711 0.727 0.711 surat 0.642 0.671 0.655 0.663 0.672 0.781 0.772 0.697 0.673 0.677 0.687 0.730 surendranagar 0.455 0.503 0.470 0.457 0.459 0.492 0.466 0.441 0.437 0.455 0.474 0.448 vadodara 0.568 0.547 0.566 0.593 0.598 0.595 0.638 0.596 0.593 0.614 0.618 0.626 valsad 0.606 0.618 0.612 0.614 0.620 0.590 0.647 0.602 0.579 0.576 0.547 0.583 source: author’s estimation. agriculture and food sciences research, 2018, 5(1): 30-46 45 appendix: b table-b1. association of slsi, esi, eei and sei with climatic factors factors slsi esi eei sei maxtwise maxtspse maxtsuse maxtause slsi 1 0.529** 0.650** 0.576** -0.115 -0.037 0.011 -0.029 esi 0.529** 1 0.259** -0.188** -0.081 -0.113 -0.122 -0.073 eei 0.650** 0.259** 1 -0.002 -0.248** -0.238** -0.132 -0.210** sei 0.576** -0.188** -0.002 1 0.082 0.216** 0.212** 0.176** maxtwise -0.115 -0.081 -0.248** 0.082 1 0.439** 0.311** 0.440** maxtspse -0.037 -0.113 -0.238** 0.216** 0.439** 1 0.341** -0.064 maxtsuse 0.011 -0.122 -0.132 0.212** 0.311** 0.341** 1 0.324** maxtause -0.029 -0.073 -0.210** 0.176** 0.440** -0.064 0.324** 1 mintwise -0.124 -0.037 -0.129 -0.057 0.591** -0.213** -0.013 0.345** mintspse 0.011 0.024 -0.192** 0.145* 0.333** -0.058 -0.015 0.316** mintsuse 0.076 -0.026 -0.036 0.160* 0.140* -0.412** 0.065 0.343** mintause -0.023 0.053 -0.068 -0.024 0.253** -0.508** -0.218** 0.382** pcpwise 0.220** 0.111 0.235** 0.061 -0.089 0.025 -0.047 -0.074 pcpspse -0.138* -0.170* -0.07 -0.02 0.260** 0.038 0.044 0.306** pcpause -0.165* -0.041 -0.206** -0.056 0.551** 0.200** -0.207** 0.022 pcpause -0.160* 0.142* -0.091 -0.278** 0.413** 0.065 0.019 -0.124 source: author’s estimation. *and** shows that correlation coefficients are statistically significant at 0.05 and 0.01 significance level. table-b1. association of slsi, esi, eei and sei with climatic factors conti… factors mintwise mintspse mintsuse mintause pcpwise pcpspse pcpause pcpause slsi -0.124 0.011 0.076 -0.023 0.220** -0.138* -0.165* -0.160* esi -0.037 0.024 -0.026 0.053 0.111 -0.170* -0.041 0.142* eei -0.129 -0.192** -0.036 -0.068 0.235** -0.07 -0.206** -0.091 sei -0.057 0.145* 0.160* -0.024 0.061 -0.02 -0.056 -0.278** maxtwise 0.591** 0.333** 0.140* 0.253** -0.089 0.260** 0.551** 0.413** maxtspse -0.213** -0.058 -0.412** -0.508** 0.025 0.038 0.200** 0.065 maxtsuse -0.013 -0.015 0.065 -0.218** -0.047 0.044 -0.207** 0.019 maxtause 0.345** 0.316** 0.343** 0.382** -0.074 0.306** 0.022 -0.124 mintwise 1 0.698** 0.729** 0.777** -0.149* 0.149* 0.391** 0.333** mintspse 0.698** 1 0.779** 0.758** -0.176** -0.132 0.123 0.129 mintsuse 0.729** 0.779** 1 0.856** -0.184** -0.091 0.009 -0.021 mintause 0.777** 0.758** 0.856** 1 -0.179** -0.083 0.173* 0.149* pcpwise -0.149* -0.176** -0.184** -0.179** 1 0.057 -0.059 -0.004 pcpspse 0.149* -0.132 -0.091 -0.083 0.057 1 0.308** 0.017 pcpause 0.391** 0.123 0.009 0.173* -0.059 0.308** 1 0.419** pcpause 0.333** 0.129 -0.021 0.149* -0.004 0.017 0.419** 1 source: author’s estimation. *and** shows that correlation coefficients are statistically significant at 0.05 and 0.01 significance level. appendix: c table-c1. association of slsi, esi, eei and sei with socio-economics factors factors slsi esi eei sei pd pgr fagca ur ci hht pcamp slsi 1 0.529** 0.650** 0.576** -0.360** -0.559** -0.229** -0.144* 0.127 0.161* -0.014 esi 0.529** 1 0.259** -0.188** -0.592** -0.491** 0.260** -0.620** 0.082 -0.408** 0.288** eei 0.650** 0.259** 1 -0.002 -0.483** -0.362** -0.296** -0.287** 0.082 -0.097 0.387** sei 0.576** -0.188** -0.002 1 0.294** -0.171* -0.313** 0.485** 0.062 0.628** -0.538** pd -0.360** -0.592** -0.483** 0.294** 1 0.235** 0.052 0.502** -0.011 0.394** -0.364** pgr -0.559** -0.491** -0.362** -0.171* 0.235** 1 0.340** 0.391** -0.002 0.192** -0.036 fagca -0.229** 0.260** -0.296** -0.313** 0.052 0.340** 1 -0.260** 0.105 -0.239** 0.147* ur -0.144* -0.620** -0.287** 0.485** 0.502** 0.391** -0.260** 1 -0.016 0.895** -0.601** ci 0.127 0.082 0.082 0.062 -0.011 -0.002 0.105 -0.016 1 0.005 -0.074 hht 0.161* -0.408** -0.097 0.628** 0.394** 0.192** -0.239** 0.895** 0.005 1 -0.557** pcamp -0.014 0.288** 0.387** -0.538** -0.364** -0.036 0.147* -0.601** -0.074 -0.557** 1 fgy 0.308** 0.106 0.262** 0.174* -0.052 -0.100 -0.019 0.156* -0.032 0.313** -0.105 giagca 0.136* -0.057 0.081 0.183** 0.082 0.044 -0.072 0.071 0.476** 0.08 -0.105 fcphl 0.045 -0.134* -0.054 0.211** 0.201** 0.093 -0.003 0.156* 0.414** 0.149* -0.166* pcddp 0.383** 0.186** 0.499** 0.041 -0.182** -0.279** -0.220** -0.03 -0.013 0.174* 0.037 aophl 0.267** 0.031 0.362** 0.093 -0.100 -0.220** -0.252** 0.032 0.411** 0.087 -0.038 lr 0.312** -0.321** -0.073 0.767** 0.511** -0.057 -0.237** 0.657** 0.009 0.717** -0.399** flr 0.291** -0.294** -0.054 0.700** 0.470** -0.026 -0.181** 0.695** 0.011 0.789** -0.348** hhea 0.271** -0.240** -0.084 0.651** 0.246** -0.127 -0.244** 0.643** 0.009 0.752** -0.646** phasw 0.403** -0.221** 0.033 0.743** 0.236** 0.015 -0.372** 0.540** 0.008 0.703** -0.442** imr -0.247** -0.497** -0.232** 0.196** 0.412** 0.276** -0.185** 0.715** 0.009 0.597** -0.513** br 0.152* 0.267** -0.086 0.085 -0.160* -0.155* 0.003 -0.346** -0.077 -0.294** 0.026 source: author’s estimation. *and** shows that correlation coefficients are statistically significant at 0.05 and 0.01 significance level. agriculture and food sciences research, 2018, 5(1): 30-46 46 table-c1. association of slsi, esi, eei and sei with socio-economics factors conti… factors fgy giagca fcphl pcddp aophl lr flr hhea phasw imr br slsi 0.308** 0.136* 0.045 0.383** 0.267** 0.312** 0.291** 0.271** 0.403** -0.247** 0.152* esi 0.106 -0.057 -0.134* 0.186** 0.031 -0.321** -0.294** -0.240** -0.221** -0.497** 0.267** eei 0.262** 0.081 -0.054 0.499** 0.362** -0.073 -0.054 -0.084 0.033 -0.232** -0.086 sei 0.174* 0.183** 0.211** 0.041 0.093 0.767** 0.700** 0.651** 0.743** 0.196** 0.085 pd -0.052 0.082 0.201** -0.182** -0.100 0.511** 0.470** 0.246** 0.236** 0.412** -0.160* pgr -0.1 0.044 0.093 -0.279** -0.220** -0.057 -0.026 -0.127 0.015 0.276** -0.155* fagca -0.019 -0.072 -0.003 -0.220** -0.252** -0.237** -0.181** -0.244** -0.372** -0.185** 0.003 ur 0.156* 0.071 0.156* -0.03 0.032 0.657** 0.695** 0.643** 0.540** 0.715** -0.346** ci -0.032 0.476** 0.414** -0.013 0.411** 0.009 0.011 0.009 0.008 0.009 -0.077 hht 0.313** 0.08 0.149* 0.174* 0.087 0.717** 0.789** 0.752** 0.703** 0.597** -0.294** pcamp -0.105 -0.105 -0.166* 0.037 -0.038 -0.399** -0.348** -0.646** -0.442** -0.513** 0.026 fgy 1 -0.05 0.008 0.417** 0.072 0.164* 0.257** 0.367** 0.210** 0.108 -0.075 giagca -0.05 1 0.870** -0.098 0.642** 0.144* 0.101 0.035 0.237** 0.088 0.054 fcphl 0.008 0.870** 1 -0.072 0.583** 0.230** 0.204** 0.13 0.219** 0.137* 0.024 pcddp 0.417** -0.098 -0.072 1 0.135* 0.053 0.145* 0.177** 0.156* -0.077 -0.041 aophl 0.072 0.642** 0.583** 0.135* 1 0.067 0.071 0.165* 0.126 0.021 0.035 lr 0.164* 0.144* 0.230** 0.053 0.067 1 0.970** 0.532** 0.584** 0.457** -0.341** flr 0.257** 0.101 0.204** 0.145* 0.071 0.970** 1 0.588** 0.585** 0.469** -0.390** hhea 0.367** 0.035 0.13 0.177** 0.165* 0.532** 0.588** 1 0.515** 0.412** -0.038 phasw 0.210** 0.237** 0.219** 0.156* 0.126 0.584** 0.585** 0.515** 1 0.394** 0.104 imr 0.108 0.088 0.137* -0.077 0.021 0.457** 0.469** 0.412** 0.394** 1 -0.332** br -0.075 0.054 0.024 -0.041 0.035 -0.341** -0.390** -0.038 0.104 -0.332** 1 source: author’s estimation. *and** shows that correlation coefficients are statistically significant at 0.05 and 0.01 significance level. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 85 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 85-88, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.85.88 © 2019 by the authors; licensee asian online journal publishing group ethylene biosynthesis inhibitors on the quality of the coffee beverage and efficiency of the mechanical harvest alisson augusto santos1 thayane leonel alves2 evandro freire lemos3 josé de arruda barbosa4 ian lucas de oliveira rocha5 gabriela mourão de almeida6 ( corresponding author) 1,5agronomist, universidade do estado de minas gerais (uemg), passos, minas gerais, brazil. 2,4,6master student, post graduated graduation in agronomy (science of the soil), universidade estadual paulista (unesp), jaboticabal, são paulo, brazil. 3agronomist, prof. dr., universidade do estado de minas gerais (uemg), passos, minas gerais, brazil. abstract the aim of the project is to evaluate the efficiency of the product based in ethephonin® the standartization of the maturation of the fruits, in an answer of the mechanical harvest and the quality of the coffee beverage. the study was conducted at aruã farm, in piumhi, southwest of brazil. the plants used in the research are from coffea arábica l, planted in 2005, in the space of 3.2m x 0.60m. the experimental design adopted had randomized blocks with four treatments: t1 – ethephon® (130ml/100ml water), t2 – controlling treatment, t3 – ethephon® plus ph reductor (130ml plus 40ml/100 l water), t4 – ethephon® plus bioprotector (130 ml plus 285.71 gr/100 l water) and 4 repetitions, totalizing, 16 parcels. to determine the percentual of the leaves fall of the coffee trees, four scores of the leaves: before applying the product, 15 days after applying, before and after the mechanical harvest. the harvest started when, the maturation of the beans got the following levels: t1 – 89.79% of cherry fruits, t2 – 72.98% of fruits, t – 86.02% of cherry fruits, t4 86.63% of cherry fruits. after the harvest the beans were taken to testes, of the region. the product ethephon influenced in the maturation of the coffee fruits, increasing the efficiency of the mechanical harvest, reducing the volume (percentual) of the pass through. the product didn’t influence the defoliation of the plants. there was no interference among the treatments to taste the beverages and the samples ranking. keywords: coffee tree, flowering, weather, physiological maturation, coffea arábica, fall of the leaves, beverage tasting. citation | alisson augusto santos; thayane leonel alves; evandro freire lemos; josé de arruda barbosa; ian lucas de oliveira rocha; gabriela mourão de almeida (2019). ethylene biosynthesis inhibitors on the quality of the coffee beverage and efficiency of the mechanical harvest. agriculture and food sciences research, 6(1): 85-88. history: received: 15 february 2019 revised: 20 march 2019 accepted: 24 april 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study was financed in part by the coordenação de aperfeiçoamento de pessoal de nível superior brasil (capes) finance code 001. the aruã farm, for the support in the development of the work. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 86 2. material and methods ..................................................................................................................................................................... 86 3. result and discussion ..................................................................................................................................................................... 87 4. conclusion ......................................................................................................................................................................................... 88 references .............................................................................................................................................................................................. 88 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.85.88&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.85.88&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/123 https://orcid.org/0000-0001-5796-9267 https://orcid.org/0000-0003-3532-2117 https://orcid.org/0000-0001-5679-6923 https://orcid.org/0000-0002-7393-8839 https://orcid.org/0000-0002-5078-550x https://orcid.org/0000-0002-8374-2290 agriculture and food sciences research, 2019, 6(1): 85-88 86 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to the existing literature by evaluating the efficiency of the product based in ethephonin® the standartization of the maturation of the fruits, in an answer of the mechanical harvest and the quality of the coffee beverage. 1. introduction the coffee trees belong to the rubiaceas family, and the most produced species in brazil are coffea arabica and coffea canefora. the c. arabica is an autogamy plant, with only 10% of cross-polination and the c. canefora is alogama, presenting incompability of the gametes of the same plant [1]. the flowering depends on the genotype and the phenotype of the plant, in the c. arabica, it occurs in diferent times, from august to november, it can go until january, due to the weather changes [2] the granulation of the coffee beans is the hardening of the endosperm, after the sleepy stage of the fruits, having the direct influence of the weather conditions as well. after the granulation, the ripening of the green to red and yellow fruits, depending on the variety starts. the senescence of the fruits and the gradative dry of the mucilage are easy phases of infection by microorganism, occuring in the fermentation of the beans and affecting their quality [3]. the manual harvest encopasses the withdraw of the beans on cloths, after this, it is made the shaking to remove the leaves. the mechanical harvest is made by autistic harvester or the pulled one, making all the derive and the fruits gathering process. due to the uneven flowering of the flowers, the maturation is uneven too, it is possible to find green, green-cane, cherry fruits in the same plant. the ideal time of the harvest occurs when the cherry fruits reach the red or yellow coloring, in the most pasrt of the species [4]. to determine the time of the harvest is necessary to have an evaluation of the maturation of the fruits, choosing four plants to represent the field, the fruits are picked and the quantity harvested is measured, after that, the sample, one liter of coffee is taken, and the green fruits, green-cane, cherry, raising and dry are separated, they are weighted and related to each percentage. the ideal measure to start the harvest is from 5% to 20% of green fruits in the samples [3]. the processing step of the coffee is very important, in the after harvest, it is fundamental the separation of the green fruits and not maturated cherry fruits to obtain a quality drink, and, for this to happen, many processing technics are used: keep the fruit intact, being this fruit processed in its integral shape, denominated natural coffee, remove only the shell, part of the mucilage, denominated peeled cherry; remove the shell and the mucilage mechanicaly; or remove the shell mechanicaly and the mucilage by the fermentation way (depleted) [5]. according to the instruction of the normative number 8, from june, 11th 2003, ministerio de agricultura, pecuaria e abastecimen to normative [6] it is defined that the benefited coffee, row bean, be classified in: cathegory (coffea arabica and coffea canefora), subcategory (flat, moca and spout); group (arabica and robusta); subgroup (arabica: extremely soft, only soft, hard, river, robust: excellet, good, regular, anormal); class: (green/blue, green cane, green, yellow. brown, gray, white and discrepant; and type. the application of the growing regulatories, based on ethylene, has an advantage with the uniform maturation and the anticipation of the fruits harvest. though, the coffee growing is made in a exquisite way, there are not many studies in this case. the product based on the active principle [7]: ethephon, registered at ministerio da agricultura, pecuaria e abastecimento – mapa n 03292, the application must be done when the fruits of the plants get 90% of physiological maturation, since they are not affected by prages and diseases. according to the standart of breathing of each fruit, they are classified as climateric and non climateric, being the breathing the main and principal process after harvest of the fruits. in the case of the climateric plants, the respiratory peak is due to the increasement of the production of ethylene (gas), occuring the ripening of the fruits after the harvest [8]. during many years, the addition of technics of the production of ethylene were added, without knowing its effects, for economical reasons, the research progresses to the final production of fruits that don´t have the interference in its quality [9]. so, the verification of the efficience of the product based on ethephon in the standardization of the coffee fruits maturation were intended, the percentage of the leaves fall and its benefits to the mechanical harvest, and its influence in the final quality of the coffee relating to the classification of the beverage. 2. material and methods the study was conducted at aruã farm, in piumhi, west region of minas gerais, brazil. the area has an average high of 840m, the average temperature is 19ºc and the precipitation is 1562 mm annually. the weather is classified as cwa (temperate climate wet in dry winter and hot summer) and the vegetation is native, typical of cerrado, according to koppen classification. the plants used at the research are of the new world variety (coffea arabica l.), planted in 2005, in the space of 3.2m x 0.60m and the average population of 5208.33 plants ha-1. the experimental adopted design was outlined in blocks used in 4 treatments, such as: t1 – ethephon® (130ml/100ml water), t2 – controlling treatment, t3 – ethephon® plus ph reductor (130ml plus 40ml/100 l water), t4 – ethephon® plus bioprotector (130 ml plus 285,71 gr/100 l water) and 4 repetitions, totalizing, 16 parcels. each parcel had 30 plants, being useful 22 central plants and 4 plants of each border. the blocks were distributed in all the coffee line. all the applications were made using the volume of 700l ha-1, in april 13th 2017, the moment that the plants presented 90% of the fruits physiologicaly mature, through the manual costal sprays, with the capacity of 20 liters and conical jet nozzle, with low output. to determine the percentual of the coffee tree leaves fall, four countings of plants leaves were made: before applying the product, 15 days after applying the product, before the mechanical harvest and after the mechanical harvest. each parcel was marked with a ribbon, four branches of three plants randon inside the work area, being two branches in each side of the coffee street. the process that takes to the leave fall is connected to the breaking of the layer abscission existing in the region of the petiole, near the brach. it occurs due to the alteration of the levels of auxine and, mainly ethylene. agriculture and food sciences research, 2019, 6(1): 85-88 87 © 2019 by the authors; licensee asian online journal publishing group the harvest started when, aproximately, the percentual of maturation was reached: t1cherry fruit: 89.79% and green fruits: 10.21%; t2 – cherry fruits: 72.98% and green fruits: 27.01%; t3cherry fruits 86.02% and green fruits13.97%; t4 – cherry fruits: 86.63% and green fruits: 13.37%. an automotive harvester from case was used, model 200, on may, 14th 2017, 31 days after the application of the product, with operational velocity of 900 meters and the vibration of the bars of 850 rotations per minute (rpm). following, the sweeping was made, to take the advantage of the fruits that fell on the floor before and during the mechanical harvest snd right after, the manual repass was made, picking the remaining fruits of the mechanical harvest. after harvested in each operation, the coffee beans were measured and passed in a sieve with air curtain, to remove the dirty parts (leaves, stone and wood). following, they were taken in a box containing water and separated by the different steps of maturation: green beans, cherry beans and dry beans. due to the weather conditions, in the first day after the harvest, the beans were dried in the shadow in the shed of the property and soon, when there was no rain forecast, the lots of beans were taken to the yard coated with concrete, until the final dry (humidity 11.5%). the solving of the layers was made, the minimum 10 times per day, to uniform the drying. after dried, the beans were submitted for a period of rest and right after they were benefited. the obtained results were put in an eletronic table, to make the statistics analyses. it was made using genes [10] to identify the treatments that were superior agonomically. 3. result and discussion by the middle of the medium squares, was realized that the effect of the treatment was significant to 5% to the percentual of the picked volume by the mechanical harvest and non significant to the percentuals of the fallen leaves, before and after the mechanical harvest table 1. table 1. analyses of the variance of the percentual riped in the mechanical harvest (vcm) and the percentual of the fallen leaves during the action of the product before the mechanical harvest (qfac), and the percentage of fallen leaves after the mechanical harvest (qfdc). passos, 2017. fv gl qm vcm qfac qfdc blocks 3 3.0696 0.1176 4.2026 treatments 3 75.6471 * 0.3736 ns 187.6138 ns waste 9 15.0070 0.1716 85.4367 total 15 average 73.91 1.71 82.51 cv (%) 5.24 24.23 11.20 note: ns;*;**: non significative, significative a 5% e 1%, respectively, by test f. it is possible to verify that the use of the product ethephon®, propitiated a superior percentage riped in the mechanical harvest, related to the controlling treatment, having na increase of the volume from 67.6% to 75.03%. there was no difference between the treatments ethephon®, ethephon® with ph reductor and ethephon with bioprotector, although, all of them were superior to the controlling treatment. similar results were obtained in studies made by silva, et al. [11] those, that the efficience of the harvest increased ifrom 65% to 77% of the pending load, for the treatment with ethephon®. related to the defoliation before and after the mechanical harvest, the average of the treatments, didn’t differ between them, the same was verified in the work of dias, et al. [12] table 2. table-2. average of the percentual got in the mechanical harvest (vcm); and the percentage of the fallen leaves before the harvest(qfac), and after the harvest(qfdc). passos, 2017. treatment average vcm qfac qfdc ethephon® 75.03 a 1.73 a 79.82 a witness 67.60 b 1.30 a 91.00 a ethephon® + reductor 77.57 a 2.04 a 74.89 a ethephon® + bioprotector 75.44 a 1.76 a 84.32 a note: average followed by the same letter of the column doesnt differ between them, by the test of scott & knott of 5% of probability. it is necessary to observe that during the period that the experiment was installed, the preciptation was 104.5 mm (collected in the place of the experimente). it is possible to verify in the literature that the coffee treee needs an annual precipitation between 1500 mm and 1800 mm, according to each region. it was observed, by the middle of the mediun squares, that the effect of the treatment was not significative to determine the quality of the beverage and all the samples (sweeping, cherry, dry and green), not even to the percentual of pva black, green and blazing); from the samples of dry and green, being significative to 5% to pva and the sweeping and cherry pva table 3. table-3. analyses of the variance of the beverage tasting of the samples of the sweeping (bva), cherry(bcd), dry (bbo), green (bve); and the percentage of pva of the sweeping sample (pvava), dry (pvabo), and green (pvave) passos, 2017. fv gl qm bva bcd bbo bve pvava pvacd pvabo pvave blocks 3 1.57 14.746 0.43 0.22 0.70 0.45 1.96 0,08 treatments 3 3.01 ns 42.480 ns 1.31 ns 0.14 ns 7.72 * 3.26 * 1.88 ns 2,41 ns waste 9 1.67 19.555 1.80 0.15 1.46 0.63 1.34 28,4 total 15 average 27.2 30.70 27.2 27.0 6.33 4.52 6.75 44.6 cv (%) 4.75 14.40 4.93 1.46 19.1 17.6 17,1 11.9 note: ns;*;**: non significative, significative to 5% and 1%, respectivaly, by test f. agriculture and food sciences research, 2019, 6(1): 85-88 88 © 2019 by the authors; licensee asian online journal publishing group it was verified that the drinking taste and the classification of the beans, didn’t suffer influence of the treatments table 4. it can be justified in function of the non occurrence of rain, during the period of the harvest. the same was observed by silva, et al. [11] in the taste of the cup, when there was no influence in the application of the mature in the beverage, soon, related to the defect of this project, was observed s discreet incensement for the treatment with ethephon®. table-4. average of the tasting beverageof the sweeping samples (bva), cherry (bcd), dry (bbo) and green (bve); and the percentage of pva of the sample of the sweeping (pvava), cherry (pvacd), dry (pvabo) and green (pvave). passos, 2017. treatment average bva bcd bbo bve pvava pvacd pvabo pvave ethephon® 27.9 a 33.0 a 27.2 a 27.0 a 4.66 a 4.25 a 6.00 a 45.0 a witness 28.0 a 27.9 a 27.3 a 26.8 a 6.66 a 4.66 a 6.33 a 43.5 a ethephon® + reductor 26.4 a 27.9 a 26.6 a 27.2 a 6.00 a 3.50 a 7.33 a 45.25 a ethephon® + bioprotector 26.5 a 34.0 a 28.0 a 27.2 a 8.00 a 5.66 a 7.33 a 44.75 a note: average followed by the same letter of the column doesnt differ between them, by the testo f scott & knott of 5% of probability. 4. conclusion the product ethephon® influenced in the maturation of the fruits of the coffee trees, increasing the efficiency in the mechanical harvest, reducing the volume (percentage) of repass. the product didn’t influence the index of defoliation, however, the coffee trees that received ethephon® associated with a bioprotector presented the tendency to remain with more leaves. there was no interference between the treatments for the beverage tasting and the classification of the samples, it was observed, though, a tendency of a better quality in the beverage in the treatmens ethephon and ethephon plus bioprotector, in the samples of cherry fruits. references [1] c. h. s. d. carvalho, coffee cultivars: origin, characteristics and recomendation. brasilia, df: embrapa coffee, 2008. [2] a. p. camargo, "flowering and fruiting of arabica coffee in the different coffee regions of brazil," brazilian agricultural research, brasília, vol. 20, pp. 831-839, 1985. [3] c. m. d. mequita, j. e. rezende, j. s. carvalho, j. m. a. fabri, n. c. moraes, p. t. dias, r. m. d. carvalho, and w. g. araújo, manual de café: harvest and prepare (coffea arabica l.). belo horizonte: emater-mg, 2016. [4] j. c. carvalho, f. m. borém, r. g. f. a. pereira, and f. silva, "influence of different harvesting systems on coffee quality (/ coffea arabica / l.)," science and agrotechnology (brazil), vol. 27, pp. 1089-1096, 2003. available at: https://doi.org/10.1590/s141370542003000500017. [5] c. h. r. reinato, f. m. borem, m. a. cirillo, and e. c. oliveira, "quality of the coffee dried in terreiros with different pavings and thicknesses of layers," coffee science, lavras, vol. 7, pp. 223-237, 2012. [6] normative, "normative instruction no. 8, june, 11 2003 of the ministry of agriculture, livestock and supply, brazil. law number 9,972, dated may 25, 2000, decree number 3,664, from november, 2000, and process number 21000.006212 / 2002-41," 2003. [7] e. purgatto, ethylene and ripening. napan: support center for research in food and nutrition, 2017. [8] g. m. felipe, ethylene. in: plant physiology 2 vol. 2. são paulo: epu, 1986. [9] a. b. rena and m. maestri, "physiology of coffee," agricultural report, belo horizonte, vol. 11, pp. 26-40, 1985. [10] c. c. d. genes, "a software package for analysis in experimental statistics and quantitative genetics," acta scientiarum, vol. 35, pp. 271-276, 2013. [11] f. m. d. silva, t. j. arre, e. d. s. tourino, t. s. gomes, and m. d. c. alves, "use of ethrel in the mechanized and selective harvesting of arabica coffee (coffea arabica l.)," coffee science, lavras, vol. 4, pp. 178-182, 2009. [12] r. e. a. a. dias, f. m. d. silva, j. p. b. cunha, r. c. avelar, and f. c. fernandes, "efficiency of mechanized harvesting of coffee with the use of ethylene biosynthesis inhibitor," coffee science, lavras, vol. 9, pp. 527-536, 2014. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn(e): 2411-6653 issn(p): 2518-0193 vol. 3, no. 2, 66-74, 2016 http://www.asianonlinejournals.com/index.php/aesr 66 enset (ensete ventricosum) production in ethiopia: its nutritional and socio-cultural values tadessa daba1  masayoshi shigeta2 1japan society for the promotion of science research fellow, kyoto university 2graduate school of asian and african area studies, kyoto university (corresponding author) abstract enset is a perennial root crop indigenous to ethiopia cultivated dominantly in the south and southwestern highlands. there are several enset varieties or clones mainly produced for their starch from the pseudostem while some are exclusively cultivated for their corm. kocho, bulla, and corm (amicho) are the main food products of enset. we studied the nutritional, socio-economic, and cultural values of this crop. the corm (neqaqa) was found to be more nutritious than bulla (gena) except in starch and zinc contents. enset is rich in starch but low in protein composition hence, dietary protein source is mandatory. it plays great role in the food security of the country and a staple food for significant ethiopian population. it has several environmental, nutritional, and socio-cultural importance however its cultivation is under disease challenges. continuous research effort is required to improve the production of enset and its food products and fiber processing. keywords: bulla, corm, enset, nutritional value, socio-culture, variety. contents 1. introduction ............................................................................................................................................................................... 67 2. research methodology .............................................................................................................................................................. 68 3. results and discussions ........................................................................................................................................................... 68 4. summary .................................................................................................................................................................................... 73 references ...................................................................................................................................................................................... 73 citation | tadessa daba; masayoshi shigeta (2016). enset (ensete ventricosum) production in ethiopia: its nutritional and socio-cultural values. agriculture and food sciences research, 3(2): 66-74. doi: 10.20448/journal.512/2016.3.2/512.2.66.74 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: contribution/acknowledgement: this work is licensed under a creative commons attribution 3.0 license we are so thankful to japan society for the promotion of science (jsps) for the financial support. we would like to thank areka agricultural research center (aarc), researchers zerihun yemataw and tesfaye dejene. we thank ms. tadelech wolde and mr. abiyot alemu for their very kind and dedicated help during sampling around areka. our gratitude extends to abebe chindi, research at holetta agricultural research center. we also appreciate mr. paskal mota for allowing his enset farm and provision of samples. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 28 december 2016/ revised: 22 march 2016/ accepted: 17 august 2016/ published: 10 november 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.2/512.2.66.74 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=tadessa%20daba https://orcid.org/orcid-search/quick-search?searchquery=masayoshi%20shigeta http://search.crossref.org/?q=10.20448/journal.512/2016.3.2/512.2.66.74 agriculture and food sciences research, 2016, 3(2): 66-74 67 1. introduction enset (enset ventricosum) is a genus of banana family and termed as false banana. it is a gigantic leafy monocarpic evergreen perennial root crop and main food source mostly for the southwestern part of ethiopian highlands. biogeographers and agronomists have considered the center of origin for enset agriculture to be the ethiopian highlands [1, 2]. other scholars have also presumed enset domestication in ethiopia to 10,000 years ago [3]. the representative enset growing areas of southwest highlands ethiopia are shown in fig. 1. this does not mean that enset is cultivated and used only in these regions as food. it is customary food in the southwest, central, towns, and cities with the ethiopian delicious food, kitfo. depending on the climate and agro-ecology, the main food crops and diet vary from region to region in the country. thus, cautious interpretation is required on general statements of ethiopian diet. more than one sixth of ethiopian population (nearly 15 million people) depends on enset [4]. fig-1. map of ethiopia showing the major enset growing geographical areas of the southwestern part. source: tsegaye and struik [5]. enset is a classic multipurpose crop that every botanical part is used for numerous material cultures. enset is a large banana like plant [3] and its all botanical parts are shown in fig. 2. the leaves and pseudostems are utilized for wrapping, sheeting to sit, thatching, making containers, shading from sunshine and protecting from rain [6]. the sheaths, fiber, and leaf of the plant are used for house construction, food wrapping, cattle feed, and ropes [7]. it is also used for various socio-economic and cultural values in the enset growing areas. some varieties are needed for certain ritual activities in big ceremonies and funerals. fig-2. matured enset at flowering stage showing all the botanical parts of the crop. source: brandt, et al. [3] the edible parts of enset vary from place to place. in general, the pulp of the pseudostem, the young shoots, and the corm are eaten. the processed pseudostem usually undergoes fermentation, which becomes flour and ultimately the basic ingredient of bread and porridge upon drying. in some parts of the country the pseudostem pulp, together agriculture and food sciences research, 2016, 3(2): 66-74 68 with the young shoots are boiled and eaten as a vegetable that tastes like irish potatoes when fresh [8]. seldom, the mealy substance of the seeds and the pith of the inflorescence stalk are boiled and eaten [9, 10] though the three most common enset derived foods are kocho, bulla, and amicho (corm). the plant is cut before flowering the pseudostem and leaf midribs are scraped (fig. 3), fermented for 10-20 days and becomes ready for various foods preparation. the juice of squeezed and dehydrated parenchymatic scrapings of the pseudostem gives a high-grade product. this product is called bulla, which is tightly packed in enset leaves and fermented in a silo [10]. the low-protein enset products are staple or co-staple foods without the required protein supplement for people [7]. fig-3. various enset varieties and enset processing. the kocho or bulla type on the left and the corm type on the right (a). processing enset samples used for nutritional analysis around areka agricultural research center (aarc) (b). the processed enset (kocho, bulla, and corm) are rich in carbohydrate and good sources of minerals. however, the starch yield and nutrient compositions vary among enset clones or varieties. according to aarc, more than 265 enset clones were identified and characterized. enset varieties are categorized to two main groups namely the kocho or psuedostem types and the corm types. there happen variations in the qualities (color, texture, aroma, etc.) of the final enset products, which are affected mainly by enset variety and processing skills. enset varieties that yield the whitest kocho and bulla color are considered to be the best quality while yield is very important trait for a family food according to farmers. the majority of enset varieties are produced mainly for processing to starch through fermentation, kocho or bulla while some other are produced entirely for their corm. detail nutritional studies of the identified varieties were not undertaken so far except little information on either general or undefined enset varieties. therefore, two best enset varieties supposed to be the representatives of the kocho type (gena) and corm type (neqaqa) were considered for their nutritional composition study. the nutritional, socio-cultural, and the opportunities and challenges of enset cultivation in the country are also described in this study. 2. research methodology the bulla, corm samples, and various information were obtained mainly from areka agricultural research center (aarc), which is the center of excellence for research on enset nationally. some preliminary information was also obtained from holetta agricultural research center (harc), root crops and temperate fruits research program. the social aspects, agronomic, and pertinent issues related to enset were studied. two enset varieties, one from the kocho type namely gena and the other from the corm type, neqaqa were processed with the help of aarc research staff in december 2014. the air dried bulla of gena and ground corm of naqaqa were brought to japan for their detail nutritional composition analysis. the samples were analyzed at japan food research laboratory (jfrl). various literatures and previous studies on different aspects of the crop were reviewed in addition to information from farmers and research institutions. 3. results and discussions 3.1. enset cultivation in ethiopia based on the review of previous studies, enset plantation in ethiopia as a food crop has long history. many scientists of different disciplines have developed theories that assume enset domestication in ethiopia to 10,000 years ago [3]. early scholars considered the indigenous hunter/gatherers of southern ethiopia may be the first ones to cultivate enset [8]. it was suggested in westphal [8] that the high land of kefa, which is the humid corner of southwest ethiopia, is the natural center of distribution of enset. others also proposed that the ethiopian highlands as the primary center of origin for enset cultivation [1, 2, 11]. it is an indigenous crop to ethiopia, where primarily cultivated as a staple food though wild enset varieties were reported to be found in other african and asian countries. agriculture and food sciences research, 2016, 3(2): 66-74 69 further research is required on the taxonomy and distribution of enset however four wild species found in subsaharan africa and madagascar and other two wild enset species distributed over asia [12, 13]. in the late 19th century, stigand [14] designated enset as "groves of what appeared to be bananas" in his first impression in the baka chief's area of ari area. based on sidamo area field study and comprehensive literatures survey, the finnish-scandinavian geographical expedition in 1953-54 described the enset cultivation in ethiopia [9]. cushitic-speaking people later introduced enset agriculture to the northern ethiopian highlands and then replaced by crops like wheat, barley, and teff following the migration of semitic-speaking groups into northern ethiopia [8]. he also reported the early cultivation of enset in the northern part of the abyssinian highland in the course of his survey on the source of the nile. enset domestication was brought to southwestern ethiopia sometime in prehistory of sidamo tribes [15]. later the agaw, central cushitic-speaking peoples of northern ethiopia, began to grow enset and other crops like wheat, barley, and cattle like goats and sheep into their economy [3]. the multipurpose use and propagation method of enset are mostly shared by the ethiopian agriculturalists that are classified as omotic and cushitic with one exception of semitic people, gurage [6]. in the mid to highlands of southern peoples nations nationalities regional state (spnnrs), western arsi-bale, and western oromia, enset is widely cultivated [7]. according to straube, the amarro people harvest enset when they are needed since they do not ferment its starch as cited in westphal [8]. in jima area, it is used during the rainy season when cereals are scanty otherwise, it is not so important like maize [8]. the aforementioned studies show the early domestication of enset in ethiopia and their gradual distribution pattern across the country and its importance as a staple and semi-staple food in various parts of the country. 3.2. agronomy and enset varieities enset (ensete ventricosum) belongs to order schistaminae and family musaceae. the musaceae family has two genera, musa and ensete [16]. farmers categorize enset and name based on several features that differentiate one from another and there are several cultivars in different agro-ecologies. the phenotype and utilization values are the basis for the naming of enset landrace accessions by farmers [17]. though scattered plants can be seen at lower altitudes, enset mainly cultivated at high elevations from 1,200 to 3,100 masl [18] with the optimum altitudes between 2,000 and 2,750 masl [19]. the existence of several different enset varieties is ascribed to the use of enset and its products for diverse purposes [20]. according to aarc, some enset cultivars like gena, merza, kucha, and silkente are preferred for their processed starch, kocho quality and productivity. others such as naqaqa, arke, and sweete are cultivated exclusively for their corm. according to aarc, the average maturity of gena is 5.07 years and that of neqaqa is 3.78 years and the origin of both varieties is walaita. the productivities of gena and neqaqa are 21.24 and 14.91 ton/hectare/year. however, the corms of the kocho type varieties are also edible in most cases. under the production of five out of seven zones, gena is the most broadly disseminated enset clone yemataw, et al. [21]. taboje [22] reported that gena provides up to 16.9 ha-1 year-1 of kocho and it is one of the high yielding clones. at aarc, 279 accessions of enset were evaluated [23]. according to negash and niehof [24] twelve distinguishing features to categorize enset accessions that are grouped into three clusters are: (1) phenotypic characteristics: pseudostem, midrib, leaf, and petiole colors, (2) agronomic characteristics: disease resistance (susceptible, intermediate, tolerant); maturity (early, intermediate, late); plant vigor (poor, medium, high), and (3) use-value characteristics: bulla quality (not good, good); corm use (not used, used); kocho yield (low, medium, high); medicinal value (not used, used); fiber quality (low, medium, high). farmers have conventionally established sophisticated way of enset propagation. it commences with cutting pseudostem of 2-3 years old from the underground corm and cover with soil when rainy season begins. up to 100 suckers emerge from one corm mother and they are separated from the corm and planted individually after maximum of 2 years. the farmers transplant them after few years in to the final pit prepared usually by removing the secondary roots from the corm and adding manure. the plants can be transplanted only once or up to four times with wider spacing. totally it takes 4-6 years for an enset to mature and get ready to be processed [3]. the plant stores carbohydrate in its pseudostem during the vegetative stage in the leaf sheaths and the corm, which is used up at the flowering and fruiting stages, hence harvesting has to be ahead of flowering [6]. the seeds can stay viable for more than 25 years [13]. the enset product yields of different clones evaluated at aarc differ greatly [21]. the sampled enset varieties in this study from aarc were of the high performing clones. shank and ertiro [7] studied the product yield and plant size and modeled yield estimation derivatives. individual plant yields vary from 114.7 kg (303 cm high plant x 298 cm circumference) to 3 kg (93 cm x 58 cm plant). they reported that the yield has a correlation of 87% with circumference and 83% with the pseudostem height. the resultant model plant yield = -36.5 + 0.23 x circumference + 0.19 x height in cm accounted for 82% of the observed variation in plant yield [7]. 3.3. the nutritional qualities of enset significant ethiopian population relay on enset for their major daily diet. however, it is limiting in protein and some mineral compositions. there are differences in the nutritive values among enset cultivars and different botanical parts used as food. gena and neqaqa, which were considered to represent the kocho/bulla and corm types, respectively, were analyzed for their nutritive values (table 1). bulla is supposed to be the best quality product obtained from the juice of decorticated leaf sheath or pseudostem. however, the corm was found to contain better nutrients except potassium, in which bulla is better as shown in table 1. the dry matter (dm) content of the air dried enset flour samples were 92% in bulla (gena) and 94% in corm (neqaqa), and the organic matter fraction on dm basis is around 99% (table 1). as fractions of dm, the crude protein contents are 0.7% in bulla and 8.8% in corm. the corm has better composition of other nutrients though bulla has higher carbohydrate. both the corm and bulla were found to contain most of the essential amino acids however bulla contains very small amounts of these nutrients. glutamine, tryptophan, and asparagine are amino agriculture and food sciences research, 2016, 3(2): 66-74 70 acids that are not found in corm mohammed, et al. [25]. hirose, et al. [26] examined the behavior of starch and storage in the enset tissues. they reported the similarities and differences of enset starch among the well utilized starches of potato, corn, and sago. they suggested the better possible use of enset starch as a binder for compressed tablets because of its better binding stability than that of potato starch. table-1. comparative nutrient compositions of enset varieties (bulla and corm). nutrient gena (bulla) neqaqa (corm) proximate (g/100 g) water 7.8 5.7 protein 0.6 8.3 fat 0.1 0.6 ash 1.0 3.2 carbohydrate 89.6 64.8 fiber 1.0 17.4 energy 363 kcal/100 g 333 kcal/100 g minerals (mg/100 g) sodium 3.7 5.2 phosphorous 30.1 80.4 iron 6.99 12.3 calcium 58.7 99.7 potassium 357 1.24 magnesium 11.9 59.6 copper 0.02 0.70 zinc 0.20 22.3 manganese 0.67 5.84 essential amino acids (mg/100 g) lysine 23 212 histidine 9 124 phenylalanine 21 238 leucine 33 334 isoleucine 18 170 methionine 8 73 valine 22 223 threonine 15 133 vitamins (mg/100 g) thiamine (b1) 0.02 0.28 riboflavin (b2) 0.01 0.05 note: this composition is not on dm basis, it is on the basis of air dried sample flours. the starch yield varies greatly with variety, management practices, and harvesting stage. shank and ertiro [7] stated that variations in the spacing and frequency of transplanting, agro-ecology, harvesting age, and variety make the assessment of enset yield difficult. about 47.0 kg of edible portion can be obtained from one mature enset and one meal of washi, food from fermented pseudostem starch, for instance is cooked for about five people on average with the share of about 600 to 800 g per person [6]. from 100 g fermented enset starch, a total of 186 ± 28 kcal energy is obtained [27]. however, 100 g of bulla and corm contain 363 and 333 kcal, respectively (table 3). hence, one person can get 2,541 kcal from bulla and 2,331 kcal from corm of roughly 700 g. this is nearly enough daily maintenance energy during low working activity but cannot be assumed out of 700 g washi for it contains a lot of moisture. one person requires roughly 8 enset plants per year based on 500 g per day [6]. the enset kocho yield ranges from 3 to 12 ton dm/ha depending on the variety and the age at which it is harvested [28]. in terms of protein, a person can get only 4.2 g from bulla and 58.1 g from corm considering 700 g air dried as an average daily intake. this shows that for an average person of 65 kg farmer gets only 8% from bulla and 112% from corm, respectively. it should be noted that protein is not enough if they live on enset food (kocho/bulla) alone. one hundred grams of washi normally contains only 0.9 g of protein [29] which means that only 6.3 g of protein can be obtained from 700 g in a day. corm (neqaqa) contains substantial amount of protein (9%) on dm basis (table 2) and provides excess of daily required protein in quantity. the names of the varieties are referred in the parentheses in this paper to explain that the nutritional composition differences may arise from the varieties not only of the botanical parts. the leaves had markedly higher protein content than that of the pseudostem and corm of the same plant [30, 31] which is in agreement with the general fact that more proteins are found in the leaves than in the stems or roots of plants. the protein content of 118 different landraces from different places in ethiopia enset leaves ranged from 10 to 23% that can be good dry season protein source for livestock [32]. the proximate and mineral contents of enset were compared with other common root and cereal crops (table 2). the corm type contains significantly higher compositions of minerals than the bulla type. the iron and calcium composition of the corm are higher than potato and its zinc content is much higher than even teff. the starch fraction of enset pseudostem was reported to be comparable to sorghum [33]. the nutrient contents of kocho and bulla is sufficient to human requirement similar in extent to cereal flours [34]. on the other hand, abebe, et al. [35] reported that the protein content and the amino acid composition of enset are relatively low. this makes legumes recommendable in a kocho-based diet, especially in cases where little animal products are consumed [25]. the facts of enset being low in protein composition and necessity of protein supplementation are in agreement with our current study shown in table 2 and 3. on the other hand, bulla contains higher carbohydrate and hence, energy than the corm. in other proximate compositions, the corm type is entirely superior. it was found to be better only in potassium among macro minerals. the corm contains more than twice of most minerals and many times higher of some minerals like zinc, magnesium, copper, and manganese. however, this variation may be attributed to the enset varieties/cultivars or botanical parts (the pseudostem and corm). bulla is known to be the best quality product from the pseudostem part. therefore, kocho is supposed to compose less of the analyzed nutrients than bulla and obviously than the corm. in order to agriculture and food sciences research, 2016, 3(2): 66-74 71 precisely evaluate the nutrient qualities of the botanical parts of enset, the samples ought to have been taken from the same plant. however, mohammed, et al. [25] have compared the compositions of the corm, above ground, and pseudostem of the same enset. in their study, the corm is higher in starch content and the above ground is better in fiber and the leaf was the best in protein content. table-2. proximate and mineral comparisons of enset with potato and teff (on dm basis) parameter bulla (gena) (ensete ventricoum) corm (neqaqa) (ensete ventricoum) potato (solanum tuberosum)* teff (eragrostis teff)** proximate g/100 g moisture 7.8 5.7 75.17 12.2 protein 0.65 8.80 6.44 14.58 fat 0.11 0.64 2.01 3.53 ash 1.08 3.39 3.34 2.16 carbohydrate 97.18 68.72 86.47 70.67 fiber 1.08 18.45 1.73 9.05 energy 393.71 353.13 391.24 minerals (mg/100 g) sodium 4.01 5.51 99.36 1.88 phosphorous 32.65 85.26 197.22 319.40 iron 7.58 13.04 7.09 22.37 calcium 63.67 105.73 67.14 146.26 potassium 387.20 1.31 1743.86 479.12 magnesium 12.91 63.20 139.63 178.82 copper 0.02 0.74 1.21 zinc 0.22 23.65 4.74 manganese 0.73 6.19 19.24 note: *elfaki and abbsher [36] * * average compositions of kuncho and key-tef on dm basis potato is the common root crop and teff is the dominant cereal crop and daily food source for ethiopians. this was the point of comparing enset with these crops. in table 3, we indicated that the corm is more nutritious than bulla in terms of essential amino acids composition. the amino acid content of kocho is similar to sweet potato and potato [35-37]. these food sources need to be supplemented in order to provide enough protein in general and specifically the deficient amino acids in kocho and the root crops. various beans are cultivated in ethiopia and common bean (phaseolus vulgaris) is commonly grown around home gardens [38] this practice must be encouraged in enset-based farming system. the clonal differences used by farmers, variation in the places of growth in different environments and soil qualities, and harvesting time differences unsurprisingly affect the nutrient qualities of enset [25]. there were no enough studies and the nutritional research on enset and its dietary analysis has so far been challenged ethiopian nutrition institute (eni) [29]. argen and gibson [27] described the nutritional composition of several enset dishes and indicated the necessity of additional protein source for an enset to be a staple food. enset fractions are good sources of some minerals like potassium, phosphorous, calcium (except corm), and magnesium [39]. in all these minerals, the corm was found to be much better than bulla except it contains much higher concentration of potassium than corm in our current study (tabe 1). most enset fractions except leaf lamina are deficient in copper but rich in iron and manganese [31]. however, all these minerals were found to be limiting both in bulla and corm (table 1 and 2). the macro and trace mineral content differences because of enset varieties can help in strategic supplementation of mineral deficiencies [31]. enset food products are usually used after fermentation. fermentation normally enhances the nutritive value of foods but similar result was not obtained in kocho, possibly because of water soluble nutrients leaching through the long incubation [40, 41]. the nutritive value of enset was compared with other common root crop, potato and ethiopian popular cereal, teff (table 2 and 3). most of the proximate composition of enset is lower than both potato and teff except the carbohydrate content of bulla (gena) is much higher than both crops and the total energy content is equivalent to teff. the corm is the highest in fiber composition. teff is superior in the total protein and fat contents than both enset and potato. in terms of mineral contents, teff comes first followed by potato and then by the corm. in this study, the corm was better in lysine content than bulla but much lower than that of potato and teff (table 3). on the contrary, steinkraus [40] and gashe [41] reported that enset contains high lysine and low methionine. generally, enset is poor in protein composition and particularly bulla (gena) contains extremely low. the essential amino acid content of the corm (neqaqa) is remarkably higher than that of bulla (gena). however, teff is the best followed by potato (table 4). in essential amino acids, the daily recommended intake (dri) is not fulfilled by depending on enset as a sole daily diet. in this case, 700 g of enset was considered for an average daily intake from shigeta [6] and brandt, et al. [3]. only phenylalanine, histidine, and threonine daily requirements are attained from corm (neqaqa) while none is fulfilled from bulla. this shows that foods from enset should necessarily be supplemented with protein rich foods as recommended also by many others. in ethiopia usually, stews are made of mainly bean powders however vegetables are used in some cases. unless the nutritional conditions are improved, basing solely on enset leads to protein deficiencies that entails poor growth and weakened immune systems. agriculture and food sciences research, 2016, 3(2): 66-74 72 table-3. comparison of amino acids (mg/100g protein) of enset with other crops on dm basis. amino acids bulla (gena) corm (neqaqa) teff* (eragrostis teff) potato (solanum tuberosum)** ada 65 kg (mg)** available in 500 g enset (mg) bulla corm lysine 24.95 224.81 378.68 227.10 1950 175 1574 histidine 9.76 131.50 319.40 129.88 650 68 921 phenylalanine 22.78 252.39 741.52 459.12 (phe+tyr) 1625 281 3199 leucine 35.79 354.19 1063.46 500.32 2535 251 2479 isoleucine 19.52 180.28 498.09 318.16 1300 137 1262 methionine 8.68 77.41 522.22 91.34 (met+cys) 975 61 542 valine 23.86 236.48 689.49 724.93 1690 167 1655 threonine 16.27 141.04 559.84 253.32 975 114 987 note: *average composition of kuncho and key-tef on dm basis ** elfaki and abbsher [36] *** fao/who/unu [42] 3.4. socio-economic and cultural values of enset enset is a perennial evergreen crop with many aesthetic values and multipurpose in the south and southwestern ethiopia. it has many material uses and socio-cultural values. a large number of enset plants around houses provide comfort, shading for people and some crops such as coffee which need only moderate sunshine [6]. the use of enset and its products for different purposes such as food (amicho (corm), bulla, and kocho), medicine, rituals, and construction purposes could attributed to the existence of various enset varieties [20]. enset garden is sometimes an indicator of the economic status of farmers; many varieties more mature and large number of enset plants are found in the gardens of wealthier households [24]. gurage people depend much on enset socially and economically to obtain their essential needs [8]. once it reaches certain stage of growth, cultivated enset can be utilized for several purposes throughout a year [8]. most botanical parts of enset are good fodder for livestock. it is drought resistant because it contains a lot of water in pseudostem and used for cattle stall feeding especially in the dry season when grass is scarce. it provides good fiber, a by-product obtained from decorticating the leaf sheaths and the dried fibers are strong enough to make high quality ropes. the pseudostem yields very strong fibers, even the unprocessed leaf sheaths is used for tying livestock, bundling harvests from fields, and fencing. according to brandt, et al. [3] the fiber has excellent structure and its strength is equivalent to the fiber of abaca, a world-class fiber crop. in rural areas the fiber is used to make sacks, bags, ropes, cordage, mats, construction materials, and sieves. about 600 tons of enset fiber per year is sent to factories for processing [3]. fresh enset leaves are used for wrapping foods, serving plates, and pit linings to store kocho for fermentation. men sometimes make cap from enset and the fresh leaves are used as clothes for women skirts and often worn in the markets and ceremonies [6]. the dried petioles and midribs are used as firewood, to make mats, and tying materials for house construction. for cleaning rags, brushes, baby cushions, pot stands, as wrappers for butter, kocho, and other items to transport to local market [3]. particular varieties and enset parts are used medicinally for both human and livestock for problems like diarrhea, birth control (as an abortifacient), and assisting to discharge placenta [3]. enset has cultural values during wedding and funeral ceremonies. during wedding, enset leaves can be used on tables for serving food and as skirts for women in some cases in addition to being the major food for the ceremony. shigeta [6] elucidated that at funeral ceremony, people beat the psuedostems of enset laid on the ground in circle like drums and they also go with enset leaves on their hands. the members of the lineage cut all the enset plants when the head of the household dies, to express their sorrow and desperation [6]. the enset plant can also be sold in some cases and the processed products like kocho and bulla are sold anytime in the rural markets and towns. therefore, it is an immediate cash income source for a family to buy their daily needs. 3.5. some health benefits of enset some enset varieties are believed to have medicinal value and used by the enset growing community. for example in areka area, a variety called sweete is strongly recommended for treating a person with bone problem. this may be because it contains high calcium and phosphorous. even in the central highlands and cities where enset is not a staple, bulla is fed to a mother who gave birth for strengthening and fast recovery. they also make atmit (gruel) and given to a person caught cold. different enset varieties were reported to have medicinal and religious (ritual) significances for prevention, healing, and other therapeutic purposes. according to tsehaye and kebebew [20] tayo is a variety with a light red pseudostem and midrib with deep green leaf. the boiled corm and starchy powder bulla of this variety is eaten with milk to cure ailments such as joint displacement and swelling, broken bone fractures, and used to cure similar disorders in domestic animals, especifically it is fed with salt to dairy cows. some enset clones like astara and tayo are believed to have medicinal values and are used to treat human and livestock [43]. the corm of enset is fed to stimulate placental discharge and reduce delivery-associated stomachache and eaten with cheese, butter, and milk. officho, a dehydrated starch suspension and bulla are used together with milk for bone related problems [20]. furthermore, some clones are used for abortion or to reduce fertility. the following are some of the enset clones that are claimed to have medicinal and ritual values by farmers in kaffa sheka zone [43]: • choro is fed to a woman immediately after baby delivery in order to stimulate the discharge of placenta. in terms of ritual significance, planting this clone in their backyard deters away the devil or evil spirits by kaffa farmers. • the bulla or corm of tayo are consumed for bone fractures, backache, and swelling displacement of joints treatment. agriculture and food sciences research, 2016, 3(2): 66-74 73 • shasi wagi (wagi beli) is recommended for a woman to prevent uterus contraction and relive stomachache after baby delivery. • the corm of ariko is boiled and fed when a person catches flu. in addition, children eat its uncooked stem to keep healthy and gain weight. however, the scientific clinical studies of these enset medical uses were not reported. in fact, its potential use as glucose source is suggested because of its rich carbohydrate content. focusing mostly on the corm (amicho) by the farmers signifies their appropriate traditional knowledge of its better nutrient composition based on our study. 3.6. opportunities and challenges of enset cultivation having a field that encompasses the homestead is considered aesthetically desirable by enset-based societies, enset beautifies the landscape by its green foliage. it also affects the macro-environment of an area in a positive manner [3]. the native soils are altered positively through the long-term application of manure in areas where enset is grown for many years. enset’s perennial leaves canopy and the abundant accumulation of litter also reduces soil erosion and organic matter depletion. it plays the same role as trees in protecting people, other plants, and animals from wind and sun. plant species like enset possessing deep roots and leaf canopies of long duration improve the hydrological dynamics of an area. they improve water in the soil and aquifers through increasing water infiltration and reducing surface runoff. as a result, this increases water availability and discharge to springs and decreasing the effective dry season length [3]. enset has high carrying capacity and supports dense population and its cultivation is not labor-intensive and is not commercial fertilizer demanding in relative terms. it also has many non-food values as mentioned in the previous sections. collectively, the severe problems facing enset cultivation are diseases. they include sheath, corm, and dead heart leaf rots caused by an unknown bacterial pathogen and fungus as well as viral diseases, lesions, nematodes, and rootknot [3]. the knowledge limitation in the implementation of preventing strategies contributed to the severity of enset plant diseases. they are caused by several organisms. bacterial wilt, caused by the bacteria xanthomonas campestris pv musacearum, is the worst to the enset farming system [3]. it attacks the plant at any stages even after full maturity. it is a serious loss for the farmers when a disease kills an enset plant late in its life cycle as they have already invested labor, land, and resources for several years. in some cases, such situations lead farmers to replace enset farming with annual crops abandoning their land. the other shortcoming of this crop is it being poor in nutrient composition particularly protein and minerals. hence, appropriate nutritional strategies need to be studied and designed to promote the processing of ense food at industrial level. 4. summary enset is a multipurpose evergreen tree crop staple food for significant ethiopian population. it is indigenous to ethiopia and believed to be originated in the southwestern part of the country. it has several socio-economic and cultural values developed by the society for many years. enset has limitations in nutritional adequacy particularly in essential amino acids and important minerals. therefore, supplementary food sources of protein and minerals are highly recommended for people who entirely depend on especially bulla/kocho. in addition, nutritious varieties need to be selected and promoted. in order to sustain and improve enset cultivation, serious and continuous work is required to control diseases, improving its productivity, and nutritional schemes. furthermore, its products need to be processed in factories with better quality and high market values. references [1] j. r. harlan, "ethiopia: a center of diversity," economic botany, vol. 23, pp. 309-314, 1969. 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[43] a. negash, "diversity and conservation of enset (ensete ventricosum welw. cheesman) and its relation to household food and livelihood security in south-western ethiopia," wageningen university, phd thesis, 2001. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 2, no. 1, 1-5, 2015 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 1 dietary fibers in grain species as sources of wholegrain foods gyurova, desislava krasteva 1* --enikova, rositsa kirilova 2 1,2 department “organic analyses”, national centre of public health and analyses, sofia, bulgaria abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ................................................................................................................................................................................. 2 2. material and methods ................................................................................................................................................................. 2 3. results and discussion ................................................................................................................................................................ 3 4. in conclusion ............................................................................................................................................................................... 4 references ........................................................................................................................................................................................ 4 the paper presents the results of an analysis of 20 lots of cereal products (grains and processed grains products) from different geographic regions. the samples were analyzed for dietary fibers content applying the enzymatic-gravimetric method aoac 985.29. the differences in the amounts of dietary fibers in the individual grain varieties cover wide intervals and vary between 19 and 41% approximately, and between particular types of flour – in the interval 9 – 25% approximately. the obtained results can be used in completing and upgrading the chemical composition tables of bulgarian foods as well as respond to the needs of medical dietology and dietetic practice in bulgaria from the viewpoint of formulation of healthy foods and recipes. those data can also be used to provide for informed choice through listing of nutritional information on food labels. keywords: wholegrain foods, flours, sources, dietary fibers, cereals, analysis. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(1): 1-5 2 1. introduction the health benefits of dietary fibers are indisputably proven by medical science and practice. a number of serious diseases with broad prevalence are favorably affected by the regular consumption of dietary fibers within the frames of implemented dietary regimes. thus, for example, it has been proven that the consumption of cereals and wholegrain foods (especially those with oats and barley fibers) is associated with protective reactions against cardiovascular diseases in adults [1-5]. they reduce the risk for coronary heart disease [1, 3-6]. a number of research studies outline that wholegrain foods intake (at least 2, 5 g daily) is associated with 21% smaller risk for cardiovascular events. the regular intake of cereals rich in soluble fibers, similar to those in oats, including betaglucans causes significant reduction of blood cholesterol levels. the consumption of wholegrain foods reduces the risk for development of colon cancer [7, 8]. this is associated with the effect of dietary fibers in accelerating the intestinal passage through the alimentary canal. the intake of 1 to 3 portions daily of foods rich in dietary fibers reduces the colorectal carcinoma risk in adults with 20% approximately. it has also been established that cereals and wholegrain foods decrease the probability for diverticulitis development [9]. the health effects of wholegrain foods on the risk for development of type 2 diabetes are quite important [1013]. studies on nurses have revealed that two portions of wholegrain foods daily could cause a 21% reduction of the risk for type 2 diabetes developments. the above mentioned diseases are currently associated with overweight and obesity issues. individuals with overweight tend to energy-dense diets. foods rich in fibers, such as cereals and wholegrain bread are an adequate effective measure for bodyweight reduction as a particular amount of food introduces much less energy than the same amount of concentrated foods with low fiber content. foods with high content of fibers need more time for digestion and create a sense of repletion which prevents surfeit. wholegrain foods are a constant part of healthy dietetic regimes as, besides fibers, they introduce other bioactive nutrients in the human organism too [2]. they have low natural levels of saturated fatty acids but contain polyunsaturated fatty acids, mainly linolenic acid. they do not supply cholesterol and are an excellent source of slow carbohydrates and protein, of soluble and insoluble fibers and stable starch. wholegrain foods are also a good source of iron, magnesium, copper, selenium, phosphorus and zinc, as well as of antioxidants and phytochemicals supporting blood cholesterol reduction. they are an important source of b-vitamins including folic acid. full meeting of folate demands is observed in population groups in fertile age on healthy diets with wholegrain foods [14]. many plant species included in the diet of the population are natural sources of dietary fibers. their edible parts, rich in fibers can be consumed directly or can be used as fiber sources added to other processed foods. some plant foods contain significant amounts of both soluble and insoluble fibers. those are grains of numerous species, whole fruits and pulses. soluble fibers are found in various amounts in all foods of plant origin, including wheat, oats, rye and barley and other grain species, pulses (beans, green peas, soybean, etc.), some fruits and fruit juices (including plums, forest fruits, bananas and the pulp of apples and pears); some vegetables as broccoli and jerusalem artichoke; tuberiferous plants, such as sweet potatoes, carrots, onions, etc. soluble fibers dissolve in water and help reduce blood glucose and cholesterol [8]. whole grains and wholegrain foods are sources of insoluble fibers [15]; wheat and corn bran; nuts and seeds, such as flax seeds; vegetables as green beans, cauliflower, vegetable marrows, celery; some fruits, including avocado and bananas; the peel of many fruits and tuberiferous plants, including tomatoes and plums – an example of fruit with thick peel [15-17]. the insoluble fibers do not dissolve in water and stimulate the peristaltic and passage of the food through the digestion tract, stimulate regular defecation and prevent constipation. the cereal species used in human diet are numerous: wheat, rye, oats, barley, corn, rice, buck-wheat, millet, triticale, einkorn wheat, spelt. wheat belongs to the polymorphous genus triticum of the family graminea/poacea. winter wheat (t. aestivum l.) and hard wheat (t. durum desf.) are cultivated in bulgaria. emmer wheat (t. turgidum l.), the hybrid between wheat and rye triticale, spelt, einkorn wheat (t. monococcum l.) and wild emmer (t. dicoccum schrank) are much less implemented. those wheat varieties grow in weaker, eroded soils, can be cultivated in semi-mountain regions and recently have gained popularity as ecologically pure and appropriate for dietetics. the grains consist of three main parts as follows:  bran and aleurone layer that in grinding separate as bran, but contain the fibers, unsaturated fatty acids, vitamins and minerals;  endosperm – the main internal part of the seed containing mainly starch;  germ – the smallest part of the seed containing vitamin e, folate, thiamin, the minerals phosphorus and magnesium. the available literature evidence on health benefits of wholegrain foods and dietary fibers in them enable constant monitoring of the content of ballast substances in cereals, popular in the individual countries and promoted as sources of such substances, adequate for dietetic purposes. the aim of this survey is to determine the content of total fibers in cereal and flour lots used by the bulgarian food industry in the production of dietetic foods. 2. material and methods samples of 20 lots of cereals (grain and grain products) of various geographic origins were delivered in 2014 to the laboratory for analysis of dietary fibers content. the analysis was made with the implementation of the enzymatic-gravimetric method aoac 985.29 for determination of dietary fibers content. agriculture and food sciences research, 2015, 2(1): 1-5 3 laboratory samples were prepared for the tasks of the analysis. the results are a mean value of three parallel samples of each type with listed standard uncertainty. 3. results and discussion the results of the analysis of total fiber content in various grain types are presented in table 1: table-1. content of total dietary fibers in cereals studied cereal products df ± su*, % wheat grain bulgaria 41,13 ± 0,41 wheat grain bulgaria 24,82 ± 0,25 wheat (soft) bulgaria 19,06 ± 0,14 wheat (hard) bulgaria 19,23 ± 0,14 wheat soft grain usa 29,46 ± 0,29 einkorn wheat grains – bulgaria i 13,50 ± 0,10 einkorn wheat grains – bulgaria ii 19,38 ± 0,14 einkorn wheat grains – bulgaria ііі (galabovo) 31,59±0,31 einkorn wheat grains – italy 25,60 ± 0,26 * su – standard uncertainty in bulgaria wheat is the major agrarian crop, cultivated on 10 – 15 million decares [18]. the most popular is the so called winter wheat used for production of bread grain and forage. southern bulgaria and the northern region near the black sea provide good conditions for cultivation of hard wheat – adequate for the production of pasta, hulled grains, etc. recently some wheat varieties also have gained popularity, especially einkorn wheat. einkorn wheat (triticum monococcum) has uncultivated and cultivated forms and subtypes. emmer wheat (triticum dicoccum) is a type of wild wheat. in the mesolite neolite period emmer wheat has been widely used as plant food, replaced later by cultivated wheat. spelt (triticum spelta) known in bulgaria also as “dinkel” is a type of ancient wheat. it has been a major food in some parts of the near east and europe since the bronze age until the middle ages [19]. nowadays it is perceived as healthy food as it has a higher content of proteins than wheat as well as more fibers, vitamins and minerals. it contains almost twice more vitamin a and b vitamins, fats, phosphorus and proteins, but minimal gluten amounts. the flour obtained from spelt is fine and the bread, biscuits and pasta made of it are very tasty. our studies refer to wheat and einkorn wheat grain. the results are an example for significant differences in the total fibers content in the various grain types. this content in the various grain varieties from bulgaria varies in a very broad interval (approximately 19 – 41%). soft wheat grain, imported from the usa is a medium fiber source. yet greater differences are shown by the individual einkorn wheat lots – the fiber content in them varies from 13 to about 31 %, and the italian lot contains 25% of ballast substances. flours are powder-like food products obtained by grinding certain grains. most often they are produced from wheat and its varieties, oats, rye, corn, millet, rice, buck-wheat, etc. flours are used for production of traditional cereal foods – bread and bakery, confectionery, pasta, cereals, etc. the technological processing of grains causes a significant loss of micronutrients. when it is ground into flour, most useful components are removed with the bran. high-quality white flours contain significantly less vitamins and minerals than dark and wholegrain flours. currently the modern mills produce various flour types with/without removal of the bran and germ. refined white flours are produced from the endosperm with elimination of the bran and germ. this process causes loss of fibers, vitamins, minerals, antioxidants and other bioactive substances. the latter can be added back in refined flours but similar actions cannot compensate for the losses fully. refined flours are used for the production of white bread, pasta, bakery, sweet or salty biscuits, wafers, pizza, cakes and pastry. those products usually contain high levels of added sugar, fats or salt, their glycemic index is higher than that of their wholegrain equivalents. the consumption of refined cereal products causes sharp elevation of blood sugar and a strong pancreatic reaction. wholegrain flours are natural products obtained by grinding of the whole grain without removal of any fractions from it duleva [20]. the bran of the grain contains great amounts of minerals, iron, copper, selenium, magnesium, bvitamins, and also has a substantial amount of ballast substances. when the whole grain is ground to wholegrain flour, we achieve a product that has all substances from the composition of wheat, corn or rye preserved [20]. from medicobiological point of view flours of whole grains, produced by the so-called simple grinding are the most valuable. they are rich in vitamins, minerals, cellulose and proteins. from technological point of view, though, they are of low quality as they are unstable for storage and have poor bakery qualities. wholegrain flours and their products contain the three parts of the grain – bran, germ, endosperm. wholegrain cereal products are very diverse – various types of wheat, rye and combined bread, pasta, wholegrain cereals, croup, oatmeal, brown rice, extruded products, variegated combined grain mixtures. the dietetic practice implements also the grain fragments rich in fibers and bioactive substances – wheat bran, germs etc. rye flours have different chemical composition and bakery qualities from those of wheat. the bread made of rye flour is dark, with small volume, poor porosity, sticky, wet inner part. the proteins in the rye flour contain basic fractions – gliadin and gluten in almost equal amounts. the amino acid composition of rye flour proteins is close to that of wheat proteins but they have greater content of the essential amino acids lysine and threonine. rye flour contains more own sugars and easily soluble polysaccharides and trisaccharides that are hydrolyzed to hexoses. corn flour contains a large amount of carbohydrates, where the amount of starch is the greatest reaching up to 85%. it is poor in proteins and amino acids and relatively richer in fats, which determines its short shelf-life. the corn flour bread has coarse, solid, non-elastic and quickly drying inner part, small volume and crackled crust. because of those bakery qualities corn flour is not used individually but as an additive (up to 15%) to wheat flour. agriculture and food sciences research, 2015, 2(1): 1-5 4 table 2 presents the results from our analysis of total fibers in various wheat, rye, einkorn wheat and spelt flour types. table-2. content of total dietary fibers in flours analyzed cereal products df ± su*, % wholegrain wheat flour bulgaria i 16,41±0,30 wholegrain wheat flour bulgaria ii 23,40 ± 0,23 wheat flour type 400 bulgaria 9,01 ± 0,07 wheat flour type “dark” bulgaria 16,12 ± 0,12 wholegrain rye flour bulgaria 20,10 ± 0,15 rye flour type 1000 bulgaria 13,47 ± 0,10 einkorn wheat flour bulgaria i 16,39 ± 0,12 einkorn wheat flour bulgaria ii 15,92 ± 0,12 wholegrain wheat flour usa 25,35±0,25 wholegrain wheat flour “pastry” usa 18,52 ± 0,14 spelt flour usa 12,41±0,09 * su – standard uncertainty the main criterion for determination of the wholegrain extent of flours is the ash content – the higher the ash content, the higher wholegrain extent the flour has. the value of the ash content specifies the flour type and is specially marked when writing the flour type. the number specifying the flour type corresponds to its ash content in mg/100 g, transposed in percents [20]. for example, flour “type 1150” has ash content of 1.15%. flours with high ash content contain a big relative rate of the grain bran, they have darker color, more vitamins, minerals and enzymes. flours with higher ash content contain more proteins but their bakery qualities are worse. the flours that are most widely used in bulgaria are:  wholegrain wheat flour, ground from whole grains, not sieved in any way – type 1850 (also called “graham”);  type 2000 – wholegrain einkorn wheat flour;  type 1750 – wholegrain rye flour;  type 1000 – partially refined rye flour;  type 1150 – partially refined wheat flour, called in bulgarian practice “tipovo”;  type 500 and type 400 are white wheat flours. the obtained results concerning wheat flours show a relatively high content of total fibers but too large differences between various lots declared as wholegrain. the differences established between the two rye lots are absolutely explicable – the wholegrain one contains a much greater percentage of fibers compared to the partially refined of type 1000. the imported usa lots of wholegrain wheat flour have high fiber content. according to international literature sources [21, 22] lower content of total dietary fibers is detected in analogous products analyzed by the same enzymatic-gravimetric method 985.29, respectively:  for wholegrain wheat flour the values vary in the interval 9,9 – 13,8% [22];  for wholegrain rye flour the fibers are from 12,9 to 16,7% [1.1]; 14,8 % [21]  for wholegrain wheat the total fibers content is 12,0% [21];  for flour made of wheat and rye, 40% wholegrain 7,5 % [21]. the differences in the quantities of fibers in those cases could be substantiated by the characteristic features of the geographic regions as well as by the type and variety of cultivated wheat types [15]. there are no specified standard requirements to the content of dietary fibers in flours. the producers of dietetic bread and dietetic cereals, though, must be well informed about the composition of flours, produced by the mills and especially about those flour assortments, declared as wholegrain. the quality certificates of the flours have to contain mandatory clauses stating the dietary fibers content. this also requires unification of the analytical methods. our opinion is that aoac 985.29, implemented in this study, is the most appropriate referent method for determination of total fibers. only in the presence of those conditions the dietetic bread and cereals assortments would be able to maintain constant composition and quality and to comply with the stated health pretensions for wholegrain dietary products. 4. conclusion we should confirm the advice of experienced specialists in dietetics:  “substitute white rice, white bread and white pasta with brown rice and wholegrain products”;  “for your breakfast chooses those cereals that have a wholegrain component as their first ingredient”. the current agrarian policy and food industry present broad possibilities for production of variable high quality cereals, rich in dietary fibers and relevant vitamins, minerals and essential bioactive substances, necessary for the healthy diet and high life quality of the contemporary people. references [1] e. b. rimm, a. ascherio, e. giovannucci, d. spiegelman, m. j. stampfer, and w. c. willett, "vegetable, fruit, and cereal fiber intake and risk of coronary heart disease among men," jama, vol. 275, pp. 447-51, 1996. [2] http://www.betterhealth.vic.gov.au/bhcv2/bhcarticles.nsf/pages/cereals_and_wholegrain_foods?open, healthy living > food and nutrition foods, cereals and wholegrain foods. australia: deakin university, 2015. [3] l. brown, b. rosner, w. w. willett, and f. m. sacks, "cholesterol-lowering effects of dietary fiber: a meta-analysis," am. j. clin. nutr., vol. 69, pp. 30-42, 1999. [4] n. m. mckeown, j. b. meigs, s. liu, p. w. wilson, and p. f. jacques, "whole-grain intake is favorably associated with metabolic risk factors for type 2 diabetes and cardiovascular disease in the framingham offspring study," am. j. clin. nutr., vol. 76, pp. 390-8, 2002. http://www.betterhealth.vic.gov.au/bhcv2/bhcarticles.nsf/pages/cereals_and_wholegrain_foods?open agriculture and food sciences research, 2015, 2(1): 1-5 5 [5] n. m. mckeown, j. b. meigs, s. liu, e. saltzman, p. w. wilson, and p. f. jacques, "carbohydrate nutrition, insulin resistance, and the prevalence of the metabolic syndrome in the framingham offspring cohort," diabetes care, vol. 27, pp. 538-46, 2004. [6] m. a. pereira, e. o. reilly, and k. augustsson, "dietary fiber and risk of coronary heart disease: a pooled analysis of cohort studies," arch. intern med., vol. 164, pp. 370-6, 2004. [7] c. s. fuchs, e. l. giovannucci, and g. a. colditz, "dietary fiber and the risk of colorectal cancer and adenoma in women," n. engl. j. med., vol. 340, pp. 169-76, 1999. [8] http://www.hsph.harvard.edu/nutritionsource/carbohydrates/fiber/, "carbohydrate, fiber, harvard school of public health, nutritional source," 2015. [9] w. h. aldoori, e. l. giovannucci, h. r. rockett, l. sampson, e. b. rimm, and w. c. willett, "a prospective study of dietary fiber types and symptomatic diverticular disease in men," j. nutr., vol. 128, pp. 714-719, 1998. [10] s. m. b. schulze, e. liu, b. rimm, j. e. manson, w. c. willett, and f. b. hu, "glycemic index, glycemic load, and dietary fiber intake and incidence of type 2 diabetes in younger and middle-aged women," am. j. clin. nutr., vol. 80, pp. 348-56, 2004. [11] s. krishnan, l. rosenberg, and m. singer, "glycemic index, glycemic load, and cereal fiber intake and risk of type 2 diabetes in us black women," arch. intern. med., vol. 167, pp. 2304-9, 2007. [12] s. liu, w. c. willett, and m. j. stampfer, "a prospective study of dietary glycemic load, carbohydrate intake, and risk of coronary heart disease in us women," am. j. clin. nutr., vol. 71, pp. 1455-61, 2000. [13] t. t. fung, f. b. hu, and m. a. pereira, "whole-grain intake and the risk of type 2 diabetes: a prospective study in men," am. j. clin. nutr., vol. 76, pp. 535-40, 2002. [14] v. birdanova, m. stoinovska, n. statev, l. boyanov, and k. petkov, "current issues of the students catering at the medical university – pleven," science & technologies, medicine, (in bulgarian), vol. 2, pp. 215-219, 2012. [15] d. k. gyurova, "dietary fibers in bulgarian foods," ph. d. dissertation, national centre of public health and analyses, bulgaria, 2013. [16] a. alvarado, e. pacheco delahaye, and p. hevia, "value of a tomato byproduct as a source of dietary fiber in rats," plant foods hum. nutr., vol. 56, pp. 335–48, 2001. [17] m. stacewicz – sapuntzakis, "chemical composition and potential health effects of prunes: a functional food," crit. rev. food sci. nutr., vol. 41, pp. 251–86, 2001. [18] http://bg.wikipedia.org/wiki/%d0%9f%d1%88%d0%b5%d0%bd%d0%b8%d1%86%d0%b, "wheat," 2014. [19] m. nesbitt, wheat evolution: integrating archaeological and biological evidence, 2001, in wheat taxonomy: the legacy of john percival, linnean, special issue. edited by p. d. s. caligari and p. e. brandham vol. 3. london: linnean society, 2001. [20] v. duleva, what are the different types of flours? how to choose balanced nutrition with wholegrain types. in bulgarian. available: http://www.edna.bg/, 2013. [21] http://www.matportalen.no/verktoy/the_norwegian_food_composition_table/, "the norwegian food composition table," the norwegian food safety authority, the directorate of health and social affairs and the department of nutrition at the university of oslo2006. [22] http://www.foodcomp.dk/v7/fcdb_search.asp, "the official danish food composition database, danish food composition databank, new version 7.01," 2009. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. http://www.hsph.harvard.edu/nutritionsource/carbohydrates/fiber/ http://bg.wikipedia.org/wiki/%d0%9f%d1%88%d0%b5%d0%bd%d0%b8%d1%86%d0%25b http://www.edna.bg/ http://www.matportalen.no/verktoy/the_norwegian_food_composition_table/ http://www.foodcomp.dk/v7/fcdb_search.asp 109 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 109-113, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.109.113 © 2019 by the authors; licensee asian online journal publishing group performance of cvl-1 seedling growth and dry weight attributes under greenhouse and field conditions as affected by seed source and seedling age md. mahbubul islam1 jannatul ferdous2 manika rani debnath3 ( corresponding author) 1chief scientific officer and agronomy division of bangladesh jute research institute, dhaka. bangladesh. 2,3scientific officer, agronomy division of bangladesh jute research institute, dhaka. bangladesh. abstract experiments were conducted to evaluate the plant establishment of white jute (corchorus capsularis l.) seeds collected from five different sources of bangladesh. the experiments were conducted at greenhouse of bangladesh jute research institute (bjri), dhaka and field of jute agriculture experimental station (jaes), bjri, manikganj during march to june 2018. the treatments consisted of five seed sources viz., i) bjri, ii) bangladesh agriculture development corporation (badc), iii), iv) farmers of two locations and v) local market. farmers source was divided into two sub-sources (farmer1 of manikganj and farmer2 of kishoreganj locations). two different conditions were i) greenhouse and ii) field. the four seedling ages were 30, 40, 50, and 60 days. the cv. cvl-1 was used as study material. randomized completely block design with three replication were followed. results revealed that seedling growth and dry weight parameters varied significantly due to seed sources. the shoot height, base diameter, root length, shoot dry weight and root dry weights varied significantly due to seed sources in both the condition of greenhouse and field. bjri and badc sources showed better performance than the others. the attributes showed significant variations for seedling ages in both the conditions also. the highest values were observed in 60 days of seedling age. the interactions of seed sources and seedling ages were highly significant for all the growth and dry weight attributes. the bjri seeds of 60 days of age performed the highest shoot height, base diameter, root length, shoot dry weight and root dry weights. seeds from bjri and badc of the cvl-1 varieties showed better performances in terms of seedling growth and dry weight compared to seeds from farmers’ and local markets. keywords: white jute (corchorus capsularis l.), seedling ages, shoot length, root length, base diameter, shoot dry weight, root dry weight. citation | md. mahbubul islam; jannatul ferdous; manika rani debnath (2019). performance of cvl-1 seedling growth and dry weight attributes under greenhouse and field conditions as affected by seed source and seedling age. agriculture and food sciences research, 6(1): 109-113. history: received: 6 march 2019 revised: 2 april 2019 accepted: 8 may 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: the research article is funded by institute (bangladesh jute research institute). competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 110 2. materials and methods ................................................................................................................................................................. 110 3. results and discussion ................................................................................................................................................................. 110 4. conclusion ....................................................................................................................................................................................... 112 references ............................................................................................................................................................................................ 112 http://www.asianonlinejournals.com/index.php/aesr/article/view/128 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.109.113&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.109.113&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2019, 6(1): 109-113 110 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by evaluating the plant establishment of white jute (corchorus capsularis l.) seeds collected from five different sources of bangladesh. 1. introduction jute (corchorus spp.) is one of the main cash crops of bangladesh. it plays an important role earning about 56% foreign exchange through exporting jute and jute goods. jute is a common term used both for plant and the fibre obtained from the bark of the plants, corchorus capsularisl. and corchorus olitoriusl. it covers about 2.86% of total cropped area. bangladesh annually covering about 0.761 millions ha of land by jute crop to produce about 1.62 millions ton of fibre. the farmers require about 5500 tons of seed to cultivate the said area [1]. bangladesh produces world’s best quality jute. the agriculture climate of bangladesh is very much suitable for quality fibre production. as a result supply of quality jute seed will ensure the fibre quality as well as quantity to a great extent. the government of bangladesh has given more thrust to promote the jute seed industry by strengthening public sectors dealing with jute seed and also by sensitizing private sectors to reduce the dependence on imported jute seed. the activities of private sectors regarding jute seed are limited on seed import only [2]. bangladesh requires about 5,000 metric tons of jute seeds of which only 12-15% is produced and distributed by bangladesh agricultural development corporation [3]. the rest of seeds is solely produced and utilized by the farmers’. the quality of farmers’ seed is not maintained during production, processing and storage. in many cases, farmers’ collect their seeds from their friends or neighbors and thus, there is no specific system to control the quality of seeds. hence, it is conceived that the quality of seeds produced by majority of the farmers’ is of low standard. also, there is very little information regarding the level of farmers’ knowledge on the production of jute seed and method of quality testing before sowing. farmers’ sometimes get good germination and good crop, but in most cases they get poor germination and a poor crop, and occasionally seeds do not germinate which results in total crop failure [4-6]. the quality status of jute seed at farm level is very poor and farmers’ are normally ignorant of seed quality and quality evaluating tests. farmers’ are not aware of the germination percentage, vigor value, pathogens associated with seeds and percentage of moisture in their seeds. although, there are many researches on the quality of the seed, no effort has yet been made to evaluate the quality of seed at farm level despite 75% of the total requirements of the jute seed are solely produced and distributed by the farmers’. if it is possible to address and appreciate the seed growers about their seed problems and help understand them to assess quality of jute seeds, they could establish it as a profitable enterprise [7, 8]. seeds from bjri and badc of both the jute varieties showed better quality in terms of vigor and pathogen incidence compared to seeds from farmers’ and local markets [9]. jute seed quality does not vary only from farmer to farmer, but also from source-to-source. information relating to viability, vigour and emergence of jute seed are very scanty under bangladesh condition. moreover, the category and extent of jute seed quality through seedling emergence and establishment of different sources are yet to be elucidated. the present study was, therefore, undertaken to evaluate the performances of seedling growth of white jute and their interactions as influenced by seed sources and seedling age. 2. materials and methods the experiments were conducted at the crop management laboratory, agronomy division of bangladesh jute research institute (bjri). the emergence experiment was conducted at crop management laboratory, agronomy division of bangladesh jute research institute (bjri), dhaka as greenhouse and at field of jute agriculture experimental station, manikgonj of bjri during march to june 2017. the treatments consisted of five seed sources viz., i. bjri, ii. bangladesh agricultural development corporation (badc), iii., iv) farmers of two locations and v. local market. the farmers seed source was divided into two sub-sources. the sub sources were farmer1 of manikganj and farmer2 of kishoreganj locations. the seed sample variety cvl-1 of corchorus capsularis l. was used as study material. primary seed samples of 250 g each from all sources and from 20 farmers of each location were collected randomly. all the primary seed samples were mixed thoroughly to make a composite sample of each location. from the composite samples 500g seeds were taken and regarded as submitted sample. the submitted seed samples were kept in brown paper bags. all the seed samples collected from the different seed sources were labeled properly and preserved in gene bank of bjri at 200 c till the samples were used for studies. working seed samples were taken from the preserved seed samples as per requirement. total procedure was maintained following the rules of ista (international seed testing association) [10]. local market seed samples were collected and processed as that of farmers source of two locations. after collection of seed samples following tests were conducted. the soil of field opened with power tiller in the month of mid february, then the land was prepared with the country spade. the laboratory greenhouse soil was prepared and managed as per recommendation of bjri. the land was made free from weeds and plant debris by hand collection. during final plot preparation npk fertilizers were applied in the form of urea, triple super phosphate and muriate of potash. the rates of fertilizers used were 83-25-30-11 kg of npks ha-1, respectively. half quantity of n and the whole quantity of p, k and s were applied as basal dose. the other half quantity of n was top dressed in two equal splits at 15 das. seeds used for the experiments had minimum of 80% germination. seeds were sown on 25 march and 26 march in greenhouse and in the field, respectively. for each unit plot 500 seeds of each sample sources were sown in broadcast method and later covered with soil. in establishment test, 30, 40, 50 and 60-day-old seedlings were randomly collected from the greenhouse laboratory and as well as from field. the data of seedling shoot length, base diameter, root length, shoot dry weight and root dry weight of the seedlings were collected. treatment mean differences were adjudged by dmrt following level of significance (0.05 and 0.01%) [11]. 3. results and discussion results revealed that at greenhouse, seedling height, base diameter, root length, shoot dry weight and root dry weights differed significantly due to seed sources for the variety cvl-1 at greenhouse conditiontable 1. the agriculture and food sciences research, 2019, 6(1): 109-113 111 © 2019 by the authors; licensee asian online journal publishing group highest seedling height, base diameter, root length, shoot dry weight and root dry weights (103.75 cm, 10.75 mm and 12.20 cm, 8.13 g and 3.02 g, respectively) were recorded in bjri seeds, which were statistically similar to badc seeds except root length. the lowest seedling height, base diameter and root length were recorded in farmers seed of manikgonj table 1. the farmers seed of manikgonj and kishoregonj; and market seed had statistically equal seedling height, base diameter, root length, shoot dry weight and root dry weights, the variation was found in seedling height table 1. irrespective of seed sources seedling height, base diameter, root length, shoot dry weight and root dry weights of cvl-1 varied significantly due to different seedling age at fieldtable 1. except root dry weight all others showed statistical similarity in bjri and badc seeds. seedling shoot dry weight and root dry weight were highest in bjri seeds followed by badc. the lowest seedling shoot dry weight of farmers seed of kishoreganj, and root dry weight of farmers seed of manikganj; and stem dry weight in farmers seed of manikganj and root dry weight in market seed were recorded. in terms of seed stem dry weight badc seed showed statistical similarity with farmers and local market seeds table 1. under field condition significant variations were found in shoot and root dry weight among the seed sources of cvl-1 table 1. statistical similarity observed in bjri and badc seeds in terms of seedling shoot and root dry weight. the shoot dry weight and root dry weights were highest in bjri seeds followed by badc. the lowest shoot and root dry weight were observed in farmers seed of manikganj. local market seeds had the third highest shoot and root dry weights among the seed sources. the seedling shoot dry weight of market seed had similarity with bjri and badc seeds table1. development of a normal seedling from the seed is an essential requirement in the maintenance of seed viability. a normal seedling essentially should have a well-developed root system, some intact hypocotyls, two cotyledonary leaves and plumule with developing green leaves [5]. table-1. effect of seed sources on seedling growth characters of cvl-1 under green house and field condition. seed sources at greenhouse at field seedling height (cm) base diameter (mm) root length (cm) shoot dry weight (g) root dry weight (g) seedling height (cm) base diameter (mm) root length (cm) shoot dry weight (g) root dry weight (g) bjri 103.75 a 10.75 a 12.20 a 8.13 a 3.02 a 99.33 a 10.18 a 11.74 a 6.94 2.83 a badc 101.25 a 10.39 a 11.65 b 7.88 ab 2.79 a 96.58 a 9.83 b 11.21 a 7.41 2.76 a farmer1 84.25 cd 8.72 c 10.79 c 7.04 c 2.39 c 83.08 c 8.27 e 9.78 c 6.13 2.29 c farmer2 86.58 c 9.03 bc 10.98 c 6.92 d 2.55 bc 86.50 bc 8.48 d 10.01 bc 6.47 2.43 bc local market 91.42 b 9.36 b 10.90 c 7.23 b 2.66 b 89.08 b 8.71 c 10.37 b 6.65 2.49 b note: in a column figures having common letter(s) do not differ significantly by dmrt at 1% level of probability. irrespective of seed sources the seedling age showed significant differences in respect of seedling height, base diameter, root length, seedling shoot and root dry weights at field of the variety cvl-1 at greenhouse condition table 2. the highest seedling heights base diameter and root length were 156.40 cm, 15.97 mm, 15.92 cm, 18.97 g and 7.94 g, respectively. the lowest were observed at 30 das. the seedling height, base diameter and root length increased with the increase of seedling age. significant differences were observed under seedling age regarding seedling height, base diameter and root length of cvl-1 due to seed sources table 2. the highest seedling height, base diameter, root lengths, shoot dry weight and root dry weights were observed at 60 days of seedling age. the lowest seedling height, base diameter and root length were 45.40 cm, 4.91 mm, 7.19 cm, 1.03 g and 0.34 g, respectively, were observed at 30 days of seedling age at greenhouse. a chronological increase was found in seedling height, base diameter and root length with the increase of seedling age from 30 to 60 days at an interval of 10 days table 2. the highest seedling height, base diameter, root length, seedling shoot and root dry weight, were found at 60 days after sowing (das) at field. the highest seedling height, base diameter and root length were found in bjri seeds, which were similar to badc seeds except base diameter. the farmer’s seed of manikganj had the lowest values for these parameters, which were similar to farmer’s seed of kishoreganj especially for seedling height at field condition table 2. local market seeds showed a little bit higher performance than farmers seeds. local market seeds produced similar seedling height as that of farmers’ seed of kishoreganj table 2. irrespective of seed sources seedling age differed significantly regarding seedling shoot dry weight and root dry weight in cvl-1 table2. gradual increases were found in shoot and root dry weight with the increase of seedling age. in terms of shoot and root dry weight the variations among the seedling age were very high. the highest seedling shoot dry weights were found at 60 days and root dry weights were recorded in both the years at 60 das. the lowest shoot dry weight and root dry weights were found at 30 das table2. seedling age significantly affected the shoot and root dry weights in cvl-1 under field condition irrespective of seed sources. gradual increases were found in seedling shoot and root dry weight with the increase of seedling age. very high variations among the duration were recorded in shoot and root dry weight at field condition also. the highest shoot and root dry weights were observed at 60 das, while it was the lowest at 30 das table 2. table-2.effect of seedling characters of cvl-1 under greenhouse and field condition. seedling age (days) at greenhouse at field seedling height (cm) base diameter (mm) root length (cm) shoot dry weight (g) root dry weight (g) seedling height (cm) base diameter (mm) root length (cm) shoot dry weight (g) root dry weight (g) 30 45.40 d 4.91 d 7.19 d 1.03 d 0.34 d 38.80 d 4.53 d 6.71 d 0.97 d 0.33 d 40 68.87 c 6.96 c 10.18 c 2.78 c 0.79 c 67.47 c 6.37 c 10.00 c 2.56 c 0.78 c 50 103.13 b 10.76 b 11.92 b 6.98 b 1.66 b 101.00 b 10.44 b 11.43 b 6.48 b 1.93 b 60 156.40 a 15.97 a 15.92 a 18.97 a 7.94 a 154.44 a 15.03 a 14.35 a 16.88 a 7.21 a note: in a column figures having common letter(s) do not differ significantly by dmrt at 1% level of probability. das= days after sowing. agriculture and food sciences research, 2019, 6(1): 109-113 112 © 2019 by the authors; licensee asian online journal publishing group the interaction between seed sources and seedling age affected the seedling height, base diameter, root length, shoot dry weights and root dry weights significantly for cvl-1 at greenhouse table 3. the highest seedling height, base diameter, root length, shoot dry weight and root dry weights were recorded (167.33 cm, 17.23 mm, 16.77 cm, 20.61 g and 8.91 g, respectively) in bjri seeds at 60 days seedling age. the lowest seedling height and base diameter (41.00 cm and 4.03 mm, respectively) were observed in farmers’ seed of manikgonj with 30 days seedling age. at 30 days seedling age, was statistically identical to farmers’ seed of manikgonj and market seed table 3. badc seeds showed statistically identical results to bjri. the seedling shoot and root dry weights were the lowest in farmers seed of manikgonj and in local market seed and in farmers seed of kishoregonj at 30 das. farmers’ seed of both manikgonj and kishoregonj; and market seeds showed statistical similarity as regard to seedling age table 3. seedling height, base diameter, root length, seedling shoot and root dry weights of cvl-1 under field condition were significantly affected by the interaction between seed sources and seedling age. the highest seedling height, base diameter, root length, seedling shoot and root dry weights were observed in bjri seeds at 60 days of seedling age followed by badc seeds. in farmers seed of manikganj the lowest values for these parameters were found at 30 das. seedling height, base diameter and root length observed under greenhouse condition were higher than those at the field table 3. badc seeds had statistical similarity with bjri seed. the seedling shoot and root dry weights were lowest at 30 das in farmers seed of manikgonj. similar root dry weight was observed in farmers’ seed of kishoregonj. farmers’ seed of both the locations and market seed’s had shoot and root dry weights statistically indifferent table 3. results are in agreement with islam [5] and islam [9]. table-3.interaction effect of seed sources and seedling age on seedling growth characters of cvl-1 under green house and field. seed sources and seedling age(days) at greenhouse at field seedling height (cm) base diameter (mm) root length (cm) shoot dry wt (g) root dry wt (g) seedling height (cm) base diameter (mm) root length (cm) shoot dry wt (g) root dry wt (g) bjri 30 51.00d 6.00d 8.13cd 1.13d 0.40d 44.67d 5.43d 7.70d 1.05d 0.39d 40 81.33c 7.97c 11.13c 2.98c 0.85c 76.62c 7.37c 10.60bc 2.98c 0.85c 50 115.33b 11.80b 12.77b 7.82b 1.92b 112.00b 11.40ab 12.61b 7.56b 2.03b 60 167.33a 17.23a 16.77a 20.61a 8.91a 164.00a 16.53a 16.07a 18.56a 8.06a badc 30 48.67d 5.83d 7.67d 1.09d 0.38d 43.32d 5.20d 7.30d 0.98d 0.37d 40 79.00c 7.67c 10.50c 2.90c 0.83c 74.00c 7.10c 10.17bc 2.88c 0.81c 50 113.00b 11.33b 12.33b 7.49b 1.85b 108.63b 11.13b 12.10b 7.12b 2.01b 60 164.33a 16.73a 16.10a 20.06a 8.08a 160.34a 15.87a 15.27a 16.94a 7.87a farmer1 30 41.00d 4.03d 6.77d 0.96d 0.29d 33.32d 3.77d 6.20d 0.88d 0.29d 40 59.33c 6.27cd 9.77dc 2.62c 0.73c 59.00c 5.63c 9.33c 2.10c 0.70c 50 94.00b 10.03b 11.40b 6.74b 1.15b 92.33b 9.70b 10.60b 5.58b 1.79b 60 142.67a 14.53ab 15.23a 17.86a 7.39a 147.65a 13.96a 13.00a 15.97a 6.39a farmer2 30 42.00d 4.20d 6.63d 0.98d 0.31d 34.67c 3.93d 5.97d 0.95d 0.29d 40 62.00c 6.30c 9.77c 2.68c 0.75c 62.00c 5.76c 9.83c 2.26c 0.75c 50 93.33b 10.17b 11.80b 6.28b 1.69b 95.67b 9.90b 10.83b 6.00b 1.89b 60 149.00a 15.47a 15.73a 17.74a 7.46a 153.63a 14.33a 13.40a 16.67a 6.81a local market 30 44.33d 4.50d 6.77d 1.00d 0.32d 38.00d 4.30d 6.40d 0.98d 0.32d 40 62.67c 6.60c 9.77c 2.74c 0.79c 65.62c 6.00c 10.07bc 2.56c 0.78c 50 100.00b 10.47b 11.30b 6.56b 1.68b 96.33b 10.07ab 11.00b 6.13b 1.93b 60 158.67a 15.87a 15.77a 18.61a 7.87a 156.31a 14.47a 14.00a 16.17a 6.94a note: in a column figures having common letter(s) do not differ significantly by dmrt at 1% level of probability. 4. conclusion cvl-1 seed’s seedling growth varied significantly due to seed sources and seedling age. the seedling height, base diameter, root length, shoot dry weight and root dry weights for seedling establishment of cvl-1 seed’s varied significantly due to seed sources in both the condition of green house and field. the interaction of seed sources and seedling ages were highly significant and the bjri seeds of 60 days age showed the highest plant height, base diameter, root length, shoot dry weight and root dry weights. seeds from bjri and badc of the cvl-1 varieties showed better performances in terms of seedling growth and dry weight compared to seeds from farmers’ and local markets. references [1] bbs, yearbook of agricultural statistics of bangladesh. dhaka, bangladesh: ministry of planning, government of the people’s republic of bangladesh, 2015. [2] m. m. islam and n. uddin, "research and development advances of jute seed in bangladesh: a review," haya: the saudi journal of life sciences, vol. 4, pp. 52-68, 2019. [3] m. salim, m. a. hannan, m. a. r. sarkar, and m. ali, "seed quality in late seed crop as affected by late sowing," bangladesh journalseed science and technology, vol. 2, pp. 11-17, 1998. [4] m. a. hossain, s. a. mannan, k. sultana, and a. l. khandakar, "survey on the constrains of quality jute seed at farm level," research report., agriculture. support serv. proj., bangladesh jute res. inst., dhaka, bangladesh, pp. 45-50. 1994. [5] m. m. islam, jute seed technology, 1st ed. mohamadpur, dhaka: pub. by md. mahmudul islam, 397, middle monipur, mirpur, dhaka-1216. college gate book binding and printing, college gate, dhaka, bangladesh. pp. 120. 2009. [6] m. m. islam, "technological advances in off-season jute seed production," journal of experimental biosciences, vol. 1, pp. 75-82, 2010. [7] m. m. islam, m. a. r. sarkar, m. ahmed, s. m. a. hossain, and s. m. moniruzzaman, "evaluation of jute seed quality attributes and their relationship as affected by seed sources," bangladesh journal jute and fib. reserces, vol. 28, pp. 31-38, 2008. [8] m. m. islam, m. a. r. sarkar, m. ahmed, s. m. moniruzzaman, and m. n. uddin, "germination, vigour and emergence indicators of corchorus olitorius l, seed and their relationship as influenced by seed sources," bangladesh journal jute and fib. reserces, vol. 29, pp. 1-8, 2009. agriculture and food sciences research, 2019, 6(1): 109-113 113 © 2019 by the authors; licensee asian online journal publishing group [9] m. m. islam, "seed quality of two corchorus spp. collected from different sources in bangladesh," journal of agricultural technology management, vol. 19, pp. 1-11, 2016. [10] ista (international seed testing association), "international rules for seed testing," seed science and technology, vol. 13, pp. 356513, 1985. [11] a. k. gomez and a. a. gomez, statistical procedures for agricultural research, 2nd ed. new york: john wiley and sons inc, pp. 124. 1984. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 2, no. 1, 25-31, 2015 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 25 a comparative evaluation of the economic contributions and uses of strychnos cocculoides and schinziophyton rautanenii fruit trees to poverty alleviation in mile 20 village of namibia selma n. elago 1* --lisias t. tjaveondja 2 1,2 okahandja national forestry research centre, okahandja, namibia abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ............................................................................................................................................................................... 26 2. materials and methods ............................................................................................................................................................. 26 3. results ........................................................................................................................................................................................ 28 4. discussion ................................................................................................................................................................................... 29 5. conclusion .................................................................................................................................................................................. 30 6. acknowledgement ..................................................................................................................................................................... 30 references ...................................................................................................................................................................................... 31 in namibia, plant foods including indigenous fruits are amongst the most important non-timber forest products. indigenous fruits constitute an important source of livelihood for the people of rundu rural west constituency and as such this study compared two species: strychnos cocculoides and schinziophyton rautanenii and assessed which species contribute more to the households‟ cash income and food source so as to encourage its promotion. the study further assessed the traditional and other uses of strychnos cocculoides and schinziophyton rautanenii fruits and products. the study focused on mile 20 village in the kavango region of namibia. the study adopted an emergent, exploratory, and inductive qualitative approach and a triangulation method comprising of self-designed household survey questionnaire, interview and observation methods were employed in gathering data. to get the target sample from the three hundred households‟ population, the village was stratified into four zones of north, south, east and west. simple random sampling was used to select thirty-one households from all the zones. the study findings indicates that strychnos cocculoides contribute to households‟ cash income only, while schinziophyton rautanenii contribute both to cash income and to food. chisquare test showed that there is significant differences (p≤0.05) in the sources of the contributions of households cash income (χ 2 = 36.516, df = 6, p=0.001) as well as the contributions of schinziophyton rautanenii to cash income and food source (χ 2 = 11.645, df = 1, p=0.001).the results further indicate a wide range of products can be obtained from these two fruit trees species. some of these products are identified as medicinal, firewood, crafts and oil. keywords: strychnos cocculoides, schinziophyton rautanenii, indigenous fruits, livelihood, cash income, namibia. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(1): 25-31 26 1. introduction rural people have been eating indigenous fruits, nuts and tubers for millennia and those foods still form a high proportion of the diet of some people, especially in poorer households. in namibia, plant foods are amongst the most important non-timber forest products. more plant foods are available in northern and north-astern namibia, mainly because this is where most species that bear fruits occur. of all the tree species in namibia, 157 species (35%) have been recorded as being used for food in one form or another [1]. namibia is a country at south of the sahara located in southern africa with a size of 852,418 km 2 and with a population of about 2 million at a rate of 1.87% from 2005 to 2010 [2]. majority (68%) live in the rural areas where most people earn a living from agriculture [3]. namibia is not a forest-rich country, most of the country‟s territory (65%) covered by sparsely wooded savannah and denser woodlands of 20% occurring in the wettest regions in the north-east of the country [4]. community forests in namibia are central to promoting sustainable and participatory forest governance and enhancing livelihoods in communal land areas like mile 20 [4]. communal land in namibia covers 26 million hectares, representing 36% of total land cover [4]. in the early 1990s the government of namibia placed great emphasis on a more sustainable and equitable approach to the exploitation of natural resources which included the forestry sector [4]. the development of forestry was one of the main strategic directions of poverty reduction and economic growth that resulted in the increase of the country‟s gross domestic product (gdp) [5, 6]. the namibian economy is dominated by mining. however, agriculture, fisheries and tourism sectors also contribute significantly to the national economy. forestry for instance contributes more to the informal and subsistence activities. non timber forest products (ntfps) play an important role in the local economy of the namibian people. the direct and indirect contribution of ntfp to national economy during 2004 was estimated to be n$ 619,459,000 (u$ 77,432.375) [1]. for these reasons commercial and personal harvest of forest products has being increasing every year because the products derived from forest and trees are important sources of cash income and employment for the rural poor. for example, strychnos cocculoides harvested in kavango have been used to produce a liqueur which has been exported to south africa [1]. indigenous fruits constitute an important source of livelihood for the people of rundu rural west constituency. the local people in this constituency depend on indigenous fruits in their day-to-day life and also contribute to household income. indigenous fruits (if) have currently received considerable interest from various stakeholders, due to their economic and nutritional values [7]. ntfps are now globally recognized for their contribution to improved rural livelihood by providing food, nutrition and medicine, and by generating employment, revenues and foreign exchange earnings. the forestry sector social inputs lies in its fuel-wood and ntfps contribution, which are the main origin of domestic energy for the majority of people, and an important source of food security and income generation. as revealed by ramadhani [8], there is a substantial amount of trading and consumption of uapaca kirkiana and strychnos cocculoides indigenous fruits in both rural and urban areas of zimbabwe and these fruits reduce poverty by 30% and generate income above the poverty line throughout the year. dagmar [9] also proved that majority of rural household benefits from consumption and sale of indigenous fruits. ekane [10] in his study on the socio economic impacts of forest products revealed that forest resources are a major source of livelihood for forest dwellers or people living in most rural areas. rural poor people live in marginal lands, far from economic growth points, such that during off-cropping season they have no other sources of income generating activities rather than harvesting and selling indigenous fruits. the government programs has focused more on timber production and overlooked commercial production of indigenous fruits. the socio-economical contribution of individual indigenous fruit trees has not been looked at properly although it is well-known that indigenous fruits play a significant role in many rural people‟s livelihoods. indigenous fruits were said to be very important to rundu rural west community especially during years of drought, when the harvest of people‟s staple food is low, they rely on indigenous fruits for their survival. there is therefore the need to assess the socio-economic impact of indigenous fruits in order to help the government to improve the status of local people. it is against this background that this study was conducted as part of a broader study in order to assess and justify if the rural community derive their main source of cash income from the sale of indigenous fruits. the study has compared two species: strychnos cocculoides and schinziophyton rautanenii and assessed which species contribute more to the households‟ cash income and food source so as to encourage its promotion. the study further assessed the traditional and other uses of strychnos cocculoides and schinziophyton rautanenii fruits. 2. materials and methods 2.1. description of indigenous fruit tree species 2.1.1. strychnos cocculoides strychnos cocculoides is a small tree ranging from 2 to 8 meters high. it is mostly found on deep sand in northeastern namibia, elsewhere occurring in northern south africa, zimbabwe, zambia, botswana, malawi and angola. the fruits are globose in shape dark green when young and yellow to orange when ripe. strychnos‟, meaning „deadly‟, is an ancient greek name given to a certain poisonous member of the solanaceae family. linnaeus, who founded the genus strychnos on the indian species s. nux-vomica, which yields strychnine, possibly associated the deadly qualities of both groups when he named the genus. the fruit of this species is popular in north-eastern namibia, and are frequently sold along the road and in informal markets in towns and villages. the fruits can also be fermented into alcoholic drink locally known as “kashipembe”. the fruit ripen between april and august and are about the size of oranges. strychnos ripen best on the tree. fruits are harvested when fruit skin turns yellow or when they taste sweet (russeted types, but most are picked earlier and ripe in storage). ripe fruit is eaten fresh or is used to prepare alcoholic drink. furthermore, it used for medicinal purposes of which the unripe fruit can be pounded and mixed with water. this mixture can be used to treat snake bites, ear pain, and swellings. the leaves of the tree can be wrapped around the neck for treating neck pain, and the roots can be eaten to treat stomach ache in children. the agriculture and food sciences research, 2015, 2(1): 25-31 27 roots from a young tree can be used for treating chest pain; coughing and stomach ache [11]. the wood of the tree is not highly valued and therefore not really used for anything. 2.1.2. schinziophyton rautanenii schinziophyton rautanenii (formally known as ricinodendron rautanenii) is a medium to large deciduous tree growing to a height of 12 meters. schinziophyton rautanenii belong to the euphorbiaceae family and were declared a protected species in namibia in 1952 [1]. it is distributed across northern namibia, growing on deep kalahari sands. it is dominant and most abundant species in the kavango region. the fruits ripe on the tree from november to february and fall down when fully matured, from march to may. it is left drying on the ground (march-may) before been collected. the fruits can be harvested from may to august. the bark is smooth, yellowish to yellow-grey, with pieces peeling off to expose a yellowish under bark. the fruits of this tree species serve multiple purposes, for example the pulp of the fruits can be made into porridge, the peel and flesh can be used for the production of hot fermented alcoholic drink locally known as “kashipembe”. the nuts can be finely crashed and added to the meat/vegetables to make a tasty soup. the nuts yield a high quality yellow oil of which about 60% is used for food and cosmetics [1]. the species is also used for medicinal purposes. the bark of the schinziophyton rautanenii tree is used for treating stomach aches, sleepless night, fever, back pain and some rare traditional diseases. if people are really sick they can sit in a bath of stamped bark and water. the wood is used for making canoes, and carving crafts and utensils. 2.2. description of the study area the study was conducted in rundu rural west constituency in kavango region, north east of namibia. kavango region is one of the thirteen political regions in namibia and lies between 18.00 o e and 22.00 o e and 17.09 o and 18.01 o s. within the region there are other constituencies namely kahenge, kapako, mashare, mpungu, mukwe, ndiyona, rundu rural east and rundu urban. the population is estimated to be 26 622 while that of the whole region is estimated at 202,691 [3]. the study specifically focuses on mile 20 village within the constituency. mile 20 is located approximately 32 km south of rundu, along the rundu-grootfontein main road. kavango region has an average annual rainfall of 550 mm, which increases slightly from southwest to northeast. the first rain falls in september or october and the late rains in may. eighty percent of the rain falls between december and march, where the maximum rainfall in 24 hours can be 100-120mm [12]. animal husbandry and horticulture is playing an increasing role in the economic lives of the kavango people [1]. another important economic activity is fishing, which provides a substantial source of protein to the people. the most important crops are pearl millet (pennisetum glaucum), sorghum and maize. ground nuts, beans, pumpkins, and tobacco are cultivated on a smaller scale [1]. the site was selected based on the highly abundant and marketing potential of strychnos cocculoides and schinziophyton rautanenii. most of the local people in the study area are engaged in indigenous fruits collection and marketing. furthermore the area is easily accessible and the community had been visited in 2003 by the fruit tree improvement specialist program in the directorate of forestry (dof) in collaboration with the food agriculture organization (fao). map of namibia showing study area 2.3. research design and data collection the study adopted an emergent, exploratory, and inductive qualitative approach. the reason for choosing this approach is that qualitative research certainly excels at generating variables that are very detailed, has special value for investigating complex and sensitive issues and is good when one really want to achieve a deep understanding of the issue under study [13]. to harness the needed information, a triangulation method comprising of self-designed household survey questionnaire, interview and observation methods were employed in gathering data from the respondents. the researchers also conducted documents review and also collected qualitative information from forestry experts in the kavango region to complement the other methods. the questionnaire sought to capture data on the use of indigenous fruit trees as well as their role within the household system in the study area. the questionnaire consisted mainly of close-ended questions which required the respondents to tick their best option. to get the target sample from the three hundred households‟ population, the village was stratified into four zones of north, south, east and west. simple random sampling was used to select thirty-one households from all the zones. the questionnaire was administered to all the residents in the household after which focus group discussions were used to gain insight into participant‟s view on the subject under consideration. household heads were purposefully agriculture and food sciences research, 2015, 2(1): 25-31 28 interviewed. in addition, physical observation was also done in order to have a clue on the socio-economic impact of indigenous fruit trees in the study area. all the households sampled were involved in indigenous fruits collection and sales. research assistants from hamoye forestry office and the authors collected the needed data. the data obtained from all the sources were evaluated using, content analysis, cross-tabulations, descriptive statistics and graphs. 3. results 3.1. demographic description from the analysis, it was observed that the respondents who represented the households, 74% were females and 26% were males. in terms of age, 14 were 18-30 years, 10 were in the age group of 30-50 years while 7 were between 50-80 years. occupation wise, the respondents were mainly farmers and their dependents were mostly children, husband and housewives. the survey also indicated that 21 respondents have 1-10 dependents in their family, while 8 had 10-20 family members. the highest family size was 20-30 which was reported by 2 respondents. 3.2. comparison of strychnos cocculoides and schinziophyton rautanenii contribution to households’ cash income and to food to compare strychnos cocculoides and schinziophyton rautanenii contribution to household‟s cash income and to food, data have been analysed quantitatively to bring out the extent of socio-economic contribution in the mile 20 village. from figure 1, all (100%) of the respondents reported that strychnos cocculoides contribute more to household cash income and not to food. in contrast 80.6% of the respondents stated that schinziophyton rautanenii contributes more to household cash income while 19.4% stated that it contributed to food. figure-1. distribution of the contribution of fruits to household cash income figure 2 shows income and livelihood change percentages. the figure shows that 51.6% of the household members stated that the income earned is enough to sustain their families while 48.4% respondents felt that the income is not enough. chi-square test showed no significant differences (p>0.05) in the respondents by the households members (χ 2 = 0.032, df =1, p 0.857). however all household respondents reported that there is a change in their livelihood. from the figure it is clear that there has been a general improvement in the socio-economic wellbeing of the respondents. figure-2. distribution of income and livelihood change 3.3. chi-square test (χ 2 ) chi square was used to expressed the probability to accept that the groups have different distributions if p < 0.05; or p < 0.01 by comparing the results obtained with the theoretical results for cross-tabulations. the chi square tells us whether the frequency distributions in the various columns are similar in shape, or are different. we determined whether the chi square value is what would be expected by chance alone, or not, and with what probability. table 1 shows results of the cross-tabulation between sources of cash income. agriculture and food sciences research, 2015, 2(1): 25-31 29 table-1. source of cash income source of cash income observed n expected n residual indigenous fruits 16 4.4 11.6 formal employment 4 4.4 -.4 casual 2 4.4 -2.4 pension 3 4.4 -1.4 farming 2 4.4 -2.4 wages & salaries 3 4.4 -1.4 others 1 4.4 -3.4 total 31 table-2. chi-square test of source of cash income source of cash income chi-square 36.516 df 6 asymp. sig. .000 chi-square test (table 2) shows that there is significant differences (p≤0.05) in the source of the contributions of households cash income (χ 2 = 36.516, df =6, p=0.001). table 3 shows results of the cross-tabulation between cash income and food source. table-3. cash income and food source contribution of schinziophtyton ratautanenii observed n expected n residual cash income 25 15.5 9.5 food source 6 15.5 -9.5 total 31 table-4. chi-square test of cash income and food source contribution of schinziophtyton ratautanenii cash income and food source contribution chi-square 11.645 df 1 asymp. sig. .001 chi-square test shows that there is significant differences (p<0.05) in the contributions of schinziophyton rautanenii to cash income and food source (χ 2 = 11.645, df =1, p=0.001). 3.4. traditional and other uses of the fruits to assess the traditional and other uses of strychnos cocculoides and schinziophyton rautanenii fruits at the household level is the third objective of this study. the data for this objective has been analysed quantitatively and presented in table 5 which revealed that 18 respondents obtain medicinal products from strychnos cocculoides tree while 24 respondents obtain from schinziophyton rautanenii. about 10 respondents use schinziophyton rautanenii for fuel wood. other 31 respondents revealed that they make crafts and tools from schinziophyton rautanenii. table-5. traditional uses of the indigenous fruits purpose tree species uses number of respondents s. cocculoides s. rautanenii m ed ic in e leaves 8 roots 8 bark 1 15 oil 9 medicine/ other 1 w o o d firewood 10 making crafts 31 making tools 31 fencing others 4. discussion 4.1. comparison of strychnos cocculoides and schinziophyton rautanenii contribution to households’ cash income and to food indigenous fruits constitute an important source of livelihood for the people of rundu rural west constituency. the local people in this constituency depend on indigenous fruits in their day-to-day life and also contribute to household income. the findings show that all the households‟ respondents were of the view that strychnos cocculoides contribute only to household cash income and not to food. in contrast 80.6% of the respondent‟s stated that schinziophyton rautanenii contributes to household cash income, while few respondents (19.4%) stated that it contributes to food. however, in terms of cash income contribution, both fruit trees species contributes significantly and there is no significant difference between the contributions of the species. the reason why all the respondents were of the view that strychnos cocculoides contribute only to household cash income may be attributed to poor agriculture and food sciences research, 2015, 2(1): 25-31 30 storability of the fruits. therefore the fruits are sold as soon as they are collected, whereas schinziophyton rautanenii has good storability hence consumed in period of food shortage. households mainly collect strychnos cocculoides fruits for marketing purposes. in addition the fruits are also sold in urban areas across the country whereas schinziophyton rautanenii fruits are mostly sold at local markets in the region. these results are contrary with the study of dagmar [9] that revealed that strychnos cocculoides is more important for home consumption during food shortage and not for household cash income. the main reason for this contradiction may be because dagmar [9] conducted the study in zimbabwe and he was comparing uapaca kirkiana, strychnos cocculoides and parinari curatefollia. in his findings uapaca kirkiana fruits were more important in generating cash income than others. this study further reveal that half of the households investigated were of the view that the income generated is sufficient enough to sustain household family members while the other half felt that it is not enough. even though some felt that the income is not enough, all the households respondents have experienced changed in their livelihood and well-being as a result of the income generated from the sale of indigenous fruits. with these cash income, some households within the village have been able to bring water pipe line in their homesteads. some respondents pointed out that from the sale of indigenous fruits; they are able to purchase various kinds of goods. these goods include new clothes for their family members, buy households needs, variety of food types, cell phones etc. these respondents that were of the view of inadequate income encounter problems during indigenous fruits sale. some of the respondents identify such problems as low pricing, competition in selling, lack of customers and lack of transport to transport the fruits to other marketing areas. one of the respondent states that the income she generate is not enough because fruits are only sold within a short season. the results show that forest resources are a major source of livelihood for people living in rural areas. the findings have proven that strychnos cocculoides contributes significantly to household‟s cash income only, while schinziophyton rautanenii contributes both to cash income and to food. 4.2. traditional and other uses of the indigenous fruit trees people in the rural areas have access to a wide range of products from the indigenous fruit trees. both strychnos cocculoides and schinziophyton rautanenii are multipurpose trees, fruits are the main products, but other products such as medicine, firewood, crafts, can also be obtained. the findings show that both species treat many different medical ailments. for example the unripe fruit of strychnos cocculoides can be pounded and mixed with water, this mixture is used to treat snake bites and swellings. the liquid of the unripe strychnos cocculoides fruit can be applied to treat ear pain and one can drink it to cure stomach ache. the bark of the schinziophyton rautanenii tree is used for treating stomach ache, and some rare traditional diseases. if people are sick they can sit in a bath of stamped bark and water. all households‟ respondents confirm that the wood for schinziophyton rautanenii can be used to make canoes, carving crafts and making utensils while the wood for strychnos cocculoides is not important, therefore is not used for anything. the results are in line with the report of fao [14] that reported that ntfps contribute to the improved rural livelihood by providing other traditional products such as medicine and wood products. the results have proven right, the hypothesis that strychnos cocculoides and schinziophyton rautanenii fruits products have other important traditional uses at household level apart from income generation and food security. currently, there is no policy implication in the utilisation of the indigenous fruits. ntfps have the potential to diversify the rural economy and improve the rural household livelihood in terms of food security. commercialisation of both species can give rural people a possibility to improve their economic situation and secure people‟s livelihood. based on the observations and informal interview with the household respondents, strychnos cocculoides and schinziophyton rautanenii fruit trees are regarded as the most important fruits trees in the area. there is a need for the ministry to improve the sale of strychnos cocculoides and schinziophyton rautanenii fruits, by encouraging the communities to supply more fruits to both rural and urban markets within the country. therefore, this action will improve cash income of the rural people not only in rundu rural west constituency, but also in other places within the country. 5. conclusion namibia‟s forest resources constitute an important heritage which provides both economic and environmental benefits. among the strategies used by the rural people to overcome food shortages, is the use of wild fruits from indigenous fruits trees. strychnos cocculoides and schinziophyton rautanenii are the most popular fruit species in rundu rural west constituency kavango region and both indigenous fruit trees contribute significantly to their livelihood and household cash income and to households‟ food. the study compared the economic contributions and traditional uses of strychnos cocculoides and schinziophyton rautanenii fruit trees. the results show that forest resources are a major source of livelihood for people living in rural areas. the findings proved that strychnos cocculoides contributes significantly to household‟s cash income only, while schinziophyton rautanenii contributes both to cash income and to food. chi-square test showed that there is significant differences (p≤0.05) in the source of the contributions of households cash income (χ 2 = 36.516, df = 6, p=0.001) as well as the contributions of schinziophyton rautanenii to cash income and food source (χ 2 = 11.645, df = 1, p=0.001). apart from the fruits, strychnos cocculoides and schinziophyton rautanenii also provide a wide variety of products. the two species provide other products such as medicine, firewood, craft and utensils. wood for schinziophyton rautanenii can be used to make canoes, carving crafts and making utensils while the wood for strychnos cocculoides is not important and therefore is not used for anything. 6. acknowledgement the authors would like to thank the namibia ministry of agriculture, water and forestry for funding this study. we are grateful to the directorate of forestry for granting us the opportunity to undertake this study. we extend our agriculture and food sciences research, 2015, 2(1): 25-31 31 appreciation to the people in the study area particularly the people of mile 20 village, rundu rural west constituency, kavango region who took great interest in the study, shared their knowledge and patiently answered many questions. references [1] j. mendelsohn and s. el obeid, "forests and woodlands of namibia," research and information service of namibia, directorate of forestry, ministry of agriculture, water and forestry, namibia, 2005. [2] un, "population growth rate of namibia in 2005-2010. population division of the department of economics and social affairs of the united nations secretariat, world population prospects: the 2010 revision." available http:esa.un.org/undp/unpp/index.htm, 2010. [3] central bureau of statistics, "population and housing census 2001. national planning commission," government of republic namibia, report, 2001. [4] a. benkenstein, s. hengari, and w. mbongo, "community forests in namibia: ensuring sustainable local-level forest management. south african institute of international affairs (saiia), saiia policy briefing 119." available www.saiia.org.za, 2014. [5] t. o. oksanen, b. pajari, and t. tuomasjukka, "forests in poverty reduction strategies: capturing the potential," in european forest institute proceedings no. 47.tuusula, finland, 2003. [6] j. e. mbaiwa, the socio-economic and environmental impacts of tourism development on the okavango delta. north-western botswana, harry oppenheimer okavango research centre. botswana: university of botswana, 2002. [7] m. chakanga, "timber trade and timber industries in namibia. ministry of environment and tourism, directorate of forestry, namibia," fao working paper fp/30f, 2003. [8] t. ramadhani, marketing if indigenous fruits in zimbabwe. socio-economic studies on rural development vol. 129. kiel, germany: wissenschaftsverlag vauk, 2002. [9] m. dagmar, economics of indigenous fruits tree crops in zimbabwe. germany: university of hannover, 2004. [10] n. b. ekane, "the socio economic impact of pronus africana management in the mount cameroon region. case study of the bukwango community," master of science thesis, department of urban planning and environment, royal institute of technology stockholm, 2006. [11] m. c. palgrave, keith coates palgrave trees of southern africa, 3rd ed. cape town: struik, 2002. [12] a. erkilla and h. siikonen, "silvica calerica 20 forestry in namibia 1850-1990. university of joensuu, faculty of forestry, finland, 1992 survey of research needs in northern and southern namibia," directorate of forestry, research division, forestry research and development project, 1992. [13] a. d. day, how to write and publish a scientific paper, 5th ed. cambridge, united kingdom, 2003. [14] fao, forest resource assessment 2005. rome: food and agriculture organistaion of the united nations, 2005. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. http://www.saiia.org.za,/ 120 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 120-126, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.120.126 © 2019 by the authors; licensee asian online journal publishing group sustainability of climate change adaptation measures in south-south, nigeria osuji, e. e.1 okwara, m. o.2 essien, u. a.3 agu, c. g.4 oguegbuchulam, m. n.5 ( corresponding author) 1department of agricultural economics, michael okpara university of agriculture umudike, nigeria. 2 3,department of agricultural economics, federal university of technology owerri, imo state, nigeria. 4department of cooperative economics and management, institute of management and technology enugu, nigeria. 5department of agricultural extension and management, imo state polytechnic umuagwo, owerri, nigeria. abstract the rampaging effects of climate change on nigeria agriculture cannot be totally undermined, hence, the sustainability of climate change adaptation measures in south-south, nigeria. a multistage sampling technique was used to select 142 farmers who were interviewed to elicit information bordering on climate change in the area. data were analyzed using descriptive statistics, ordinary least square multiple regression technique and the scaling model. results showed that the socio-economic factors investigated influenced the decisiveness of the farmers in embracing adaptive strategies to checkmate the overindulgence of climatic variations in the area. age of the farmers, educational level, household size, farming experience, farm size, extension contacts were all positive and significant factors. result also shows that there is increasing rate of temperature, rainfall volume, sunshine hours and number of rainy days and decreasing rate of light intensity and wind. the adaptation techniques of the farmers were classified into highly sustainable adaptation strategies which include; use of pest and disease resistant varieties, use of organic fertilizers, tree planting, early information seeking on climate change etc and slightly sustainable; planting of different crop varieties, mulching, mixed cropping, etc. all the farmers in the area perceived inadequate capital as a major constraint in coping with climate change effects. hence, this study recommends government all levels to sensitize the crop farmers on the devastating effects of climate change on agriculture as well as provide supports where necessary to cushion its negative impacts. keywords: agriculture, adaptation, climate change, mitigation, sustainability, variability, farmers’ perception, mean scores, ols regression, nigeria citation | osuji, e. e.; okwara, m. o.; essien, u. a.; agu, c. g.; oguegbuchulam, m. n. (2019). sustainability of climate change adaptation measures in south-south, nigeria. agriculture and food sciences research, 6(1): 120-126. history: received: 12 march 2019 revised: 8 april 2019 accepted: 14 may 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 121 2. materials and methods ................................................................................................................................................................. 121 3. results and discussion ................................................................................................................................................................. 122 4. conclusion and recommendations ............................................................................................................................................. 126 references ............................................................................................................................................................................................ 126 http://www.asianonlinejournals.com/index.php/aesr/article/view/130 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.120.126&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.120.126&domain=pdf&date_stamp=2017-01-14 https://orcid.org/0000-0001-8605-834x http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2019, 6(1): 120-126 121 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this paper affirmed the adaptation measures of crop farmers’ to climate change variability using sustainable procedures which enhances farm outputs, yields and productivity, hence ensuring food security and improved income of the crop farmers in nigeria. 1. introduction in recent time, climate change is notably one of the major challenges confronting nigeria agriculture. its impacts have been felt both at the national, regional, and local levels.nigeria is currently under pressure from climate change stresses and its adverse consequences. many areas in south-south region of nigeria are recognized as having climates that are among the most variable in the world on seasonal time scales [1]. floods and droughts occur in these areas times without number leading to famine, hunger and widespread disruption of socio-economic well-being. nigeria agriculture is largely weather-sensitive and particularly vulnerable to climate change [1]. this vulnerability has been demonstrated by the devastating effects of recent flooding in the south-south region of the country and the various prolonged droughts that are currently witnessed in some parts of northern region. moreover, the over exploitation of land resources (forests), desertification, land degradation and increase in population pose additional threats to climatic variations and changes in nigeria [2].thus, understanding farmers’ responses to this climatic variation is very crucial as this will help in designing appropriate sustainable and coping strategies to ameliorate this ugly climatic trend. also the spatial and erratic rainfall and temperature distribution patterns being experienced in the south-south region of nigeria have influenced the farm productivity of the farmers leading to loss of food crops, income, low outputs, etc, thereby increasing food insecurity in the region [3]. conventional approaches to understanding climate change were restricted to identifying and quantifying the potential long-term climate impacts on different ecosystems and economic sectors. although, essential in depicting general trends and dynamic interactions between the atmosphere, biosphere, land, oceans and ice. literatures have shown that science driven approach has failed to address the regional and local impacts of climate change and the farmers’ abilities to adapt to climate-induced changes [4]. though, this impact-driven approachhashelped in assessing past and present current vulnerability, existing sustainable adaptation strategies, and how these might be modified to suit recent climatic trend. adaptation to climate change is a new process for both developed and developing nations and concrete experience in applying an integrated approach to adaptation is limited [5]. adaptation is a process through which societies make themselves able to cope with future uncertainties. adapting to climate change entails taking the right measures to reduce the negative effects of climate change (or exploit the positive ones) by making the appropriate adjustments and changes. however, sustainable adaptation measures in south-south region of nigeria range from technological options such as improved farming methods, timely cultivations and reducing usage of water during drought seasons. other strategies include early warning systems for extreme events, better water management, improved risk management, various insurance options and biodiversity conservation. considering the speed at which climate is changing due to global temperature rise, it is obvious that the vulnerability of developing countries to climate change is increasing and their capacity to adapt has also increased [6]. both mitigation and adaptation measures must be seriously pursued to tackle the climate change problem and to create an effective and inclusive international climate change regime. sensitivity to the issue of adaptation has grown over the last couple of years, particularly after the ipcc (intergovernmental panel on climate change) and tar (third assessment report) [7]. adaptation has now emerged as an urgent policy priority, prompting action both within and outside the climate change negotiations [8]. adaptation calls for natural resource management, buttressing food security, development of social and human capital and strengthening of institutional systems. such processes, besides building resilience of farmers, communities, regions and countries to all shocks and stresses, including climate variability and changes are good development practice in themselves. however, the risks associated with climate change call for a broad spectrum of policy responses and strategies at the local, regional, national and global level. hence the essence of this study which seeks to examine the sustainability of climate change adaptation measures in south-south, nigeria. 2. materials and methods this study was carried out in rivers state, nigeria due to the devastating effects of climate change in the area. it has a total land mass of 11,077 sq km and is located on latitudes 40 32’ and 50 53’ north and longitudes 70 25’ and 80 25’ east of the equator. it is bounded on the south by the atlantic ocean, to the north by imo and abia states, to the east by akwaibom state and to the west by bayelsa and delta and states. the inland part of rivers state consists of tropical rainforest; towards the coast the typical niger delta environment features many mangrove swamps. temperature range is between 23-31ºc and vegetation found in the state includes the saline water swamp, mangrove swamp and the rain forest. major seasons are the dry (november-february) and wet seasons (october – march). the climatic and soil condition of the study area favour the extensive production of various food crops such as yam, cassava, maize, vegetables plantains and cocoyam. multistage sampling technique was used to select two local government areas each out of the three senatorial zones of the state. three communities were picked from each of the selected local government areas hence; ten registered crop farmers were randomly selected from the various communities giving a sample size of 180 farmers. questionnaire, in-depth scheduled interviews and direct personal field interactions were used to extract information from the respondents but after sorting out the data instruments, only 142 questionnaires were found useful for data analysis. data were analyzed using descriptive statistics such as the (mean, frequency counts, relative frequency), ordinary least square multiple regression technique as well as the scaling model. in this study, 4 functional forms were fitted into the ordinary least square multiple regression model expressed as follows; eqn. (1) linear function eqn. (2) agriculture and food sciences research, 2019, 6(1): 120-126 122 © 2019 by the authors; licensee asian online journal publishing group semi-log function eqn. (3) doublelog function eqn. (4) exponential function eqn. (5) where; y = ith ratio of adaptation strategies/techniques of farmers’ x1 = age (years) x2 = gender (male=1; female=0) x3 = marital status (married=1; single=0) x4 = household size (number of pesons) x5 = level of education (years spent in school) x6 = farming experience (years) x7 = farm size (hectare) x8 = extension contacts (number of contacts) e = error term 3. results and discussion 3.1. socio-economic factors influencing farmers’ adaptation to climate change table 1; below shows the socio-economic factors influencing adaptation of farmers to climate change. majority of the farmers aged between 41-50 years with a corresponding mean age of 50 years. this implies that the farmers are not too old to embrace adaptive measures in mitigating climate change incidence. age is known to influence adaptive capabilities of farmers as it relates to climate change outcomes [9]. female farmers in the area accounted for 54.2% as against 45.8% of male farmers. table-1. farmers’ socio-economic factors influencing adaptation strategies to climate change variability. socio-economic factors frequency relative frequency mean age 50 31 – 40 24 17.0 4150 64 45.1 51-60 20 14.1 61 and above 34 24.0 gender male 65 45.8 female 77 54.2 marital status single 44 31.0 married 98 69.0 household size 7 1-5 45 31.7 6-10 93 65.5 11-15 4 2.8 15 and above 0 0 educational level 7 0 (no formal) 31 21.8 1 – 6 36 25.3 7 13 67 47.1 14 and above 8 5.6 farming experience 10 1-5 19 13.4 6 – 10 59 41.5 11 – 15 44 31.0 16 and above 20 14.1 farm size (ha) 2 less than 1 55 38.7 2 – 3 81 57.0 4 – 5 4 2.8 6 and above 2 1.4 extension contacts 10 1-5 41 28.9 6 – 10 46 32.4 11 – 15 39 27.5 16 and above 16 11.3 total 142 100 source: field survey, (2018). this implies the dominance of female farmers in the area and support the claim that women engaged more in climate change mitigation. african women are seen as climatic change agents and thus, contribute more in agriculture and food sciences research, 2019, 6(1): 120-126 123 © 2019 by the authors; licensee asian online journal publishing group domestic food production [10]. again, married farmers dominated the entire area accounting for 69% with logical implication that marriage contributes to household size which plays a major role in combating climate change. furthermore, marriage is sine qua non to family labour which helps in adapting to climatic conditions. almost 66% of the farmers had household sizes which range from 6-10 with a mean household size of 7. this implies that farmers tend to utilize more labour which is indispensable in mitigating climate change occurrences. it is believed that the higher the household size, the higher the farmers’ adaptation to climate change since more hands will be needed to mitigate the negative effects of climate change. large sizes of households encourage adaptive potentials of farmers as well as position them for improved outputs [11]. the study revealed the mean educational level of the farmers as 7 years which implies that the farmers attempted post primary education which exposes them to innovative ideas in handling climatic outcomes. education is believed to equip farmers with the right knowledge needed to overcome adverse climatic changes and weather conditions farmers in the area had 10 years of farming experience. farming experience instills in farmers the ability to cope with adverse climatic conditions and adaptive measures. farming experience enhances farmers’ knowledge and understanding of climatic events and conditions as well as equips farmers to adapt favorably to changing climate. experienced farmers are more skilled and organized in handling climatic issues and variations. about 57% of the farmers had farm sizes ranging between 2 to 3 hectares with a mean farm size of 2 hectares. small farm sizes influences the perception of farmers in mitigating climatic change. farm sizes in rural areas is often fragmented and scattered making it unsuitable to adopt adaptation strategies/measures especially crop rotation, etc to mitigate climatic effects. farmers in the area had reasonable extension contacts (10, contacts/annum). this implies that farmers in the area were exposed to various adaptation strategies by the extension agents that visited the area. extension contacts expose rural farmers’ to innovative ideas and adaptation techniques and further equip them with a good understanding and knowledge of climatic changes and variations which helps in mitigating adverse climatic conditions. 3.2. farmers’ sources of information on climate change variability table 2 reveals the sources of information on climate change variability available to the farmers. from the table, extension contacts proved to be the major source of information to the farmers in the area. extension agents’ are information channels and change agents. they expose farmers to adaptive measures used in mitigating adverse climatic variations. about 89% of the farmers learnt about climate change through the viewing of television and listening to radio. these medium offers a wider dissemination of information on climate change variations. while less than 86% of the farmers learnt about the effects of climate change through their friends/relatives. farmers often learn new things much faster and much easier from their colleagues (friends/relatives). 73% and 47% of the farmers got to know of climate change through the non-governmental organizations (ngo’s) as well as other government agencies. in recent times non-governmental organizations and government agencies have simultaneously engaged in the dissemination of information bordering on climate change variations [12]. also 18% of the crop farmers knew about climate change through the print media. print media such as (newspapers, magazines, literatures, journals, articles, dossiers on climate change, etc) helps to diffuse information on climatic variation and / or conditions prevalent in an area, while 33% and 70% of the crop farmers discovered of climate change through seminars/conferences and local workshops. these medium exposes farmers to information bordering on climate change. farmers who are privileged to attend seminars and workshops are better informed on climatic variations as well as on other agricultural packages and concerns. furthermore, other sources of information on climate change variability indicated by the farmers include; self observations and personal experience (55%), interactions with other enlightened farmers (64%), internet/web services (16%) and others (14%). these medium are effective and proven sources of information gathering of farmers. table-2.farmers’ source of information on climate change variability. sources of information **frequency relative frequency ngo’s 103 72.5 government agency 67 47.2 self-observation and experience 78 54.9 interaction with enlightened farmers 91 64.1 radio 126 88.7 television 129 90.8 friends/relatives 124 87.3 extension agents 132 92.5 seminars/conferences 47 33.1 internet/web 23 16.2 local workshops print materials others 99 25 20 69.7 17.6 14.1 source: field survey, (2018) **multiple responses recorded. 3.3. farmers’ perception of climate change variability table 3 shows farmers’ perception of climate change variability in the area. a mean index of 2.0 was estimated using scaling techniques. results show that there is increasing rate of temperature (2.41), rainfall volume (3.05), sunshine hours (2.45), and number of rainy days (2.91). this implies a high variability occurrence of climate change prevalent in the area. the standard deviation values of the aforesaid climate variables were all less than 1.0., except rainfall volume which implies that the volume of rainfall exceeded its previous records in the area. furthermore, farmers in the area perceived a decreasing rate of light intensity and wind while relative humidity and soil ph in the area remained unchanged. this further indicates vagaries of climatic atmospheric conditions dominant in the area. hence these atmospheric variations undoubtedly influenced agricultural operations in the area. farmers in a bid to agriculture and food sciences research, 2019, 6(1): 120-126 124 © 2019 by the authors; licensee asian online journal publishing group maximize farm utility engaged in multiple adaptation strategies to cushion the negative effects of climate change variations [13]. table-3.farmers’ perception of climate change variability. climatic/soil index mean score s.d remark temperature 2.41 0.96 increasing rainfall volume 3.05 1.88 increasing sunshine hours 2.45 0.82 increasing number of rainy days 2.91 0.98 increasing humidity 2.00 0.77 unchanged light intensity 1.93 0.09 decreasing wind 1.81 1.58 decreasing soil ph 2.00 0.59 unchanged note: m > 2.0 = increasing, m < 2.0 = decreasing, and m = 2.0 = unchanged. source: field survey, (2018) 3.4. farmers’ adaptation strategies in mitigating climate change variability table 4 shows the adaptation strategies/techniques practiced by the farmers in mitigating climate change effects. these adaptation techniques are classified into degrees of sustainability with respect to the mean score obtained. the highly sustainable adaptation strategies mostly practiced by the farmers include; use of pest and disease resistant varieties (3.90), use of organic fertilizers (3.01), tree planting (2.67), early information seeking on climate change (2.25), soil conservation practices (2.09), cover cropping (2.90), increase use of family labour (2.01), cultivation of early maturing varieties’ (1.99), early and late planting (1.98), regular weeding to avoid breed of insect pests (1.98), use of improved crop varieties (1.54) and insurance options (1.90). use of pest and disease resistant varieties are widely used by farmers in combating the adverse effects of climate change. farmers’ often make good use of crop varieties that are free from pest and disease infestations in mitigating climatic variation. organic fertilizers are often used by farmers to checkmate adverse climatic conditions as well as replenish and / or restore the fertility of the soil probably due to easy accessibility and cheap source of the manure [2]. the planting of trees is a vital adaptive measure used to withstand uncertainties in regards to climatic change. trees are planted to prevent undue destruction of the vegetative cover of the soil and other nefarious activities of climate change. early information seeking on climate change enables farmers to avert the consequences of climate change on time and also forestall adverse effects of climatic variations. soil conservation practices such as cover cropping helps to prevent unforeseen variations of climatic change. cover cropping is an adaptive technique that increases soil fertility which in turn enhances the productivity of the farmers [14].family labour improves agricultural activities in the sense that more hands are hired to facilitate farm operations. cultivation of early maturing varieties is a widely mitigation strategies mostly adopted by farmers in coping with the demands of changing climates. this strategy assist farmer to improve on crop yield and avert ugly incidences of climate change. furthermore, farmers also engaged in early and late planting to checkmate variations of climate change. early and late planting is frequently practiced by crop farmers to mitigate and overcome excessive incidence of climate change. table-4.farmers’ adaptation strategies in mitigating climate change variability. farmers’ adaptation strategies/techniques mean score s.d remark tree planting (afforestation) 2.67 0.76 highly sustainable soil conservation practices 2.09 0.98 highly sustainable planting different crop varieties 1.45 0.62 slightly sustainable early and late planting 1.98 0.88 highly sustainable use of improved varieties 1.54 0.67 highly sustainable irrigation practices improved farming methods 1.23 2.09 1.46 0.40 slightly sustainable slightly sustainable use of organic fertilizers 3.01 0.58 highly sustainable crop–livestock diversification 1.05 0.69 slightly sustainable mixed cropping 1.44 0.87 slightly sustainable crop rotation 1.43 2.98 slightly sustainable mulching 1.45 0.66 slightly sustainable change in planting dates 1.67 0.54 highly sustainable cultivation of early maturing varieties 1.99 0.61 highly sustainable cover cropping 2.90 0.87 highly sustainable relocation of farm lands 1.05 0.63 slightly sustainable increase use of family labour 2.01 0.84 highly sustainable use of pest and disease resistant varieties 3.09 0.09 highly sustainable avoidance of farm areas susceptible to flooding 1.40 0.59 slightly sustainable regular weeding to avoid breed of insects pests 1.98 0.77 highly sustainable breeding of drought and heat resistant crop varieties 1.35 0.60 slightly sustainable avoidance of deforestation early information seeking/warning on climate change improved risks management insurance option better water management biodiversity conservation 1.09 0.64 2.25 0.73 1.21 0.91 1.90 0.31 1.01 0.57 1.45 0.69 slightly sustainable highly sustainable slightly sustainable highly sustainable slightly sustainable slightly sustainable note: m ≥ 1.5 =highly sustainable and m < 1.5 = slightly sustainable. source: field survey, (2018). agriculture and food sciences research, 2019, 6(1): 120-126 125 © 2019 by the authors; licensee asian online journal publishing group regular weeding on the farm is usually carried out to avoid breeding of insects’ pests which alters yields on the farm. use of improved crop varieties helps farmers to reduce losses and improve the productivity of the land. farmers often use improved crop varieties to avert total crop failure as a result of the changing climate. in recent times, farmers’ insure their farmlands and other valuable farm assets to cushion the negative effects of certain uncertainties emerging from climate change. however, the values obtained of the standard deviations in highly sustainable techniques are all less than one, implying that these adaptive techniques do not vary much with the mean scores estimated hence agrees with the result. similarly, quite a number of the adaptive measures of the farmers were slightly sustainable and these include; planting of different crop varieties (1.45), mulching (1.45), mixed cropping (1.44), avoidance of farm areas susceptible to flooding (1.40), crop rotation (1.43), irrigation practices (1.23), breeding of drought and resistant crop varieties (1.35), improved farming methods (1.46), avoidance of deforestation (1.09), crop-livestock diversification (1.05), improved risk management (1.21), insurance options (1.90), better water management, relocation of farmlands (1.05) biodiversity conservation (1.45). of all the adaptive measures that were slightly sustainable only crop rotation and irrigation practices had a high standard deviation signifying a slight variation from the mean scores obtained, probably due to the technicalities associated with these two techniques. 3.5. factors’ influencing farmers adaptation to climate change variability table 5 reveals the factors influencing farmers’ adaptation to climate change variability. four functional forms of the ordinary least square multiple regression models were fitted in to assess the factors influencing farmers’ adaptation to climate change variability in the area. from the result, the linear functional form was isolated as the lead equation based on the significance of the explanatory variables, highest r2, and f-statistics and was further used for interpretations. the coefficient of multiple determination (r2) was0.8722. this suggests that about 87% of the total variation in the endogenous variable were explicitly explained by the exogenous variables investigated. the f-value of 42% further indicated the fitness of the model. the coefficient of age was positive and significant at 5% level. this implies that as farmers’ advances in age, their ability to adapt to climatic changes increases per time. this is probably as a result of accumulated farming experiences which leverage their acceptance of adaptive techniques to forestall the despicable incidence of climate change[15]. household size was also positive and significant at 5% level. this reveals increase in family labour via expansion of household size. as household size increases, more family labour (more hands) is effectively utilized in mitigating the negative impacts of climate change in the area. the coefficient of level of education was significant at 1% level and was positive too. this implies that education increases the farmers’ knowledge and understanding of climate change and as well as reposition the farmers’ mindset in adopting new scientific strategies with respect to combating the incidence of climatic variations. the farming experience of the farmers has a positive relationship, and was significant at 5% level. this implies that 1% increase in farming experience of the farmers will lead to a corresponding increase in farmers ability in adapting to climatic changes. farming experience enhances the farmers’ drive in adapting to climatic changes which ensures improved farrn outcomes. farm size was also significant at 1% level having a positive sign. this implies that larger farm sizes motivate farmers to practice adaptive strategies that repel climatic changes. farmers with large farm sizes are likely to adopt anti climatic techniques than farmers with lesser farm sizes. the coefficient of extension contacts was positive and also significant at 5% level. this further implies that 1% increase in extension contacts of the crop farmers will equally lead to a corresponding increase in adaptative abilities of the farmers. the relevance of extension agents in championing the impacts of climate change can not be undermined as they are evidently known as change agents in exposing the farmers to understand the phenomenon of climate change and adaptive ways to mitigate and combat its adverse effects[16]. however, other variables such as gender and marital status was not significant at all and this might be due to statistical errors encountered during the process of investigation. table-5. factors influencing farmers’ adaptation to climate change variability. variables coefficients t-values std. error sign. levels age (x1) 2.0935 2.5091 0.8343 ** gender (x2) 1.8853 1.0981 1.7168 ns marital status (x3) 0.3657 1.0071 0.3631 ns household size (x4) 0.9074 2.5671 0.3534 ** level of education (x5) 2.5680 4.0912 0.6276 *** farming experience (x6) 0.3785 2.6109 0.1449 ** farm size (x7) 1.0987 3.0902 0.3555 *** extension contacts (x8) 1.9041 2.6710 0.7128 ** constant 2.7862 1.2091 2.3043 ns r2 0.8722 f-value n 41.631 142 source: field survey, (2018). 3.6. farmers’ perceived constraints to climate change variability table 6 shows the farmers’ perceived constraints to climate change variability. from the table, 100% of the farmers; which implies that all the farmers in the area perceivedinadequate capital as a major constraint to climate change? this could be true because capital is majorly required to acquire a piece of land, purchase farm inputs, hire labourers, and above all practice anti climatic adaptive strategies and/ or techniques [2]. about 90% of the farmers indicated inadequate farmlands and inadequate information on climate change. most times, inadequate farmlands and information on climate change makes it difficult for farmers to adopt climatic techniques. again, 92% and 87% of the farmers indicated land fragmentations and erosion issues as their major constraints. this duo makes it practically impossible for farmers to adopt climatic adaptive techniques. furthermore, other farmers in the area indicated high cost farm inputs (62%), constant deforestation (70%), inadequate extension agriculture and food sciences research, 2019, 6(1): 120-126 126 © 2019 by the authors; licensee asian online journal publishing group agents (16%), high cost of labour (24%) and long distance of farmlands (86%). it could be seen from the result that all these factors played a role or the other in influencing the farmers’ ability in mitigating adverse climatic effects. table-6.farmers’ perceived constraints to climate change variability. perceived constraints **frequency relative frequency inadequate farmlands 140 98.6 inadequate capital 142 100 inadequate information on climate change 134 94.4 high cost farm inputs 88 61.9 land fragmentations 131 92.3 erosion issues 123 86.6 constant deforestation 99 69.7 inadequate extension agents 22 15.5 high cost of labour 34 23.9 long distance of farmlands 122 85.9 note: **multiple responses recorded. source: field survey, (2018). 4. conclusion and recommendations the findings of the study revealed that the socio-economic factors played a major role in influencing the decision of the farmers in embracing adaptive strategies to checkmate the excesses of climatic variations in the area.the mean age of the farmers in the area is 50 years. this implies that the farmers are not too old to embrace adaptive measures in mitigating climate change incidence. the coefficient of age was also positive and significant at 5% level. this implies that as farmers’ advances in age, their ability to adapt to climatic changes increases per time. the mean educational level of the farmers is 7 years which implies that the farmers attempted post primary education which exposes them to innovative ideas in handling climatic outcomes. extension contacts proved to be the major source of information to the farmers on climate change. results show that there is increasing rate of temperature, rainfall volume, sunshine hours and number of rainy days. this implies a high variability occurrence of climate change prevalent in the area. the adaptation techniques of the farmers were classified into highly sustainable adaptation strategies which include; use of pest and disease resistant varieties, use of organic fertilizers, tree planting, early information seeking on climate change etc and slightly sustainable; planting of different crop varieties, mulching, mixed cropping, etc. all the farmers in the area perceivedinadequate capital as a major constraint in combating climate change effects. hence this study recommends government all levels to sensitize the crop farmers on the devastating effects of climate change on agriculture as well as provide supports where necessary to cushion its negative impacts. references [1] t. g. apata, "effects of global climate change on nigerian agriculture: an empirical analysis," cbn journal of applied statistics, vol. 2, pp. 31-45, 2008. [2] e. e. osuji, "impacts of sustainable soil management techniques on land productivity and poverty levels of arable crop farmers in imo state, nigeria," unpublished ph.d dissertation, department of agricultural economics, michael okpara university of agriculture umudike, nigeria, 2017. [3] o. e. ayinde, o. ajewole, i. ogunlade, and m. adewumi, "empirical analysis of agricultural production and climate change: a case study of nigeria," journal of sustainable development in africa, vol. 12, pp. 275-283, 2010. [4] t. g. apata, k. d. samuel, and a. o. adeola, "analysis of climate change perception and adaptation among arable food crop farmers in south western nigeria," presented at the contributed paper prepared for presentation at the international association of agricultural economists’ 2009 conference, beijing, china, august 16-22, 2009.pp. 12-19 [5] j. b. robinson and d. herbert, "integrating climate change and sustainable development," international journal of global environmental issues, vol. 1, pp. 130-149, 2001. [6] building nigeria’s response to climate change (bnrcc), presented at the 2008 annual workshop of nigerian environmental study team (nest): the recent global and local action on climate change, held at hotel millennium, abuja, nigeria; 8-9th october, 2008, pp. 14-23. [7] j. a. schidhuber, "global food security under climate change," in proceedings of the national academy of sciences, nigeria, 2007. [8] m. munasinghe, climate change and sustainable development linkages: points of departure. france: oecd 2001: poverty reduction: the dac guidelines, 2003.pp. 29-34. [9] e. e. osuji, u. g. anyanwu, n. c. ehirim, e. u. eze, and a. tim-ashama, "economics of processed cassava products in imo state, nigeria," journal of research in business and management, vol. 5, pp. 9-19, 2017. [10] a. s. oyekale, "vulnerability of peasant cocoa farmers to climate change in south-west nigeria," journal of human ecology, vol. 40, pp. 33-41, 2012. available at: https://doi.org/10.1080/09709274.2012.11906521. [11] p. obasi, a. henri-ukoha, i. ukewuihe, and n. chidiebere-mark, "factors affecting agricultural productivity among arable crop farmers in imo state, nigeria," american journal of experimental agriculture, vol. 3, pp. 443-454, 2013. available at: https://doi.org/10.9734/ajea/2013/2030. [12] c. a. nhemachena, "micro level analysis of farmers adaptation to climate change in southern africa," ifpri discussion paper no. 00714, environment and production technology division, washington d.c, pp. 23-79, 2007. [13] s. k. juana, "farmers perception of climate change and adaptation strategies," journal of geography and regional planing, vol. 1, pp. 138143, 2013. [14] a. henri-ukoha and e. e. osuji, "determinants of arable crop farmers’ use-levels of sustainable soil management techniques in imo state, nigeria," nigerian journal of agriculture, food and environment, vol. 13, pp. 163-168, 2017. [15] a. egeru, "role of indigenous knowledge in climate change adaptation," indian journal of traditional knowledge, vol. 11, pp. 217 219, 2012. [16] fme, national adaptation strategy and plan of action for climate change in nigeria (naspa-ccn). abuja: federal ministry of environment, climate change department, 2011.pp.22-27. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 3, no. 1, 12-18, 2016 http://www.asianonlinejournals.com/index.php/aesr 12 achievements of cassava agronomy research in southern ethiopia in the last two decades daniel markos1  legesse hidoto2 fiseha negash3 1,2 awassa agricultural research center, p.o.box 06, awassa, ethiopia 3 areka agricultural research center, p.o.box79, areka, ethiopia ( corresponding author) abstract the paper describes research results of obtained in the process of generation of appropriate cultural practices, cropping systems and other agronomic studies of cassava in ethiopia. based on plant population studies carried out for three consecutive years on major cassava growing areas of snnpr, plant spacing of 100 cm x 80, 120 cm x 80 and 80 cm x 80 was recommended for awassa, amaro kele and amaro jijola areas, respectively. the moisture content of cassava roots were invariably higher in varieties qule, kele and local in 12th and 15th map; but, decreases there after markedly in a linear way. conversely, viscosity and root yield of the cassava increases significantly (p < 0.05) starting the 12th map. results of intercropping maize and cassava showed that alternate rows (1c:1m) had ler advantage of 71%. similarly, growing cassava intercropped with haricot bean, cow pea, soy bean and mung bean resulted in ler of 1.82, 1.49. 1.48 and 1.62 compared to sole beans in amaro area. in one study that investigated planting positions and planting parts, slant and vertical plantings are suitable in awassa sandy soils compared to horizontal planting. similarly, this study revealed that planting materials shall be taken from middle and top part of the main stem of cassava compared to its branches. in another study that investigated land preparation methods, it was found that furrow and ridge planting produced comparable results of 48t/ha and 43t/ha compared to 38t/ha produced by flat plantings. planting date trial carried out for three consecutive seasons elaborated that rainfed cassava planting could be carried out from mid april to early may based on optimum marketable and total tuber yield obtained. thus, the crop responded very well to crop management practices like plant density, land preparation methods, planting positions, intercropping, etc.and farmers or investors could optimize production by using these recommendations. keywords: plant density, intercropping, ler, planting date, starch content. contents 1. introduction .................................................................................................................................................................................13 2. objectives ....................................................................................................................................................................................13 3. methodologies..............................................................................................................................................................................13 4. results and discussion ...............................................................................................................................................................14 5. planting dates .............................................................................................................................................................................16 6. planting position and planting materials .................................................................................................................................16 7. land preparation methods ........................................................................................................................................................17 8. conclusions ..................................................................................................................................................................................17 9. future research directions .......................................................................................................................................................17 references ........................................................................................................................................................................................17 citation | daniel markos; legesse hidoto; fiseha negash (2016). achievements of cassava agronomy research in southern ethiopia in the last two decades. agriculture and food sciences research, 3(1): 12-18. doi: 10.20448/journal.512/2016.3.1/512.1.12.18 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: contribution/acknowledgement: this work is licensed under a creative commons attribution 3.0 license all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 12 january 2016/ revised: 23 january 2016/ accepted: 29 january 2016/ published: 4 february 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group https://orcid.org/orcid-search/quick-search?searchquery=daniel%20markos https://orcid.org/orcid-search/quick-search?searchquery=legesse%20hidoto https://orcid.org/orcid-search/quick-search?searchquery=fiseha%20negash http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.12.18 http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2016, 3(1): 12-18 13 1. introduction cassava is a stable food of more than 600 million people all over the world and is grown in as many as 90 countries [1]. it is the most important root crop in africa. the next is yam. it is a hardy crop that grows reasonably well in poor soils and in areas well with erratic or low or unpredictable rainfall. it has been common among the rural poor mainly because it required fewer fertilizer inputs. “cassava is a staple food crop cultivated in several developing countries, largely by small farmers. it is a source of subsistence and of cash income to poor farmers as well as a source of rural employment, particularly of women manly because it tolerant to poor soils and harsh climatic conditions producing reasonably yield poor management conditions. it is a perennial crop native to tropical america with its center of origin in north-eastern and central brazil. it is cultivated mainly for its enlarged starchy root. globally, it is the sixth most important source of calories in the human diet. in africa, it has expanded as a food security crop. in snnpr, the average total area planted to the crop and production of cassava per annum is 4942 ha and 53036.2 tones respectively. during the past 20 years, production of cassava expanded rapidly in asia, especially in thailand, in response to expanded demand for imports by the european community, where it is used as livestock feed. there are concerns, however, about the likely decline in demand for cassava as food as incomes rise in developing countries and also about the stability of the european demand. cassava (manihot esculenta crantz) is sixth most important source of calories in the human diet-world,fourth most important food energy source in the tropics, is food security crop, because of its ability and capacity to yield well in drought prone, marginal wasteland under poor management where other crops would fail, the average total coverage and production of cassava per annum in southern region is 4942 ha and 53036.2 tones, respectively and in ethiopia, cassava is grown in southern and south eastern parts as staple crop. fig-1. cassava in local markets in ethiopia source: picture taken in konso market, southern ethiopia cassava based production systems include small scale rain-fed farming in pathes of plots in backyard eg. amaro, bele, gofa, yeki and tepi areas, medium scale production in wide private farms eg. amaro, metu and gofa areas, irrigated cassava production particularly in dry periods eg. amaro and finally cassava is grown intercropped together with taro, sorghum, teff, safflower, maize, enset, common beans , etc problems in cassava production include limited availability of additional land for crop production, decreased soil fertility and declining yield for major food crops have been cited as the major concerns for agriculture’s ability to provide nourishment for the increasing population, cassava is a long duration crop that takes more than 18 months for harvest and is planted at intra and inter row spacing of 80-120 x 60-100 and takes 3-4 months to develop enough canopy. 2. objectives  to sensitize and document the findings of cassava agronomy research in southern ethiopia  to draw conclusions based on major findings of cassava management research and subsequently recommend the necessary technologies for further use 3. methodologies cassava agronomy research has been carried out in the last twenty years in southern agricultural research institute. summary of research findings was prepared and presented by authors in the national first cassava research review workshop from 17-18 may 2012 at awassa, sari southern ethiopia. data of different experiments were agriculture and food sciences research, 2016, 3(1): 12-18 14 gathered from various research centers and authors. the gathered data were systematically compiled in logical manner that makes sense where conclusions were drawn accordingly using standard methodologies. agronomic data like plant height, weight and number of roots/plant and roots/plot, root length and root diameter were measured at harvest where as canopy diameter was estimated at mid flowering in all experiments. the ler is calculated as ler = l a + l b + …… + l n = aasy + bbsy + …… + nnsy = σ=ninnsy1 where l a , l b … l n is the ler for the individual crops y a , y b … y n are the individual crop yields in intercropping. s a , s b … s n is their yields as sole crops when ler is greater than 1 or more, it signals yield advantage and a ratio of less than 1, is yield disadvantage [2]. 4. results and discussion 4.1. plant population density at awassa an experiment was conducted in 2004 and 2005 cropping seasons in fluvisols of awassa at 7 0 04’n and 38 0 31’e longitudes at an altitude of 1700 m a.s.l. with the objective of determining appropriate plant and row spacing that maximizes cassava root yield in sub-humid region of ethiopia. the experiment was laid in randomized complete block design with three replication where each replication contain factorial combination of four levels of plant spacing (60 cm, 80 cm, 100 cm and 120 cm) and three levels of row of spacing (80 cm, 100 cm and 120 cm). significant positive association was observed between dry root yields (dry) and root diameter, root size, root number, root length, canopy diameter and dry biomass yield (dby). results showed that significantly higher root diameter (p<0.05) and significantly wider canopy diameter were observed due to row spacing of 120 cm than that of 80 cm row spacing. however, responses of root and canopy diameter were not significantly different (p<0.05) among 100 cm and 120 cm row spacing. significantly higher (p<0.01) root yield/plant was measured due to 120 cm row spacing than other row spacing levels. however, cassava root yield per unit area was significantly higher (p<0.05) due 100 cm and 120 cm row spacing than 80 cm row spacing. but, the effects of row spacing were not significant on number of roots/plant and length of roots. when plant spacing was considered, canopy diameter (p<0.01) and number of roots/plant (p<0.05) were significantly higher due to 120 cm plant spacing than other plant spacing levels. yield/plant due to plant spacing levels of 100 and 120 cm was not significantly different (p<0.05). also, effects of plant spacing were not significantly different (p<0.05) on cassava root diameter, root length and root yield/unit area. however, 60 cm plant spacing produced significantly lower (p<0.05) number of roots/plant, canopy diameter and root yield/plant than that of 80 cm. hence, use of 100 cm row spacing and 80 cm plant spacing is optimum for cassava production in fluvisols of awassa and similar agro-ecologies [3]. 4.2. plant population density at amaro kele, jijola, bele and loma factorial experiment was laid in rcbd with three replications at jijola district, amaro to determine optimum inter and intra row spacing for cassava. the treatments involved 80, 100 and 120 cm row spacing and 60, 80, 100 and 120 cm plant spacing. aboveground biomass was significantly (p<0.05) affected by spacing and the highest biomass was observed at spacing of 100 cm x 120cm. similarly, the root yield of cassava was statistically higher due to 80 cm x 80 cm spacing. in amaro kele area 120 cm x 80 cm spacing produced significantly higher yield compared to other interaction levels. non significant results were obtained in the plant population study trials at loma and bele [2, 4, 5]. 4.3. harvesting stage and cultivars effects on starch quality and quantity the trial was carried out during 2009-2010 at gofa woreda, snnpr. a factorial combination of five level of harvesting stage (9, 12, 15, 18, 20, 24 months after planting) and three cassava varieties (‘kelo’, ‘qulle ’ and a local variety) were laid out in randomized complete block design with three replications. data on tuber yield, yield components were recorded and some starch quality parameters from cassava tubers were analyzed in laboratory and all the data were statistically analyzed. statistically analyzed quality parameters results indicated that moisture of the tuber starch was affected significantly by harvesting stage and variety of cassava. ph and viscosity of the tuber starch were significantly affected by harvesting stage. the highest viscosity of the starch was recorded by harvesting the crop at 18 months after planting. moisture content was significantly reduced after 15 months after planting. ph of the starch was significantly reduced 15 months after harvesting. neither the harvesting stage nor the variety did not affect significantly the color and dry appearance of the cassava starch. tuber yield and yield components of the crop were significantly affected by the interaction of variety and harvesting stage. the highest tuber yield (151q/ha) was obtained when variety ‘qul'e’ was harvested at 21 months after planting. generally significantly better tuber yield of all varieties were recorded by harvesting cassava starting 18 months after planting. table-1. moisture content of different cassava varieties as affected by harvesting stage variety harvesting stage (months after planting) mean 9 12 15 18 21 kelo 63.5 82.4 84.0 67.8 55.1 70.56 qulle 63.3 81.4 77.1 56.2 64.5 68.49 local 65.1 83.9 80.1 50.6 64.9 69.49 mean 63.99b 82.57a 80.39a 58.18c 60.53bc cv (%) 7.6 source: unpublished data agriculture and food sciences research, 2016, 3(1): 12-18 15 table-2. viscosity and ph of cassava as affected by harvesting stage harvesting stage (months after planting) viscosity ph 9 7.7ab 6.7ab 12 4.97c 6.9ab 15 6.52bc 7.4a 18 9.42a 6.7ab 21 8.17ab 6.1b cv (%) 32.3 11.2 source: unpublished data table-3. tuber yield of different cassava varieties as affected by harvesting stage variety harvesting stage (months after planting) mean 9 12 15 18 21 kelo 36.14 49.38 45.25 113.7 105.7 70.03 qulle 19.9 47.5 72.1 138.5 161.8 86.0 local 15.9 48.34 71.63 51.17 140 65.5 mean 23.99 48.41 63.01 101.3 135.8 lsd(5%)variety= 17.63, harvesting stage=22.76, variety x harvesting stage = 39.43 cv (%) = 33.5 source: unpublished data 4.4. cassava / maize intercropping this study was conducted in 2005 and 2006 at goffa woreda, gamo goffa zone, to determine optimum ratios of component crops that will improve land use system of intercropping system. there were four cassava/maize intercrop row arrangements: 1row cassava: 1 row maize, 2rows cassava: 1 row maize, 2rows cassava: 2 rows maize, 1row cassava: 2 rows maize, and two sole crops of maize and cassava were incorporated. maize hybrid variety ‘bh140’ and ‘quele’ variety of cassava were used in the trial. other agronomic practices were applied equally for all plots. the experiment was lied out in randomized complete block design with four replications. the combined analysis over years showed that different maize plant densities intercropped with cassava had significant (p<0.001) effect on grain yield and yield components of maize. the highest grain yield of maize was obtained when maize and cassava was intercropped in alternate rows (1row cassava: 1 row maize). on the other hand cassava tuber yield was not affected by the intercropped maize. the analysis of land equivalent ratio showed that the advantage of intercropping was 71 % over sole cropping. among the intercropping pattern the highest yield advantage were also obtained when cassava and maize were intercropped in alternate rows. the economic analysis also indicated that the highest net benefit (13,715.4 birr/ha) were obtained when one maize row was grown between 2 cassava rows. table 4. grain yield of maize and tuber yield of cassava (q/ha) under different intercropping pattern (mean 2005-2006) treatments maize cassava stand percent of maize at harvest 1c:1m 53.11a 450.5 91.2a 1c:2m 47.32ab 474.5 60.6b 2c:1m 21.40c 464.5 94.7a 2c:2m 30.08bc 470 92.4a sole cassava 502 sole maize 66.28a 65.1b pr cv (%) 0.0002 26.1 <.0001 14.6 means followed by the same letter(s) with in a are not statistical different by dmrt (p< 5%) source: awarc [2] 4.5. cassava-legumes inter-cropping the trial was carried out for two successive cropping seasons from 2005 to 2008 at amaro clay loam textured soil with a ph of 6.5 and located in an altitude of 1400 m a.s.l. the site has bimodal rainfall with very short rain season starts from last week of february to end of march and the second season from september to october. the cassava variety used was kelo while the four legume varieties were haricot bean (omo-95), mung bean (boroda -1), soybean(awassa 95), and cow pea ( maze). two rows of 40cm apart were made between the two cassava rows to plant legumes with the spacing of 40cmx 10cm haricot bean, 40cmx 5cm for soy bean, 40cmx 10 for cow pea and 40cmx 5 cm for mung bean. both crops were planted at a time during the first shower of rainfall, february –may. data of each crop were taken from 5 randomly tagged plants per experimental unit (plots). root yield of cassava were weighed using spring balance during cassava harvest and grain yield of legumes also weighed using ordinary balance. land equivalent ratio of cassava is calculated as intercrop yield of cassava/pure stand yield of cassava and that of legumes is calculated as intercrop yield of legumes/pure stand yield of legumes; the overall land equivalent ration (ler) is simply the sum of ler of cassava and ler of legumes. the competitive value is simply the ratio of the individual ler’s of the two crops. the collected data were subjected to anova using sas computer software. table-5. effect of intercropped grain legumes with cassava on yield of legumes kg/ha) treatments sole yield (kg/ha) intercrop yield (kg/ha) ler competitive value haricot bean 3510 3222 1.09 1.49 cow pea 3302 2844 0.86 1.37 soybean 2010 2010 1.00 2.08 mung bean 1740 1948 1.12 2.24 source: unpublished data agriculture and food sciences research, 2016, 3(1): 12-18 16 intercropping cassava with haricot bean, cowpea, soybean and mung bean, reduces cassava yield by 27%, 37%, 52% and 50% respectively. however, intercropping cassava with haricot bean, cowpea, soybean and mung bean resulted in 82, 49, 48 and 62% greater land use efficiency than for either crops grown alone. table-6. agronomic performance of cassava under intercropped and sole production conditions treatments roots/ plant root length(cm) root diameter (cm) biomass (ton/ha) root yield (ton/ha) lerc ler cassava + haricot bean 2.7 32 2.98 49.38b 14.81b 0.73 1.82 cassava + cow pea 2.7 36 3.13 35.99b 12.88c 0.63 1.49 cassava+ soybean 2.9 38 2.87 49.29b 9.71de 0.48 1.48 cassava+ mung bean 2.4 29 2.30 44.25b 10.26d 0.50 1.62 sole cassava 2.9 37 3.48 72.50a 20.33a cv% 50.8 27.69 36.09 22.24 31.57 lsd5% ns ns ns 13.69* 7.24* note: ns = not significant, letters in the same column= significant at 5% probability. source: unpublished data the highest value of ler was obtained by growing cassava with haricot bean (1.82) followed by cassava with mung bean (1.62) whereas relatively the lowest was cassava grown with soybean (1.48). overall land equivalent ratio (ler) was greater than one when cassava intercropped with legumes. this suggested that the actual productivity was higher than expected when cassava was intercropped with grain legumes. this is so because early slow growth of cassava was effected by grown additional legume, cassava can maintain its maximum growth after component crop removal and the growth one these crops under cassava has no adverse effect on growth performance of each crops. land use efficiency improved by 82% when cassava intercropping with haricot bean and that of mung bean was by 62% and the lowest was with soybean and is by 48% which indicated that the actual productivity was higher than expected when cassava was intercropped with legumes. the benefit of obtaining additional legume grain would have positive advantage on food security and land use efficiency, especially in weed suppression and soil conservation in the first six months of cassava growth were slow growth of cassava could lead to such problems 5. planting dates cassava can be planted in any time of the year if moisture is available either in the soil through rainfall or irrigation. however, planting date trial carried out for three consecutive years showed that march to may plating is better than the other planting in the major cassava growing areas of ethiopia. at planting there should be enough moisture to secure 80 to 90% of germination; however, if soils are waterlogged aeration and root formation were usually hindered. the harvesting date study revealed that the released qule and kele varieties could be harvested in 18 months time. however, starch contents are higher if cassava is harvested in dry months. table-7. mean of cassava root yield as affected by planting dates for three consecutive growing seasons at amaro planting dates mean root yield (t/ha) marketable yield total root yield april 7/2000,01,02 14.85 20.52 may 2/2000,01,02 13.42 23.07 may 17/2000,01,02 8.91 17.83 june 1/2000,01,02 9.77 17.41 june 16/2000,01,02 5.25 16.05 july 1/2000,01,02 9.51 21.48 july 16/2000,01,02 5.78 14.7 july 31/2000,01,02 6.53 18.92 aug. 15/2000,01,02 4.81 15.9 aug. 30/2000,01,02 2.61 14.55 sept. 14/2000,01,02 3.79 14.46 sept. 29/2000,01,02 2.46 13.04 cv 10.85 16.32 lsd (5%) 4.19 5.07 source: awarc [1] 6. planting position and planting materials the orientation of cuttings during planting depends on plant variety and environmental conditions, and influences numerous growth characteristics and yield. the study compared different planting methods. results showed that slant and vertical plantings are suitable in awassa sandy soils compared to horizontal planting. similarly, the study revealed that planting materials shall be taken from main stem of cassava compared to its branches. in this study it was confirmed that the middle and top portion of the cassava stem produced quality planting materials that resulted in superior yield. agriculture and food sciences research, 2016, 3(1): 12-18 17 table-8. mean of yield components and yield of cassava at awassa position planting materials root length cm root diameter, cm root yield (t/ha) no. stems/hill slant main stem top 29 4.5 27.5 3.2 main stem middle 25.5 3.8 24.0 3 main stem bottom 28.2 3.3 21.7 2.8 branch stem top 35 3.8 16.7 1.8 branch stem bottom 26.7 3.5 13.3 2.2 vertical main stem top 27 3.7 20.5 2.3 main stem middle 25 3.5 18.7 2.8 main stem bottom 26 3.7 14.4 3 branch stem top 28.7 3.7 15.5 2 branch stem bottom 24.8 3 16.7 2 horizontal main stem top 21.5 3.2 9.1 3.8 main stem middle 21.5 2.7 11.4 3.5 main stem bottom 22.2 2.8 6.9 3 branch stem top 26.8 3.8 11.5 2.3 branch stem bottom 25.2 3.8 9.9 2.2 lsd pp x pm (0.05) ns ns 6.64 ns planting position (0.05) 2.99 ns 2.97 0.33 planting parts (0.05) 3.86 3.4 3.83 0.43 cv (%) 22.07 25.38 46.18 24.09 source: legesse, et al. [6] 7. land preparation methods flat, furrow and ridge planting were compared under different plant density scenarios at amaro. results showed that furrow and ridge planting produced comparable results of 48t/ha and 43t/ha compared to 38t/ha produced by flat plantings. this might be due to reaped advantage of moisture conservation that subsequently manifested in wider canopy diameter, root length and diameter of cassava. however, manual harvesting required deeper digging, more labor and time in furrow planting compared to others. therefore, planting cassava in a ridge was recommended in amaaro kele area. table-9. mean of root yield of cassava as affected by land preparation methods at amaro kele source: gobeze, et al. [3] 8. conclusions cassava is produced in diverse ecological systems for multiple purposes in ethiopia. this diversity and multiplicity coupled with site specificity of soils and climate related parameters makes generalization of agronomic recommendations difficult. however, specific recommendations were made on similar edaphic and agro-ecological conditions. results showed that cassava can be grown easily under variable climatic and edaphic conditions. when cassava is intercropped with legumes, the cassava root yield generally decreases compared with when cassava is planted alone. this is due to the competition of the component crops for light, water and nutrients. however, cassava-legume intercropping systems usually increase the land use efficiency and economic return over solely grown cassava. these advantages can occur as a result of complementary use of growth resources such as nutrients, water and light by the component crops. the yield advantage may be in terms of higher yield or higher net income. it could further be explained that the yield can be quantified in terms of dry matter production, grain or root yields, nutrient uptake, energy or protein production and market value. 9. future research directions strategic cropping system integration should be sought particularly with climbing beans, cow pea and mung beans to allow green manuring, residue cover, residue incorporation, atmospheric nitrogen fixation and enhance cassava and component crop productivity besides breaking disease and insect pest cycles. herbicides should be screened for early stage of cassava growth for sedges management as cassava does not tolerate weed effects at early stage of growths. further studies on fertilizer requirements, plant density, land preparation, planting positions and harvesting methods shall be carried out under irrigated and mechanized farming conditions. references [1] awarc, progress report of horticultural section division for the year 2003. awassa, ethiopia: awarc, 2005. [2] awarc, progress report of agronomy & crop physiology division for the year 2004. awassa, ethiopia: awarc, 2005. [3] l. gobeze, h. legese, and m. daniel, "effect of land preparation methods and spacing in growth and yield of cassava," in proceedings of african society of crop sciences, 2007, pp. 68-72. land preparation methods cassava yield (t/ha) ridge 27 a furrow 21 ab flat 19 b lsd (5%) 7 agriculture and food sciences research, 2016, 3(1): 12-18 18 [4] m. daniel, l. gobeze, and h. legesse, "effects of inter and intra row spacing on growth and yield of cassava (manhot esculenta cranz) in fluvisols of southern ethiopia," in proceedings of horticultural science society of ethiopia, 2009. [5] l. gobeze, h. legesse, and m. daniel, "effect of inter and intra row spacing on yield and yield components of cassava in moisture stress areas," in proceedings of african crop science society, 2005, pp. 147-150. [6] h. legesse, l. gobeze, a. shegro, and n. geleta, "impact of planting position and planting material on root yield of cassava," journal of agricultural science and technology usa, vol. 5, pp. 448-454, 2011. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 15 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 15-21, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.15.21 © 2019 by the authors; licensee asian online journal publishing group proportions of organic waste in the process of composting thayane leonel alves1 evandro freire lemos2 josé de arruda barbosa3 gabriela mourão de almeida4 antônio michael pereira bertino5 ( corresponding author) 1,3,4master student, postgraduated graduation in agronomy (science of the soil), universidade estadual paulista (unesp), jaboticabal, são paulo, brazil. 2agronomist, prof. dr., universidade do estado de minas gerais (uemg), passos, minas gerais, brazil. 5phd in agronomy (plant production), universidade estadual paulista (unesp), jaboticabal, são paulo, brazil. abstract the objective of this study was to evaluate the composting process made from different combinations of organic residues and to gain agronomic knowledge about the final product. the experimental design was randomized blocks, with five treatments: t1 50% sawdust, 35% fruit / vegetable husks and 15% coffee grounds and filter; t2 50% sawdust, 30% fruit / vegetable peels and 20% coffee grounds and filter; t3 60% sawdust, 30% fruit / vegetable peel and 10% coffee grounds and filter; t4 60% sawdust, 20% fruit / vegetable peels and 20% coffee grounds and filter; and t5 70% sawdust, 20% fruit / vegetable peels and 10% coffee grounds and filter and four replicates. the parameters monitored daily were temperature and humidity. after 140 days of initiation, the sample was collected for analysis. the material was analyzed in the soil and leaf fertility laboratory of the uemg unit passos to determine the organic matter content, c / n ratio, ph and macronutrient and micronutrient content. treatments 1 and 2 presented the best values in relation to the other treatments, due to the lower percentage of sawdust. the low ambient temperatures in the composting process, the high c / n ratio of the sawdust, the proportion of residues used in the formation of the compost and the volume of the material may have had a negative influence on the composting process. keywords: biological process, decomposition, degradation, mineralization, nutrients, organic fertilizer, organic waste. citation | thayane leonel alves; evandro freire lemos; josé de arruda barbosa; gabriela mourão de almeida; antônio michael pereira bertino (2019). proportions of organic waste in the process of composting. agriculture and food sciences research, 6(1): 15-21. history: received: 25 october 2018 revised: 2 december 2018 accepted: 7 january 2019 published: 20 march 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study was financed in part by the coordenação de aperfeiçoamento de pessoal de nível superior brasil (capes) finance code 001. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 16 2. material and methods ..................................................................................................................................................................... 16 3. results and discussion ................................................................................................................................................................... 17 4. conclusion ......................................................................................................................................................................................... 20 references .............................................................................................................................................................................................. 20 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.15.21&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.15.21&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/1745 https://orcid.org/0000-0003-3532-2117 https://orcid.org/0000-0001-5679-6923 https://orcid.org/0000-0002-7393-8839 https://orcid.org/0000-0002-8374-2290 https://orcid.org/0000-0002-1870-0027 agriculture and food sciences research, 2019, 6(1): 15-21 16 © 2019 by the authors; licensee asian online journal publishing group 1. introduction every day in brazil, 260000 tons of solid waste are produced, 53% of which consists of organic waste, 25% paper and cardboard, 3% plastic, 2% glass, 2% metal and 15% others, that only 2% of this total is reused. in order to increase the percentage of reuse, it is important to encourage the recycling of waste because of its environmental impact [1]. in brazil, almost 50% of municipalities still use "open dumps". the volume of garbage produced by the brazilian population increases annually, which is largely discarded incorrectly [2]. in 2016, calixto [3] affirms that selective collection is fundamental to reduce the environmental impact caused by the country's dumps, on average, each brazilian produces 1 kg of garbage per day, being 365 kg per year, he announced that 47% of the rural population of the country does not have any type of garbage collection. the bad management of garbage, besides causing environmental problems, can cause social problems: many collectors guarantee the sustenance of their families by selling what is found in garbage dumps and even minors doing this type of work. the treatment of organic waste is an excellent alternative from the environmental point of view, in order to achieve a solution for this excess, being an option to produce organic fertilizers [4]. due to the large amount of organic waste produced by brazilian cities, the work theme becomes relevant because it addresses the use of these residues through the composting process, in the formation of organic fertilizer, being an alternative for the use of waste and a fertilizer option for farmers. composting is the procedure performed by several organisms where the biodegradable organic fraction of the waste is decomposed under controlled aerobic conditions and other parameters [5]. for pereira [6] composting is an aerobic biological process, obtained by the controlled biodegradation of organic waste, generating an organic compound. the composting process is carried out by the microorganisms present in the residues, provided that the conditions of temperature, aeration and humidity are ideal [7]. the final product of the composting is a mass of fine texture and homogeneous, without characteristic smell of the residues that gave rise to it (excellent conditioner and improver of the properties of the soil). it also results in a dark liquid of bad smell called slurry, which is caused if the composting is conducted improperly nunes [8]. nunes [9] mentions that the use of organic fertilizer brings with advantage an increase in the cation exchange capacity; aggregation of particles to the soil, reducing the erosive process; stores more water because it increases water retention capacity; decreases temperature oscillations during the day; and increases the availability of phosphorus. organic fertilizers are specified according to ordinance nº 1 of march 4th, 1983. in the compost it is necessary to have a minimum of 40% of organic matter, a minimum of 1% of nitrogen, a maximum of 40% of humidity, a maximum of 18 / 1 c / n ratio, ph minimum of 6.0, minimum of 1% of calcium, minimum of 0.5% of magnesium, minimum of 1% of sulfur, minimum of 0.02% of boron, minimum of 0.1% of iron, minimum of 0.05% of manganese, minimum of 0.1% of molybdenum and minimum of 0.1% of zinc [10]. custodio, et al. [11] define that the organic materials that can be used to form the compound are practically all kitchen waste and garden waste, being: rest of vegetables and fruits; coffee powder and its filter; eggshell; tea bags; dry straw and grass; branches, leaves, bark and tree pruning, sawdust; among others. and what cannot be put are materials that may have some kind of contaminating or polluting material and non-puttable materials of difficult decomposition. examples: feces of domestic animals; toilet paper; diapers; ash and cigar stump; wood; coal; barbecue meat; fish; fat; cheeses; sick plants; weeds; glass; metals; plastics; leather; eraser; fabric; etc. organic materials discarded in the trash are classified as greens (rest of raw vegetables, rest and fruit peels, coffee grounds and filter, rest of bread, eggshell, tea bag, etc.) and brown (sawdust, leaves, bark, dried straw, hay, grass trimming, tree pruning, etc.). the greens have in their composition a higher proportion of nitrogen, c / n less than 30/1, has a high moisture content and decomposes faster; and browns that have the highest proportion of carbon in relation to nitrogen, c / n higher than 30/1. examples: coffee husk c / n: 30; coffee grounds, fine c / n: 32; coffee grounds, coarse c / n: 33; shaving c / n: 391; sawdust c / n: 482; green coconut c / n: 78; coconut fiber c / n: 144; of coal c / n: 186 [12]. the decomposition is where all the organic material will be placed. it can be done in various ways being they forming a pile or pyramid in a yard, digging a hole in the ground, wooden or plastic container, among others. this will depend on the creativity of each person and availability of material, because the ideal is that you do not buy the decomposer but rather you create it. this decomposer should be placed in an easily accessible, low slope, protected from wind, direct sunlight and animals [13]. the decomposer can be classified into three types, according to ishimura, et al. [14]: composting in piles, composting in beds and compost in cube or boxes. composting in piles uses small volumes with measures smaller than three meters in diameter. on the other hand, the composting in rows uses large volumes, being the trapezoidal format, with two meters in the lower base, one in the upper base and indeterminate length. cube or box composites use small volumes, can be constructed of wood, masonry or other material, and in varying sizes. the three phases of the composting process described by fetti [15] are defined as mesophilic, thermophilic and the final stage of maturation. the first phase, mesophilic, lasts approximately 15 days, bacteria survive at temperatures around 40ºc, and they metabolize the simplest nutrients. the second phase, thermophilic, is the longest, extends for two months, bacteria and fungi called thermophiles, act at high temperature (65-70 ° c), degrading more complex molecules. the last stage, maturation phase, can persist for one to two months, in which the temperature, microbiological activity and acidity decrease. therefore, the purpose of this study was to evaluate the composting process made from different combinations of organic waste and to gain agronomic knowledge about the final product. 2. material and methods this study was conducted at the experimental farm of the state university of minas gerais (uemg) unit passos, in passos, mg (20ºs latitude and 46ºw longitude of greenwich). the study area is located at an average elevation of 700 m, with average annual temperature: maximum 22.8 ° c and minimum of 21.7 ° c; and average annual rainfall of 1709.4 mm, dry winters and rainy summers, classified climate, according to köppen, as cwa, subtropical climate / tropical altitude climate, with dry winter and hot summer (tropical rainy). agriculture and food sciences research, 2019, 6(1): 15-21 17 © 2019 by the authors; licensee asian online journal publishing group in the composting process the following wastes were used: sawdust, fruit / vegetable waste and coffee grounds and filter. these residues were purchased from the restaurant and departments of uemg unidad passos, from the various furniture factories in passos, fruit and vegetable markets, supermarkets and private residences. the treatments were defined by mixing the organic compounds in combinations in the following proportions table 1: table-1. description of the treatments of the project. passos, 2017. treatments description t1 50% sawdust, 35% fruit / vegetable peel and 15% coffee grounds and filter. t2 50% sawdust, 30% fruit / vegetable peel and 20% coffee grounds and filter. t3 60% sawdust, 30% fruit / vegetable peel and 10% coffee grounds and filter. t4 60% sawdust, 20% fruit / vegetable peel and 20% coffee grounds and filter. t5 70% sawdust, 20% fruit / vegetable peel and 10% coffee grounds and filter. the compost was produced in reusable plastic gallons with a volume of 20 liters, which were used to commercialize mineral water, making a longitudinal cut in them to remain lying down, and holes in the bottom for the slurry to run. the parameters temperature and humidity were monitored during the maturation period of the compost. the temperature was determined with the aid of a thermometer, introducing it into the compound at three points. for the monitoring of moisture, the hand test was performed figure 1 [16] according to the three parameters of the figure the wetting of the compost was done and when wet, it was also stirred so that the process occurred under ideal conditions. figure-1. hand moisture test. note: legend: a) when you open your hand, the sample crumbles, it is too dry. b) when you shake your hand, a liquid runs between your fingers, it is too wet. c) when opening the hand, the sample forms an acorn, it is in the ideal humidity. source: moraes [16]. after the composting process was completed (temperature reduction and no odor, after 140 days of the start of the process), the sample was collected from the recycled waste to perform the analysis. each sample was dried in greenhouses at 60ºc, until constant weight, to determine dry matter and humidity. they were ground in a mill and the material was analyzed in the soil and leaf fertility laboratory of uemg unit passos for determination of organic matter content, c / n ratio, ph and macronutrient and micronutrient content. the experiment was conducted in a causal block design (dbc) with five treatments (five different waste compositions) and five replicates. the results were tabulated in a spreadsheet, to perform the statistical analyzes that were carried out with the help of the genes computer application to identify the treatments that generated superior agronomic compound [17]. 3. results and discussion the results of the characterization of the different residues are shown in table 2. it is observed that the humidity of the residues is within the expected, since the fruit and vegetable skins have a large amount of water (86.8%), sawdust is a material (2.18) and the coffee powder was not too humid (51.38%), since it had been dried before composting. macros and micronutrients presented fruit and vegetable peels as nutrient richer material than other residues. it can be observed that sawdust presents a c / n ratio (114.84), which is much higher than that referred to as limiting the composting process. regarding organic matter, sawdust and coffee powder had a higher value than fruit and vegetable peels, this may be due to the fact that these materials are already in the most advanced decomposition stage. as for the ph, the fruit / vegetable peels presented better value in relation to coffee powder and sawdust. the temperature variation in the composting process figure 2 didn’t occur as described in the literature, where it shows that in the first stage, the temperature is around 40ºc, then in the second phase, thermophilic, it acts between 65º-70ºc and final stage of maturation, the temperature gradually decreases. this may have been influenced by the proportions of the residues, since even in winter (may to october), when the temperature ranged from 8 to 33 °c figure 3, arrigoni, et al. [18] in argentina, where the average annual temperature is 8.4 °c, performed a work with composting and the temperature of the process varied according to the literature. several authors have observed different behavior of temperature in the composting process, this indicates that average values of temperature should not be considered as standard [19]; [20]; [21]; [22]; [23]; [24]. agriculture and food sciences research, 2019, 6(1): 15-21 18 © 2019 by the authors; licensee asian online journal publishing group table-2. characterization of the residues used in the composting process. passos, mg. characteristics residues fruit / vegetable peel coffee grounds and filter sawdust humidity (%) 86.81 51.38 2.18 m.s. (%) 13.19 48.62 97.82 n (g/kg) 20.37 21.42 4.13 p (g/kg) 2.54 1.37 0.07 k (g/kg) 20.95 11.31 0.21 ca(g/kg) 10.15 0.91 0.36 mg (g/kg) 1.50 1.81 0.78 s (g/kg) 1.88 1.31 0.30 b (mg/kg) 22.87 9.07 5.34 cu (mg/kg) 8.57 19.14 0.58 fe (mg/kg) 431.42 533.77 155.18 mn (mg/kg) 45.42 36.16 46.99 zn (mg/kg) 17.05 8.40 8.30 total carbon (%) 37.26 49.09 47.43 carbon / nitrogen ratio 18.29 22.92 114.84 m.o. (%) 64.24 84.63 81.77 ph 5.64 4.95 4.52 note: legend: m.s. (dry matter); n (nitrogen); p (phosphorus); k (potassium); ca (calcium); mg (magnesium); s (sulfur); b (boron); cu (copper); fe (iron); mn (manganese); zn (zinc); c / n (carbon / nitrogen ratio); m.o. (organic matter). figure-2. temperature variation during the composting process. figure-3. temperature data according to the station of passos, mg. source: instituto nacional de meteorologia (inmet) [25]. at the end of the process, treatment 1 showed a 67.7% humidity on average, treatment 2 with 67.4%, treatment 3 with 64.7%, treatment 4 with 65.3% and treatment 5 with 65.2%. according to the brazilian legislation, the compost must have a humidity of at most 40%, therefore, these results are not in accordance with the legislation, being necessary to let the compost dry. by means of the statistical analysis, it can be verified that in treatments 1 and 2 the average values for macronutrients: nitrogen, phosphorus, potassium and sulfur were higher than the other treatments, treatments 3 and 4 presented intermediate values for these macronutrients and treatment 5 resulted in the lowest values for agriculture and food sciences research, 2019, 6(1): 15-21 19 © 2019 by the authors; licensee asian online journal publishing group these same variables. however, for calcium and magnesium, there was no difference between treatments table 3. treatment 5 presents in its composition a greater amount of sawdust (70%), the material being poorer chemically presented in table 2 previously, which explains the worse final performance of the compound. treatments 3 and 4 present intermediate sawdust content (60%) and treatments 1 and 2 have the lowest amount of sawdust in their composition (50%), which is in agreement with the results of macronutrients found, that is, a correlation negative relationship between the amount of sawdust of the compound and the nitrogen, phosphorus, potassium and sulfur contents of the final compound. information on the chemical composition of the domestic compound is very scarce, there are no reference values. table-3. means of macronutrients. passos, 2017. treatment mean (g/kg) nitrogen phosphorus potassium calcium magnesium sulfur t1 (50% s; 35% cfh; 15% bcf) 13.65 a 0.856 a 7.928 a 1.870 a 1.150 a 1.088 a t2 (50% s; 30% cfh; 20% bcf) 12.57 a 0.816 a 7.640 a 1.986 a 1.114 a 0.966 a t3 (60% s; 30% cfh; 10% bcf) 9.70 b 0.610 b 6.192 b 2.638 a 1.024 a 0.772 b t4 (60% s; 20% cfh; 20% bcf) 10.27 b 0.536 b 5.328 b 1.488 a 1.210 a 0.724 b t5 (70% s; 20% cfh; 10% bcf) 7.93 c 0.432 c 3.976 c 1.250 a 1.002 a 0.596 b note: averages followed by the same letter in the column do not differ from each other by the scott & knott test at 5% probability. caption: s (sawdust); cfh (fruit peel / vegetables); bcf (coffee grounds and filter). these treatments received the highest percentage of coffee grounds (15%, 20% and 20%, respectively), which according to the table 5 shows very high values of this micronutrient. treatment 1 was higher than the others for the final manganese content table 4. the boron, iron and zinc elements showed no difference between the means of the evaluated treatments. table-4. means of micronutrient. passos, 2017. treatment mean (mg/kg) boron copper iron manganese zinc t1 (50% s; 35% cfh; 15% bcf) 12.422 a 7.962 a 311.58 a 70.052 a 11.708 a t2 (50% s; 30% cfh; 20% bcf) 10.634 a 8.228 a 284.16 a 62.618 b 9.858 a t3 (60% s; 30% cfh; 10% bcf) 11.344 a 5.634 b 373.57 a 57.986 b 12.187 a t4 (60% s; 20% cfh; 20% bcf) 9.514 a 7.072 a 301.27 a 55.258 b 11.654 a t5 (70% s; 20% cfh; 10% bcf) 9.440 a 4.630 b 262.76 a 56.868 b 10.688 a note: averages followed by the same letter in the column do not differ from each other by the scott & knott test at 5% probability. caption: s (sawdust); cfh (fruit peel / vegetables); bcf (coffee grounds and filter). it was verified that for the ph variable, treatments 1, 2 and 3 (6.582, 6.492 and 6.610 respectively) were superior to treatments 4 and 5 (6.194 and 6.238 respectively), which did not differ among them table 5. the treatments 1, 2 and 3 presented the highest proportion of fruit / vegetable peels (35%, 30% and 30%, respectively), and this residue presented the ideal ph value. according to barroso, et al. [26]; souza, et al. [27]; kampf [28] ideal values for ph of a substrate rotates around 6.0 to 6.5, symptoms of nutritional deficiency of plants are found on substrates with ph below 5.0. for the variables: carbon and organic matter, there was no difference between the treatments. however, for the carbon / nitrogen ratio, treatment 5 was superior to the other treatments. the treatments 3 and 4 were equal to each other and were higher than treatments 1 and 2, which did not differ between them. the higher the c/n ratio, the slower the mineralization of the material, which in this case is negative table 5. this result is in agreement with the proportion of sawdust used in each treatment, since this residue presented a high c/n ratio. it was observed that treatments with lower c/n ratio (treatments 1 and 2) presented the lowest proportion of sawdust (50%), treatments 3 and 4 presented intermediate c/n ratio and received ratios of sawdust also intermediate (60%), while treatment 5 presented the highest c / n ratio and the highest proportion of sawdust (70%). table-5. averages of the percentage of carbon, the c/n ratio, the percentage of organic matter (m.o.) and ph. passos, 2017. treatment mean % carbon c/n % m.o. ph t1 (50% s; 35% cfh; 15% bcf) 40.348 a 29.884 c 69.560 a 6.582 a t2 (50% s; 30% cfh; 20% bcf) 42.264 a 34.348 c 73.210 a 6.492 a t3 (60% s; 30% cfh; 10% bcf) 39.978 a 42.294 b 68.920 a 6.610 a t4 (60% s; 20% cfh; 20% bcf) 42.502 a 41.518 b 73.274 a 6.194 b t5 (70% s; 20% cfh; 10% bcf) 40.982 a 51.874 a 70.654 a 6.238 b note: averages followed by the same letter in the column do not differ from each other by the scott & knott test at 5% probability. caption: s (sawdust); cfh (fruit peel / vegetables); bcf (coffee grounds and filter). the high value of the c/n ratio in the treatments may be due to the raw material used (sawdust, high c/n ratio), unfavorable to the process, making degradation difficult. this type of residue is usually bulky and difficult to be degraded, unless it is ground, which could not be achieved in this work. this can increase the composting period. the results obtained in this study show that all treatments comply with the brazilian legislation on the organic matter content, which requires that the minimum amount of organic matter for the compound is 40%.composting can be an efficient technology for the treatment and recycling of organic solid waste. however, the low ambient temperatures in the composting process, the high c/n ratio of the sawdust, the proportion of residues used in the formation of the compost and the volume of the material may have influenced the composting process. the factors that influence composting are: microorganisms present in the compost, temperature, humidity, aeration, granulometry, c/n ratio and ph factor. the microorganisms are responsible for the decomposition of the material being composted, without them there is no way to do the composting. the temperature is related to the agriculture and food sciences research, 2019, 6(1): 15-21 20 © 2019 by the authors; licensee asian online journal publishing group thermal destruction of the microorganisms, temperatures above 65 °c, the process is delayed. the humidity of the compound is important; you cannot have too little or too much moisture in it, because it can disrupt the decomposition of organic matter. without oxygen, there is no life in the compound, so decomposition is slower and unpleasant odors will appear. in the beginning it is necessary to stir more frequently and then from time to time. when preparing the material it is necessary that the particle size is the smallest possible, because the smaller the particle size, the faster the composting process. the ideal c/n ratio to make the compound is in the range 25/1 to 35/1, that is 25 parts of carbon for each of the nitrogen. as composting is done by the aerobic method, this causes the compound to become alkaline throughout the process [11]. the diversity and concentration of the nutrients present in the compound are related to the intensity of the microbiological activity of the decomposing microorganisms and their population. for the metabolic activities of the microorganisms to occur, it is necessary to have several macro and micronutrients in the organic residues. among them, the most important are nitrogen and carbon, their presence is a limiting factor in the composting process, carbon is a basic source of energy for vital activities and nitrogen is necessary for protoplasmic reproduction of microorganisms. palious waste is a source of carbon and green materials are sources of nitrogen [6]. 4. conclusion there are no reference values for macronutrient and micronutrient results, as far as c/n results are concerned, the data were outside the standards established by brazilian legislation, but organic matter and ph variables met the requirements. in general, treatments 1 and 2 (50% of sawdust + 35% of organic residue + 15% of coffee grounds and 50% of sawdust + 30% of organic residue + 20% of coffee grounds, respectively) presented the best nutritional values in relation to the other treatments, influenced by the lower percentage of sawdust. therefore, it is necessary to carry out more studies with smaller ratios of sawdust, to see the viability of the composting process. since the proportion of 30% of green material and 70% of brown material can not be used as a standard for composting. references [1] recycle, "composition of brazilian garbage in regard to veja. recicloteca information center on recycling and environment." available: http://www.recicloteca.org.br/coleta-seletiva/a-composicao-do-lixo-brasileiro/ [accessed 31 may. 2016], 2011. 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[28] a. n. kampf, commercial production of ornamental plants. guaíba: agropecuária, 2000. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn(e) : 2411-6653 issn(p) : 2518-0193 vol. 3, no. 2, 59-65, 2016 http://www.asianonlinejournals.com/index.php/aesr 59 morphological characterization of deepwater rice genotypes m.m. emam ahmed1 md. maksudul haque2  a.b.m arif hasan khan robin3 mohammad anwar hossain4 1scientific officer, plant breeding division, bangladesh rice research institute, gazipur 1701, bangladesh 2scientific officer (golden rice), plant breeding division, bangladesh rice research institute, gazipur 1701, bangladesh 3,4department of genetics and plant breeding, bangladesh agricultural university, mymensingh 2202, bangladesh ( corresponding author) abstract the experiments was conducted to study the early establishment morphological characters of advanced deep water rice.the parameters such as plant height, number of leaves, and number of effective & noneffective tillers were studied with different das as treatment in field condition. the genotype br2242b-2-5, br5915-b-7, bazzail-65 &gabura showed more plant height at 30 das & 60 das. but hbj.a.iv & bazail-65 showed more plant height at 90 das & 120 das. br224-2b-2-5 & br5915-b7 showed more leaves and tillers at early stage (60 das). but hbj.a.iv &birpala showed better performance for same characters at later stage (90 das, 120 das & 150 das). the advanced genotype br224-2b-2-5 is able to establish significantly at early stage as it showed vigorous growth against stress condition present at early stage. hbj.a.iv and birpala have potential of producing more leaves and tillers throughout the growth stages and hbj.a.iv and birpala is the tall type dwr cultivar as it produced more seedling height and plant height than others. the yield of the genotype br224-2b-2-5 and br5915-b-7 are 3.07 t ha-1 and 2.71 t ha-1 respectively. finally, the advanced genotype br224-2b2-5 may be a good source to meet the future challenge. keywords: morphological, characterization, deepwater, rice (oryza sativa l.). contents 1. introduction ............................................................................................................................................................................... 60 2. materials and methods ............................................................................................................................................................. 60 3. result and discussion................................................................................................................................................................ 61 4. conclusion .................................................................................................................................................................................. 64 references ...................................................................................................................................................................................... 64 citation | m.m. emam ahmed; md. maksudul haque; a.b.m arif hasan khan robin; mohammad anwar hossain (2016). morphological characterization of deepwater rice genotypes. agriculture and food sciences research, 3(2): 59-65. doi: 10.20448/journal.512/2016.3.2/512.2.59.65 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: this work is licensed under a creative commons attribution 3.0 license contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 16 august 2016/ revised: 28 september 2016/ accepted: 3 october 2016/ published: 10 october 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.2/512.2.59.65 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=m.m.%20emam%20ahmed https://orcid.org/orcid-search/quick-search?searchquery=md.%20maksudul%20haque https://orcid.org/orcid-search/quick-search?searchquery=a.b.m%20arif%20hasan%20khan%20robin https://orcid.org/orcid-search/quick-search?searchquery=mohammad%20anwar%20hossain http://search.crossref.org/?q=10.20448/journal.512/2016.3.2/512.2.59.65 agriculture and food sciences research, 2016, 3(2): 59-65 60 1. introduction rice (oryza sativa l.) is the most important and extensively cultivated cereal crops in bangladesh. rice has been considered as staple food and about 80% of the total cultivated lands in bangladesh are used for rice cultivation [1]. bangladesh is the world’s fourth largest rice producer. in the last three to four decades, great efforts in rice research and farming innovations were made to boost rice production. the country’s total production has also increased to about 33.3 million tons (milled rice) in 2011 whereas indonesia, india, and china produced 44.31, 103.8 and 135.1 million tons respectively [2]. furthermore, rice alone contributes about 9.5 % of the total agricultural gdp in the country. among all crops, rice is the driving force of bangladesh agriculture. in fact, food production in bangladesh is dominated by a single crop (rice) and a single season (boro, which accounts for over 60% of total rice production) [3]. rice is considered as major crop in bangladesh. it is not only the main source of carbohydrate but also provides 69.61% of calories and 56.15% of the proteins in the average daily diet of the people [4]. the rice production also increased steadily along with net food demands (rice & wheat) and reached 32.1 and 33.3 million tons in 2009 and 2010 [2]. complete submergence annually affects about 16 million ha of rice in south and southeast asia, and about one third of the total rice growing area in africa. the effect of damage caused by transient submergence is dependent on the characteristics of flood waters, including temperature, turbidity, concentration of dissolved gases, and extent of light penetration [5]. deepwater rice (dwr) is grown in flooded conditions with water more than 50 cm (20 inch) deep. more than 100 million people in south and southeast asia rely on deepwater rice for their sustenance. many districts of bangladesh are flooded during the rice cultivation season every year and thus, curtail the national rice yield by causing severe damage to the rice cultivated field. therefore, it is high time to select potential rice cultivars for breeding program to develop submergence tolerant as well as flash flood resistant rice variety [6]. the topographical situation along with availability of water and sub tropical climate constitutes an excellent habitat for rice cultivation in bangladesh. in fact, there are different rice ecosystems, namely upland (direct seeded pre-monsoon aus), irrigated (mainly, dry season boro), rainfed lowland (mainly monsoon season transplanted aman, medium-deep stagnant water up to 50-100cm) [7]. rice is a semi-aquatic plant and one of the most important crops cultivated in both tropical and temperate regions. dwr is grown in more than 50 cm water for one month or longer during the cropping season. based on stature and depth of water, these are of two types: (i) traditional tall, and (ii) floating. traditional tall cultivars are tall with long leaves, and grown at water depths between 50 and 100 cm; floating rice is grown in 100 cm or deeper situations. in bangladesh most of the rice grown in the low lying areas during monsoon are floating rice, generally called as deepwater rice, locally known as broadcast aman, jolidhan, poushdhan etc. [8]. in bangladesh deepwater rice covers an area of 0.48 million ha [7] where there is no other option to cultivate modern t. aman varieties. in haor & beel areas like sunamganj, sylhet, habiganj, b. baria, faridpur, gopalganj and pabna, local deepwater rice varieties are cultivated which have low yield potential [8]. the depth of water in some areas can exceed 4 m as in floating rice areas. apparently each of these types of floods requires specific adaptive traits, which necessitates the development of unique varieties [9]. though dwr is cultivated in small areas with low yield, attention should be given to achieve breakthrough in yield potential. many advanced lines of dwr having better yield potential have been developed by bangladesh rice research institute (brri). screening genotypes at seedling stages have several benefits, such as low cost, ease of handling, less laborious and getting rid of susceptible genotypes at earliest [10]. in vitro selection techniques involving the use of polyethylene glycol (peg), is one of the reliable methods for screening desirable genotypes and to study further the effects of water scarcity on plant germination indices [11, 12]. peg is a non-penetrate and non-toxic osmotic substance which can be used to lower the water potential of culture medium [13]. the selected abiotic stress tolerance rice cultivars have a potential of direct introduction in to farmer fields [14] or utilize them in breeding programs to develop abiotic stress tolerance rice cultivars [14, 15]. early establishment of crop is necessary for dwr to face the flood water throughout the growing season (typically one or two months). the crop established before monsoon begins or floodwater enters can withstand better in raising water level situation. thus, effective crop growth can boost up total grain production at the end of the season. crop improvement against moisture stress has been difficulty mainly for 1) lack of suitable screening technique that allows large population and 2) complexity in its arrival times and extents, since it occurs with different intensity and extent in different years. growth study along with different plant structures is of utmost necessity for crop improvement under stress condition. though dwr area is comparatively low compared to total rice area, there is necessity of working on it because there exist low lying rice areas where it is one and only option during aman season. if high yield potential dwr varieties are introduced, there will be significant increase in national rice production. hence there is need of advanced breeding lines to select for a rice variety suitable for dwr areas. considering the above facts, the present research studies was undertaken to fulfill the following objectives: to study the osmotic stress tolerance of newly developed advanced dwr genotypes through peg treatment and to study the extent of variation of growth status of advanced rice genotypes under osmotic stress and actual field conditions. 2. materials and methods the experiment was conducted at the deepwater rice field of brri, regional station, habiganj during the period of may, 2014 to november, 2014 to characterize seven advanced dwr genotypes morphologically. the experiment was carried at the field of brri, regional station, habiganj from april to november in 2014. the site is at 245 n latitude and 913 e longitude having an elevation of 14 m from the sea level. the soil of the area is acidic agriculture and food sciences research, 2016, 3(2): 59-65 61 (ph 4.5) and clay type. texture of the soil is heavy with high organic matter content (3.9%). on average yearly rainfall of that place is 2330 mm. in 2014, the highest temperature recorded in may is 33 c and the lowest in january is 12 c. the range of the on average flooding height is 250-300 cm that occurs in between july to august. the experiment was laid out in a randomized complete block design (rcbd) with three replications. the unit plot size was 5 x 5 m. flash flood occurs from the last week of april to may and it is one of the highest risks of the haor (depressed) areas. 2.1. experimental treatments the experimental treatments were as follows: a. genotypes: 7 i. br224-2b-2-5b ii. br5915-b-7b iii. bazail-65 iv. gabura v. lal-khama vi. hbj.a.iv vii. birpala b. morphological data on different days after sowing (das): 5 i. 30 das ii. 60 das iii. 90 das iv. 120 das v. 150 das 2.2. design of the experiment 2.2.1. collection of experimental data before harvesting according to treatments (30 das, 60 das, 90 das, 120 das and 150 das) following data was taken: i. plant height (cm) ii. number of leaves plant -1 iii. number of effective tillers plant -1 iv. number of non-effective tillers plant -1 i. plant height (cm) the height of five rice plants were measured from the ground level to the tip of flag leaf and mean plant height was recorded in cm. i. number of leaves plant -1 all the leaves present on same sample plants were counted and average data kept on data sheet. ii. number of effective tillers plant -1 all the effective tillers of five randomly selected plants were counted and were averaged. iii. number of non-effective tillers plant -1 all the non-effective tillers of same selected plants were counted and were averaged. iv. yield (t ha-1) the grain was harvested when the plant reached 80% maturity and yield of each individual plot was estimated at 14% moisture. 2.3. statistical analysis of data the recorded data were compiled and tabulated in proper form for statistical analysis. analysis for variance was done following the rcbd with the help computer package mstat software. the mean differences were compared with dmrt [16]. 3. result and discussion 3.1. response among the genotypes/varieties for different plant characters the mean squares of genotype for plant height, number of leaves, number of effective tillers and number of noneffective tillers are significant indicating the presence of adequate variability among the genotypes for these parameters (table 1).the results of genotypic responses to different das are presented in table 2 and figure 1, 2, 3 and 4.the yield data is also presented here to make a justified conclusion to present study (table 2 and figure 5). table-1. summary of analysis of variance on some plant characters in field condition sources of variance degree of freedom mean square plant height number of leaves number of effective tillers number of non effective tillers yield factor a (genotype) 6 161.39** 438.35** 26.95** 1.02** 0.035** factor b (treatment) 4 80786.10** 9685.06** 669.68** 15.82** 31.851** ab 24 255.02** 224.77** 12.73** 0.60** 0.035** error 68 33.22 26.02 3.52 0.31 0.002 note: **significant at 1% level of probability agriculture and food sciences research, 2016, 3(2): 59-65 62 table-2. mean performance of different genotypes/varieties for different plant characters name of the genotype/variety plant height (cm) number of leaves no. of effective tillers no. of noneffective tillers yield (t ha-1) br224-2b-2-5 127.67a 30.07bc 7.0b 0.87 bc 3.07 a br5915-b-7 124.53 ab 26.53 c 7.13 b 0.87 bc 2.71 c bazail-65 124.73 ab 26.53 c 6.60 b 0.67 c 2.97 b gabura 120.53 bc 31.53 b 6.93 b 1.00 abc 2.74 c lal-khama 118.73 c 36.33 a 8.07 b 1.2 ab 2.73 c hbj.a.iv 119.87 c 39.13 a 9.80 a 1.4 a 2.75 c birpala 120.53 bc 38.73 a 9.67 a 1.27 ab 2.30 c lsd (0.05) 3.95 3.69 1.39 0.39 0.07 s.e() 1.40 1.30 0.49 0.13 0.01 note: * the values bearing different letters in the same column are significantly different at 5% level of probability the plant height varied from 118.73 to 127.67 cm and maximum plant height (127.67 cm) was found in the genotype br224-2b-2-5 whereas minimum plant height (118.73 cm) for lal-khama (table 2). in case of number of leaves, the data showed a variation from 26.53 to 39.13 and maximum number of leaves was found in hbj.a.iv whereas minimum number of leaves in br5915-b-7 & bazail-65.the number of effective tillers varied from 9.80 to 6.93 and maximum number of effective tillers was recorded for the genotype hbj.a.iv whereas minimum number effective tillers for bazail-65. br224-2b-2-5, br5915-b-7, bazail-65, gabura, lal-khama showed no significant difference. the number of non effective tillers, varied from 0.67 to 1.4 and the highest value 1.4 was found in hbj.a.iv and the lowest value was found for the genotype bazail-65. in case of yield, the data varied from 2.30 to 3.07 t ha-1 and the highest yield was obtained from br224-2b-5-2 (3.07 t ha-1) and lowest yield from birpala (2.30 t ha-1) (table 2). 3.2. response of genotypes/varieties at different das mean squares due to different treatments (t1, t2, t3, t4, and t5) were significant (table 2). the effect of different das is shown in table 3. among the five treatments t5 (150 das) showed best result as the day increase plant responses significantly with das. plant height and number of leaves were highly significant. in case of number of leaves, number of effective and non-effective tillers there were no significant difference between t4 and t5 (table 3). table-3. mean effect of different das on plant characters different das plant height (cm) number of leaves number of effective tillers number of noneffective tillers t1= 30 das t2= 60 das t3= 90 das t4= 120 das t5= 150 das 35.52 e 100.62 d 119.91 c 153.43 b 202.38 a 4 d 16.72 c 39.28 b 50.76 a 52.67 a 0.0 d 4.52 c 8.85 b 12.62 a 13.42 a 0.0 c 0.28 c 1.19 b 1.81 a 1.91 a lsd (0.05) 3.34 3.12 1.17 0.33 s.e. () 1.12 1.12 0.41 0.12 note: * the values bearing different letters in the same column are significantly different at 5% level of probability 3.3. effects of genotype x das treatment on plant characters mean squares due to interaction (variety x treatment) were significant (table 4.5). interaction effects of genotype and treatment (das) on plant characters are presented in figures 1, 2, 3, and 4. the genotypes br224-2b-2-5, br5915-b-7, bazail-65, and gabura showed maximum plant height at t1 and t2 but hbj.a.iv and bazail-65 showed maximum plant height at t3 and t4 (figure 4.8). in case of number of leaves all the genotypes were same at t1 but br224-2b-2-5, and br5915-b-7 recorded the highest number of live leaves at 60 das (t2) (figure 4.9). hbj.a.iv and birpala showed more leaves at t3, t4, and t5 (figure 2). no tiller was observed at 30 das (t1). br224-2b-2-5, and br5915-b-7 showed maximum effective tillers at 60 das (t2) (figure 4.10). like number of leaves hbj.a.iv and birpala showed maximum effective tillers at t3, t4, and t5 (figure 2 & 3). the cultivar gabura showed more non-effective tillers at t3 but hbj.a.iv showed more number of non-effective tillers at t4 and t5 (figure 4). the genotypes br224-2b-2-5, br5915-b-7, bazail-65, and gabura showed maximum plant height at 30 dasand 60 das.hbj.a.iv and bazail-65 showed maximum plant height at 90 das and 120 das (figure 4.8). in case of number of leaves all the genotypes were same at 30 das. br224-2b-2-5, and br5915-b-7 showed maximum leaves at 60 das (t2) but hbj.a.iv and birpala showed maximum leaves at later stages (120 das and150 das) (figure 4.9). no tiller was observed at 30 das (t1), br224-2b-2-5 and br5915-b-7 showed maximum effective tillers at 60 das (t2) (figure 4.10). like number of leaves hbj.a.iv and birpala showed maximum effective tillers at later stages (120 das and 150 das) as shown in figure 4.9 and 4.10. sanchez, et al. [17] reported that taller rice plants produce fewer tillers than shorter plants which are similar to the present study. ashfaq, et al. [15] also observed that plant height and number of tillers are essential traits in boosting yield performance. 3.4. yield performance after yield estimation of seven deepwater genotypes it was found that br224-2b-2-5 has the highest yield performance (3.07 t ha-1) and birpala showed the lowest yield (2.30 t ha-1) among all the genotypes (table 3 and figure 5). the yield also varied significantly in relation to different genotypes and the highest yield was found for the genotype br224-2b-2-5(3.07 t ha-1) and lowest for birpala (2.30 t ha-1) (table 3). similar results were reported by several authors [18, 19] to describe yield potentiality of rice in a continuously flooded ecosystem. agriculture and food sciences research, 2016, 3(2): 59-65 63 figure-1. plant height of different rice genotypes at different treatments. mean (se) was calculated from three replicates from each treatment. t1, t2, t3, t4 and t5 indicate 30 das, 60 das, 90 das, 120 das and 150 das respectively. bars with different letters are significantly different at p0.05 applying lsd test. figure-2. number of leaves of different rice genotypes at different treatments. other details as in figure 4.8 figure-3. number of effective tillers of different rice genotypes at different treatments. other details as in figure 4.8 agriculture and food sciences research, 2016, 3(2): 59-65 64 figure-4. number of non-effective tillers of different rice genotypes at different treatments. other details as in figure 4.8. figure-5. mean yield of different rice genotypes. mean (se) was calculated from three replicates. bars with different letters are significantly different at p<0.05 applying lsd test. 4. conclusion in the study, seven advanced dwr genotypes were grown at field condition to evaluate their yield and yield contributing characters. the results indicated that advanced genotypes br224-2b-2-5, br5915-b-7, and bazail-65 showed early growth status in case of plant height, number of leaves, and even number of tillers. but later on, hbj.a.iv and bazail-65 recorded the highest plant height at t3 (90 das) and t4 (120 das). in case of number of leaves br224-2b-2-5 and br5915-b-7 has maximum number of leaves and tillers at 60 das. but hbj.a.iv and birpala showed more leaves and tillers at later stages (t3, t4 and t5). the cultivar gabura showed more noneffective tillers at early (t3) but hbj.a.iv showed more number of non-effective tillers at t4 and t5. the highest yield was found for the genotype br224-2b-2-5(3.07 t ha-1) and the lowest for birpala (2.30 t ha-1). we summarize that the advanced genotype br224-2b-2-5 is able to establish significantly at early stage as it showed vigorous growth against stress condition present at early stage. hbj.a.iv and birpala have potential of producing more leaves and tillers throughout the growth stages and hbj.a.iv and birpala is the tall type dwr cultivar as it produced more seedling height and plant height than others. the yield of the genotype br224-2b-2-5 and br5915-b-7 are 3.07 t ha1 and 2.71 t ha-1 respectively. finally, the advanced genotype br224-2b-2-5 may be a good source to meet the future challenge. references [1] bbs, "statistical pocket book of bangladesh," bangladesh bureau of statistics. statistics division, ministry of planning, govt. of the people’s republic of bangladesh. chapter v, 2011. [2] fao, production year book for 2011. rome, italy: fao. un, 2011. [3] ministry of food and disaster management (mofdm), "monitoring report of the national food policy plan of action and country investment plan," fpmu, food division, mofdm, khadday bhaban, dhaka, 2012. [4] fao, production year book for 2011. rome, italy: fao. un, 2009. [5] k. das, d. panda, r. sarkar, j. reddy, and a. ismail, "submergence tolerance in relation to variable floodwater conditions in rice," environmental and experimental botany, vol. 66, pp. 425-434, 2009. [6] s. matin, m. ashrafuzzaman, m. islam, s. sikdar, and n. zobayer, "molecular marker based (ssr) genetic diversity analysis in deepwater rice germplasms of bangladesh," international journal of biosciences, vol. 2, pp. 64-72, 2012. agriculture and food sciences research, 2016, 3(2): 59-65 65 [7] bbs, "statistical pocket book of bangladesh," bangladesh bureau of statistics. statistics division, ministry of planning, govt. of the people’s republic of bangladesh. chapter v, 2009. [8] banglapedia, "national encyclopedia of bangladesh." retrieved from: http://www.banglapedia.org/, 2012. [9] r. lafitte, a. ismail, and j. bennett, "abiotic stress tolerance in tropical rice: progress and the future," oryza, vol. 43, pp. 171-186, 2006. [10] s. rauf, "breeding sunflower (helianthus annuus l.) for drought tolerance," communication in biometry and crop science, vol. 3, pp. 29-44, 2008. [11] k. kocheva and g. georgiev, "evaluation of the reaction of two contrasting barley (hordeum vulgare l.) cultivars in response to osmotic stress with peg 6000," bulgarian journal of plant physiology, vol. 1, pp. 290-294, 2003. [12] g. sakthivelu, m. devi, p. giridhar, t. rajasekaran, g. ravishankar, t. nedev, and g. kosturkova, "drought induced alterations in growth, osmotic potential and in vitro regeneration of soybean cultivars," genetics and applied plant physiology, vol. 34, pp. 103112, 2008. [13] z. khodarahmpour, "effect of drought stress induced by polyethylene glycol (peg) on germination indices in corn (zea mays l.) hybrids," african journal of biotechnology, vol. 10, pp. 18222-18227, 2011. [14] d. djilianov, t. georgieva, d. moyankova, a. atanassov, k. shinozaki, s. smeeken, d. verma, and n. murata, "improved abiotic stress tolerance in plants by accumulation of osmoprotectants gene transfer approach," biotechnology and biotechnological equipment, vol. 19, pp. 63-71, 2005. [15] m. ashfaq, a. khan, s. khan, and r. ahmad, "association of various morphological traits with yield and genetic divergence in rice (oryza sativa l.)," international journal of agricultural biology, vol. 14, pp. 55–62, 2012. [16] k. gomez and a. gomez, statistical procedures for agriculture research, 2nd ed. new york: john wiley and sons, 1984. [17] o. sanchez, e. arrieta, and e. espinoza, "vegetative and reproductive development of costa rican weedy rice compared with commercial rice (oryza sativa)," planta daninha, vol. 25, pp. 13-23, 2007. [18] b. miller, j. hill, and s. roberts, "plant population effects on growth and yield in water seeded rice," agronomy journal, vol. 83, pp. 291–297, 1991. [19] t. sakamoto and m. matsuoka, "generating high yielding varieties by genetic manipulation of plant architecture," current opinion in biotechnology, vol. 15, pp. 144-147, 2004. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.banglapedia.org/, agriculture and food sciences research issn(e) : 2411-6653 issn(p) : 2518-0193 vol. 3, no. 1, 25-28, 2016 http://www.asianonlinejournals.com/index.php/aesr 25 the role of agricultural education in ensuring national security in nigeria francis, a. angbre1 1department of agricultural education, college of education, akwnaga, nasarawa state abstract nigeria is presently facing serious security challenges and instability that threaten our existence as people of one destiny. this article examines the important roles agricultural education can play in ensuring our national security. vocational agriculture is a skill acquisition training programme for youths to be self-employed and self-reliance in agricultural production for empowerment. in addition, democratic principles and the rule of law must be enshrined in our political life for equity, fairness and justice. the roles of agricultural education in ensuring national security are in the areas of reduction in rural-urban migration, youth empowerment and reduction in youth unemployment and growth in industry. in conjunction with this, government needs to take pro-active security measures for the security and safety of the nation. keywords: role, agricultural education, ensuring, national security. contents 1. introduction ............................................................................................................................................................................... 26 2. conclusion .................................................................................................................................................................................. 27 3. recommendations ..................................................................................................................................................................... 27 references ...................................................................................................................................................................................... 28 citation | francis, a. angbre (2016). the role of agricultural education in ensuring national security in nigeria. agriculture and food sciences research, 3(1): 25-28. doi: 10.20448/journal.512/2016.3.1/512.1.25.28 issn(e) : 2411-6653 issn(p) : 2411-6653 licensed: this work is licensed under a creative commons attribution 3.0 license funding: this study received no specific financial support. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. history: received: 26 april 2016/ revised: 18 may 2016/ accepted: 21 may 2016/ published: 26 may 2016 ethical: this study follows all ethical practices during writing. publisher: asian online journal publishing group http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512/2016.3.1/512.1.25.28 http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=francis,%20a.%20angbre http://search.crossref.org/?q=10.20448/journal.512/2016.3.1/512.1.25.28 agriculture and food sciences research, 2016, 3(1): 25-28 26 1. introduction nigeria is at present facing security challenges which threaten the corporate existence and stability of the country. this comes into force in the wake of the increasing terrorism, kidnapping armed robbery, cultism, political thugery, and above all, ethnic, fulani herdsmen/farmers and religious clashes. the level of insecurity is on the increase despite the huge sums of money budgeted on yearly basis as security vote by the three tiers of government. security is a collective responsibility of all citizens of a country in ensuring a secure and peaceful environment for optimal performance in chosen careers for the economic growth and development of this nation. a nation’s stability is a sine-qua non for foreign investment as well as economic growth and development in all facets of our economy. adabara [1] affirmed that youth unemployment and poverty are on the increase in nigeria, with an army of unemployed youths parading the streets in search of limited jobs. he further stressed that this has been responsible for the present security challenges the country is facing. it has also been opined that countries where government is a major employer of labour is bound to face high unemployment rate, nigeria being a typical example [2]. majority of the labour force in nigeria are unskilled coupled with the mass rural-urban migration that tend to compound the unemployment situation and further aggravate security challenges. the present 6-3-3-4 educational system is faulty because it has not been able to address the present youth unemployment, as most graduates are white collar job seekers instead of being job creators. this also points to the fact that the current school curriculum for agricultural education at nce level is faulty and fails to address the present realities on the ground. in the past, informed agricultural education where children were exposed to in their early life. they accompanied their parents to the farm, where they were taught the art and rudiments of farming with much emphasis on practical training in basic skills acquisition. this strong emphasis on practical has gradually given way to strong theoretical work in the modern formal agricultural education thereby creating challenges to agricultural education. development can only be achieved under a peaceful and stable environment which is a major pre-requisite for an increase in crop yield and sustained growth, especially in the efficient use of scarce resources for optimal production. political instability, most especially in africa, has been attributed to the undemocratic nature of our elections and complete absence of internal democracy within the political parties, thereby leading to discontentment among politicians [3]. this has plunged many african countries into instability that threatens national security with attendant destruction of lives and properties. in nigeria where illiteracy level is high, there is the tendency of mutual suspicious of dominance which can culminate into security threat, thereby, jeopardizing security and national integration. 1.1. education for national security education is a long time investment by the three tiers of government and private individuals for its continued existence, preservation of culture and traditions, national integration and improvement of the society. education is therefore, an instruction for national development and self-reliance. suleiman [4] sees educationas a fulcrum of societal civilization and development. he further stressed that a civilized person relates and interacts with one another irrespective of ethnic, religious and cultural differences for national integration. at individual level, people need education for a variety of reasons: firstly, they need it for knowledge acquisition, this will enable them to understand the dynamics of the world and scientific innovations. secondly, education is desired by many as a medium through which they can acquire professional and vocational skills to qualify for paid employment. thirdly, education may also be desired for the social status it confers on its recipients. at the national level, education is required for development, integration and national security. the products of our school system should be able to think for themselves, participate meaningfully in the act of decision making on important national issues and in passing mature judgment on all such matters that affect the security of the country. this is clearly what education attempts to build in the mind of nigerian youths. this is most important and very necessary in a time when nigeria is facing security challenges and instability, as well as the polarization of politics along ethnic and religion divides or issues. the system of education should be able to produce citizens who are alive their social, political and economic responsibilities to their fatherland. education should be geared toward imparting the basic principles of modern knowledge on its citizens. the products of the schools should be trained to acquire the ability to adopt such principles in solving their own problems and those of the society at large. development which is an off shot of education can only be achieved under a peaceful and stable environment, when natural and manmade resources are effectively utilized in the production of goods and services that impact positively on the people. the present 6-3-3-4 education curriculum at nce level should be reviewed, so that it would be more practically oriented and be geared towards skill acquisition. this aspect of education is most crucial to the needs of our youths in this era of violence, militancy, terrorism, kidnapping, political thuggery, poverty and diseases. the only way out of this quagmire is to equip our youths with such skills that will help them to appreciate the opinion of others as people with right to life, as well as be able to cope with problems surrounding them. mohammed [5] sees vocational education as systematic learning experiences which are designed to fit individuals for gainful employment in recognized occupations as semi-skilled workers. vocational education is therefore, that part of attitudes, work habit and application needed by workers to enter and make progress in employment on a useful productive basis. vocational education is any form of education whose primary aim is to prepare persons for self employment in recognized occupation [6]. this implies that graduates from vocational education will be self-reliant and even become employer of labour (job creators) rather than job seekers. alabi [7] sees vocational agriculture as the training in broad fields of agriculture in order to provide knowledge and skills which will enable the individual make a living. vocational education as a skill acquisition programme will therefore facilitate good quality education, improve health condition, empowerment, as well as prepare students against the hazards and hardship of the environment they might find themselves. it also contributes to the quality of life and the promotion of peaceful co-existence. skills acquisition in vocational education is aimed at facilitating the agriculture and food sciences research, 2016, 3(1): 25-28 27 development of psycho-social skills that the youths required to deal with the demands and challenges of everyday life as well as coping with emotions and stress [8]. skills acquisition for self employment is the focus of vocational education. it is aimed at eradicating poverty and youth unemployment in the society. when the youths are gainfully employed and self reliant, they cannot be used or manipulated by greedy politicians to perpetuate violence in the society or be involved in acts of terrorism, kidnapping, armed robbery, ethnic and religious conflicts. security challenges are the direct effect and consequences of a misplaced educational system and vocational programme. a well articulated vocational education programme that is free from political gimmick and manipulated will go a long way in reducing security challenges and further rekindle the hope and aspiration of foreign and local investors. vocational education gives training and impacts the necessary skills to individuals who shall be self-reliant economically. idleness is a base for laziness whose end result is involvement in criminal activities. a man who profitably engages in one productive exercise or the other is likely to be able to meet his basic needs as a responsible citizen [3]. 1.2. role of agricultural education in national security agricultural education as a vocational course is an integral part of our educational system. its main focus is on manpower training in skill acquisition in all the disciplines of agriculture. it is aimed at exposing, inculcating and developing knowledge, practical skills, competences and attitude. it provides the right type of skills and knowledge that will make an individual function effectively in any agricultural related job. agricultural education roles in national security are as follows: • reduction in rural-urban migration one major cause of insecurity and instability in a nation is the mass movement of unemployed youths to the urban centres in search of jobs. they easily become available tools for use in promoting insecurity. if these youths are well trained in vocational agriculture, with basic skills acquisition in the various disciplines of agriculture and given the necessary assistance, they can be self-reliant. this will drastically reduce rural-urban migration, thereby creating an enabling environment for effective participation in nation building and development: therefore security is guarantee. • youth empowerment poverty and hunger have been identified as major factors that make youths take to crime and other social vices. when youths are given the right manpower training in skills acquisition, especially in agriculture, they can become self-reliant in their fields of specialization. this will improve their economic power/income and be able to meet their nutritional requirement for healthy living and physical development. with improvement in economic power and social status as well as enough food on the youth’s table, the rate of crime and involvement in violence will be a thing of the past. • reduction in youth unemployment in nigeria, agriculture is a major employer of labour when compared with other sectors of the economy. training in vocational agriculture allows for skill acquisition in all the disciplines of agriculture which further makes the youths to be self-employed. when they are fully engaged in their chosen fields, they no longer have the time to be involved in violence and other criminal activities. this will reduce or eliminate insecurity in our society. • self reliance in food production when youths put into practical use the acquired skills in vocational agriculture, there would be abundance of food for the teaming population. the increase in food production would be sustained and the country can boast of self-reliance in agricultural production. poverty and hunger would be eradicated from the society, thereby paving the way for peace and stability to prevail in the country. • industrial growth availability of essential agricultural raw materials and its effective utilization can only be achieved when the right skills are acquired through training. the skilled manpower goes into mass production, which are essential pre-requisites for industrial growth. the multiplier effect of this is the creation of more job opportunities for the unemployed youths, reduction in poverty level and crime. the overall benefit of this to the society is the reduction in security threats. 2. conclusion education is a veritable tool in the fight against security challenges in nigeria. to effectively achieve these objectives, the educational system must be such that inculcates skills acquisition through effective training in vocational agriculture and entrepreneurship. this is to meet up with the challenging demands of our society, which has been characterized by different challenges like terrorism, violence, kidnapping, armed robbery, political thuggery, etc. agricultural education as a vocational course can effectively play a major role in ensuring national security. in addition to skills acquisition programme, government needs to take pro-active measures that would further enhance the security and safety of the people. this can be achieved through strict adherence to democratic principles and rule of law in our polity and proper screening of people at various government and private agencies. 3. recommendations the important roles agricultural education play in ensuring national security are very glaring and cannot be overlooked in the present dispensation. for vocational training in agriculture to succeed in combating the present security challenges, the following recommendations are proffered: • the government should as a matter of urgent importance put in place ways on how to create employment opportunities through agricultural educational by using the principles and practice of agriculture create wealth and jobs. agriculture and food sciences research, 2016, 3(1): 25-28 28 • curriculum in vocational agriculture at nce level should be reviewed, so that it will be more practically oriented. • vocational education should be made compulsory at all levels of our tertiary institutions. • teachers need to be equipped with necessary skills that are most needed to inculcate such knowledge into our youths through short in-service training. • funds should be made available at the right time to make our schools functional and efficient in impacting knowledge and skills on the students. • government should urgently put in place policies that would fully engage every citizen in one form of economic activity or the other that would enable them to earn a living. this will go a long way in ensuring national security. • there should be serious public awareness complain as to the importance of peaceful coexistence among the general public. • government also need to take proactive security measures for the security and safety of its citizens. • security is join responsibility of all citizens, therefore, everyone should join hands with the government to fight this security challenges. • security is a joint responsibility of all citizens. they should join hands with government to fight this security challenges. references [1] i. adabara, "vocational agricultural education production means for strengthening nigerian economy," oju journal of vocational and technical education, vol. 2, pp. 32 -37, 2013. [2] p. m. kolade, "agricultural education as panacea for national security," belt journal of education in nigeria, vol. 3, pp. 40 – 45, 2012. [3] d. j. ishaya, "the role of vocational and technical education in national development. the challenges of the new millennium," oju journal of vocational and technical education, vol. 2, pp. 165170, 2014. [4] a. d. suleiman, "role of vocational agriculture in equipping youths for self employment in nigeria," benue state university journal of education, vol. 7, pp. 53 – 60, 2013. [5] m. mohammed, foundation of vocational education, 5th ed. enugu: cape publishers, 2010. [6] a. u. jimoh, principles and methods in vocational and technical education. nsukka: university trust publishers, 2012. [7] m. b. alabi, "vocational and technical education in nigeria: challenges and prospects," nigerian vocational journal. university of nigeria, nsukka, vol. 5, pp. 35 -40, 2011. [8] k. i. audu, "achieving security and stability in nigeria through employment generation potentials in agricultural education," belt journal of education in nigeria, vol. 4, pp. 9 – 13, 2013. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research vol. 4, no. 1, 24-29, 2017 issn(e) 2411-6653/issn(p) 2518-0193 doi: 10.20448/journal.512.2017.41.24.29 24 assessment of the secondary metabolite patulin and lycium barbarum fruit on ins-1 rat pancreatic β-cells madeha n.al-seeni1 ahmed m.ali2 nagwa m. elsawi3 asma s.abdo4  ( corresponding author) 1biochemistry department, faculty of science, king abdel-aziz university, jaddah, saudi arabia. 2chemistry department, faculty of science, al –azhar university, assiut branch, egypt 3,4chemistry department, faculty of science, sohag university, sohag, egypt abstract patulin has been shown to have diabetogenic effects in mice. the effects on pancreatic β-cell viability and function of secondary metabolite patulin were investigated. using forty adult albino male rats which divided into 4 groups. control group was injected subcutaneously daily with distilled water for one week; group i was injected subcutaneously daily with patulin (0.2 mg/kg/day) for two weeks. group ii was injected with the same toxin and dose for two weeks after that they were treated by goji extract (2 ml/kg /day.) for two weeks. group iii was treated by goji for two weeks after that they were injected with patulin for two weeks. some biochemical parameters of blood samples for experimental rodents were evaluated. patulin group shows significant increase in amylase and glucose levels. but level of c-peptide and insulin decreased significantly in this group. however, there is no significant alteration in lipase level between patulin group and control. groups treated with goji extract in groups two and three (therapeutic and prophylactic) illustrated non significant alteration in biochemical parameters compared to control. keywords: patulin, pancreas, goji extract, insulin. citation | madeha n.al-seeni; ahmed m.ali; nagwa m. elsawi; asma s.abdo (2017). assessment of the secondary metabolite patulin and lycium barbarum fruit on ins-1 rat pancreatic βcells. agriculture and food sciences research, 4(1): 24-29. history: received: 4 april 2017 revised: 3 july 2017 accepted: 6 july 2017 published: 11 october 2017 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 25 2. materials and methods ................................................................................................................................................................... 25 3. results ................................................................................................................................................................................................ 26 4. discussion .......................................................................................................................................................................................... 27 references .............................................................................................................................................................................................. 28 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2017.41.24.29&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=madeha%20n.al-seeni https://orcid.org/orcid-search/quick-search?searchquery=ahmed%20m.ali https://orcid.org/orcid-search/quick-search?searchquery=nagwa%20m.%20elsawi https://orcid.org/orcid-search/quick-search?searchquery=asma%20s.abdo agriculture and food sciences research, 2017, 4(1): 24-29 25 1. introduction mycotoxins are reported to be important for infection and possibly growth under natural environmental conditions. a known example of mycotoxins is patulin [1]. patulin has been used as a model for exposure to secondary metabolites via contaminated food; it is produced by fungi, in particular penicillium and aspergillus, which may contaminate food products like apples and apple juice. the effects of patulin have been studied extensively, where it has been shown to be both genotoxic and neurotoxic through its covalent binding to essential sulphydryl groups in proteins and peptides. this toxicity of patulin has already been demonstrated against various mammalian cells and has been attributed to its ability to inhibit rna and protein synthesis. it is also interfere with the activity of many enzymes such as glycogen phosphorylase, glucose6-phosphatase, hexokinase and aldolase that are considered crucial for cell survival [2]. o oh o o fig-1. chemical structure of patulin(pat) [ref ] lycium barbarum fruit, generally called goji berry or wolfberry, is a well-known traditional chinese medicine. goji berry has shown a wide range of health benefits such as the functions and activities associated with the liver, kidney, eyesight, sex, immune system, and longevity [3]. also, it improves the insulin resistance of the cells and prevents damage from diabetes [4]. in addition, lycium barbarum may help in lowering glucose level in diabetic person. in vitro hypoglycaemic effect assay indicated that lbp may decrease high glucose level by protecting pancreatic β-cells from oxidative damages and alleviating the insulin resistance in liver cells. hypoglycaemic activity of lycium barbarum fruit may be correlative with polysaccharides, a mainly functional composition, considered as potential candidate for developing a new beneficial food or new anti-diabetic agent [5]. it has long been used to treat diabetes mellitus and related hyperlipidemia [6]. fig-2. lyciumbarbarum (goji berry) [7] in this study, the biochemical profile of fungal secondary metabolite, patulin, on the ins-1 rat β-cell was examined, in order to evaluate the possible role of goji extract in reversing patulin toxicity. 2. materials and methods 2.1. chemicals patulin (4-hydroxy-4h-furo[3, 2-c]pyran-2 (6h)one) was obtained from sigma chemical company. 2.2. extraction of goji wine goji berry was extracted according to method described by yang and baojun [8]where, 5 g of goji fruits, 25 ml alcohol and 75 ml distilled water were put into erlenmeyer flask (the ratio of goji berry and alcohol is referred to the ratio of making commercial goji wine). the erlenmeyer flask was sealed with parafilm and tinfoil and put in refrigerator for 5 days. filtration was made to separate the solid and liquid phases after finishing steeping process, solvent is removed from filtrate by rotatory evaporator to obtain crude goji extract. 2.3. animals and experimental design 2.3.1. animals forty male adult albino rats weighing 240-300 g at the age of 3.0-4.0 months were obtained from the animal house, faculty of medicine, assiut university. they were housed in standard environmental conditions (22± 3°c, and a 12 h light/ dark cycle as percpcsea guidelines) and fed with standard rodent diet. the rats were randomly assigned to four groups of 10 rats each (5 rats in each cage) and treated as follows: control group: injected subcutaneously daily with distilled water; group i: injected subcutaneously daily with patulin (0.2 mg/kg/day) for two weeks respectively. agriculture and food sciences research, 2017, 4(1): 24-29 26 group ii: injected by patulin as a previous dose for two weeks after that they were treated with goji extract (2 ml/kg/day) for another two weeks. group iii: treated by goji extract for two weeks then, they were injected subcutaneously with the same dose of the toxin also for another two weeks. 2.4. biochemical studies animals were anesthetized under light diethyl ether and blood samples were collected from the heart into plain tubes and stored at -20°c until analyses were performed. sera were used for the measurement of amylase, lipase, glucose, c-peptide and insulin. level of amylase was measured according to ranson [9] level of lipase was measured according to rick [10] level of glucose was measured according to trinder [11] level of c-peptide was measured by elisa kit that was purchased from immunospec company and level of insulin was measured by elisa kit that was purchased from demeditec. 2.5. statistical analysis the data were presented as mean ± sem and the level of significance between groups were *p<0.05, ** p<0.01, *** p<0.0001. they were analysed using one-way anova followed by newman-keuls test. statistical graph pad prism 5 was used for the analysis of the data. 3. results the biochemical changes were studied in blood of male albino rats as a result of administration of a dose from patulin (0.2 mg/kg b. w.) and observed over a period of two weeks (gi) compared to the control group. gii treated by patulin as previous dose for two weeks after that they were injected subcutaneously with goji extract 2 ml /kg /day for two weeks. in addition giii treated by goji for two weeks then, they were injected subcutaneously with the same toxin for two weeks. table. 1. and fig. 3. represent the mean value of amylase concentration (μkat/l) of g i treated animals for two weeks to be (40.46±0.4) which was highly significantly increased from normal value of control group (33.75 ± 0.32), where p<0.001. on the other hand, non-significant changes in the mean value of amylase concentrations were observed between gii (32.18 ± 0.4) and giii (30.76 ± 0.8) compared to control.the mean value of lipase concentration (μkat/l) table.1.and fig.4. of g1 (0.185 ± 0.007), g ii and g iii showed no significant change (p>0.05) compared with control group. table.1. and fig.5.represent the mean value of glucose concentration (mmol/l) of g i was significantly elevated to (10.82±1.33) (p<0.05)than normal control samples (7.604 ±0.383). this level was still slightly elevated to 8.31±0.77 in gii (p>0.05), and 9.77±0.64 in giii (p<0.05), but it is still within normal range with control group. the mean value of c-peptide concentration (ng/ml) in table.1. and fig.6.showed significant decrease (p > 0.05) compared with control. however, no significant changes in the mean value of c-peptide were observed between gii and giii compared to control. the mean value of insulin concentration (μlu/ml) in table.1and fig.7. showed significant decrease (p > 0.05) compared with control. however, this decrease returned again back to normal value in gii and giii respectively with non-significant change compared to control group. table-1. effect of patulin (0.2 mg/kg b. w.) and goji extract (2 ml/kg /day) on some biochemical parameters of male rats: groups control gi gii giii amylase(μkat/l) 33.75 ± 0.32 40.46 ± 0.4*** 32.18 ± 0.4 n.s 30.76 ± 0.8 n.s lipase(μkat/l) 0.217±0.04 0.189±0.03 n.s 0.207±0.01 n.s 0.241±0.007 n.s glucose(mmol/l) 7.604 ±0.383 10.82±1.33* 8.31±0.77 n.s 9.77±0.64 n.s c-peptide(ng/ml) 0.341±0.02 0.197 ± 0.01* 0.297 ± 0.01 n.s 0.338 ± 0.03 n.s insulin(μl u/m) 0.58±0.101 0.383±0.13* 1.35±0.10*** 1.54±0.13*** note: values are expressed as the mean ± se for 10 animals in each group. gi = treated group with patulin. gii , giii = treated groups with goji significant change in comparison between groups: *p<0.05; ***p<0.001 n.s non significant (p > 0.05) amylase contro l gi gii giii 0 10 20 30 40 50 groups co nc .  ka t/l fig-3. α-amylase levels in control and treated groups with patulin (2 mg/kg.b.w) and goji extract (2 ml/kg.b.w) agriculture and food sciences research, 2017, 4(1): 24-29 27 lipase contro l gi gii giii 0.0 0.1 0.2 0.3 groups co nc .  ka t/l fig-4. lipase levels in control and treated groups with patulin (2 mg/kg.b.w) and goji extract (2 ml/kg.b.w) glucose contro l g i g ii g iii 0 5 10 15 groups co nc .m m ol /l fig-5. glucose levels in control and treated groups with patulin (2 mg/kg.b.w) and goji extract (2 ml/kg.b.w) c-peptide contro l gi gii giii 0.0 0.1 0.2 0.3 0.4 0.5 groups ng /m l fig-6. c-peptide levels in control and treated groups with patulin (2 mg/kg.b.w) and goji extract (2 ml/kg.b.w) insulin contro l g i g ii g iii 0.0 0.5 1.0 1.5 2.0 groups l u/ m l fig-7. insulin levels in control and treated groups with patulin (2 mg/kg.b.w) and goji extract (2 ml/kg.b.w) 4. discussion the present study is a biochemical analysis on the effect of microbial product, patulin on ins-1 pancreatic beta cells (β-cells) and the possible role of goji extract as a therapy on pancreas dysfunction. the destruction of βcells and development of type 1 diabetes have been linked with exposure to environmental factors [12] which may contribute up to 50% of the risk of developing this type of diabetes [13]. the most important environmental factors are thought to be infection (viral infection), diet (lack of particular food during childhood) and some inorganic compounds (arsenate ions) [14, 15]. to study pancreatic function of rats treated with patulin, α-amylase, agriculture and food sciences research, 2017, 4(1): 24-29 28 lipase, glucose, c-peptide and insulin have been measured as biochemical indicators. alpha-amylase catalyzes the hydrolysis of α[1, 4]-d-glycosidic linkages of starch and other glucose polymers. inhibitors of this enzyme could be of use in the treatment of diabetes, resulting in inhibition of starch breaking and decreases levels of blood glucose [16]. because of the harmful side-effects of synthetic products and the fact that plants are easily accessible, interest in drugs of herbal origin has significantly increased. our present biochemical data in the patulintreated rats showed an obvious increasing (p<0.001) in pancreatic levels of amylase as it is shown in table. 1 and fig.3. these elevation levels attributed to acute pancreatitis caused by patulin [17]. also, many reports have dealt with the relationship between blood amylase and carbohydrate metabolism [18-20]or hormones that affect carbohydrate metabolism [20-22].these studies showed that high levels of amylase result from administration of hormones that decreases utilization of carbohydrate or in physiological states of diminished carbohydrate utilization. another parameter is lipase, which is triacylglycerol acylhydrolase and an omnipresent enzyme with physiological significance and industrial potential. it catalyzes the hydrolysis of triglycerides to glycerol and free fatty acids. it is soluble in water and hydrolyzes insoluble substrates to more polar lipolytic products. lipase is released from pancreatic acinar cells and reabsorbed by the kidney tubules so, it remains elevated for prolonged period which may be helpful in late presenting cases [23]. in this study, there is non-significant (p>0.05) change between patulin treated group and control as it is shown from table. 1. and fig.4. where, many studies have reported a negative predictive value of serum lipase in diagnosing acute pancreatitis to be between 94 -100 % even if amylase increases significantly [24, 25]. the digestive tract breaks down carbohydrates such as, starch and sugars found in many foods to glucose, a form of sugar that enters the bloodstream. with the help of the hormone insulin, cells found in the body absorb glucose and use it for energy. when the body doesn't make enough insulin (pancreas dysfunction) leading to accumulation of sugar in the blood and diabetes. in our study level of glucose in the blood was measured after administration of patulin for two weeks (g i) and it shows highly significant increase (p<0.05) in the mean value in table 1 and fig 5. this leads to damaging nerves and blood vessels and thus complications such as heart disease, stroke, kidney disease, blindness, dental disease, and amputations. other complications of diabetes may increase susceptibility to other diseases and loss of mobility with aging [26]. in addition, c-peptide and insulin levels in serum were measured. c-peptide is produced in equal amounts to insulin and can therefore be used to assess endogenous insulin secretion in type-1-diabetes. the physiology of cpeptide makes it appropriate for assessing insulin secretion. insulin is produced in the pancreatic b-cells by enzymatic cleavage of the prohormone precursor proinsulin to produce insulin and c-peptide in equimolar amounts. its half-life is longer than insulin (20–30 vs. 3–5 min) and it therefore circulates at concentrations approximately five times higher in the systemic circulation [27, 28]. from table.1 and fig.6 it is cleared that mean value of c-peptide in rats treated with patulin shows significant decrease (p > 0.05) compared with control. this scarce value confirms previous results obtained from amylase and glucose that is patulin induced type 1 diabetes in male rats after subcutaneous injection for 2 weeks. because dropped levels of c-peptide means significant destruction of β-cells and only 10 % of normal cell mass remains [29]. insulin is known to facilitate the absorption of glucose through the specialized membrane of the insulin sensitive cells which invariably results in an increase in blood glucose level due to delayed glucose uptake [30]. data from table.1 and fig.7. show that there is significant decrease in the mean value (p > 0.05) compared with control. insulin and glucagon have distinct actions in diabetes where, they show that the impairment of pancreatic exocrine functions is due to insulin deficiency and relative glucagon predominance [31]. this reducing of insulin caused by patulin in g i affect on pancreatic function resulting in changes in the composition and quality of pancreatic juice as well as its decreased production leading to accumulation of glucose in the blood and this agreed with the increasing in serum glucose showed in fig.5. [32]. from table 1 we observe that levels of amylase, glucose and c-peptide return to normal values of control in group ii and group iii (treatment with goji extract after and before patulin respectively) and also, observe that insulin levels show highly significant increase compared to control. the most effective methods for treatment of diabetes are inhibition of digestion and absorption of carbohydrates in small intestine. that is happening in two manners, the first is inhibition digestion of carbohydrates with inhibition of alpha amylase and the second is inhibition of absorption via sodium/glucose co-transporters [16, 33]. goji berries contain many nutrients with high biological activity, such as polysaccharides, carotenoids, phenylpropanoids and others. the polysaccharide complex is the most important and the most abundant group of compounds present in goji fruits where it comprises 5-8 % of dried matter of the fruits. luo, et al. 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https://scholar.google.com/scholar?hl=en&q=effects%20of%20the%20microbial%20secondary%20metabolites%20pyrrolnitrin,%20phenazine%20and%20patulin%20on%20ins-1%20rat%20pancreatic%20β-cells https://scholar.google.com/scholar?hl=en&q=effects%20of%20the%20microbial%20secondary%20metabolites%20pyrrolnitrin,%20phenazine%20and%20patulin%20on%20ins-1%20rat%20pancreatic%20β-cells http://dx.doi.org/10.1111/j.1574-695x.2011.00844.x https://scholar.google.com/scholar?hl=en&q=a%20review%20of%20the%20anticancer%20and%20immunomodulatory%20effects%20of%20lycium%20barbarum%20fruit https://scholar.google.com/scholar?hl=en&q=a%20review%20of%20the%20anticancer%20and%20immunomodulatory%20effects%20of%20lycium%20barbarum%20fruit https://scholar.google.com/scholar?hl=en&q=an%20evidence-based%20update%20on%20the%20pharmacological%20activities%20and%20possible%20molecular%20targets%20of%20lycium%20barbarum%20polysaccharides http://dx.doi.org/10.2147/dddt.s72892 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https://scholar.google.com/scholar?hl=en&q=hypoglycemic%20and%20hypolipidemic%20effects%20and%20antioxidant%20activity%20of%20fruit%20extracts%20from%20lycium%20barbarum http://dx.doi.org/10.1016/j.lfs.2004.04.056 https://scholar.google.com/scholar?hl=en&q=protective%20effect%20of%20lycium%20barbarum%20polysaccharides%20on%20streptozotocin-induced%20oxidative%20stress%20in%20rats http://dx.doi.org/10.1016/j.ijbiomac.2006.11.002 68 agriculture and food sciences research vol. 5, no. 2, 68-72, 2018 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2018.52.68.72 forecasting waste agricultural plastics generation in the republic of korea and its policy implications youngjae chang1 geumsoo kim2  ( corresponding author) 1professor, division of management science, inje university, south korea 2professor, department of global commerce, hoseo university, south korea abstract the generation of waste plastics mulch is predicted to decline after 2019 and then stabilize at around 200,000 tons per year. this is within the current gross treatment capacity of private and public facilities. however, the korean waste agricultural plastics (wap) management system is vulnerable to market uncertainty, which might lead to uncontrollable accumulation of untreated waste plastics as experienced in 2005. shifting from the keco (korea environment corporation)led system to a more flexible market-oriented system is recommended. keywords: waste agricultural plastics, recycling, waste management. citation | youngjae chang; geumsoo kim (2018). forecasting waste agricultural plastics generation in the republic of korea and its policy implications. agriculture and food sciences research, 5(2): 68-72. history: received: 7 june 2018 revised: 13 august 2018 accepted: 18 september 2018 published: 15 october 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 69 2. korean wap management system .............................................................................................................................................. 69 3. forecasting wap generation ........................................................................................................................................................ 70 4. conclusion ......................................................................................................................................................................................... 71 references .............................................................................................................................................................................................. 72 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.52.68.72&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.52.68.72&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/1507 https://orcid.org/0000-0003-2132-2570 https://orcid.org/0000-0002-1207-6083 agriculture and food sciences research, 2018, 5(2): 68-72 69 1. introduction modern farming takes advantage of a wide range of plastics and large amounts of wap are inevitably generated. vinyl-house horticulture and insulated rice seedbed preparation are practiced nationwide and, in 2015, approximately 320,000 tons of wap was generated. this figure has remained almost the same over the last decade. the collection rate has stagnated at just under 60%. therefore, collection and proper treatment of wap, including recycling, is a serious social concern in korea. in korea, wap are classified as municipal solid waste, the final treatment of which is the responsibility of the basic administrative districts (i.e., eup, myon, dong)1. therefore, in principle, these districts have to collect and recycle or conduct final disposal (landfilling and incineration) of wap. however, farmers do not have sufficient incentives to take waste plastics to private collectors or public collection points voluntarily and the basic administrative districts do not have the resources to deal with the waste fully. waste vinyl-house plastic coverings are relatively clean and thus well-collected and almost recycled through market. however, the most prevalent type of waste plastics, mulching low density polyethylene(ldpe) and high density polyethylene(hdpe), which accounted for 76% of the total 322,964 tons of waste generated in 2015, would create a serious problem if left intact in fields. uncollected and improperly treated waste plastics are detrimental to the environment and also undermine agricultural land productivity. for this reason, the keco has established collection and treatment networks and subsidizes farmers and treatment firms in cooperation with local governments. in 2010, on average, 100 won per kg of collected waste came from subsidies of which 14 won was provided by the provincial government, 76 won by the city governments, and the remaining 10 won by the central government. however, this subsidization scheme has not been strong or flexible enough, resulting in insufficient and stagnated collection rates. as urbanization has advanced, now in farms only the old people are left. this trend is making it more expensive to bring waste plastics to collection points, but the level of subsidies has remained almost the same for a long time. on the treatment side, there has sometimes been overcapacity and insufficient capacity at other times. this disequilibrium in market may be resolved by price adjustment. however, this takes time and there are highly likely to be negative impacts such as a backlog of untreated waste plastics. to ease this fluctuation in demand and supply keco operates treatment companies and subsidized private treatment companies. the focus of this paper is not on the design of this system but only on the provision of materials to aid discussion by estimating and predicting wap generation and collection. the paper is organized as follows. in the next section, we provide an overview of the korean wap management system, including time trends of wap generation and collection, wap-related materials flow, and so on. in section 3, we forecast wap generation and collection for the next 5 years and compare this with the current treatment capacity. we then discuss the policy implications of our empirical results. finally, section 4 contains some concluding remarks. 2. korean wap management system 2.1. legal position of wap in the korean waste management system wap is municipal solid waste and the collection and final treatment of it is the responsibility of the basic administrative districts in korea. therefore, farmers here do not have as strong an incentive for collecting and properly treating as farmers in japan and taiwan, who are responsible for the proper final treatment and thus almost all wap is collected and treated via proper channels [1, 2]. 2.2. materials flow in wap management and keco as shown table 1 the total amount of generated wap has remained around 320,000 tons per year for the last decade. mulching ldpe is the most prevalent type of waste (approximately 40%), followed by hdpe (35%) and vinyl-house ldpe (20%). table-1. wap generation, collection, and treatment in korea unit: tons year generation collection collection rate (%) treatment untreated inventory 2009 310,009 189,238 61.04 236,259 72,973 2010 324,101 176,849 54.57 208,377 41,444 2011 331,490 181,609 54.79 185,424 37,628 2012 337,877 178,130 52.72 180,950 34,809 2013 332,575 189,306 56.92 174,181 49,934 2014 329,239 188,279 57.19 193,065 45,168 2015 322,964 186,965 57.89 211,143 21,141 source: keco internal data farmers are advised to bring any generated wap to community collection points. at this stage local governments subsidize farmers to increase their collection rate, combining the subsidies from the keco (or central government) and the provincial government. designated private collectors then bring the collected wap to keco-operated storage facilities. (see figure 1). the collection rate has not exceeded 60% for the last decade. what has contributed the most to the low collection rate was house ldpe (see table 2), but this one was almost recycled voluntarily through market. a total of 72,000 tons of mulching ldpe, hdpe, pvc and other waste plastics went uncollected, which is a real problem. 1this does not mean that farmers are legally allowed to leave wap in fields. they are legally prohibited from disposing waste improperly. agriculture and food sciences research, 2018, 5(2): 68-72 70 table-2. collection rate by type (2015) unit: tons per year, % generation(a) collection(b) collection rate((b/a)*100) house ldpe 69,414 4,964 7.15 mulching ldpe 128,241 88,275 68.84 hdpe 118,715 92,064 77.55 pvc and others 6,594 4,914 74.52 total 322,964 186,965 57.89 source: keco internal data keco has two options for the treatment of collected wap: keco itself treats or gets it done by private firms. as of 2016, keco operates seven treatment facilities with a treatment capacity of 80,000 tons per year. in 2015, just over 100 thousands tons should be left for private firms. six of treatment firms have the capacity to process 6,000-8,000 tons each per year. keco auctions off some part of the remaining wap to private firms and for the other part of the remaining wap, keco subsidizes private firms to treat. the treatment subsidy was introduced in 2005 when the volume of untreated wap became a serious social concern. the subsidy was at its highest level in 2008 but has been declining since then. figure-1. flow diagram of wap generation, collection, and treatment source: keco internal data 3. forecasting wap generation 3.1. methodology forecasting is a two-state process. first, we attempt to identify the determinants of wap generation (mpe) through a regression analysis. specifically, wap generation is regressed on potential determinants such as paddy size (paddy), field size (field) and vinyl-house farming (house) as follows : mpe = β0 + β1paddy + β2field + β3house + ε (1) based on this estimation, we then forecast the future mpe. to do this, we need to estimate the future values of explanatory variables: we regress each of the explanatory variables on the respective previous values and time. we use the estimated value of one coefficient as its future value if it is statistically significant. if there is no statistical significance, we will average the previous 5 years' values to use as future values for each variable over 5 years. 3.2. data we use panel data covering nine regions across 12 years (2004 through 2015)2. as our dependent variable, we use only waste mulching plastics generation, which accounts for approximately 80% of the total, because the remainder, vinyl house coverings, is almost recycled voluntarily through the market. table 3 provides summary statistics for our data. table-3. summary statistics mpe paddy field house mean 26,289.56 106,379.23 4,439.76 80,904.06 stand. dev. 12,146.60 61,947.29 4,729.77 27,252.56 min. 1,319.00 18.00 0.00 52,100.00 max. 59,634.00 219,337.00 18,287.00 152,095.00 note: mpe, paddy, field, and house stands for mulching plastics generation, size of paddy field, size of dry field, and size of vinyl house, respectively. mpe is measured in tons and the other variables are measured in hactares. source: keco internal data and the korea statistical information service(kosis.kr). 2 as of 2018, there are 16 so-called wide autonomous administrative units in korea. however, due to the availability of data, we have merged these units into nine regions. for example, seoul and its surrounding kyunggi province were merged into an area referred to as the captial area. agriculture and food sciences research, 2018, 5(2): 68-72 71 3.3. empirical results we first checked the poolability of the time-series and cross-sectional data. based on an f-statistic (0.42652) we could reject the null hypothesis that the coefficients are not stable at the 5% level. we then checked for autocorrelation in error terms using the breusch-godfrey/ wooldridge test, which has detected the presence of autocorrelation. we have added the lag term for dependent variable to set our final regression model as follows: mpe = β0 + β1paddy + β2field + β3house + β4lag(mpe) + ε (2) we have found no autocorrelation in error terms after testing this new model using the breusch-godfrey/ wooldridge test (chi-square statistic=2.1548). in addition, the honda test showed no individual effect (standard normal statistic=0.024989), but a time effect (standard normal statistic=4.6167). finally by using the hausman test we selected a random effects model (chi-square statistic=5.6698) [3, 4]. table 4 presents the results of regression analysis. house (size of vinyl house farming) and lag(mpe) are significant at the 5% and 1% level, respectively. the effect of lag(mpe) is statistically significant and is strong. almost 67 percent of wap generated in the last year is again generated any of this year in inertia. notice that the effect of field (size of dry field) is almost the same as that of house (size of vinyl house farming), but it is not statistically significant. table-4. results of regression analysis coefficient standard error t-value p-value intercept 574.9600 1901.1000 0.3024 0.7630 paddy 0.0189 0.0116 1.6276 0.1070 field 0.0893 0.1403 0.6361 0.5263 house 0.0737 0.0302 2.4409 0.0165 * lag(mpe) 0.6715 0.0749 8.9676 0.0000*** note: *,**,and *** represent the 10%, 5%, and 1% significance level, respectively. we now turn to predicting the future values of explanatory variables. we then insert them into equation (2) to obtain the future values of mpe. table 5 shows a summary of the future values of explanatory variables obtained using the aforementioned method. table-5. future values of explanatory variables year paddy field house 2016 998,953.6 273,912.4 989,065.2 2017 992,768.4 274,119.1 973,989.8 2018 985,095.7 274,911.6 984,324.6 2019 940,259.2 258,131.9 992,162.6 2020 914,167.4 234,499.8 968,638.2 the 95% confidence interval for the predicted values of mpe is summarized in table 6. table-6. 95% confidence interval for the prediction of mpe year lower bound point estimate upper bound 2016 221,466.6 247,363.4 273,260.2 2017 224,616.4 247,827.1 271,037.8 2018 226,404.7 248,955.6 271,506.6 2019 190,964.2 232,544.9 274,125.6 2020 174,274.4 209,293.8 244,313.2 as shown in table 6, the generation of mpe is predicted to decline after 2019. it is also predicted to stabilize at around 200,000 tons per year after that. although there is a little increase in the mpe collection rate, for example 80%, there would be no repeat of the huge accumulation of wap that occurred on 2005 because the present treatment capacity (approximately, private 50 thousand and public 80 thousand annually) would suffice. however, due to the uncertainty in the recycled products market (for example, china’s recent suspension of recycled vinyl imports and fluctuations in petroleum markets) private wap recycling firms may go bankrupt, causing a supply shortage for treatment services, which occurred in 2005. 4. conclusion the amount of wap collected by keco is predicted to stabilize at around 160,000 tons per year. given the current treatment capacity, this would not lead to issues if the amount of untreated wap significantly increased, as it did in 2005. however, this keco-led system is vulnerable to various market uncertainties regarding recycled products and petroleum markets. unlike korea, japan and taiwan define wap as industrial waste. therefore, farmers in these countries are responsible for the proper treatment of wap. in both countries the central government does not play such active a role as it does in korea. collection and treatment occur in response to market mechanisms [5-7]. in light of the situation in these countries, korea also needs to strengthen the legal responsibility of producers and consumers (farmers) of agricultural plastics to ensure proper wap management. specifically, we can consider the alternatives that by putting agricultural plastics into epr(extended producer’s responsibility) items, producers are made agriculture and food sciences research, 2018, 5(2): 68-72 72 responsible for collecting and treating wap or that by reclassifying wap into industrial waste farmers are made responsible for proper treatment [8]. we do not exclude the role of governments in wap management. governments may adopt the role of facilitating the smooth operation of markets. taiwanese government compensates farmers for sorting and bringing waste plastic containers to treatment firms, but not for general wap. japanese government also compensates farmers for bringing waste plastics to collection points. in addition, it also operates a minimum level of public treatment facilities as a buffer for market uncertainty [6, 7]. references [1] e. hosoda, waste policies and related legislation in japan. ch.1 in the economics of waste management in east asiaed. by m. yamamoto and e. hosoda. ny, usa: routledge, 2016. [2] s. c. lee, an introduction to taiwan's waste management and recycling policy. ch.6 in the economics of waste management in east asiaed. by m. yamamoto and e. hosoda. ny, usa: routledge, 2016. [3] b. h. baltagi, econometric analysis of panel data, 2nd ed. uk: john wiley & sons, 2001. [4] w. h. greene, econometric analysis. ny, usa: macmillan, 1990. [5] h. takeya, "collection, disposal and exchange of waste agricultural plastics in japan, the republic of korea and china," tech monitor, pp. 50-55, 2009. [6] nier(national institute of environmental research), "japanese resource circulation institutions and status of 3r reference book. nier, rok, 2015.(in korean)," 2015. [7] tepa (taiwanese environmental protection administration), retrieved: www.epa.gov.tw, 2017. [8] r. porter, the economics of waste. washington dc, usa: resources for the future, 2002. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.epa.gov.tw,/ 89 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 89-97, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.89.97 © 2019 by the authors; licensee asian online journal publishing group sensory evaluation of snacks produced from wheat-breadfruit flour and nutritional composition of the flour blends folorunso, adekunle a.1 habeeb, aisha s. 2 ajayi, odunola t.3 ( corresponding author) 1,3department of family, nutrition and consumer sciences, faculty of agriculture, obafemi awolowo university, ile-ife, nigeria. 2department of hospitality management, federal polytechnic, bida, niger state, nigeria. abstract this study investigated sensory nutritional evaluation of snacks produced from wheatbreadfruit flour blends. two different snacks, meat pie and chinchin were produced using three different mixing proportions (50:50,30:70 & 70:30) of wheat-breadfruit flour blends for each making a total of six snack samples respectively. fresh breadfruits, wheat flour and other ingredients were obtained from in ile-ife, osun state. the samples were presented to a 50 member panelists to rate them on a 9-point hedonic scale and proximate analysis was also conducted on the flour blends using standard methods to determine their nutritional composition. analysis of variance (anova) was performed on the data gathered. the result showed that the highest value of moisture content, ash content and crude fiber content were observed in flour sample a (50:50), highest value of protein content, and fat content were observed in flour sample b (30:70) and highest value of carbohydrate content was observed in flour sample c (70:30). results revealed that there are significant difference at (p<0.05) in the protein, fat, crude fiber, and carbohydrate content of the blends of flour. there are significant difference at (p<0.05) in the results for sensory evaluation of chin chin and meat pie samples. the highest values for chinchin samples are 7.60, 7.74, 7.76, 7.62, and 7.84 for colour, flavor, taste, texture and overall acceptance respectively. and for the meat pie samples, the highest values are 7.56, 7.42, 7.52, 7.54, and 7.68 for colour, flavor, taste, texture and overall acceptance respectively. it was concluded that snack samples produced from 30% breadfruit flour with 70% wheat flour blend are the most preferable by the panelists. keywords: nutritional composition, post-harvest loss, imported wheat, locally crops, sensory analysis, breadfruit. citation | folorunso, adekunle a.; habeeb, aisha s.; ajayi, odunola t. (2019). sensory evaluation of snacks produced from wheat-breadfruit flour and nutritional composition of the flour blends. agriculture and food sciences research, 6(1): 89-97. history: received: 19 february 2019 revised: 25 march 2019 accepted: 29 april 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ....................................................................................................................................................................................... 90 2. materials and methods .................................................................................................................................................................... 90 3. results and discussion .................................................................................................................................................................... 95 4. conclusions ........................................................................................................................................................................................ 96 5. recommendations ............................................................................................................................................................................ 96 references .............................................................................................................................................................................................. 97 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.89.97&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.89.97&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/124 https://orcid.org/0000-0002-9890-765x agriculture and food sciences research, 2019, 6(1): 89-97 90 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this contributes to existing literature by examining the diversification of breadfruit, a locally grown crop in terms of nutritional composition and consumer acceptability using wheat-breadfruit flour blends in snacks production. 1. introduction snacks are important foods which come in a variety of forms including packaged snack foods and other processed food as well as items made from fresh ingredients at home, the consumption of which is steady and increasing in nigeria. it is a portion of food, smaller than a regular meal, generally eaten between meals. it has been hitherto produced from wheat as a major raw material. in nigeria, wheat production is limited and wheat flour is imported to meet local flour needs for bakery products as such huge amount of hard earned foreign exchange is used every year for importation of wheat [1]. efforts have been made to promote the use of composite flours in which flour from locally grown crops and high protein seeds replace a portion of wheat flour for use in snacks, thereby decreasing the demand for imported wheat and producing protein-enriched bread [2]. breadfruit (artocarpus altilis) is a member of the moraceae family and is normally consumed after cooking. it’s a rich source of carbohydrates, fiber, vitamin (vitamin c) and mineral (potassium). this plant plays a main function to ensure food safety in the rural communities where they are predominant due to its high carbohydrate content (76.71 to 79.24 %). it has been used as an important source of energy for years. in africa, breadfruits are found in some country (senegal, guinea-bissau, cameroun, sierra leone, nigeria, liberia and ghana). however, the current usage, particularly, in developing countries, is limited by the poor fresh fruit storage properties. a few days after harvesting (3 days), the deterioration of the fruit settles and because of their high water content they are easily susceptible to microbial attack as well as their bulky nature makes their transportation difficult. it is therefore reasonable to maximize the potential of this precious nutritious fruit by transforming them into various finished products easily storable. this production of flour from breadfruit is a useful technique to extend its storage and consumption. different conventional flours are important and have precious functional roles in global food systems. however, their increasing price led to the search of other flours less expensive unconventional as an alternative for replacement partially or completely the traditional conventional flours in foods as a means against malnutrition [3]. many developing nations spend huge amount of their foreign exchange for the importation of food especially wheat, rice and sugar. for instance, in 2011 africa spent more than $ 50 billion on food imports. nigeria spends $ 4.2 billion yearly for the importation of wheat. according to amusa, et al. [2] in 2010 alone, nigeria spent n 635 billion ($ 4.2 billion) on the importation of wheat, it has been reported that wheat importation is growing at the rate of 13% per annum. it has been estimated that at this growth rate, nigeria wheat importation could reach 17 million metric tons (mmt) by 2020, which is equivalent to the entire wheat production by canada (the third largest wheat producing country in the world [2]. the use of composite flours has been reported by many authors. oloye [4] investigated the use of supplementation of flours of soybean and plantain in wheat in the production of bread. also, mepba, et al. [3] produced composite breads and biscuits from mixed flours of wheat and plantain, with 30% supplementation of plantain flour. though breadfruit has been made into flour and evaluated in bakery products not much has been done in the area of composite flour blends of breadfruit and beniseed. therefore, it becomes pertinent to evaluate the composite flours aiming at reducing wheat flour inflation and curtail excessive losses of breadfruit during season. breadfruit is a valuable food resource, but its usage is limited by poor storage properties of the fresh fruit. therefore, there is need for conversion into flour to provide a more stable storage form as well as increasing its versatility [5]. hence, this study was aimed to determine the nutritional composition of wheat-breadfruit flour blends and assess the sensory qualities of snacks produced from wheat-breadfruit flour .the specific objectives of this study include: i. to assess the nutritional properties of flour produced from wheat breadfruit flour blends ii. produce the flour blends of different proportions. iii. assess the sensory evaluation of products produced from wheat-breadfruit flour blends the study will be of utmost importance to the catering establishments as the study will enlighten them on the benefit of making use of blend of wheat-breadfruit flour in snacks production. also, it will help the government to minimize importation activities by reducing the quantity of wheat flour importation as the study ascertain the importance of blending wheat flour with breadfruit flour for snacks production. 2. materials and methods 2.1. collection of materials fresh breadfruit fruits were obtained at gbongan, osun state, while wheat flour and other ingredients were obtained from lagere market in ile-ife, osun state. the production of the snacks took place at food nutrition and dietetics laboratory in the department of family, nutrition and consumer science, faculty of agriculture, obafemi awolowo university. tools and equipment include frying pan, chopping board, rolling pin, bowl, colander, draining spoon and knife. 2.2. production of breadfruit into flour the process for making the breadfruit flower is as follow; the breadfruit was washed and peeled. then, it was halved and the heart was removed. the halves was then cut into very thin slices, then the slices were laid on drying rack in the sun to dry. breadfruit must be brittle when properly dried. the dried breadfruit was agriculture and food sciences research, 2019, 6(1): 89-97 91 © 2019 by the authors; licensee asian online journal publishing group grinded using attrition mill and was sieved several times until a powdery substance was formed. the breadfruit flour was then stored in clean air tight jars. figure-1. production of breadfruit flour. source: oloye [4]. 2.3. mixing proportions sample 1 (chin-chin): a: 50% breadfruit flour with 50% wheat flour, b: 30% breadfruit flour with 70% wheat flour, c: 70% breadfruit flour with 30% wheat flour sample 2 (meat pie): a: 50% breadfruit flour with 50% wheat flour, b: 30% breadfruit flour with 70% wheat flour, c: 70% breadfruit flour with 30% wheat flour 2.4. nutritional determination of the blends nutritional determination of the flour blends were carried out using methods of association of analytical chemists [6]. moisture content one gram of sample in preweighed crucible was placed in an oven (1050c) for 24 hours, cooled, and reweighed. the percentage moisture was calculated as follows: moisture (%) = 𝑊2− 𝑊3 𝑊2− 𝑊1 𝑋 100 where w1 is the weight of the crucible, w2 is the weight of the crucible after drying at 105oc and sample, and w3 is the weight of the crucible and the sample after cooling in airtight desiccators. crude protein crude protein content was determined using the microkjeldahl methods as prescribed by pearson [7]. a volume of 10ml h2s04 added to 3g of sample was digested with a kjeldahl digestor (model bauch 4390) for one and half hour. a volume of 40ml water was added and distilled using a kjeldahl distillation unit (model unit b-316) containing 40% concentrated sodium hydroxide and millipore water. liberated ammonia was collected in 20ml boric acid with bromocresol green and methyl red indicators and titrated against 0.04n h2s04. a blank (without sample) was likewise prepared. percent protein was calculated as: crude protein (%) = 𝑆𝑎𝑚𝑝𝑙𝑒 𝑡𝑖𝑡𝑒𝑟−𝑏𝑙𝑎𝑛𝑘 𝑇𝑖𝑡𝑒𝑟 𝑥 14 𝑥 6.25 𝑆𝑎𝑚𝑝𝑙𝑒 𝑊𝑒𝑖𝑔ℎ𝑡 𝑋 100 where 14 is the molecular weight of nitrogen and 6.25 is the nitrogen factor where 14 is the molecular weight of nitrogen and 6.25 is the nitrogen factor. crude fiber a weighed crucible containing 1g of defatted sample was attached to the extraction unit (in kjeldahl, d 40599, behr labor-technik gmbh, düsseldorf, germany) and into this 150ml of hot 1.25% h2s04 was added and digested for 30minutes,the acid was drained and sample washed with hot distilled water for 1/2 hour. the crucible was removed and oven dried overnight at 1050c, cooled, weighed and incinerated at 550oc in a muffle furnace ( mf1-02;pcsir labs, lahore, pakistan) overnight and reweighed after cooling. percentage extracted fiber was calculated as: crude fiber (%) = 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝐷𝑖𝑔𝑒𝑠𝑡𝑒𝑑 𝑆𝑎𝑚𝑝𝑙𝑒 − 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑎𝑠ℎ𝑒𝑑 𝑠𝑎𝑚𝑝𝑙𝑒 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑆𝑎𝑚𝑝𝑙𝑒 𝑋 100 agriculture and food sciences research, 2019, 6(1): 89-97 92 © 2019 by the authors; licensee asian online journal publishing group lipid lipid content was estimated using tecatorsoxtec (model 2043[20430001];hilleroed, denmark). a quantity of 1.5g sample was mixed with 2.3g anhydrous sulfate was weighed into a thimble and covered with absorbent cotton, while 40ml of petroleum ether (40-60c bpt) was added to a pre-weighed cup. both thimble and cup were attached to the extraction unit. the sample was extracted using ethanol for 30min and rinsed for one and half hour. thereafter, the solvent evaporated from the cup to the condensing column. extracted fat in the cup was then placed in an oven at 105oc for 1hour and cooled and weighed. percent fat was calculated as: lipid (%) = 𝐼𝑛𝑖𝑡𝑖𝑎𝑙 𝑐𝑢𝑝 𝑤𝑒𝑖𝑔ℎ𝑡−𝐹𝑖𝑛𝑎𝑞𝑙 𝑐𝑢𝑝 𝑤𝑒𝑖𝑔ℎ𝑡 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑆𝑎𝑚𝑝𝑙𝑒 𝑥 100 ash ash and mineral contents were determined according to aoac (association of analytical chemists) numbers 923.03 and 984.27 [6]. two grams of sample was added into a pre-weighed crucible was incinerated in muffle furnace at 600oc ash (%) = 𝑊2− 𝑊3 𝑊2− 𝑊1 𝑥 100 where w1 is the weight of cleaned, dried, ignited, and cooled cruciblew2 the weight of the crucible and sample after incinerating at 600oc, and w3 the weight of the crucible and sample after cooling in an airtight homogenized vessel. carbohydrate the carbohydrate content was determined by difference, that is, addition of all the percentages of moisture, fat, crude protein, ash, and crude fiber was subtracted from 100%. this gave the amount of nitrogenfree extract otherwise known as carbohydrate. % carbohydrate = 100(% moisture + %fat + %ash + %crude fiber +%crude protein) energy value the sample energy value was estimated (in kcal/g) by multiplying the percentage of crude protein, crude lipid, and carbohydrate with the recommended factors (2.44, 8.3, and 3.57, respectively) as proposed by martin and coolidge [8]. 2.5. production of snacks two snacks were produced with the blends of wheat-breadfruit flour at different proportions making a total of six snack samples: 1. production of chin-chin with 50% breadfruit flour and 50% wheat flour ingredients (1) 100g of breadfruit flour and 100g of wheat flour (2) 20ml evaporated milk (peak milk) / 20g powdered milk (3) 100g granulated sugar (4) 100g margarine (5) 1 medium nutmegs | 1/2 teaspoons ground nutmeg (6) 200ml vegetable oil (tasteless and odourless) the milk and sugar was mixed in a bowl and left to soak. in a separate bowl, the ground nutmeg was mixed with 200g of composite flour, then the margarine was rubbed into the flour till well mixed with the flour, leaving no lumps. the sugar/milk mixture was then added to the margarine/flour mixture and was mixed by hand till smooth dough was achieved. the dough was then placed on the chopping board and was kneaded gently with a dough roller. then the dough was cut into desire size flat. then, the dough was deep fried in hot vegetable oil till when a golden brown colour was obtained. the fried pieces were set onto a flat tray to dry and cool down quickly. when the chinchin had cooled down completely, it was stored in a dry, airtight container. 2. production of chin -chin with 30% breadfruit flour and 70% wheat flour ingredients (1) 60g of breadfruit flour and 140g of wheat flour (2) 20ml evaporated milk (peak milk) / 20g powdered milk (3} 100g granulated sugar (4) 100g margarine (5) 1 medium nutmegs | 1/2 teaspoons ground nutmeg (6) 200ml vegetable oil (tasteless and odorless) the milk and sugar were mixed in a bowl and left to soak. in a separate bowl, the ground nutmeg was mixed with 60g of breadfruit flour and 140g of wheat flour, then the margarine was rubbed into the flour till well mixed with the flour, leaving no lumps. the sugar/milk mixture was added to the margarine/flour mixture and was mixed by hand till smooth dough was achieved. the dough was then placed on the chopping board and was kneaded gently with a dough roller. then the dough was cut into desire size flat. then, the dough was deep fried in hot vegetable oil till when a golden brown colour was obtained. the fried pieces were set onto a flat tray to dry and cool down quickly. when the chinchin had cooled down completely, it was stored in a dry, airtight container. 3. production of chinchins with 70% breadfruit flour and 30% wheat flour ingredients (1) 140g of breadfruit flour and 60g of wheat flour (2) 20ml evaporated milk (peak milk) / 20g powdered milk (3) 100g granulated sugar (4) 100g margarine (5) 1 medium nutmegs | 1/2 teaspoons ground nutmeg http://www.allnigerianrecipes.com/images/peak-milk.jpg http://www.allnigerianrecipes.com/images/peak-milk.jpg http://www.allnigerianrecipes.com/images/peak-milk.jpg agriculture and food sciences research, 2019, 6(1): 89-97 93 © 2019 by the authors; licensee asian online journal publishing group (6) 200ml vegetable oil (tasteless and odourless) the milk and sugar was mixed in a bowl and left to soak. in a separate bowl, the ground nutmeg was mixed with 140g of breadfruit flour and 60g of wheat flour, then the margarine was rubbed into the flour till well mixed with the flour, leaving no lumps. the sugar/milk mixture was then added to the margarine/flour mixture and was mixed by hand till smooth dough was achieved. the dough was then placed on the chopping board and was kneaded gently with a dough roller. then the dough was cut into desire size flat. then, the dough was deep fried in hot vegetable oil till when a golden brown colour was obtained. the fried pieces were set onto a flat tray to dry and cool down quickly. when the chin chin had cooled down completely, it was stored in a dry, airtight container. 4. production of meatpie with 50% breadfruit flour and 50% wheat flour ingredients (1) 100g of breadfruit flour and 100g of wheat flour (1) irish potatoes (2) 200g minced beef (3) 1 medium onion (4) 1 teaspoon baking powder (5) carrot (6) 2 medium eggs (7) 100g margarine 100g of breadfruit flour and 100g of wheat flour was sifted into a bowl, then baking powder was added and mixed with the flour. the margarine was then added and mixed with the dough until a crumb like texture is formed. then small amount of water was added. the dough was then kneaded thoroughly. onion and irish potatoes was chopped and was fried together with the minced beef in vegetable oil and appropriate seasoning was added. the rolling pin was used to flatten the dough and was cut into required sizes. then the fillings were filled inside the pie and the edges of the dough enclosed by applying egg while on the edges to prevent the filling from coming out of the dough during baking. the pie was then baked at 700c for 30 minutes till a golden brown colour was obtained. 5. production of meat -pie with 30% breadfruit flour and 70% wheat flour ingredients 1) 60g of breadfruit flour and 140g of wheat flour 2) irish potatoes 3) 200g minced beef 4) 1 medium onion 5) 1 teaspoon baking powder 6) carrot 7) 2 medium eggs 8) 100g margarine 60g of breadfruit flour and 140g of wheat flour was sifted into a bowl, then baking powder, was added and mixed with the flour. the margarine was then added and mixed with the dough until a crumb like texture is formed. then small amount of water was added. the dough was then kneaded thoroughly. onion and irish potatoes was chopped and was fried together with the minced beef in vegetable oil and appropriate seasoning was added. the rolling pin was used to flatten the dough and was cut into required sizes. then the fillings were filled inside the pie and the edges of the dough enclosed by applying egg while on the edges to prevent the filling from coming out of the dough during baking. the pie was then baked at 700c for 30 minutes till a golden brown colour was obtained. 6. production of meat pie with 70% breadfruit flour and 30% wheat flour ingredients 9) 140g of breadfruit flour and 60g of wheat flour 10) irish potatoes 11) 200g minced beef 12) 1 medium onion 13) 1 teaspoon baking powder 14) carrot 15) 2 medium eggs 16) 100g margarine 140g of breadfruit flour and 60g of wheat flour was sifted into a bowl, then baking powder, was added and mixed with the flour. the margarine was then added and mixed with the dough until a crumb like texture is formed. then small amount of water was added. the dough was then kneaded thoroughly. onion and irish potatoes was chopped and was fried together with the minced beef in vegetable oil and appropriate seasoning was added. the rolling pin was used to flatten the dough and was cut into required sizes. then the fillings were filled inside the pie and the edges of the dough enclosed by applying egg while on the edges to prevent the filling from coming out of the dough during baking. the pie was then baked at 700c for 30 minutes till a golden brown colour was obtained. agriculture and food sciences research, 2019, 6(1): 89-97 94 © 2019 by the authors; licensee asian online journal publishing group figure-2. flow chart for the production of chinchin. figure-3. flow chart for the production of meat pie. 2.6. sensory analysis test the two different snacks were made using three different mixing proportions (a, b and c) for each snack to make a total of 6 snack samples were evaluated using a 9-point hedonic scale. the samples were presented to a 50 member panelist to rate them on the hedonic scale where 9 was maximum and 1 was minimum. the samples were scored for the appearance, colour, taste, flavor and overall acceptability. the panelists were instructed to rinse their mouth thoroughly with water after testing any of the samples before proceeding to the next sample. this is to prevent the taste of the samples from interfering with one another. agriculture and food sciences research, 2019, 6(1): 89-97 95 © 2019 by the authors; licensee asian online journal publishing group 2.7. statistical analysis analysis of variance (anova|) was performed on the data gathered from the sensory evaluation to determine differences between the samples. 3. results and discussion 3.1. sensory evaluation table 1 summarizes the sensory mean scores for colour, flavour, taste, texture, overall acceptance for snack sample 1 and sample 2 respectively. snack sample 1: a: 50% breadfruit flour with 50% wheat flour, b: 30% breadfruit flour with 70% wheat flour, c: 70% breadfruit flour with 30% wheat flour snack sample 2: d: 50% breadfruit flour with 50% wheat flour, e: 30% breadfruit flour with 70% wheat flour, f: 70% breadfruit flour with 30% wheat flour the snack sample b was rated highest (7.56±1.15) in terms of colour followed by snack sample a with (7.48±0.10) and sample c (7.00±1.54) was rated least out of the three chin chin samples. the snack sample e was rated highest (7.60±1.12) in terms of colour followed by snack sample d (6.44±1.63) and snack sample f (6.12±1.76) was rated least out of the three meat pie samples. the snack sample b was rated highest (7.42±1.18) in terms of flavor followed by snack sample a (6.88±1.55) and snack sample c (6.48±1.71) was rated least out of the three chin chin samples. the snack sample e was rated highest (7.74±1.26) in terms of flavor followed by snack sample d (6.06±1.85) and snack sample (5.64±2.09) was rated least out of the three meat pie samples. the snack sample b was rated highest (7.52±1.36) in terms of taste followed by snack sample a (6.82±1.72) and snack sample c (6.48±1.91) was rated least out of the three chin chin samples. the snack sample e was rated highest (7.76±1.42) in terms of taste followed by snack sample d (5.86±2.01) and snack sample f (5.54±2.22) was rated least out of the three meat pies considered. the snack sample b was rated highest (7.54±1.01) in terms of texture followed by snack sample a (7.36±1.14) and snack sample c (6.00±1.83) was rated least out of the three chin chin samples. the snack sample e was rated highest (7.62±1.37) in terms of texture followed by snack sample d (6.16±1.74) and snack sample f (5.70±1.88) was rated least out of the three meat pies samples. the snack sample b was rated highest (7.68±1.44) in terms of overall acceptance followed by snack sample a (7.36±1.14) and snack sample c (6.00±1.83) was rated least out of the three chin chin samples. the snack sample e was rated highest (7.84±1.28) in terms of overall acceptance followed by snack sample d (6.30±1.69) and snack sample f (5.88±1.80) was rated least out of the three meat pies samples. table-1. sensory evaluation of snacks from blends of breadfruit flour and wheat flour. chin -chin meat pie parameter a b c d e f colour 6.44±1.63b 7.60±1.12a 6.12±1.76c 7.48±0.10b 7.56±1.15a 7.00±1.20c flavour 6.06±1.85b 7.74±1.26a 5.64±2.09c 6.88±1.55b 7.42±1.18a 6.48±1.71c taste 5.86±2.01b 7.76±1.42a 5.54±2.22c 6.82±1.72b 7.52±1.36a 6.48±1.91c texture 6.16±1.74b 7.62±1.37a 5.70±1.88c 7.36±1.14b 7.54±1.01a 6.00±1.83c overall acceptance 6.30±1.69b 7.84±1.28a 5.88±1.80c 7.24±1.19b 7.68±1.44a 6.60±1.83c note: *different superscript in the same column indicates significant difference (p<0.05) * mean of standard deviation of fifty panelists (n=3) key: a: 50% breadfruit flour with 50% wheat flour chinchin b: 30% breadfruit flour with 70% wheat flour chin -chin c:70% breadfruit flour with 30% wheat flour chin -chin d: 50% breadfruit flour with 50% wheat flour meat pie e:30% breadfruit flour with 70% wheat flour meat pie f: 70% breadfruit flour with 30% wheat flour meat pie according to table 1, there is significant difference in the colour, flavour, taste, texture and overall acceptance at (p<0.05) of snacks a and b, a and c, b and c respectively. also, there is significant difference in the colour, flavour, taste, texture and overall acceptance at (p<0.05) of snacks d and e, d and f, e and f respectively. snack b was rank highest in the colour, flavour, taste, texture and overall acceptance out of all the three chinchin samples, thereby making it to be the most preferable by the panelists and snack e was rank highest in the colour, flavour, taste, texture and overall acceptance out of all the three meat pie samples, thereby making it to be the most preferable by the panelists, snack c of the chin -chin sample was ranked lowest in term of colour, flavour, taste, texture and overall acceptable and snack f of the meat pie samples was ranked lowest in term of colour, flavour, taste, texture and overall acceptable. based on the above illustrations, it can be concluded that there is significant difference in the colour, taste, flavour, texture and overall acceptance of all the snacks and that snack b is the most preferable sample from the chinchin samples and snack e is the most preferable sample from the meat pie samples by the panelists while snack c and f is the least preferable by the panelists from the chinchin and meat pie samples respectively. agriculture and food sciences research, 2019, 6(1): 89-97 96 © 2019 by the authors; licensee asian online journal publishing group 3.2. proximate analysis table 2 summarizes the proximate composition scores for protein, moisture content, fat, ash, crude, cho and dry matter of wheat-breadfruit flour blends. sample b is the richest in protein (12.35±0.01) followed by a (9.83±0.03), then c (6.35±0.03). sample a has the highest moisture content (8.48±0.04) followed by c (8.39±0.01) then b (8.33±0.01). sample b has the highest fat content (1.77±0.01) followed by a (1.21±0.0) then c (0.90±0.00). sample a has the highest ash content (3.38±0.02) followed by b (3.28±0.00) then c (3.21±0.01). sample a has the highest crude fiber content (2.63±0.01) followed by b (2.26±0.02) then c (1.78±0.02). sample c is the richest in carbohydrate (79.37±0.05) followed by a (74.47±0.01) then b (71.97±0.06). sample c has the highest dry matter (91.61±0.01) followed by b (91.67±0.0) then a (91.52±0.04). table-2. proximate composition of blends of breadfruit flour and wheat flour. nutrient a b c protein 9.83±0.03b 12.35±0.01a 6.35±0.03c moisture content 8.48±0.04a 8.33±0.01a 8.39±0.01a fat 1.21±0.01b 1.77±0.01a 0.90±0.00c ash 3.38±0.02a 3.28±0.00a 3.21±0.01a crude fiber 2.63±0.01a 2.26±0.02b 1.78±0.02c cho 74.47±0.01b 71.97±0.06c 79.37±0.05a dry matter 91.52±0.04a 91.67±0.01a 91.61±0.01a note: *different superscript in the same column indicates significant difference (p<0.05) * mean of standard deviation of three determinations (n=3) key: a: 50% breadfruit flour with 50% wheat flour b: 30% breadfruit flour with 70% wheat flour c: 70% breadfruit flour with 30% wheat flour from table 2 there is significant difference at (p<0.05) in the protein, fat, crude fiber, and cho of the flour samples and there is no significant difference at (p>0.05) in the moisture content, ash content and dry matter of the flour samples. the flour sample a has the highest moisture content, ash content and crude fiber content while flour sample b has the highest protein content, fat content and dry matter content and sample c has the highest carbohydrate content. flour sample a has the lowest dry matter content, while flour sample b has the lowest carbohydrate and moisture content and sample c has the lowest protein, fat, ash and crude fiber content. 3.3. discussion the results for proximate composition of flours from blends of breadfruit flour and wheat flour of different proportions showed that there was significant difference at (p<0.05) in the protein, fat, crude fiber, and carbohydrate content. the result showed that values ranging from 6.35 to 12.35%, 8.33 to 8.48%, 0.90 to 1.77%, 3.21 to 3.38%, 1.78 to 2.63%, 71.97 to 79.37%, 91.52 to 91.67% for protein, moisture content, fat, ash, crude fiber, carbohydrate and dry matter respectively. the result showed that the highest value moisture content, ash content and crude fiber content were observed in flour sample a while highest value of protein content , fat content and dry matter content were observed in flour sample b. also, highest value of carbohydrate content was observed in flour sample c. the lowest value of dry matter content was observed in flour sample a while the lowest value of carbohydrate and moisture content were observed in flour sample b and the lowest value of protein, fat, ash and crude fiber content were observed in flour sample c. these results are in line with similar study on nutritional contents of processed local food flours by isong [9]. the data reviewed that the protein contents for the samples were higher than the protein content (5.4%) of maize biscuit reported by adeyeye and akingbala [10]. also, there were significantly different (p<0.05) in the results for sensory evaluation of both chin chin and meat pie samples with values ranging from 6.12 to 7.60, 5.64 to 7.74, 5.54 to 7.76, 5.70 to 7.62, 5.88 to 7.84 for color, flavor, taste, texture and overall acceptance respectively of the chinchin samples and from 7.00 to 7.56, 6.48 to 7.42, 6.48 to 7.52, 6.00 to 7.54, 6.60 to 7.68 for color, flavor, taste, texture and overall acceptance respectively of the meat pie samples. the highest value of color, flavor, taste, texture and overall acceptance was observed in snack b for the chin-chin and snack e for the meat pie sample, thereby making snacks with formulation of 30% breadfruit flour with 70% wheat flour to be the most preferable by the panelists. the result showed that all the sensory parameters of the snacks samples were all acceptable by the panelists. 4. conclusions the study showed that all sensory attributes of snacks produced from the various formulations comprising of blend of breadfruit flour and wheat flour were acceptable by the panelists with sample b from the chinchin samples and sample e from the meat pie samples containing 30% breadfruit flour with 70% wheat flour having the highest overall acceptance by the panelists. the study also showed that flour sample b (30% breadfruit flour with 70% wheat flour) had higher nutritional contents in terms of protein, fat and dry matter content followed by flour sample a in terms of moisture content, ash and crude fiber then flour sample c in term of carbohydrate content. in conclusion, sample b could be very suitable for the production of chin-chin and and sample e could be very suitable for the production of meat pie due to the dominance of the samples in both nutritional and sensory qualities over other samples. 5. recommendations it is therefore recommended that the production of snacks by mixing breadfruit flour with wheat flour using the right mixing proportion should be encouraged for the nutritional benefit of consumers. increase in the agriculture and food sciences research, 2019, 6(1): 89-97 97 © 2019 by the authors; licensee asian online journal publishing group proportion of breadfruit flour from 30% to 40% and a decrease in the proportion of wheat flour from 70% to 60% in the production of snacks can also help to increase the nutritional and quality factors of the snacks by improving the carbohydrate content. from this study, in accordance with the results gotten from the analyses, the best blend that could be recommended for the consumers is 30% breadfruit flour with 70% wheat flour due to its high nutritional factors. also, sample a and d which is a blend of 50% breadfruit flour with 50% wheat flour could also be recommended for making snacks due to its high carbohydrate content but consumers need to be enlightened on its richness in carbohydrate in order to enhance the level of acceptance of the snack. references [1] o. olaoye, a. onilude, and o. idowu, "quality characteristics of bread produced from composite flours of wheat, plantain and soybeans," african journal of biotechnology, vol. 5, pp. 1102-1106, 2006. [2] n. amusa, i. kehinde, and o. ashaye, "bio-deterioration of breadfruit (artocarpus communis) in storage and its effects on the nutrient composition," african journal of biotechnology, vol. 1, pp. 57-60, 2002. available at: https://doi.org/10.5897/ajb2002.000-010. [3] h. d. mepba, l. eboh, and s. nwaojigwa, "chemical composition, functional and baking properties of wheat-plantain composite flours," african journal of food nutrition developmen, vol. 7, pp. 1-22, 2007. [4] o. a. oloye, "sensory evaluation of and acceptance of composite bread from wheat, cassava starch and cassava flour blends," unpublished phd thesis in food science and technology department, federal university of technology, akure, 2006. [5] a. j. ajani, b. a. osundahunsi, m. o. akinsola, s. a. arowora, e. a. abiodun, and f. a. pessu, "chemical analysis of breadfruit (artocarpus communis forst) from southern western nigeria," journal of food technology, vol. 1, pp. 29 – 35, 2012. [6] aoac, official method of analysis: association of official of analytical chemists. gaithersburg, maryland, usa: international (18 ed), 2005. [7] s. o. pearson, food analysis: food science text series new york: springer, 2010. [8] e. a. martin and a. a. coolidge, nutrition in action, 4th ed. new york: holt, r and wilson co, 1978. [9] n. c. isong, "processing and preservation of nigerian fruits and green vegetable," in proceedings of the launching and first annual conference of nigeria institute of food science and technology, 1977, pp. 49-55. [10] s. a. adeyeye and j. o. akingbala, "evaluation of nutritional and sensory properties of cookies produced from sweet potato maize flour blends," researcher, vol. 6, pp. 9-19, 2014. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 134 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 134-144, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.134.144 © 2019 by the authors; licensee asian online journal publishing group supplementation of rice flour with carrot, date palm and defatted soybean flours for enhanced nutritional, antioxidants and physicochemical properties olugbenga olufemi awolu1 temitope esther olabiran2 ( corresponding author) 1department of food science and technology, federal university of technology, akure, nigeria. 2central research laboratory, federal university of technology, akure, nigeria. abstract the nutritional, antioxidants and physicochemical properties of rice flour is being improved by the addition of carrot, date palm and defatted soybean flours. the effect of the addition of defatted soy bean (5 15%), carrot (5 15%) and date palm (10 20%) flours on the chemical compositions; antioxidants and functional properties; and pasting characteristics of rice-based composite flour were investigated. while defatted soy beans (15%) resulted in the highest protein content (17.86%), blend with highest carrot flour inclusion had the highest ash (1.88%) and crude fiber (1.93%) contents. the sample with highest carrot inclusion (15%) had the best antioxidant and functional properties followed by the sample with soy bean incorporation from 10 to 15%. the same sample with 15% carrot inclusion had the second best pasting characteristics, next to that of 100% rice flour sample. it was noted that the samples consisting date palm (80% rice and 20% date palm flours; 80% rice, 5% soybean, 5% carrot and 10% date palm flours) had the least chemical composition together with the least functional and antioxidant properties. however, the sample with date palm incorporation had the third best pasting properties while 100% rice flour had the best pasting properties. overall, carrot flour incorporation resulted in rice based composite flour with the best fibre and minerals contents; functional and antioxidants properties closely followed by sample with 10-15% soybean incorporation. keywords: antioxidants, pasting properties, functional properties, rice flour, defatted soya bean flour, carrot flour, date palp flour. citation | olugbenga olufemi awolu; temitope esther olabiran (2019). supplementation of rice flour with carrot, date palm and defatted soybean flours for enhanced nutritional, antioxidants and physicochemical properties. agriculture and food sciences research, 6(1): 134-144. history: received: 19 march 2019 revised: 26 april 2019 accepted: 29 may 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 135 2. material and methods ................................................................................................................................................................... 135 3. results and discussion ................................................................................................................................................................. 138 4. conclusion ....................................................................................................................................................................................... 142 references ............................................................................................................................................................................................ 143 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.134.144&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.134.144&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/132 https://orcid.org/0000-0002-5499-1888 agriculture and food sciences research, 2019, 6(1): 134-144 135 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to the existing knowledge by enhancing utilization of rice flour as an alternative to wheat flour, and also to promote gluten-free flour for baked products production. vital nutritional compositions of the rice flour such as dietary fibre, protein contents, and bioactive components were enhanced by the incorporation of defatted soybean and carrot flours, while the functional and rheological properties were also enhance by the addition of date palm, carrot and soybean flours. 1. introduction researches into viable alternatives to wheat flour in the preparation of baked foods is currently predominant among researchers working on baked food materials. wheat has proof to be a perfect food materials for preparing baked foods. however, some health challenges in some individuals with gluten intolerance, and economic considerations [1] has led to diversification of researches into viable alternatives to the utilization of wheat as a sole source of raw materials for baked foods. while successes are being recorded in non-wheat baked food materials, there are however, some challenges which has hindered the full utilization of non-wheat materials for baked foods, especially in production of bread. non-wheat materials are lacking in gluten, which is responsible for the viscoelastic behavior of dough. present researches are therefore focused on evolving mechanism that will mimic gluten in non-wheat food materials in order to produce non-wheat dough with adequate viscoelastic properties. several techniques have been adopted to enhance the rheological characteristics of non-wheat dough, including the use of hydrocolloids, starch, enzymes, other binders and flour modification [1, 2]. it has also been found that incorporation of legumes, dietary fibre and sweeteners into non-wheat flour would enhance the rheological properties [2, 3]. cereals have been used as a viable alternative to wheat in production of wheat flour [4]. several cereals and pseudo-cereals including, rice, amaranth grain, maize, tigernut and pearl millet [5, 6] had been used as an alternative to wheat. rice (oryza sativa) is staple food and a popular cereal worldwide. the rice grain consists about 75-80 % starch, 12 % water and about 7 % protein with a full complement of amino acids. its protein is highly digestible with excellent biological value and protein efficiency ratio owing to the presence of higher concentration of lysine [7]. rice flour contains low levels of sodium and a high amount of easily digested carbohydrates, thus, making it desirable in celiac diets [8]. soybeans (glycine max) have a great potential as human food because of their high level of good quality protein, unique functional and nutritional properties. supplementation with soybeans had resulted in baked products with improved nutritional properties, and especially amino acids that are complimentary to most cereals [2, 9]. soybeans supply all nine essential amino acids and have cholesterol reducing and anticarcinogenic properties [10]. dates palm (phoenix dactylifera) is popular in northern nigeria. it is rich in several vitamins, minerals, dietary fiber but low in calories. dates provides relief from constipation, intestinal disorders, heart problems, anemia, sexual dysfunction, diarrhea, abdominal cancer, and many other health relating conditions. carrot (daucus carota subsp. sativus) is an important root vegetables rich in bioactive compounds including carotenoids, and dietary fibers; providing appreciable functional characteristics with significant healthpromoting properties. the consumption and usage of carrot and its products is increasing steadily due to its natural antioxidants and anticancer activities [11, 12]. this work was conducted to assess the nutritional, functional and pasting properties of rice based composite flour. soya beans, date palm and carrot flours were added to enhance the nutritional and rheological characteristics of the flour. 2. material and methods 2.1. materials rice grains, soya beans, date palm and fresh carrots were obtained from erekesan market, akure, ondo state, nigeria. all reagents used were of analytical grade. 2.2. samples preparation of rice flour rice was processed into flour by method described by awolu, et al. [5]. rice grains were cleaned, sorted, weighed. the grains were milled using blender (scanfrost, sfkab409), cooled and sieved through 150 µm mesh size sieve. the flour was later stored in an air tight polyethylene at room temperature. 2.3. preparation of defatted soya beans flour defatted soya bean flour was produced using the method described by ihekoronye and ngoddy [13] with slightly modification. two kilograms of soybean seeds which were manually sorted and cleaned washed with water 3 times, manually dehulled and dried in cabinet dryer at 45 oc. the dried seeds were milled using blender (scanfrost, sfkab409) and made to pass through 150 µm mesh sieve in order to obtain cooked full-fat soy bean flour. the full-fat soybean flour was defatted using soxhlet extraction with hexane as the solvent. the defatted soy bean flour was packaged in an air tight container for further analysis 2.4. preparation of date palm flour date palm flour was prepared according to the method described by shaba, et al. [14]. date palm fruits were sorted, cleaned and oven-dried at 40 oc for 72 h. the dried date palp fruits were milled using household grinder and sieved (150 µm mesh size) to obtained the date palm flour. the flour obtained was stored in an air tight polyethylene pack until use. agriculture and food sciences research, 2019, 6(1): 134-144 136 © 2019 by the authors; licensee asian online journal publishing group 2.5. preparation of carrot flour carrot flour was prepared according to the method described by gazalli, et al. [15]. about 1 kg of carrots were washed with distilled water for cleaning and removal of extraneous materials. the cleaned carrots were peeled manually, cut into slices and ground to paste in home mixer. the paste was spread evenly on trays and dried in an oven at 40 ± 2°c. the dried paste was removed from oven when constant weight was attained. dried carrot was milled in a grinder to form powder; then sieved and stored in air tight food grade plastic containers until used. 2.6. proximate composition proximate composition, including moisture, ash, protein, fat and fiber contents of the flours were determined using aoac, 2005 methods. the carbohydrate content was determined by difference. 2.7. determination of moisture content the moisture content was determined according to aoac [16]. a sample of 5 g was weighed into a known weight of a clean empty petri dish w1. the weight of the petri dish and the sample was taken and recorded as w2. the petri-dish was placed into a preset oven of 105 oc for 3 h in order to reduce the moisture. after 3 h, the petri dish was taken out and placed inside desiccators for about 30 min so that it can cool. after cooling, the sample was brought out and weighed on a weighing balance. the weight was recorded as w3. % moisture content = 𝑊2 – 𝑊3 𝑊1 × 100 (1) where: w1. = weight of empty petri dish w2 = weight of the petri dish and the sample before moisture removal w3 = weight of the petri dish and sample after the removal of moisture 2.8. determination of fat content the fat content was determined using soxhlet apparatus as described by aoac [16]. a sample of 5 g was weighed into a pre-weighed filter paper, weighed, dried in an oven and tied with thread. the filter paper containing the sample was placed in the receiver of the soxhlet apparatus. normal hexane of boiling point range 60-68 oc was used as solvent for the extraction; a 500 ml round bottom flask was filled to 3/4 with the solvent. the flask was fitted to the soxhlet apparatus with a reflux condenser and placed in an electro mantle heater. extraction began as the solvent refluxed several times and continued for 4 h until the condenser was detached. the filter paper containing the defatted sample was removed and dried to a constant weight in an oven at 50 oc. the difference in weight before extraction and after extraction was recorded in order to obtain the value of the fat extracted. % 𝐹𝑎𝑡 𝐶𝑜𝑛𝑡𝑒𝑛𝑡 = weight of fat extracted weight of sample × 100 (2) 2.9. determination of ash content the ash content of the sample was determined as described by aoac [16]. about 2g of the sample was weighed in a clean pre-weighed crucible and weight was recorded as w2. the crucible with the sample was placed in a muffle furnace and the temperature was increased to 500 oc for 3 h in order to allow the sample to burn (ash). after ashing, the crucible with the ash was cooled in desiccators and then weighed. % 𝐴𝑠ℎ 𝐶𝑜𝑛𝑡𝑒𝑛𝑡 = 𝑊3 – 𝑊1 𝑊2 – 𝑊1 × 100 (3) where: w1 = weight of empty crucible w2 = weight of sample and crucible before ashing w3 = weight of crucible and the ashed sample. 2.10. determination of crude fibre content crude fibre was determined as described by aoac [16]. a defatted sample of about 2g was weighed and transferred into a 500 ml conical flask where 200 ml of 1.25 % h2so4 was added and the sample were boiled for 30min using cooling fingers to maintain constant temperature. after boiling, the mixture was poured into filter cloth under gentle suction using a butcher funnel, rinsed well with hot distilled water. the material was transferred into a conical flask containing 200 ml of 1.25 % naoh and boiled for another 30 min while shaking gently to avoid spillage. the sample solution was filtered, washed with hot distilled water and with 1% hcl respectively. the washing was repeated twice with ethanol and trice with petroleum ether to remove any remaining fat. the residue was transferred into a clean dried crucible, oven dried, cooled in the desiccators and weighed (w2). the crucible was placed in the muffle furnace at 450 oc for 2 h, cooled in a desiccators and reweighed (w3). % 𝐶𝑟𝑢𝑑𝑒 𝑓𝑖𝑏𝑟𝑒 = weight of crude fibre weight of sample × 100 (4) 2.11. determination of crude protein content the crude protein was determined using the kjeldahl method. exactly 0.5g of sample was digested using 10ml of sulphuric acid and 0.5g selenium catalyst at 400oc for 1 h until there was a complete digestion which is indicated by a blue green colouration. the flask was allowed to cool after which the digest was diluted with distilled water into a 100 ml standard flask. about 20 ml of 40 % naoh solution was added to 10 ml of the digest and three drops of mixed indicator was added to the receiving flask containing 10 ml of 2 % boric acid solution to give a pink colour solution. the sample was distilled until about 50 ml of the distillate was collected agriculture and food sciences research, 2019, 6(1): 134-144 137 © 2019 by the authors; licensee asian online journal publishing group in the receiving flask. the resulting solution in the conical flask was titrated against 0.1m hcl solution until a colour change from green to red wine is obtained indicating the end point. nitrogen = titre value x molarity of acid used x 0.014 x dillution factor × 100 weight of the sample (5) % crude protein = % nitrogen × 6.25 2.12. determination of carbohydrate content carbohydrate content was determined by difference. the summation of protein, fat, moisture content, ash, crude fibre minus 100. % 𝑐𝑎𝑟𝑏𝑜ℎ𝑦𝑑𝑟𝑎𝑡𝑒 = 100 − (𝑚𝑜𝑖𝑠𝑡𝑢𝑟𝑒 + 𝑓𝑎𝑡 + 𝑐𝑟𝑢𝑑𝑒 𝑓𝑖𝑏𝑟𝑒 + 𝑎𝑠ℎ + 𝑝𝑟𝑜𝑡𝑒𝑖𝑛) (6) 2.13. determination of mineral elements determination of zinc, iron, calcium, magnesium, sodium, potassium and phosphorus were done according to the method of aoac [16]. the analysis of sodium and potassium were carried out using flame photometer. calcium, magnesium, iron and zinc were analyzed using atomic absorption spectrometer (buck scientific, model 210vgp). phosphorus was determined by using phosphovanado-molybdate method. 2.14. determination of functional properties the standard analytical procedures [16] for food analysis as described below were used. 2.15. bulk density firstly, a dried and empty measuring cylinder was weighed (w1). the sample was filled gently into the weighed measuring cylinder and then gently tapped at the bottom on a laboratory bench several times until there was no further diminution of the sample level, the volume was noted (v). after this, the filled measuring cylinder was weighed and recorded (w2). this process was repeated two times. bulk density was calculated using equation 7. 𝐵𝑢𝑙𝑘 𝐷𝑒𝑛𝑠𝑖𝑡𝑦 (𝑔/𝑚𝑙) = 𝑊2 – 𝑊1 𝑉 (𝑚𝑙) (7) 2.16. water/oil absorption capacity about 1.0 g of the ground sample was weighed into a conical graduated centrifuge tube and 10 ml of water or oil was added to the weighed sample. a warring whirl mixer was used to mix the sample for 30 s. the sample was allowed to stand at room temperature for 30 min and then centrifuged at 5000 rpm for 30 min. the mixed sample was transferred from the graduated centrifuge tube into a 10 cm3 measuring cylinder to know the volume of the free water or oil. the absorption capacity was expressed as grammes of water or oil absorbed per gramme of sample equation 8. 𝑤𝑎𝑡𝑒𝑟 𝑜𝑖𝑙⁄ 𝑎𝑏𝑠𝑜𝑟𝑝𝑡𝑖𝑜𝑛 𝑐𝑎𝑝𝑎𝑐𝑖𝑡𝑦 = (𝑡𝑜𝑡𝑎𝑙 𝑤𝑎𝑡𝑒𝑟 𝑜𝑖𝑙⁄ 𝑎𝑏𝑠𝑜𝑟𝑏𝑒𝑑 − 𝑓𝑟𝑒𝑒 𝑤𝑎𝑡𝑒𝑟 𝑜𝑖𝑙⁄ ) × 𝑑𝑒𝑛𝑠𝑖𝑡𝑦 𝑜𝑓 𝑤𝑎𝑡𝑒𝑟 ∕ 𝑜𝑖𝑙 (8) 2.17. foaming capacity about 2.0 g of the powdered sample were weighed and blended with 100 cm3 of distilled water using warring blender (binatone blg555), the suspension was whipped at 1600 rpm for 5 min. the mixture was then poured into a 100 cm3 measuring cylinder and its volume was recorded after 30 s. foam capacity was expressed as equation 9. 𝑓𝑜𝑎𝑚𝑖𝑛𝑔 𝑐𝑎𝑝𝑎𝑐𝑖𝑡𝑦 = (𝑣𝑜𝑙𝑢𝑚𝑒 𝑎𝑓𝑡𝑒𝑟 𝑤ℎ𝑖𝑝𝑝𝑖𝑛𝑔−𝑣𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑓𝑜𝑟𝑒 𝑤ℎ𝑖𝑝𝑝𝑖𝑛𝑔)×100 𝑣𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑓𝑜𝑟𝑒 𝑤ℎ𝑖𝑝𝑝𝑖𝑛𝑔 (9) 2.18. least gelation capacity about 5 cm3 of 2-20% (w/v) suspended samples in test tubes were heated for 1 h in a boiling water bath followed by rapid cooling under running cold tap water. the test tubes were further cooled for 2 h at 4 °c. the gelation capacity was the least gelation concentration determined as the concentration when the sample from the inverted test tube did not fall or slip. 2.19. solubility and swelling power well blended sample (1.0 g) was weighed into a centrifuge tube. about 10 ml of distilled water was added, properly mixed and boiled in a water bath at 80 °c for 30 min. the tubes were centrifuged at 2200 rpm for 15min, while the supernatant was decanted into a previously weighed petri dish (w1). it was then evaporated and dried in the oven, the dish was re weighed (w2). solubility index was calculated using equation 10. % 𝑆𝑜𝑙𝑢𝑏𝑖𝑙𝑖𝑡𝑦 = (𝑤2− 𝑤1)×100 𝑠𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝑔ℎ𝑡 (𝑊2 – 𝑊1) × 100 (10) the gel or paste from the solubility was weighed and calculated for swelling power equation 11. % 𝑠𝑤𝑒𝑙𝑙𝑖𝑛𝑔 𝑝𝑜𝑤𝑒𝑟 = (𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑔𝑒𝑙 ×100) 𝑠𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝑔ℎ𝑡 (11) 2.20. determination of pasting properties the pasting characteristics of the composite flours were evaluated using rapid visco analyser (rva). the amount of the sample to be used was determined from the instrument by inserting the moisture content (14% in this case). the mixture were vigorously but carefully stirred until there was no more lumps. the solution was carefully transferred into the canister, inserted into the paddle coupling properly. the measurement cycle was initiated by depressing the motor tower of the instrument. the test was then allowed to proceed and terminate agriculture and food sciences research, 2019, 6(1): 134-144 138 © 2019 by the authors; licensee asian online journal publishing group automatically according to the pre-set time and temperature regime. the pasting properties were recorded at the end of the experiment. 2.21. quantification of antioxidant properties of the flour blends 2.21.1. sample extraction for antioxidant assay extracts from the samples were prepared using acidified methanol (1 % hcl in methanol). the samples were extracted with 50 ml of solvents in three different phases. about 10 ml solvent was added to 0.5 g of the sample in a conical flask and completely covered with aluminum foil. the sample was stirred for 4 h using a shaker, transferred to 40 ml plastic centrifuge tubes, and centrifuged at 3500 rpm for 10 min (25 oc). it was later decanted and the supernatants were stored in a glass bottle container covered with aluminum foil and kept in the cold room prior to analysis. 2.22. determination of total phenolic content the total phenolic content was determined by using the method of waterman and mole [17]. a tannic acid standard solution of concentration of 0, 0.2, 0.4, 0.6, 0.8 and 1.0 mg/ml by serial dilution from 1mg/ml standard stock solution was prepared. about 0.5 ml of methanolic phenolic acid extract/tannic acid (standard) was added into a 50 ml volumetric flask containing 10 ml distilled water. folin-ciocalteu’s phenol reagent (2.5 ml) was added and mixed. after 2 min, 7.5 ml of sodium carbonate (naco3) solution (20 g/100 ml) was added. the content was mixed and made up to volume with deionized water. the volumetric flask was then stopped and thoroughly mixed by inverting several times while the flask was allowed to stand for 2 h from the addition of sodium carbonate after which the absorbance was measured at 760 nm at the different standard solution concentrations. 2.23. determination of total flavonoid content the total flavonoid content of the extract was determined using a slightly modified method reported by meda, et al. [18]. about 0.5 ml of appropriately diluted sample was mixed with 0.5 ml methanol, 50 μl of 10 % alcl3, 50μl of 1mol l-1 potassium acetate and 1.4 ml water, and allowed to incubate at room temperature for 30 min. thereafter, the absorbance of the reaction mixture was subsequently measured at 415 nm. the total flavonoid was calculated using quercetin as standard by using a seven-point standard curve (0-100 μg/ml), the total flavonoids content of samples was determined in triplicates. 2.24. determination of 2, 2azino-bis (3-ethylbenzo-thiazoline6-sulfonate (abts)) radical scavenging ability the reduced (abts) concentration by a certain amount of antioxidant is related to that of trolox, the water-soluble vitamin e analogue and this gives the trolox equivalents (tea) value of the antioxidants awika, et al. [19]. the working solution (2.9 ml) was added to trolox standard (0.1 ml) in test tubes and mixed. the test tubes were allowed to react for 30 min for both trolox and the samples and the absorbance of the standard and samples was measured at 734 nm. the antioxidant activity was calculated by using equation 12. antioxidant activity ( μmte g⁄ sample) = m x ∆atbs +c sample (12) where m = slope of the standard curve ∆abs = absorbance of blank-absorbance of sample c = y-intercept and te is trolox equivalent. 2.25. determination of ferric reducing antioxidant property (frap) the reducing property of the extracts was determined by assessing the ability of the extract to reduce fecl3 solution as described by oyaizu [20]. appropriate dilution of the extract (2.5 ml) was mixed with 2.5 ml (200 mm) sodium phosphate buffer (ph 6.6) and 2.5 ml (1%) potassium ferricyanide. the mixture was incubated at 50 °c for 20 min and then 2.5 ml (10%) trichloroacetic acid was added. this mixture was centrifuged at 650 rpm for 10 min. supernatant of about 5 ml was mixed with an equal volume of water and 1 ml of 0.1 % ferric chloride. the absorbance was measured at 700 nm. 2.26. 1,1-diphenyl–2 picrylhydrazyl (dpph) free radical scavenging ability the free radical scavenging ability of the extracts against dpph (1,1-diphenyl–2 picrylhydrazyl) free radical was evaluated as described by gyamfi, et al. [21]. appropriate dilution of the extracts (1 ml) was mixed with 1 ml, 0.4 mm methanolic solution containing dpph radicals, the mixture was left in the dark for 30 min and the absorbance was taken at 516 nm. the dpph free radical scavenging ability was subsequently calculated. 2.27. statistical analysis the data for the analysis was generated in triplicate and was subjected to one way analysis of variance (anova), spss 19.00 while means was separated using duncan multiple range test (dmrt) at p<0.05 3. results and discussion 3.1. proximate compositions of the flours the proximate compositions of the rice based flour blend are presented in table 1. the moisture content value ranged between 4.31 and 7.23%. the flour blends generally had low moisture content which implied that the samples could have an extended shelf life. the reduction in the moisture content can be as a result of extensive drying during processing of the flours. agriculture and food sciences research, 2019, 6(1): 134-144 139 © 2019 by the authors; licensee asian online journal publishing group there are significant (p ≤ 0.05) differences in the ash contents of the samples; the ash content ranged between 0.71 and 1.88 g/100g. carrot and soybean flours addition had the highest positive effect on the ash content. it may however affirm that, while the duo of carrot and soybean flours contributes to the minerals contents, carrot flour seemed to have greater input into the ash contents as could be deduced from sample 2 with 15% carrot flour and 5% soybean flour (with highest ash content) in contrast to sample 3 with 5% carrot flour and 15% soybean flour coming second behind sample 2 in ash content. table-1. proximate composition (%). samples moisture (g/100g) ash (g/100g) protein (g/100g) fat (g/100g) fibre (g/100g) cho (g/100g) 1 4.31±0.015f 0.71±0.000f 11.04±0.030f 0.54±0.025e 0.24±0.006f 82.83±0.015a 2 6.51±0.005b 1.88±0.010a 13.25±0.000d 0.58±0.000d 1.93±0.010a 75.89±0.006d 3 6.01±0.005e 1.45±0.015b 17.86±0.075a 0.68±0.010b 1.15±0.010c 74.65±0.080e 4 7.23±0.010a 0.83±0.000e 11.35±0.010e 0.83±0.000a 0.52±0.005e 79.50±0.025b 5 6.46±0.025c 1.37±0.010c 15.94±0.035b 0.61±0.015c 1.48±0.020b 74.00±0.055f 6 6.36±0.015d 0.86±0.025d 13.60±0.015c 0.68±0.010b 0.82±0.005d 77.55±0.056c note: mean ± standard deviation of three triplicates. mean values of the same sample on the same column with the different superscript are significantly different (p<0.05) keys: 1: 100% rice flour, 2: 80% rice flour, 5% defatted soya beans flour and 15% carrot flour 3: 80% rice flour, 15% defatted soya beans flour and 5% carrot flour, 4: 80% rice flour and 20% date palm flour, 5: 80% rice flour, 10% defatted soya beans flour and 10% carrot flour 6: 80%rice flour, 5%defatted soya beans flour, 5%carrot flour and 10% date palm flour. rice flour alone had the least ash content, while inclusion of only date palm flour alone had 2nd to the last ash content. the crude fat content ranged between 0.54 and 0.83 g/100g. date palm flour inclusion increased the fat contents of the sample. sample 4 (with 20% date palm flour) had the highest fat content, followed sample 6 (with 10% date palm flour). sample with 15% soybean flour inclusion (sample 3) also had second highest fat content. these results clearly showed that inclusion of date palm and soybean flours to the samples had positive impact of their fat contents. however, carrot flour inclusion did not contribute strongly to the fat contents. dietary fat provides essential fatty acids which enhanced the taste and acceptability of foods, slowed gastric emptying and intestinal motility thereby prolonging satiety and facilitating the absorption of liquid soluble vitamins [22]. the crude fibre content increased with increased level of carrot flour. the control (100 % rice flour) had the least crude fibre content (0.24 g/100g). carrots have been reported to contain high dietary fibers [23] dietary fibers play an important role in human health [24]. dietary fiber is important for the removal of waste from the body thereby preventing constipation and many health disorders. the protein content of the flours varies considerably with the addition of soybean, carrot and date palm flours. sample 3 (80% rice, 15% defatted soya bean and 5% carrot flour) had the highest protein values of 17.86% while the control (100% rice flour) had the least value of 11.35%. the contribution of defatted soya beans to protein is very high and this justifies the addition of soya beans which was intended to increase the protein content of the flour. samples 5 and 6, with second and third best protein contents respectively, showed that incorporation of carrot flour also contributed to the protein contents of the composite flour. the same trend were not reported in composite flour with only date palm flour (sample 4) with protein content almost equal to that of 100% rice flour. the carbohydrate content of the composite flours were reduced with the incorporation of soybean, carrot and date palm flours. control sample had the highest value of 82.83% while sample 5 (80% rice flour, 10% defatted soya beans flour and 10% carrot flour) had the least value of 72.07%. this might not be unconnected with the high protein contents in both soybean and carrot flours which resulted in high protein content of sample 5, and hence, a lower carbohydrate content. 3.2. minerals composition of the flours the mineral compositions of the composite flour are presented in table 2. sample 5 (80% rice flour, 10% defatted soya beans flour and 10% carrot flour) was the richest in terms of minerals composition, followed by sample 2 (80% rice flour, 5% defatted soya beans flour and 15% carrot flour), and then sample 4 (80% rice flour and 20% date palm flour). it could be deduced that soybean and carrot flours contribute most to minerals composition (as reflected in samples 5 and 2). a substantial addition of date palm flour (at about 20%) came third in terms of its contribution to overall minerals composition of the composite flour. the sodium content ranged between 97.69 and 189.98 mg/100g. the sodium content increased with the addition of defatted soya beans, carrot and date palm flours. the highest value of sodium (189.98mg/kg) was observed in the flour blend that has rice, defatted soya beans and carrot in 80:10:10 respectively. composite flour sample with lower soybean but higher carrot content (sample 2) had the best potassium content. increasing k intake is associated with lowering of blood pressure and the effects of increasing k intake are additive to the effects of lowering na intake [25]. it is however noted that all the samples had lower na to higher k contents ratio, which suggest that the samples would serve as appropriate meals for patients with high blood pressure. carrot and soybean flours are veritable sources of calcium as samples containing about 20% (total composition of soybean and carrot flours) had calcium contents between 160 and 192 mg/100g. calcium, however, showed a better contribution to the calcium content as the sample with 10% carrot flour incorporation had the highest calcium content; sample with 15% soybean flour incorporation had lower calcium content compared to the sample with 10% carrot content. rice flour (100%) had 116.50 mg/100g calcium content. the least value (83.74mg/100g) was observed in sample 4 (80% rice flour and 20% date palm flour). calcium is agriculture and food sciences research, 2019, 6(1): 134-144 140 © 2019 by the authors; licensee asian online journal publishing group needed for bone formation while zinc is essential for protein and nucleic acid synthesis, carbohydrate metabolism, successful pregnancy, delivery and normal development [26]. table-2. mineral composition (mg/100g) of rice flour and flour blends. sample na k ca p mg zn fe 1 97.87±0.015f 392.93±0.005f 116.50±0.000d 9.55±0.015f 151.25±0.265f 10.81±0.005f 11.78±0.015f 2 182.8±0.060d 1383.80±0.015a 174.77±0.071b 15.75±0.020c 194.30±0.056c 13.11±0.015e 21.11±0.015a 3 186.7±0.000c 1080.40±0.006c 160.58±0.508c 17.70±0.020a 174.05±0.017d 13.81±0.005c 16.81±0.005c 4 117.69±0.020e 710.41±0.020e 83.74±0.006f 11.25±0.020e 226.42±0.011a 15.33±0.000a 12.55±0.000e 5 189.98±0.015a 1364.50±0.015b 192.01±0.015a 16.35±0.000b 187.40±0.015e 13.61±0.005d 20.39±0.005b 6 159.98±0.000b 928.52±0.015d 101.12±0.115e 11.64±0.020d 207.06±0.000b 16.28±0.005b 13.00±0.000d note: values represent means of triplicate (n=3). values with the same alphabet along the same column are not significantly different (p>0.05). keys: 1: 100% rice flour, 2: 80% rice flour, 5% defatted soya beans flour and 15% carrot flour 3: 80% rice flour, 15% defatted soya beans flour and 5% carrot flour, 4: 80% rice flour and 20% date palm flour, 5: 80% rice flour, 10% defatted soya beans flour and 10% carrot flour 6: 80%rice flour, 5%defatted soya beans flour, 5%carrot flour and 10% date palm flour. soybean flour showed the best contribution to phosphorus content of the composite flour, followed by carrot flour (samples 3, 5 and 2 in that order). date palm flour contribution also came a distant fourth position to samples 3, 5 and 2. the phosphorus content of rice (100%) was the least. the results of magnesium and zinc table 2 indicated that date palm is a good source of both. the higher the incorporation of date palm flour to the composite flour, the higher the values of magnesium and zinc. magnesium is required for lowering blood pressure [27]. taking magnesium supplements has been shown to significantly reduced fasting blood sugar levels in participants with type 2 diabetes [28]. on the other hand, zinc is required by human body to activate t lymphocytes (t cells). t cells help the body in controlling and regulating immune responses and attacking infected or cancerous cells [29]. carrot also had positive effect on iron content of the composite flour, as the higher the carrot flour incorporation, the higher the iron content of the composite flour. iron is an important component of blood and enzymes involved in electron transfer. its deficiency results in fatigue, headache and sore tongue in addition to anaemia. the overall results indicated that 100% rice flour had the least minerals composition. this showed that rice flour needs to be supplemented with minerals rich food materials in order to it nutritionally beneficial. 3.3. antioxidants properties of the flour blends the presence of protein, fibre and minerals contents only in food is no longer attractive indices for consumers’ acceptance of the food. this is as a result of the widespread degenerative diseases confronting humanity, and which had not been successfully controlled with the knowledge of medical and pharmaceutical sciences. consumption of food rich in antioxidant is being propounded as possible mitigation and cure for these diseases. food rich in antioxidant have been found to be adequate in the maintenance of health and protection from coronary heart diseases and cancer [30]. the antioxidant properties of the composite flour consisting rice, carrot and soybean flours are presented in table 3. the addition of carrot and soybean flours in the composite flour resulted in composite flours with greatly enhanced antioxidant properties as observed in samples 2, 3, 5 and 6. date palm flour addition only resulted in the best frap (sample 4). the sample with 100% rice sample had the least antioxidant values. this is an evident that nutritional capacity of rice flour which is currently consumed throughout the world, and especially, in several asia countries, needs to be enhanced as established in this study in order to properly serve as nutritious food. table-3. antioxidants in flour blends. samples frap mg/g abts mm/g dpph % tpc mg/g tfc mg/g 1 7.06±0.040f 0.018±0.000d 77.40±0.400e 0.09±0.005f 0.06±0.008cd 2 21.36±0.225c 0.010±0.001b 83.50±0.000c 4.59±0.015a 0.15±0.013a 3 16.27±0.040e 0.02±0.000b 89.55±0.150a 4.53±0.004b 0.13±0.000ab 4 54.28±0.010a 0.019±0.001c 76.25±0.250f 4.49±0.005d 0.03±0.004d 5 18.24±0.085d 0.03±0.000a 85.07±0.040b 4.51±0.000c 0.10±0.057abc 6 50.06±0.345b 0.02±0.000b 80.35±0.075d 4.09±0.004e 0.09±0.000bc note: results are expressed as mean ± standard deviations (sd) of three determinations. averages followed by different letters in the same column differ by tukey’s test at p < 0.01. keys: 1: 100% rice flour, 2: 80% rice flour, 5% defatted soya beans flour and 15% carrot flour 3: 80% rice flour, 15% defatted soya beans flour and 5% carrot flour, 4: 80% rice flour and 20% date palm flour, 5: 80% rice flour, 10% defatted soya beans flour and 10% carrot flour 6: 80%rice flour, 5%defatted soya beans flour, 5%carrot flour and 10% date palm flour phenols are used as an important indicator of antioxidant capacity as well as a preliminary screen for any product that is intended to be used as a natural source of antioxidants in functional foods [31]. there were significant (p ≤ 0.05) differences in the phenolic content of the flour blends. carrot flour contributes more to the increase in the phenol content of the flour blend followed by defatted soya beans flour and date palm flour. carrot is a significant source of phytonutrients including phenolic [32] polyacetylenes [33, 34] and carotenoids [35]. the flour blends can serve as a good source of phenolic content. abts assay has been shown to be more sensitive to identifying the antioxidant activity since it has faster reaction kinetics and a heightened response to antioxidants [36]. agriculture and food sciences research, 2019, 6(1): 134-144 141 © 2019 by the authors; licensee asian online journal publishing group reducing power is a measure of the ability of the food extracts to reduce fe3+ to fe2+. substances which have reduction potential react with potassium ferric-cyanide to form potassium ferrocyanide which then reacts with ferric chloride to form ferric ferrous complex that has an absorption maximum at 700nm. reducing power has become one of the antioxidant capacity indicator of medicinal plants [37]. the ferrous reducing antioxidants property of the flour blends ranged between 7.02 to 54.28mg/g. significant differences were observed among all the flour samples. sample 4, consisting rice and date palm flour (in ratios 80:20, respectively) had the highest frap content of 54.28mg/g. though defatted soy bean and carrot flours increased the frap content of the flour blends, it is observed that date palm increased it better. several researches have established that dates are rich in frap [38]. this might explain the significant increase in the frap content of rice flour when supplemented with date palm flour. the flavonoid content followed the same pattern as the phenolic content. carrot flour was responsible for the increase in the flavonoid content of the flour blends. a range of 4.50-7.42 mg/100g total flavonoids had been reported in raw and processed carrot. flavonoids are important due to their anti-oxidative, anti-inflammatory, anti-mutagenic and anti-carcinogenic properties coupled with their capacity to modulate key cellular enzyme function. research on flavonoids received an added impulse with the discovery of the low cardiovascular mortality rate [39]. the dpph content of the composite flour indicated that soybean flour addition resulted in the highest increase of the dpph. in addition to soybean addition, carrot flour addition also defatted soya beans and carrot flour are responsible for the increase in the dpph content. dpph assay is used to predict antioxidant activities by mechanism in which antioxidants act to inhibit lipid oxidation, so scavenging of dpph radical and therefore determinate free radical scavenging capacity [40]. 3.4. functional properties of the flour blends the functional properties of the flour blends are presented in table 4. as observed in the previous analyses, samples with carrot and soybean flours incorporation had the best functional properties. in this case, sample 5 (80% rice flour, 10% defatted soya beans flour and 10% carrot flour) had the best wac and, second best oac and sc. sample 2 (80% rice flour, 5% defatted soya beans flour and 15% carrot flour) had the best oac and sc. the sample with only date palm flour incorporation to rice flour (80% rice flour and 20% date palm flour) had the least wac, oac and sc. sample 4 only increased the bulkiness of the composite flour. a wac of 140% had been reported for 100% wheat flour by [41] at the same time, incorporation of rice flour into wheat flour reduced the wac of the composite flour. reduction in wac in composite flours had been attributed to the less availability of polar amino acids in the flours [42] while more availability of hydrophilic constituents (polysaccharides) resulted in flours with high water absorption [41]. the decrease in wac with incorporation of datepalm flour agrees with the work of [43] who reported that high fibre and starch contents resulted in high water binding capacity. similarly, mbofung, et al. [44] reported that the ability of flour to absorb water had a significant correlation with its starch content. wac is essential in bulking and consistency of products, as well as in baking application [45]. the oac of wheat flour was reported to be 146% [41]. sample 5 had the same oac as wheat flour table 4, while the oac of sample 2 was 155.33%. samples 2 and 5, therefore, had oac around that of 100% wheat flour. other samples aside 2 and 5, had oac significantly (p ≤ 0.05) lower than 146%. the oac of rice flour was 123.57%, and, samples 4 and 6 with date palm inclusion had very low oac. presence of high fat had been reported to adversely affect oac, while food with high oac are better for flavour retention, improve palatability, and enhanced shelf life [41, 46]. swelling capacity (sc) is the ability of a sample to absorb water undisturbed under a room temperature. the swelling power shows the degree of the water absorption of the starch granules in the flour [47]. supplementation of the composite flour with carrot and soy bean flours resulted in high swelling capacity whereas samples with date palm flours had lower swelling capacity than rice flour. the sample with date palm flour incorporation had the best solubility power. sample 4 (80% rice flour and 20% date palm) had the highest solubility power of 13.10% while the control sample, rice flour, had the least value of 1.60%. samples 2 and 4 also had foaming capacities similar to 12.92% reported for 100% wheat flour by chandra, et al. [41]. increase in the protein content (as demonstrated by the increase in soybean content of some of the samples from 5 to 15%) could be responsible for increasing foaming capacity. table-4. functional properties of rice flour and flour blends. s/n bd g/ml wac % oac % sol % sc % fc % lg % 1 1.03±0.000b 78.34±0.390d 123.57±0.00d 1.60±0.000e 259.83±0.046d 5.64±0.073f 2.00±0.000 2 0.66±0.000f 86.67±0.000c 155.33±0.680a 7.60±0.000c 294.00±0.000a 12.96±0.000d 2.00±0.000 3 0.91±0.095c 89.57±0.045b 131.88±0.010c 6.50±0.000d 262.00±0.000c 18.12±0.051c 2.00±0.000 4 1.21±0.006a 71.76±0.255f 100.29±0.005f 13.10±0.000a 234.80±0.000f 11.37±0.045e 2.00±0.000 5 0.73±0.000e 101.45±0.000a 146.32±0.165b 7.80±0.000c 274.20±0.000b 20.35±0.020b 2.00±0.000 6 0.80±0.000d 76.59±0.100e 112.86±0.030e 11.20±0.000b 244.20±0.000e 29.62±0.005a 2.00±0.000 note: values represent means of triplicate (n=3). values with the same alphabet along the same column are not significantly different (p>0.05) keys: 1: 100% rice flour; 2: 80% rice flour, 5% defatted soya beans flour and 15% carrot flour 3: 80% rice flour, 15% defatted soya beans flour and 5% carrot flour; 4: 80% rice flour and 20% date palm flour, 5: 80% rice flour, 10% defatted soya beans flour and 10% carrot flour 6: 80%rice flour, 5%defatted soya beans flour, 5%carrot flour and 10% date palm flour bd: bulk density, wac: water absorption capacity, oac: oil absorption capacity, sol: solubility, sc: swelling capacity, fc: foaming capacity, lg: least gelation. while sample 4 with 20% date palm flour incorporation had considerable foaming capacity, the combination of date palm with soybean had astronomically increased the foaming capacity of the sample. the control sample has the least foaming capacity of 5.64%. agriculture and food sciences research, 2019, 6(1): 134-144 142 © 2019 by the authors; licensee asian online journal publishing group there was no significant (p > 0.05) difference in least gelation capacity of the flour blends. all the composite flour samples gelled at 2%. a least gelation capacity (lgc) of 8% was reported for 100% wheat flour [41]. lgc was defined as the lowest protein concentration at which gel remained in the inverted tube. samples with a lower lgc has been shown to have better protein gelling ability [48] and also enhance the swelling capacity of the sample [42]. it can be therefore inferred that the samples all had good lgc (the values are lower than values reported for 100% wheat flour). the bulk density of flour is the density measured without the influence of any compression [41]. a bulk density of 0.762 g/cc was reported for 100% wheat flour according to chandra et al. (2015). the samples with bd closest to that of 100% wheat flour are samples 5 (-0.03), sample 6 (+0.04), sample 2 (-0.10) and sample 3 (+0.14) in that order. on the other hand, samples 1 (100% rice flour) and 4 (80% rice flour and 20% date palm flour) had the highest bd. flour samples with high bd are good as thickeners, while those with lower bd are better for complementary foods. 3.5. pasting property of the flour blends the pasting property of the flour blends is presented in table 5. there was significant (p ≤ 0.05) difference in the pasting temperature of the flour blends. the pasting temperature is an indication of the minimum temperature required to cook or gelatinize the flour [49]. in general, sample 4 and 5 had the highest and same pasting temperature of 92.05 o c. the control sample has the least pasting temperature of 88.75 oc. pasting temperature is the temperature at which the first detectable increase in viscosity is measured and it is an index characterized by the initial change due to the swelling of starch [50]. table-5.pasting property of rice flour and the flour blends. sample pv cp t cp bd cp fv cp sb cp pat min pt oc 1 2543.00 2145.00 398.00 4025.00 1880.00 5.93 88.75 2 1520.00 1429.00 91.00 2622.00 1193.00 6.13 90.45 3 1186.00 1155.00 31.00 2261.00 1106.00 6.87 91.20 4 1249.00 1192.00 57.00 2261.00 1069.00 6.20 92.05 5 1282.00 1249.00 33.00 2374.00 1125.00 6.40 91.30 6 1312.00 1250.00 62.00 2354.00 1104.00 6.33 92.05 keys: 1: 100% rice flour, 2: 80% rice flour, 5% defatted soya beans flour and 15% carrot flour 3: 80% rice flour, 15% defatted soya beans flour and 5% carrot flour, 4: 80% rice flour and 20% date palm flour, 5: 80% rice flour, 10% defatted soya beans flour and 10% carrot flour 6: 80%rice flour, 5%defatted soya beans flour, 5%carrot flour and 10% date palm flour pv; pasting viscosity, t: trough, bd: breakdown, fv: final viscosity, sb: setback, pat: pasting time, pt: pasting temperature the results of the pasting characteristics indicate that the higher level of defatted soybean flour, carrot flour and date palm flour reduced the peak viscosity (pv), trough (t), break down viscosity (bd), final viscosity (fv), and setback viscosity (sb) of composite flour. this is due to the existence and interaction of components like fat and protein from soybean flour with carrot flour and date palm flour that decrease the viscosity [51]. in general, the viscosity of composite flour was lower than rice flour. the pv of composite flours ranged from 1186.00 cp to 2543 cp. it was found that the peak viscosity decreased as defatted soybean flour supplementation increased, but the ratio of carrot and date palm flours only had little effect on peak viscosity of composite flours. awolu, 2018 reported 3290.50 rvu – 5232.50 rvu for optimized based flour. the differences in the starch and protein composition in the flours could affect pasting viscosity and properties [52, 53]. the peak viscosity attained during the heating portion of tests indicates the water binding capacity of starch mixture. this often correlates with final product qualities [54] found that starch from yam (dioscorea alata) varieties with higher peak viscosities produced pounded yam with good textural quality whereas the d. alata varieties, with lower peak viscosities, did not give pounded yam of the acceptable textural quality. breakdown measures the ease with which the swollen granules can be disintegrated [49]. the break down ranged between 31.00 cp and 398.00 cp. the lower breakdowns (bd) were found in composite flours, as compared with the rice flour. the breakdown viscosity also decreases as soybean flour level increases in composite flour. the final viscosity (fv) indicated the re-association of starch granules especially amylose during cooling time after gelatinization and the formation of gel network [55]. the final viscosity ranged between 2261.00 cp and 4025 cp. setback value has been reported to correlate with ability of starches to gel into semi solid paste. the setback value ranged between 1104.00 to 1880.00 cp. the control sample had the highest set back of 1880.00cp. the lower setback viscosity was observed in samples 3 and 6 and it can be associated with increase in soybean flour level. lower set back is an indication of starch stability [3]. also the peak time ranged between 5.93mins to 6.87mins. the control sample had the least peak time of 5.93 while the flour that was complemented with much defatted soya beans had the highest peak time of 6.87mins. it is observed that soya beans is responsible for the increase in the peak time. 4. conclusion this present study showed that all the flour blends exhibited potent antioxidant property and increased the nutritional and functional quality of the flour, but amongst all, carrot exhibited the highest potency. this study has shown that nutritious gluten free flour can be produced from blends of local products such as rice, soya beans, carrot and date palm flour. the low nutritional quality of cereals can be improved through supplementation with blends of defatted soya beans and carrot flour and date palm flours. date palm has great agriculture and food sciences research, 2019, 6(1): 134-144 143 © 2019 by the authors; 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[55] f. e. ortega-ojeda, h. larsson, and a.-c. eliasson, "gel formation in mixtures of high amylopectin potato starch and potato starch," carbohydrate polymers, vol. 56, pp. 505-514, 2004. available at: https://doi.org/10.1016/j.carbpol.2004.03.021. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://dx.doi.org/10.1155/2015/165457 57 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 57-65, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.57.65 © 2019 by the authors; licensee asian online journal publishing group prediction of climate change effects on plantain yield in ondo state, nigeria olotu yahaya1 atanda e.o.2 rodiya a.a.3 okafor m.c.4 ( corresponding author) 1,2,4department of agricultural engineering & bio-environmental, auchi polytechnic, auchi, nigeria. 3department of agricultural engineering, federal polytechnic, ado, ado-ekiti, nigeria. abstract this study investigated the effects of climate change on plantain (musa spp) for three agro-ecological zones (aezs) in ondo state. climate projections of six selected general circulation models (gcms) under climate scenarios-representative concentration pathways (rcp 4.5) were applied for different periods; baseline (1975–2005); future periods 2035–2065 [2050s] and 2070–2100 [2080]). the results of regression analysis showed a positive relationship between precipitation and plantain yield, while maximum (tmax) and minimum temperature (tmin) indicated no significant relationship with plantain yield at p˂ 0.01 in all the aezs. the output of trend analysis indicated an increase in tmax of 0.046oc/year for ondo north agro-ecological zone (onaez), while ondo south agro-ecological zone (osaez) has the lowest increment of 0.003oc/year. tmin for ondo central agro-ecological zone (ocaez) increased by 0.007oc/year and decreased with 0.004oc/year and 0.030oc/year for onaez and osaez. however, analysis of precipitation events in the study areas from 1975-2005 showed that ocaez received the highest increase of 7.47 mm/year and decreased by 13.48 mm/year and 2.84 mm/year for onaez and osaez respectively. largest plantain yield reduction compared to the control-period for cccma model was -30.3% and -38.1% for the 2050s and 2080s whereas ichec model predicted an average lowest reduction of -7.5% and -12.5% for the short time and long periods in onaez. in osaez, plantain yield decreases varied from -6.3% to -8.4% for cnrm model, -6.1% to 6.7% (mpi) and -36.1% to -37.7% for cccma. in conclusion, overall climate change simulations in ocaez showed that projected climate may have relatively small negative effects on plantain yield compared to onaez and osaez respectively. keywords: general circulation models, rcp 4.5, climate change, plantain, precipitation, maximum temperature, minimum temperature, aez, ondo state citation | olotu yahaya; atanda e.o.; rodiya a.a.; okafor m.c. (2019). prediction of climate change effects on plantain yield in ondo state, nigeria. agriculture and food sciences research, 6(1): 57-65. history: received: 29 january 2019 revised: 5 march 2019 accepted: 15 april 2019 published: 11 june 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: this study contributes to the existing literature by investigating the effects of climate change on plantain (musa spp) for three agro-ecological zones (aezs) in ondo state, nigeria. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 58 2. study area ......................................................................................................................................................................................... 58 3. results and discussion ................................................................................................................................................................... 60 4. conclusion ......................................................................................................................................................................................... 64 references .............................................................................................................................................................................................. 65 http://www.asianonlinejournals.com/index.php/aesr/article/view/1836 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.57.65&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.57.65&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2019, 6(1): 57-65 58 © 2019 by the authors; licensee asian online journal publishing group 1. introduction climate change due to increased ‘greenhouse gases’ atmospheric concentrations is expected to have an important impact on the different economy (e.g. agriculture, forestry, energy consumptions, tourism, etc.) [1]. changes in the climate system and land cover have important consequences on regional and global water resources management and conservation [2]. in particular, for agriculture, such a change in climate may have significant impacts on crop growth and yield, since these are largely determined by the weather conditions during the growing season [1]. according to robinson [3] plantain is the fourth most important starchy staple after cereals, cassava and yam and based on the food value, the second most expensive starchy staple in the urban market after yam. extreme climate events, such as high-temperature stress, drought, and flooding, could result in a severe reduction in plantain production. in nigeria, banana and plantain farming sector are small in relative to its contribution to the national gross domestic product, and yet remains significant with respect to food production, rural employment and livelihoods particularly in the south-western region of the country. plantain production in these regions is during the wet season and the dry mostly under rainfed cultivation. the optimal temperature for growing plantain is 28°c, from 28oc to 20°c, growth will gradually slow down and will become negligible around 16-18°c [4]. plantain needs a lot of water. it should get around 200 mm per month or effective rainfall between 1650-1700 mm throughout its life cycle [4]. however, considering the increasing pressure that climate change may likely place on plantain cultivation; it is not yet clear how future precipitation and air temperature patterns will change and how such changes will affect plantain water requirements, irrigation water demands, and yields. however, current studies on this subject matter are still very few in nigeria, but more climate change studies had been conducted on cereal and tuber crops. global circulation models (gcms) are the appropriate tools currently employed in providing climate projections of both present-day and distant future climate variables [5]. gcms outputs cannot be applied directly because their current resolution is too large to be used on local scales. based on this, the outputs of global climate models often subjected to downscaling processes in order to be compatible with the local scales. there are two types of downscaling approaches; dynamic and statistical downscaling methods. dynamic techniques, often viewed as a miniature global climate model they have not gained popularity because they tend to use complex processes and approaches in order to capture local-scale variations [6]. statistical methods are mostly used in climate studies because they are cheap to apply and provide site-specific climate information [7]. however, this climate change study is based on the latest greenhouse gas emission scenarios called representative concentration pathways (rcps), consisting of rcp 2.6, 4.5, 6.0 and 8.5; measured in watt per metre square ((wm-2) [8]. furthermore, most of the previous climate change studies in nigeria were investigated based on the earlier version of climate change scenarios (a1, a2, b1, and b2) from fourth report on special report on emission scenario in the intergovernmental panel on climate change (ipcc). therefore, this current study will be one of the few climate change kinds of research using the newly released rcp emission scenarios in nigeria. in this study, the effects of climate change on plantain yield using an ensemble set of six gcms under rcp 4.5 for the future periods 2035-2065(the 2050s) and 2070-2100 (2080s) relative to baseline period 1975-2005 is investigated for the agro-ecological zones (aezs) in ondo state. 2. study area 2.1. location of study area ondo state is one of the states in the south-western region of nigeria. it has a total population of 3,440,024 of which 65% resides in fertile areas. it has a density of 236 persons per square meter and land square area of 15,500km2 [9]. the state has eighteen local government areas which are divided into three geographic zones and population concentration as follows: ondo-north with a population of 1,064,900; ondo-south has a population of 1,054,675 and population of 1,320, 449 ondo central [9]. generally, agriculture is the mainstay of the economy, and the chief products are cotton and tobacco from the north, cacao from the central part, and rubber and timber (teak and hardwoods) from the south and east; palm oil, kernels, banana, and plantain are cultivated throughout the state. other crops include rice, yams, corn (maize), coffee, cassava (manioc), vegetables, and fruits. the study area is bounded by the states of kwara and kogi on the north, edo on the east, delta on the southeast, and osun and ogun on the west and by the bight of benin of the atlantic ocean on the south. ondo state includes mangroveswamp forest near the bight of benin, tropical rain forest in the centre part, and wooded savannah on the gentle slopes of the yoruba hills in the north. fig.1 shows the map of ondo state indicating the three agro-ecological areas under study. 2.2. baseline climate and plantain yield historical monthly climate data (maximum temperature, minimum temperature, precipitation,) for 1975-2005 were taken from the cru ts2.1 database through the department of agro-climatological, ministry of agriculture & natural resources, ondo-state, nigeria with a spatial resolution of 30 arc-minutes. baseline plantains (1975– 2005) were collected from ondo state ministry of agriculture and natural resources. plantain yield was collected as the fiscal year basis, such as 1975–1976, 1976–1977, etc. then, these fiscal year data were converted to yearly data, for example, 1975–1976 was considered as 1976. the plantain yield data are consistent with the statistical data from the food and agriculture organization of nigeria [11]. http://www.britannica.com/place/nigeria http://www.britannica.com/place/kwara http://www.britannica.com/place/kogi http://www.britannica.com/place/edo-state-nigeria http://www.britannica.com/place/delta-state-nigeria http://www.britannica.com/place/osun http://www.britannica.com/place/ogun-state-nigeria http://www.britannica.com/topic/yoruba agriculture and food sciences research, 2019, 6(1): 57-65 59 © 2019 by the authors; licensee asian online journal publishing group figure-1. shows the map of ondo state indicating the three agro-ecological areas. source: osmhud [10]. 2.3. statistical downscaling model statistical downscaling model (sdsm) was applied to simulate 31-year historical dataset (1975-2005) from the study regions using different general circulation models (gcms) and project future climate (2035-2065 and 20702100) under representative pathway concentration (rcp 4.5). the climate models included canadian centre for climate modeling & analysis (cccma), max planck institute for meteorology (mpi), met office hadley centre (mohc), and model for interdisciplinary research on climate (miroc), irish centre for high-end computing (ichec) and national centre for meteorological research (cnrm). in this study, these models were used to produce climate variables as one stochastic set of data for the baseline period and for future periods under a climate change scenario (rcp 4.5). the outputs of simulated climate variables were used to develop climate-plantain model useful to predict the climate change effects of future climate on plantain reduction yield in the three agro-ecological zones. table 1 and table 2 show the study regions and selected gcms respectively. table-1. categorization of agro-ecological zone in ondo-state. s/n osaez ocaez onaez 1 odigbo akure south akoko n.w 2 okitipupa akure north akoko n.e 3 irele ileoluji/okeigbo akoko s.w 4 ilaje ifedore akoko s.e 5 ese-odo ondo-east owo 6 idanre ondo-west ose source: odsma [9]. table-2. gcms selected for the research study. models emission scenarios spatial resolution cccma rcp 4.5 48×96 cells, 3.750 ×3.750 mpi rcp 4.5 96×192 cells, 1.90 ×1.90 mohc rcp 4.5 88×176 cells, 2.00 × 2.00 miroc rcp 4.5 67× 134 cells, 1.120 × 1.120 cnrm rcp 4.5 64×128 cells, 2.80 ×2.80 ichec rcp 4.5 60 × 120 cells, 2.90 × 2.90 source: soden and held [5]. 2.4. data analysis 2.4.1. mann kendall and sen’s slope analysis mann kendall (m-k) test is a non-parametric statistical test widely used for the analysis of the trend in climatologic and other fields of science and engineering. one advantage of this test is that the data need not conform to any particular distribution [12]. second, the test has low sensitivity to abrupt breaks due to inhomogeneous time series [13]. using m-k test, the null hypothesis h0 assumes that there is no trend (the data is independent and randomly ordered), the observations of xi are randomly ordered in time, against the alternative agriculture and food sciences research, 2019, 6(1): 57-65 60 © 2019 by the authors; licensee asian online journal publishing group hypothesis, h1 which indicates increasing or decreasing trends. the estimation technique procedure for m-k test applies the time series of n data points and xi and xj as two subsets of data where i = 1,2,3,…, n-1 and j = i+1, i+2, i+3, …, n as indicated in equation 1. the data values are evaluated as an ordered time series. each data value is compared with all subsequent data values. if a data value from a later time period is higher than a data value from an earlier time period, the statistic s is incremented by 1. on the other hand, if the data value from a later time period is lower than a data value sampled earlier, s is decremented by 1 as expressed in equation 2. the net result of all such increments and decrements yields the final value of s [14]. (1) = (2) where xj and xi are the annual values in years j and i, j > i, respectively [15]. to derive an estimate of the slope s, the slopes of all data pairs are calculated as follows: 2.5. climate-crop yield relationship the impact on climate change on plantain (musa spp) yield was investigated and evaluated using developed newly empirical-statistical models of current and future time periods. this model does not attempt to capture details of plant physiology or crop management; they do capture the net effect of the entire range of processes by which climate affects yields, including the effects of poorly modeled processes [16]. correlation coefficient and multivariate regression analyses were performed to determine the climate-crop yield relationship using the statistical package for social sciences (spss). pearson’s correlation coefficient was used to measure the strength of the association between crop yield and climatic variability [16]. this produced a linear association. the range of correlation coefficients is -1 to +1. the complete dependency between two variables is expressed by either -1 or +1, and 0 represents the complete independence of the variables as shown in equation 3. a correlation coefficient is mathematically performed as follows: (3) the x represents the independent (climate) variable and y (plantain yield) represents the dependent variable. multivariate regression analysis of climate and crop yield was performed to estimate the percentage of the response of variations (plantain yield) from the predictor variables (minimum and maximum temperature and precipitation). baseline plantain yield was estimated using a regression model in equation 4, while equations 5,6,7 8 were used to estimate projected plantain yields for periods 2035-2065 and 2070-2100 under cccma and mohc models respectively. the multiple linear regressions with first differences in yield (∆yield) as the response variable, and first differences of minimum temperature (∆tmin), maximum temperature (∆tmax) and precipitation (∆ppt) as predictor variables. (4) (5) (6) (7) (8) 3. results and discussion 3.1. current climate and plantain yield 3.1.1. temperature and precipitation trends current minimum and maximum temperature from 1975 to 2005 for the three agro-ecological zones (onaez, osaez, and ocaez) in ondo-state were analyzed using non-parametric mann-kendall and sen’s slope estimates. the results of the trend analysis are showed in table 3. the rate of increase in maximum temperature was highest in onaez with 0.046oc/year while ondo south agro-ecological zone (osaez) has the lowest increment of 0.003oc/year. similar results were obtained by soden and held [5].this region is characterized as swamp rainforest zone with high rainfall duration, intensity, and depth. the outputs of sen’s slope estimates for minimum temperature (tmin) followed a different pattern. the tmin for ocaez increased by 0.007oc/year where the remaining two regions (onaez and osaez) decreased by 0.004oc/year and 0.030oc/year as indicated in table 3. the maximum temperature increased across the studied agro-ecological zones indicating warming throughout the period of consideration figures 2-4. in addition, an increasing trend in maximum air temperature was found in ondo state [17]. agriculture and food sciences research, 2019, 6(1): 57-65 61 © 2019 by the authors; licensee asian online journal publishing group table-3. statistical calibration for minimum and maximum temperature for three agro-ecological zones. study regions tmax det.corr corr. sig. val tmin det. corr corr. sig. val aez sen's slope (mm) r² r p-value sen's slope (mm) r² r p-value onaez 0.046 0.151 0.388 0.031* -0.004 0.020 -0.390 0.833b osaez 0.003 0.020 0.045 0.812b -0.030 0.044 -0.210 0.256b ocaez 0.004 0.001 0.025 0.890b 0.007 0.006 0.079 0.674b source: simulation output, 2018. figure-2. annual minimum and maximum temperature trends at onaez from 1975-2005. source: simulation output, 2018. figure-3. annual minimum and maximum temperature trends at ocaez from 1975-2005. source: simulation output, 2018. figure-4. annual minimum and maximum temperature trends at osaez from 1975-2005. source: simulation output, 2018. the analysis of precipitation events in the study areas from 1975-2005 showed that ocaez received the highest increase of 7.47 mm/year table 4, figure 5. however, total annual precipitation at onaez and osaez decreased by 13.48 mm/year and 2.84 mm/year. agriculture and food sciences research, 2019, 6(1): 57-65 62 © 2019 by the authors; licensee asian online journal publishing group table-4. statistical metrics for annual precipitation for the agro-ecological zones. study regions sen's slope (mm) r² r p-value alpha-value sig. onaez -13.475 0.138 -0.409 0.022* 0.050 yes osaez -2.837 0.067 0.314 0.086b 0.050 no ocaez 7.470 0.016 -0.126 0.500b 0.050 no source: simulation output, 2018. figure-5. annual precipitation trends at onaez-osaez-ocaez from 1975-2005. source: simulation output, 2018. 3.2. plantain (musa spp) yield trend plantain yield trend showed fluctuation over the period of consideration in all the studied agro-ecological zones. the yield increase of 0.039 ton/ha yearly is highly significant (p< 0.05) in osaez again, the yield reduced by -0.022 ton/ha in onaez and -0.033 ton/ha in ocaez table 5. these reductions were not significant (p > 0.05) within the baseline period (1975-2005). figure 6 depicts the plantain trend analysis. the trend of plantain yield in osaez is similar to the production pattern in nigeria for more than two decades with the country harvested 2.103 million tons from 389,000 ha [18]. however, the country experienced a great depression in plantain production between 1987–1988 and 1990 [18]. this reduction pattern is very similar to plantain yield trends in onaez and ocaez respectively. table-5. statistical metrics for plantain yield trends for the agro-ecological zones. study regions sen's slope (ton/ha) r² r p-value alpha-value sig. onaez -0.022 0.044 -0.211 0.256 0.050 no osaez 0.039 0.136 0.368 0.042* 0.050 yes ocaez -0.033 0.108 -0.329 0.071 0.050 no source: simulation output, 2018. figure-6. plantain yield trends in three agro-ecological zones from 1975-2005. source: simulation output, 2018. 3.3. plantain yield-climate simulation relationship between the control-base plantain yield and climate variables in all the three agro-ecological zones were performed using correlation and regression models. the results of sensitivity analysis in table 6 reveal that positive and strong correlation values (r = 0.80; 0.63 and 0.68) for onaez, osaez, and ocaez respectively. shahid [14] reported the same results in bangledesh. conversely, an estimated relationship between precipitation and plantain yield is significant (p < 0.01) in all the study regions. these results indicate that an increase in annual precipitation could lead to an increase in plantain yield per hectare in all the agro-ecological zones (onaez, ocaez, and osaez). coefficient of determination between plantain yields and minimum temperature for three agro-ecological is very weak with r2 = 0.005(onaez), r2 = 0.042 (osaez) and r2 = 0.053 (ocaez) with insignificant association at 95% confidence interval (p < 0.05). while testing the effects of annual maximum temperatures, no significant relationship was observed in the yield of plantain in all the studied agro-ecological agriculture and food sciences research, 2019, 6(1): 57-65 63 © 2019 by the authors; licensee asian online journal publishing group zones. however, the coefficient of maximum temperature was very weak with r2= 0.07, r2 =0.04 and 0.01 for onaez, osaez and ocaez table 6. therefore, plantain yield decreases with increasing maximum and minimum temperatures. the findings are in agreement with the studies [2, 3]. table-6. statistical for plantain yield-temperature-precipitation for the aezs(1975-2005). study regions precipitation tmax tmin aez r² r std.dev p-value r² r std.dev pvalue r² r std.dev pvalue onaez 0.65 0.80 299.6 0.00001* 0.005 0.07 1.00 0.699b 0.07 0.26 1.07 0.165b osaez 0.40 0.63 216.4 0.00001* 0.042 0.21 1.27 0.268b 0.04 0.20 0.70 0.277b ocaez 0.46 0.68 252.8 0.00001* 0.053 0.23 1.19 0.149b 0.01 -0.11 1.23 0.510b source: simulation output, 2018. 3.4. estimation of climate change impacts on plantain yield direct impacts of climate change on plantain yield for the three agro-ecological districts were estimated using a developed multiple linear regression model under control period (1975-2005) and the future periods (the 2050s and 2080s). average baseline plantain yields considered were 8.5 tons/ha, 8.6 tons/ha and 8.7 tons/ha for onaez, osaez and ocaez respectively. the results of multivariate regression analysis reveal the estimated potential impacts of climate change on plantain yield across the aezs as shown in tables 7-9. simulation outputs indicate that all the gcms predicted future reductions in plantain yield in all the three agro-ecological zones. cccma model has the highest decrease in plantain yields. yield reductions of 2,567.0 kg/ha and 3,085.5 kg/ha were predicted for the time periods of the 2050s and 2080s in onaez table 8 whereas plantain yield decrease of 3,068.5 kg/ha and 3,204.5 kg were projected in osaez table 9 using cccma model. smit and skinner [6] showed similar observation with cccma simulations however, for all the future time periods (the 2050s and 2080s) plantain yield reduction is marginally predicted in ondo central agro-ecological zone table 7. largest plantain yield reduction percentage compared to the control-period across 1975-2005-2065 for cccma model was -30.3% and -38.1% for the 2050s and 2080s respectively in onaez table 8. however, ichec model predicted an average lowest reduction of -7.5% and 12.5% for a short time and long periods. table-7. projected climate change effects on plantain yield (%) relative to baseline period for ocaez. gcms plantain yield reduction (%) plantain yield reduction (kg/ha) period (2080s) period (2050s) period (2080s) period (2050s) models 2035-2065 2070-2100 2035-2065 2070-2100 cnrm 0.3 0.7 25.5 59.5 mpi 0.6 0.8 51.9 65.5 miroc 0.7 1.0 62.9 82.5 mohc 0.7 1.0 57.0 80.8 cccma 1.0 1.2 86.7 102.0 ichec 0.1 0.2 2.6 17.0 source: simulation output, 2018. table-8. projected climate change effects on plantain yield (%) relative to baseline period for onaez. gcms plantain yield reduction (%) plantain yield reduction (kg/ha) period (2080s) period (2050s) period (2080s) period (2050s) models 2035-2065 2070-2100 2035-2065 2070-2100 cnrm 14.5 22.4 1232.5 1904.0 mpi 19.4 24.2 1649.0 2057.0 miroc 3.6 33.9 306.0 2881.5 mohc 35.1 33.9 2983.5 2881.5 cccma 30.2 36.3 2567.0 3085.5 ichec 7.3 12.1 620.5 1028.5 source: simulation output, 2018. table-9. projected climate change effects on plantain yield (%) relative to baseline period for osaez. gcms plantain yield reduction (%) plantain yield reduction (kg/ha) period (2080s) period (2050s) period (2080s) period (2050s) models 2035-2065 2070-2100 2035-2065 2070-2100 cnrm 6.3 8.4 535.5 714.0 mpi 6.1 6.7 518.5 569.5 miroc +1.8 17.1 153.0 1453.5 mohc 1.5 -1.9 127.5 161.5 cccma 36.1 37.7 3068.5 3204.5 ichec 1.8 2.4 153.0 204.0 source: simulation output, 2018. in osaez region, a substantial reduction in plantain yield is estimated by 2050s and 2080s compared with baseline yield. yield decrease varied from -6.3% to -8.4% for cnrm model, -6.1% to -6.7% (mpi), -1.5% to -1.9% (mohc), -36.1% to -37.7% (cccma), -1.8% to -2.4% (ichec). however, an increase of plantain yield of 1.8% is estimated by the miroc model in 2050s and reduction of -17.1% in 2080s as indicated in figure 7b. it could be deduced that plantain will largely decrease in osaez and onaez figure 7a and 7b. a possible explanation for this could be related to the projected increase in minimum and maximum temperature in the regions. higher agriculture and food sciences research, 2019, 6(1): 57-65 64 © 2019 by the authors; licensee asian online journal publishing group temperature influences increase in evapotranspiration (eto) and increase plantain water requirement. in ondo central agro-ecological zone (ocaez), predicted outputs from selected gcms indicate decrease in plantain (musa.spp) yield under two future scenarios figure 7c. however, miroc and mohc simulations indicate equal reduction values of 0.6% and 0.8%; ichec predicted least decrease of 0.02% and 0.2% for the periods the 2050s and 2080s respectively figure 7c. overall simulations of impacts of climate change in ocaez showed that projected climate will have relatively small negative effects on plantain yield compared to the results predicted in onaez and osaez respectively. figure-7a. predicted plantain yield (%) for period (2035-2065) and (2070-2100) relative to 1975-2005 over onaez. source: simulation output, 2018. figure-7b. predicted plantain yield (%) for period (2035-2065) and (2070-2100) relative to 1975-2005 over osaez. source: simulation output, 2018. figure-7c.predicted plantain yield (%) for period (2035-2065) and (2070-2100) relative to 1975-2005 over ocaez. source: simulation output, 2018. 4. conclusion the estimated relationship between baseline precipitation and plantain yield is significant (p < 0.01) in all the studied regions indicating an increase in seasonal precipitation could lead to increase in plantain yield per hectare in all the agro-ecological zones (onaez, ocaez, and osaez). plantain yield was reduced under projected future climate (the 2050s and 2080s) and the decrease was different based on different gcms outputs. cccma model has the highest decrease in plantain yields in all agro-ecological zones compared to other global climate models. this could be related to the projected increase in minimum and maximum temperature which influence evapotranspiration (eto) and increases plantain water requirement. in osaez region, a substantial reduction in plantain yield is estimated by 2050s and 2080s compared with baseline yield. overall simulations of impacts of climate change in ocaez showed that projected climate may likely have relatively small negative effects on plantain yield compared to the results predicted in onaez and osaez respectively. agriculture and food sciences research, 2019, 6(1): 57-65 65 © 2019 by the authors; licensee asian online journal publishing group references [1] j. hanson, r. ruedy, m. sato, and k. lo, "global surface temperature change. rev. change impacts on agriculture: an integrated assessment, 1990-2080," philosophical transactions of the royal society b, vol. 360, pp. 2067–2083, 2010. [2] p. g. oguntunde, b. j. abiodun, and l. gunnar, "spatial and temporal temperature trends in nigeria, 1901-2000," meteorology and atmospheric physics, vol. 118, pp. 95-105, 2012. available at: https://doi.org/10.1007/s00703-012-0199-3. [3] j. c. robinson, systems of cultivation and management. in: gowen s (ed), bananas and plantains vol. 10. london: chapman and hall, 2015. [4] e. n. s. lionell, "improved plantain productionthe technical centre for agricultural and rural co-operation (cta)," journal of environmental research letters, vol. 3, pp. 45-69, 2015. [5] b. j. soden and i. m. held, "an assessment of climate feedbacks in coupled ocean–atmosphere models," journal of climate, vol. 19, pp. 3350–3369, 2013. [6] b. smit and m. w. skinner, "adaptation options in agriculture to climate change," mitigation and adaptation strategies for global change, vol. 7, pp. 80-85, 2009. 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[17] o. a. ogunrayi, f. m. akinseye, v. goldberg, and c. bernhofer, "descriptive analysis of rainfall and temperature trends over akure, nigeria," journal of geography and regional planning, vol. 9, pp. 195-202, 2016. available at: https://doi.org/10.5897/jgrp2016.0583. [18] fao, the state of food and agriculture. rome: food and agriculture organization of the united nations, 2006. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 71 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 71-78, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.71.78 © 2019 by the authors; licensee asian online journal publishing group effect of calcium chloride dipping and beeswax coating on the shelf life and quality of nectarine (prunus persica (l.) batsch var. nucipersica) fruits getaneh seleshi1 kebede woldetsadik2 mulualem azene3 ( corresponding author) 1department of temperate and indigenous fruits research, holeta agricultural research center, holeta, ethiopia. 2college of agriculture and environmental sciences, school of plant sciences, haramaya university, dire dawa, ethiopia. 3college of agriculture and veterinary science, department of horticulture, ambo university, ambo, ethiopia. abstract this research was conducted to evaluate the effect of cacl2 dipping and beeswax (bw) coating on the shelf life and quality of nectarine fruits. the experiment was conducted under holeta condition during the off-season of 2018. fruits of ‘89-16n’ nectarine variety were harvested from harc orchard. the treatment consisted of a combination of four levels of cacl2 (0%, 1.5%, 3.0% and 4.5%) and three levels of bw application (0%, 3% and 6%). the experiment was arranged in completely randomized design with factorial arrangement in three replications. nectarine fruits were stored at ambient condition after being treated with the different treatment combinations of bw and cacl2. various physico-chemical parameters were assessed. the result revealed that the plw of nectarine fruits was significantly reduced. the tss and ph values as well as ta and aa contents were better maintained in cacl2 dipping with bw coatings compared with the control. the highest percentage of marketable fruits with the lowest decay percentage were also retained by cacl2 dipping and bw coatings. overall, the best result was consistently obtained at 3.0% bw with 4.5% cacl2 treated fruits for most of the parameters assessed. hence, cacl2 dipping and bw coatings, particularly 3% bw with 4.5% cacl2 treatment, could be considered for extending the shelf life and better quality of nectarine fruits. keywords: ambient condition, beeswax coating, calcium chloride, nectarine, physico-chemical, quality, shelf life. citation | getaneh seleshi; kebede woldetsadik; mulualem azene (2019). effect of calcium chloride dipping and beeswax coating on the shelf life and quality of nectarine (prunus persica (l.) batsch var. nucipersica) fruits. agriculture and food sciences research, 6(1): 71-78. history: received: 12 february 2019 revised: 15 march 2019 accepted: 19 april 2019 published: 4 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: the authors would like to thanks to holetta agricultural research center (harc), national temperate and indigenous fruits research program, researchers and staff members for their strong support and facilitation of all the necessary things to accomplish this research. besides, authors would like to acknowledge all field workers and technical assistants working in temperate fruits experimental site that helped them to get nectarine fruits used for the experiment. funding: this research is funded by the ethiopian institute of agricultural research (eiar) management. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 72 2. materials and methods ................................................................................................................................................................... 72 3. results and discussion ................................................................................................................................................................... 73 4. conclusion ......................................................................................................................................................................................... 77 references .............................................................................................................................................................................................. 77 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.71.78&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.71.78&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/120 https://orcid.org/0000-0002-3254-0533 agriculture and food sciences research, 2019, 6(1): 71-78 72 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by evaluating the effect of cacl2 dipping and beeswax (bw) coating on the shelf life and quality of nectarine fruits. 1. introduction postharvest losses in fresh fruits and vegetables are estimated in the range of 20 to 50% in developing countries [1] which are also estimated as high as 50% for perishable crops in ethiopia [2]. minimizing postharvest food losses can help to conserve resources and improve human well-being by contributing to food and nutrition security. main factors responsible for the postharvest losses of most fruits are mechanical damage, microbial spoilage and physiological deterioration [1, 3]. moisture loss is one of the main post-harvest problems that affect the quality of nectarines during long-term storage. to ensure optimum post-harvest quality, stone fruits like nectarines should be protected from excessive post-harvest moisture loss [4]. decay and the incidence of internal defects such as woolliness, pulpiness and over-ripeness are other problems that are associated with long periods of storage [5]. edible coatings with different composition have been tested and used to prolong storage life of fruits. lipids including beeswaxes are among the protective coatings used to prolong storage life of fruits [6, 7]. edible coatings or edible films contribute to enhancing the shelf life of fruits by reduction of moisture loss, solute migration and gas exchange as well as by reducing the physiological disorders. edible coatings have high potential to control browning, off flavor, microbial activity of fruits and thus extending shelf life of treated fresh commodities [8]. postharvest cacl2 application is also receiving considerable attention in recent times due to its positive effects on shelf life whilst maintaining quality of many fruits and vegetables. calcium chloride delays ripening and senescence, reduces respiration, extends shelf life, maintains firmness, and reduces physiological disorders of many fruits and vegetables [9, 10]. calcium chloride has also been reported to affect the marketing and storage of fruits through inhibition of ethylene synthesis, protein breakdown, weight loss and rotting [11]. however, there is no information on the combined effect of calcium chloride dipping and beeswax coating on the quality and shelf life of nectarines. therefore, the present study was initiated to evaluate the effect of calcium chloride dipping and beeswax coating on the shelf life and quality of nectarine fruits. 2. materials and methods 2.1. experimental site the experiment was conducted at holeta agricultural research center, which is situated at 9°00` n latitude and 38°29` e longitude at an altitude of 2400 m.a.s.l., 40 km west of addis ababa along the ambo road. the area receives a mean annual rainfall of 1100 mm and has a relative humidity of 60.6%. the main rainy season is from june to september, which accounts for 70% of the rainfall while the remaining 30% is from february to april. the average annual maximum and minimum temperatures are 22.1 oc and 6.2 oc respectively. the soil type in the area is predominantly nitosol, which is characterized by an average organic matter content of 1.8%, nitrogen 0.17%, phosphorous 4.55 ppm and potassium 1.12 meq/100 g of soil and ph of 5.24 [12]. 2.2. treatments and experimental design the treatments consisted of 4 x 3 factorial combinations of four levels of cacl2 (0%, 1.5%, 3.0% and 4.5%) and three levels of beeswax coatings (0%, 3% and 6%). there were 12 treatment combinations assigned in a completely randomized design with three replications. 2.3. experimental procedures nectarine fruits of variety ‘89-16n’ were collected at uniform firm ripe stage from 12 years old full bearing orchard of holeta agricultural research center, temperate fruits research program experimental site. hand harvesting was done and fruits were taken to the laboratory immediately after harvest. fruits were sorted for equal size, uniform color and free of disease and blemishes. the fruits were then washed with tap water to cool the field heat down, to avoid soil particles and to reduce microbial load on the fruits and then dried with muslin cloth. nectarine fruits were once dipped with 1.5%, 3.0% and 4.5% calcium chloride solutions for 15 minutes leaving the control. on the other hand, beeswax emulsions at the rates of 3% and 6% were prepared by dissolving with 90 °c heated 1000 ml water then 20 ml oleic acid was added to the molten beeswax and slowly cooled with gentle stirring [13]. for fruits that received a combination of both treatments, they were first dipped in calcium chloride solution and drained for 10 minutes before coating with beeswax. all the fruits were then stored under ambient environmental condition. one hundred sixty five fruits were allotted to each treatment. data were collected at every 5 days interval. five fruits were taken and marked from each treatment for non-destructive measurements and five fruits were taken from each treatment randomly for destructive measurements. 2.4. methodology 2.4.1. physiological loss in weight fruits samples tagged for non-destructive parameter measurements were pre-weighed by using physical balance. the physiological loss in weight were measured after each storage intervals and expressed in percentage by the following formula [14]. plw (%) = weight loss (g) weight of the fruits at the beginning of storage (g) x 100 agriculture and food sciences research, 2019, 6(1): 71-78 73 © 2019 by the authors; licensee asian online journal publishing group 2.4.2. total soluble solids the total soluble solid contents of fruits were determined by hand refractometer (0-32 °brix). the refractometer was calibrated with distilled water before use and then a drop of fruit juice from each sample were placed on the prism and readings were recorded. the total soluble solids were expressed in degree brix (°brix) [15]. 2.4.3. titratable acidity the titratable acidity of nectarine fruits was determined according to garner, et al. [16]. in a 100 ml beaker, 6 g of fruit juice with 50 ml distilled water were added and titrated against 0.1n sodium hydroxide solution to an end point of 8.2, measured with the ph meter. the milliliters (ml) of naoh used were recorded. then, the titratable acidity was calculated in terms of malic acid and was expressed as percent on pulp; acid (%) = [mls naoh used] × [0.1 n naoh] × [milliequivalent factor] grams of sample x 100 2.4.4. ascorbic acid ascorbic acid content of nectarine fruits was determined by titrimetric method using 0.5% oxalic acid, 2, 6dichlorophenol-indophenol (dcpip) solutions and ascorbic acid standard solution. the ascorbic acid content of nectarine fruit juice was computed by the following formula and the result was expressed as mg per 100 g of sample [17]; ascorbic acid = dcpip (ml)used to titrate the juice dcpip (ml) used to titrate the standard ascorbic acid x 1 mg/ml 2.4.5. decay the percentage of disordered fruits including all of the spoiled fruits that resulted from fungus, bacterial and pathogens rots were assessed with visual observation. decay loss was calculated from observation made on number of fruits infected on each day to the numbers of fruits initially stored and then percentage of decay loss was worked out by the following formula [18]; decay (%) = no_of fruits infected no− of sample fruits initially stored x 100 2.4.6. percentage of marketable fruits the percentage of marketable fruits was subjectively assessed according to the procedure of mohammed, et al. [19]. the descriptive quality attributes were determined by observing the level of decay, color, surface defects and shriveling. a 1–9 rating scales of which; 1= unsalable, 3 =unmarketable, 5 = fair, 7 = good and 9 = excellent were used to evaluate the marketable fruits. fruits receiving a rating of five and above were considered marketable, while those rated less than five were considered unmarketable. the number of marketable fruits was used as a measure to calculate the percentage of marketable fruits during storage and converted by the following formula; percentage of marketable fruits = number of marketable fruits total number of fruits x 100 2.5. data analysis the data were subjected to analysis of variance (anova) in crd with factorial arrangement to determine differences between the treatments [20]. the results were analyzed using statistical analysis system (sas) version 9.0. comparisons of the treatment means was done by the least significant difference (lsd) test at 5% significance level. 3. results and discussion 3.1. physiological loss in weight highly significant (p≤0.001) difference in the physiological loss in weight (plw) of nectarine fruits was observed due to the interaction effect of cacl2 dipping and beeswax (bw) coatings table 1. the highest rates of plw, 5.38% and 11.79%, were observed in control fruit samples while the lowest values of 2.44% and 5.47% were recorded in 6% bw with 0% cacl2 treatment after 5th and 10th days of storage, respectively. while fruits not treated with beeswax and cacl2 were discarded after 15 days of storage, the highest plw (19.30%) was recorded from 0% bw with 1.5% cacl2 among the remaining treatments, whereas treatment of 6% bw with 0% cacl2 had the lowest loss in weight. however, 6% wax combined with 3.0 and 4.5% cacl2 equally reduced the plw of fruits, which were in the range of 10.26 to 10.3%. at day 20, the plw for 3% bw with 4.5% cacl2 treated fruits was reduced nearly by half as compared to the highest plw of 27.55% for 0% bw with 3.0% cacl2 treatment. fruits not treated with beeswax as well as those treated with 3% beeswax combined with no and 1.5% cacl2 were all discarded on day 25. among the remaining treatments, the highest loss (23.36%) was observed from 6% bw with 1.5% cacl2 while the lowest plw (19.30%) was recorded in 3% bw with 4.5% cacl2 treatment. there was a general increase of plw as the storage period advanced for all of the treatments. treating nectarine fruits with 3% bw with 4.5% cacl2 kept the plw low up to the end of shelf life though 6% bw combined with 0% cacl2 treatment better reduced plw up to day 15 table 1. this result is in agreement with reports of el-badawy [21] who observed reduced weight loss in ‘florida prince’ peach treated by the combination of cacl2 and chitosan coating. weight loss of fresh fruits is mainly due to water loss because of evaporation and transpiration while dry matter is lost by respiration. hence, the lowest plw of the fruits could be related to modified atmosphere created by bw coatings and effect of cacl2 dipping on respiration, which probably lowered the rate of transpiration, water loss and oxidation reactions when the treatments are combined. however, waxed barrier must not be so complete as to block essential movement of o2 gas in to the fruit or co2 out of it kader [22]. agriculture and food sciences research, 2019, 6(1): 71-78 74 © 2019 by the authors; licensee asian online journal publishing group 3.2. total soluble solids the interaction effect of cacl2 dipping and beeswax (bw) coatings had highly significant (p≤0.001) influence on the tss content of nectarine fruits table 2. the control fruit samples had the highest tss of 10.73 °brix, whereas fruits treated with 6% bw with 4.5% cacl2 had the lowest tss of 8.4 °brix on day 5. similarly, the highest tss value was recorded in the control fruit samples while the lowest was observed in 6% bw with 0% cacl2 treatment on day 10. on day 15, tss value of 13.80 °brix was recorded in 0% bw with 3.0% cacl2 treatments while fruits treated with 6% bw combined with 0% cacl2 had 2.6 °brix less tss value. on day 25, fruits dipped in 6% bw in combination with 0% cacl2 recorded the highest tss value (13.27 °brix), which did not vary statistically with those treated with 6% bw combined with 1.5 and 4.5% cacl2. whereas, the lowest tss value (12.33 °brix) was observed both in 3% bw with 4.5% cacl2 and 6% bw with 3.0% cacl2 treated fruits. in general, there was a gradual increase in tss value of nectarine fruits for most of the treatments over time. however, cacl2 dipping and bw coatings had significantly maintained the tss of nectarine fruits higher towards the end of storage period. the increase in tss was probably due to the hydrolysis of polysaccharides and concentrated juice content because of dehydration with the passage of storage time [9, 23]. the higher tss content in control fruits might be due to faster changes of ripening resulting in breakdown of complex carbohydrates into simple sugars at a faster rate. this might also be due to high weight loss, which is mainly due to the water loss that leads to higher concentration of sugars in fruits during the storage [24, 25]. the lower respiration rate for cacl2 dipping and bw coatings might, therefore, reduce the use of metabolites resulting in slow conversion of carbohydrates to sugars. table-1. effect of calcium chloride dipping and beeswax coating on the physiological loss in weight (%) of nectarine fruits under holeta condition, 2018. note: bw: beeswax; data after ± are standard deviations (n=3); means with the same letter (s) in a column are not significantly different at p ≤ 0.05 (lsd test); ns: non-significant; ***: p ≤ 0.001. table-2. effect of calcium chloride dipping and beeswax coating on the total soluble solids content (°brix) of nectarine fruits under holeta condition, 2018. note: bw: beeswax; data after ± are standard deviations (n=3); means with the same letter (s) in a column are not significantly different at p ≤ 0.05 (lsd test); ns: non-significant; ***: p ≤ 0.001. 3.3. titratable acidity nectarine fruits showed highly significant (p≤0.001) difference in titratable acidity (ta) due to the interaction effect of cacl2 dipping and beeswax (bw) coatings up to 20 days of storage table 3. the highest ta values (1.55% and 1.42% malic acid) were recorded in 6% bw with 0% cacl2 treated fruits while the lowest ta values (1.16% and 0.68% malic acid) were observed in control fruit samples on day 5 and 10, respectively. however, at day 15 and 20, the highest ta values were maintained at 3% bw with 4.5% cacl2 treated fruits whereas fruits treated at 0% bw combined with 1.5% and 3.0% cacl2 on day 15 and 20, respectively, recorded the least (0.78%) ta value. treatment storage period (days) bw (%) cacl2 (%) 5 10 15 20 25 0 0 5.38a ±0.07 11.79a±0.39 0 1.5 5.18a±0.25 11.05b±0.10 19.30a±0.29 0 3.0 4.57b±0.09 10.28c±0.07 17.76b±0.18 27.55a±0.21 0 4.5 4.78b±0.16 9.74d±0.05 17.46b±0.29 25.47b±0.56 3 0 3.06cd±0.10 7.49f±0.26 12.35c±0.32 18.60c±0.45 3 1.5 3.32c±0.38 7.98e±0.16 12.65c±0.80 18.39c±1.04 3 3.0 2.92de±0.39 6.47h ±0.18 10.74de±0.66 16.08de±0.65 22.18b±0.26 3 4.5 2.58ef±0.15 5.76ij±0.10 9.35fg±0.93 14.08g±0.26 19.30e±0.73 6 0 2.44f ±0.34 5.47j±0.23 9.11g±0.62 14.51fg±0.50 20.00de±0.95 6 1.5 3.07cd±0.12 7.01g ±0.28 11.68cd±0.51 17.07d±0.90 23.36a±0.32 6 3.0 2.76def±0.08 5.96i±0.28 10.30ef±0.66 15.81e±0.28 21.93bc±0.78 6 4.5 2.60ef±0.13 5.98i±0.21 10.26ef±0.44 15.35ef±0.59 21.02cd ±0.11 significance bw (a) *** *** *** *** ns cacl2 (b) *** *** *** *** *** a * b *** *** *** *** *** treatment storage period (days) bw (%) cacl2 (%) 5 10 15 20 25 0 0 10.73a±0.12 13.57a±0.15 0 1.5 10.50ab±0.10 12.67b±0.23 11.80e±0.20 0 3.0 10.47b±0.12 12.73b±0.12 13.80a±0.20 13.53ab±0.12 0 4.5 9.63de±0.21 11.93c±0.12 12.47b±0.23 13.80a±0.20 3 0 9.47e±0.12 11.40d±0.20 11.73e±0.12 13.47b±0.12 3 1.5 9.20f±0.20 10.87e±0.12 12.40bc±0.20 12.93de±0.12 3 3.0 9.80cd±0.17 10.80e±0.20 11.80e±0.20 12.73e±0.12 12.80b±0.35 3 4.5 8.70g±0.10 10.37f±0.15 11.87de±0.23 12.20g±0.20 12.33c±0.12 6 0 9.40ef±0.00 9.50g±0.10 11.20f±0.20 13.27bc±0.12 13.27a±0.23 6 1.5 9.47e±0.12 10.93e±0.12 11.93de±0.12 12.67ef±0.31 13.07ab±0.12 6 3.0 10.00c±0.20 11.03e±0.15 12.13cd±0.23 12.40gf±0.20 12.33c±0.12 6 4.5 8.40h±0.20 10.33f±0.23 11.93de±0.12 13.07cd±0.12 13.00ab±0.00 significance bw (a) *** *** *** *** ns cacl2 (b) *** *** *** *** *** a * b *** *** *** *** *** agriculture and food sciences research, 2019, 6(1): 71-78 75 © 2019 by the authors; licensee asian online journal publishing group generally, the ta of nectarine fruits declined as storage period prolonged for all of the treatments. while ta value of non-treated fruits were low at all storage periods, treatments that involved cacl2 dipping and beeswax coatings, particularly 3% bw with 4.5% cacl2, maintained relatively higher ta levels. this result agreed with reports of sahar [26] who detected a decrease in acidity of nectarine fruits during 28 days of storage. during fruit ripening in storage, fruit utilizes carbohydrates and acids for energy requirement of cells and synthesis of other compounds such as flavor compounds [25, 27]. titratable acidity (ta) is often related to maturity [28]. titratable acidity of fruits is an indicator of potential storage quality, which declines gradually over the storage period [29]. the faster decrease in acidity gives rise to a faster senescence [30]. the highest reduction and lowest value of titratable acidity could, therefore, be due to higher respiration rate and related metabolic activity, which enhances the consumption of organic acids. table-3. effect of calcium chloride dipping and beeswax coating on the titratable acidity (%) of nectarine fruits under holeta condition, 2018. note: bw: beeswax; data after ± are standard deviations (n=3); means with the same letter (s) in a column are not significantly different at p ≤ 0.05 (lsd test); ns: non-significant; ***: p ≤ 0.001. 3.4. ascorbic acid highly significant (p≤0.001) variation in the ascorbic acid (aa) content of nectarine fruits was recorded due to the interaction effect of cacl2 dipping and beeswax (bw) coatings except for day 25 table 4. the lowest aa value of 8.44 mg/100 g was recorded in control fruit samples while the highest of 12 mg/100 g was observed in 6% bw + 4.5% cacl2 and 6% bw + 0% cacl2 treated fruits on day 5. however, non-significant difference between 0% bw with 1.5% cacl2 and 0% bw with 1.5% cacl2 as well as between 0% bw with 4.5% cacl2, 3% bw with 0% cacl2, 3% bw with 4.5% cacl2 and 3% bw with 4.5% cacl2 was observed on the same day. on day 10, fruits treated at 6% bw with 0% cacl2 had the highest aa value (9.78 mg/100 g) while it dropped to 4.44 mg/100 g in control fruits. whereas, fruits treated with 0% bw with 4.5% cacl2 and 6% bw with 1.5% cacl2 showed nonsignificant variation. table-4. effect of calcium chloride dipping and beeswax coating on the ascorbic acid contents (mg/100 g) of nectarine fruits under holeta condition, 2018. note: bw: beeswax; data after ± are standard deviations (n=3); means with the same letter (s) in a column are not significantly different at p ≤ 0.05 (lsd test); ns: non-significant; *: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001. the aa content of nectarine fruits in general had substantially decreased as storage period advanced irrespective of the treatments. however, relatively higher aa content of nectarine fruits was maintained with cacl2 dipping and beeswax coating, particularly 3% bw + 4.5% cacl2 treated fruits, compared with the control table 4. sahar [26] reported similar result on nectarine fruits using essential oils and davarynejad, et al. [31] on treatment storage period (days) bw (%) cacl2 (%) 5 10 15 20 25 0 0 1.16h ±0.08 0.68h±0.02 0 1.5 1.29g±0.02 1.17g±0.08 0.78d±0.07 0 3.0 1.43cde±0.03 1.23cdef±0.02 1.12c±0.00 0.78f±0.02 0 4.5 1.49ab±0.02 1.28c±0.02 1.23b±0.09 0.91e±0.04 3 0 1.36f±0.07 1.19efg±0.03 1.12c±0.00 0.92de±0.00 3 1.5 1.29g±0.02 1.20defg±0.02 1.15bc±0.07 0.92de±0.05 3 3.0 1.38ef±0.03 1.25cd±0.04 1.19bc±0.06 0.94cde±0.05 0.72±0.02 3 4.5 1.44bcd±0.01 1.36b±0.00 1.33a±0.03 1.12a ±0.00 0.75±0.03 6 0 1.55a±0.02 1.42a±0.02 1.24b±0.03 1.01b± 0.00 0.72±0.06 6 1.5 1.42def±0.03 1.19fg±0.06 1.18bc± 0.03 0.98bcd±0.03 0.69±0.03 6 3.0 1.48bc±0.02 1.24cdef±0.03 1.19bc±0.03 0.95bc±0.03 0.69±0.02 6 4.5 1.47bcd±0.00 1.25cde±0.02 1.22b±0.09 0.98bc±0.02 0.75±0.03 significance bw (a) *** *** *** *** ns cacl2 (b) *** *** *** *** ns a * b *** *** *** *** ns treatment storage period (days) bw (%) cacl2 (%) 5 10 15 20 25 0 0 8.44d±0.77 4.44d±0.77 0 1.5 9.33bcd±0.00 7.33c±0.67 4.89e±0.77 0 3.0 9.33bcd±0.00 7.56c±0.77 5.78cde±0.77 4.00d±0.00 0 4.5 10.67ab±0.00 8.89ab±0.77 7.00ab±0.00 4.00d±0.00 3 0 10.67ab±1.34 8.00bc±0.67 5.78cde±0.77 4.00d±0.00 3 1.5 8.89cd±0.77 7.78c±0.39 5.33de±0.00 4.00d±0.00 3 3.0 10.22bc±1.54 8.00bc±0.00 6.22bcd±0.77 4.00d±0.00 4.00±0.00 3 4.5 10.67ab±0.00 9.55a±0.39 8.00a±0.00 5.78a±0.77 4.44±0.77 6 0 12.00a±0.00 9.78a±0.77 7.11ab±1.54 5.33ab±0.00 4.44±0.77 6 1.5 9.78bcd±0.77 8.00bc±0.00 6.78bc±0.19 4.44cd±0.77 4.00±0.00 6 3.0 10.67ab±1.34 8.22bc±0.39 6.89abc±0.19 4.89bc±0.77 4.00±0.00 6 4.5 12.00a±0.00 8.89ab±0.38 6.22bcd±0.77 4.22cd±0.39 4.00±0.00 significance bw (a) *** *** * ** ns cacl2 (b) *** *** ** ns ns a * b *** *** *** *** ns agriculture and food sciences research, 2019, 6(1): 71-78 76 © 2019 by the authors; licensee asian online journal publishing group apricot fruits. the lower level of aa in control might be due to increased respiration, which accelerates the deteriorative oxidation reaction and loss of ascorbic acid [32, 33]. ascorbic acid is an important nutrient and is very sensitive to degradation due to its oxidation compared to other nutrients during food processing and storage [34]. 3.5. decay highly significant (p≤0.001) difference on the occurrence of nectarine fruit decay was observed due to the interaction effect of cacl2 dipping and beeswax (bw) coatings table 5. there was a highest and massive decay of fruits in control fruit samples. at day 5, only control fruit samples had decayed fruits (40%), which increased to 73.33%, followed by fruits treated with 1.5% cacl2 with no beeswax coating on day 10. all fruits that received 6% bw, irrespective of cacl2 treatments, as well as fruits treated by 3% bw with 3.0 and 4.5% cacl2 and 0% bw with 4.5% cacl2 did not show sign of decay till day 15. while, it reached 66.67% in fruits treated with 1.5% cacl2 alone and discarded in the control treatment (table 5). however, a relatively reduced fruit decay amounting to 20% to 60% was observed in fruits receiving 6 and 3% bw combined with higher concentrations of cacl2 during the subsequent storage periods. at day 25, the highest decay (60%) was observed from 6% bw with 0% cacl2 and 6% bw with 1.5% cacl2 treated fruits whereas the lowest (20%) was from 3% bw with 3.0% cacl2 and 3% bw with 4.5% cacl2 treated fruits table 5. table-5. effect of calcium chloride dipping and beeswax coating on the percentage decay of nectarine fruits under holeta condition, 2018. note: bw: beeswax; data after ± are standard deviations (n=3); means with the same letter (s) in a column are not significantly different at p ≤ 0.05 (lsd test); ns: non-significant; ***: p ≤ 0.001. the percentage of decayed fruits gradually increased as storage period advanced for all of the treatments. however, cacl2 and beeswax coating significantly reduced fruit decay loss for relatively longer period. fruits treated either at 3% bw + 3.0% cacl2 or 3% bw + 4.5% cacl2 better reduced the decay occurrence up to shelf life termination. gayed, et al. [35] reported similar finding where cacl2 and chitosan application reduced decay percentage of ‘early swelling’ peach fruit. it has been observed that coating has the ability to prevent the growth of fungi in wide horticultural produces [36]. the use of edible coating combined with natural antimicrobials is a good strategy to increase shelf life of fruits [37]. the reduction of decay in cacl2 dipping and bw coatings could be due to the coatings film property which acted as a barrier for the growth of microbes. the difference in decay percentage might also be due to the modified atmospheric difference created by different levels of bw coatings. fruit decay might also be reduced due to the combined role of cacl2, which act as a reducer of fruit softening by strengthening of the cell walls, and beeswax by covering cuticle and lenticels of the fruits. hence, reduction of respiration rate and ripening process by the treatments probably minimized the occurrence of fruit decay. 3.6. percentage of marketable fruits the percentage of marketable nectarine fruits was highly significantly (p≤0.001) influenced due to the interaction effect of cacl2 dipping and bw coatings (table 6). beeswax (3 and 6%) with all levels of cacl2, and 3 and 4.5% cacl2 alone kept fruits 100% marketable up to 10 days of storage while it dropped to 26.7 and 80% in the control and 1.5 % cacl2 treated fruits alone, respectively. treatment combinations of 3% bw + 4.5% cacl2 and 6% bw + 0% cacl2 maintained fruits marketability to 100% till day 15 while it dropped to nil in the control to 33.33% in 0% bw + 1.5% cacl2 treated fruits. at day 20, fruits in the 0% bw + 1.5% cacl2 treatment were discarded, while the highest percentage of marketable fruits (80%) was recoded for 3% bw with 4.5% cacl2 treated fruits and the lowest (33.33%) was from 0% bw with 3% cacl2 treated fruits. similarly, at day 25, all treatment combinations consisting of 0% bw with all levels of cacl2 and 3% bw with 0 and 1.5% cacl2 were discarded. among the remaining treatments, the highest percentage of marketable fruits (53.33%) was observed from 3% bw + 4.5% cacl2 and 6% bw + 3.0% cacl2 treated fruits while the lowest (20%) was recorded for 6% bw + 1.5% cacl2 treated fruits table 6. in general, the marketability of nectarine fruits gradually declined over storage time. decay and over ripening were the reason of shelf life termination for control fruits, while shriveling as well as little decays were the cause of shelf life termination and un-marketability for treated fruits. however, cacl2 dipping and beeswax coatings had significantly maintained the marketability of fruits. particularly, treatment with 3% bw + 4.5% cacl2 had consistently kept the highest percentage of marketable fruits up to the last date of storage. this result was comparable with reports of eryani-raqeeb, et al. [38] who noticed that 2.5% calcium combined with chitosan treatment storage period (days) bw (%) cacl2 (%) 5 10 15 20 25 0 0 40.00±0.00 73.33a±7.31 0 1.5 0.00±0.00 20.00b±0.00 66.67a±7.31 0 3.0 0.00±0.00 0.00c±0.00 20.00b±0.00 40.00b±0.00 0 4.5 0.00±0.00 0.00c±0.00 0.00c±0.00 20.00c±0.00 3 0 0.00±0.00 0.00c±0.00 20.00b±0.00 53.33a±6.66 3 1.5 0.00±0.00 0.00c±0.00 20.00b±0.00 40.00b±0.00 3 3.0 0.00±0.00 0.00c±0.00 0.00c±0.00 20.00c±0.00 20.00c±0.00 3 4.5 0.00±0.00 0.00c±0.00 0.00c±0.00 20.00c±0.00 20.00c±0.00 6 0 0.00±0.00 0.00c±0.00 0.00c±0.00 33.33b±7.32 60.00a±0.00 6 1.5 0.00±0.00 0.00c±0.00 0.00c±0.00 40.00b±0.00 60.00a±0.00 6 3.0 0.00±0.00 0.00c±0.00 0.00c±0.00 20.00c±0.00 40.00b±0.00 6 4.5 0.00±0.00 0.00c±0.00 0.00c±0.00 20.00c±0.00 33.33b±7.32 significance bw (a) ns *** *** *** cacl2 (b) *** *** *** *** a * b *** *** *** *** agriculture and food sciences research, 2019, 6(1): 71-78 77 © 2019 by the authors; licensee asian online journal publishing group coating extended the storage life of papaya fruits. similarly, extended shelf life period was observed in plum fruits [39] and strawberries treated with edible coatings [40]. the lowest firmness loss, shriveling, decay occurrence and better appearance due to delayed metabolic rates might be the reason for the highest percentage of marketable fruits of cacl2 + bw coating treatments. however, the causes of shelf life termination for cacl2 dipping and bw coating treated fruits were fruit shriveling, decay and over ripening while decay was the main cause of shelf life termination for control fruits. table-6. effect of calcium chloride dipping and beeswax coating on the percentage of marketable nectarine fruits under holeta condition, 2018. note: bw: beeswax; data after ± are standard deviations (n=3); means with the same letter (s) in a column are not significantly different at p ≤ 0.05 (lsd test); ns: non-significant; ***: p ≤ 0.001. 4. conclusion calcium chloride dipping and bw coatings had substantially reduced the plw and significantly maintained the tss, ta and aa of nectarine fruits. it also significantly minimized nectarine fruit decay and maintained marketability as well. hence, calcium chloride dipping and beeswax coatings, particularly treatment with 3% bw + 4.5% cacl2, could be considered as an alternative technology for shelf life extension and better quality. references [1] a. a. kader, fruits in the global market. in: knee, m. 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[40] n. b. gol, p. r. patel, and t. r. rao, "improvement of quality and shelf-life of strawberries with edible coatings enriched with chitosan," postharvest biology and technology, vol. 85, pp. 185-195, 2013. available at: https://doi.org/10.1016/j.postharvbio.2013.06.008. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 22 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 22-23, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.22.27 © 2020 by the authors; licensee asian online journal publishing group genetic variability in tomato (lycopersicon esculentum mill) genotypes in the central rift valley, ethiopia dasta tsagaye1 andargachew gadebo2 shimelis aklilu3 ( corresponding author) 1kulumsa agricultural research center, horticulture research program, assella, ethiopia. 2shool of plant and horticultural sciences, hawassa university college of agriculture hawassa, ethiopia. 3melkassa agricultural research center, vegetable breeding program, adema, ethiopia. abstract investigation of variability existing in crop cultivars is important to crop improvement. the study was conducted at melkassa agricultural research center (marc) to assess the genetic variability among 36 tomato genotypes using 6*6 simple lattice designs. there were significant differences among genotypes all traits. high phenotypic and genotypic variances were recorded for number of fruits per plant, average fruit weight, fruit shape index and juice volume indicating the existence of substantial variability. high heritability along with high genetic advance as percent of the mean was exhibited by characters viz. plant height, number of fruits per plant, average fruit weight, fruit shape index and juice volume, reflecting the presence of additive gene action for the expression of these traits and selection based on them would be ideal. generally, the present study implied the presence of genetic variability among the test genotypes with respect to the traits considered indicating an opportunity to bring about fruit yield improvements in snap bean either through direct selection or hybridization. keywords: tomato, genotype, gcv, pcv, genetic advance, genetic variability, heritability. citation | dasta tsagaye; andargachew gadebo; shimelis aklilu (2020). genetic variability in tomato (lycopersicon esculentum mill) genotypes in the central rift valley, ethiopia. agriculture and food sciences research, 7(1): 22-27. history: received: 14 november 2019 revised: 23 december 2019 accepted: 27 january 2020 published: 18 march 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: the first author acknowledges melkassa agricultural research center (marc) for providing the test genotypes and for their invaluable support in field trial implementation. funding: the authors are grateful to ethiopian institute of agricultural research (eiar) for offering financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 23 2. materials and methods ................................................................................................................................................................... 23 3. statistical data analysis................................................................................................................................................................. 24 4. result and discussion ..................................................................................................................................................................... 24 5. conclusion ......................................................................................................................................................................................... 26 references .............................................................................................................................................................................................. 26 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.22.27&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.22.27&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1417 agriculture and food sciences research, 2020, 7(1): 22-27 23 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the study was conducted at melkassa agricultural research center (marc) to assess the genetic variability among 36 tomato genotypes using 6*6 simple lattice designs. 1. introduction tomato (lycopersicon esculentum mill.) is one of the most known and widely grown vegetables in the world. tomato fruit is traded on the fresh market and used in the processing industry for making ketchup, juice, and soups. it is a valuable component of human diet with high content of important nutrients like β-carotene, lycopene and vitamin [1]. in ethiopia, tomato is one of the most popular vegetables produced by small farmers and commercial growers for both local uses as well as processing industries. it has become an important cash crop. tomato provides employment in production and processing industries [2]. considering the importance of tomato as one of the beneficial vegetables for both domestic consumption and export markets, it is important to increase its productivity along with desirable attributes through its genetic character [3] the crop is cultivated by small scale farmers under irrigation and rain fed condition and large scale commercial vegetable growers. in central rift valley where the study area is conducted the crop is mainly cultivated by small scale farmers and commercial vegetable producers. on the basis of its wide use and expansion potential the need for developing varieties that suite specific agroecological conditions and specific end use is clear. such improvement works, require knowledge on the nature and magnitude of variability in the genotypes [4] and the relative contribution of various component traits to yield [5]. variability in tomato is expected to be immense as the fruits vary greatly in shape and size [6]. studies on genetic parameters provide to select and help to develop optimum breeding procedure. considering all the facts described above the present investigation was undertaken to estimate genetic variability of tomato genotypes. 2. materials and methods the experiment was conducted at melkassa agricultural research center (marc) in 2017/2018 (at the end of october to february) using furrow irrigation. the site is situated in major snap bean growing belts which is 117 km south east of addis ababa with geographic co-ordinate of 8o 24’n latitude and 39o 12’e longitude at an altitude of 1550 m.a.s.l. the mean annual rainfall of the area is 763 mm and the mean annual maximum and minimum temperature is about 28.6 0c and 13.8 0c, respectively. the soil texture is dominantly loam and clay loam and is slightly alkaline ranging from 7.4 to 7.6 ph an optimum range for availability of major nutrients [7]. the experimental materials in the present study consisted of thirty six tomato genotypes obtained from melkassa agricultural research center. simple lattice design (6x6) was employed where each plot consisted of two rows with length of 4m and width 2m that makes a total area of 8m2. the spacing was 100cm and 30 cm between rows and plants respectively. fertilizer rate of 200kg per ha of nps and 150kg per ha of urea was applied. all other necessary cultural practices were applied to all plots uniformly. 2.1. data collected 2.1.1. data collected on plot basis days to first flowering (days): the number of days from the date of transplanting to the day when the first flower bud opens in a plot. days to 50 percent flowering (days): the number of days from the date of transplanting to when about 50% of the plants produced flowers. days to first fruit set (days): the number of days was counted from the date of transplanting to when first fruit appeared in a plot. 2.1.2. data collected on plant and fruit basis plant height (cm): the plant height was measured using measuring tape from ground level to the tip of the plant after 60 days of transplanting and expressed in centimeters and the mean was computed. number of branches per plant: number of branches arising from the main stem above the ground was counted at harvesting on individual plant basis. number of flowers per cluster: three clusters in each plant were taken at random and the number of flowers in each cluster was counted. then the average number of flowers per cluster was calculated. number of clusters per plant: fine plants were taken at random and the number of clusters in each plant was counted. then the average number of cluster per plant was calculated. harvesting continued for about one month because fruits of different genotypes matured progressively at different dates and over longtime. fruits were picked on the basis of horticultural maturity, size color and age being determined for the purpose of consumptions as the fruit grew rapidly and soon get beyond the marketable stage frequent picking was done throughout the harvesting period. number of fruits per plant: the total number of fruits harvested from the five plants was counted and the average number of fruits per plant was calculated. number of fruits per cluster: three clusters in each plant were taken at random and the number of fruits in each cluster was counted. then the average number of fruits per cluster was calculated. fruit yield per plant: the weight of fruits from each picking was from each experimental plot. total yield per plant was worked out by adding yield of all harvests and was expressed in gram (g) per plant. fruit length (cm): average fruit length of five fruits from five plants was measured using vernier caliper from tip to the base of the fruit and expressed in centimeters. fruit diameter (cm): average diameter of five fruits from each sampled plants was measured using venire caliper at the highest bulged portion of the fruit in centimeters. agriculture and food sciences research, 2020, 7(1): 22-27 24 © 2020 by the authors; licensee asian online journal publishing group fruit weight (g): average weight of five fully ripe fruits taken during harvesting was taken using sensitive balance. pericarp thickness (mm): it was recorded from five randomly selected fruits at second harvest by cutting the cross section of a fruit and measuring the pericarp thickness using vernier caliper expressed in centimeter. fruit shape index: ration of fruit length to width were recorded from five randomly selected fruits at second harvest. ph of tomato fruit: tomato juice was extracted from red ripe fruits, was determined from extracted juice, by using ph meter at room temperature. total soluble solids content of fruit (brix): brix percentage was measured by portable refractometer at room temperature. single fruit was blend and juice was collected to measure brix. juice volume (ml): tomato juice was prepared by selecting five typical and red ripe fruits from sample plants and juice was measured in cylinder and expressed in milliliter (ml). 3. statistical data analysis data were subjected to analysis of variance as per the procedure for simple lattice design as described by gomez and gomez [8] using sas version 9.2 [9]. duncan’s multiple range test was used for mean separation procedures. 3.1. variability analysis a. estimation of variance components the phenotypic and genotypic variances were estimated according to the method suggested by singh and choudhary [10] as follows: environmental variance (2e), 2e=mse (error mean square) genotypic variance (2g) 2g= msg−mse r phenotypic variance (2p) 2g + 2e/r where, r=number of replication, msg=mean square due to genotypes, mse=mean square of error (environmental variance) b. heritability in broad sense broad sense heritability values for all parameters (h2) were estimated based on the formula given by falconer and mackay [11] as follows: h2= σ2g σ2p ∗ 100 where, h2= heritability in the broad sense, 2g=genotypic variance and 2p =phenotypic variance. estimated heritability values was classified according to singh [12] that heritability values greater than 80% were very high, values from 60–79% were moderately high, values from 40–59% were medium and values less than 40% were low. c. genetic advance the genetic advance expected under selection assuming selection intensity of the superior 5% of the plants was estimated in accordance with the methods illustrated by allard [13]. ga=k*(√2 p)*h2 where, ga = expected genetic advance, k = the standardized selection differential at 5% selection intensity (k = 2.063), 2 p = is phenotypic standard deviation on mean basis and h2=heritability in the broad sense. d. genetic advance as percent of means (gam) genetic advance as percentage over mean was worked as suggested by johnson, et al. [14]. gam = ga �̅� ∗ 10where, ga = genetic advance, �̅� = grand mean; genetic advance as percent of mean was categorized as 0-10% = low, 10-20% = moderate, >20% = high e. genotypic and phenotypic coefficient of variation genotypic and phenotypic coefficients of variation were estimated according to burton and devane [15] based on the estimate of genotypic and phenotypic variance. genotypic coefficient of variation (gcv %) = √2 g �̅� *100 phenotypic coefficient of variation (pcv %) = √2 p �̅� *100 where, vg = genotypic variance, vp = phenotypic variance, �̅� = grand mean of the character. pcv and gcv were categorized as following: 0 – 10 %: low, 10 20%: moderate, 20% and above high [16]. 4. result and discussion 4.1. analysis variance the mean square due to genotypes were highly significant for all traits (p < 0.01) except for days to 50% flowering, plant height, number of branches per plant, number of fruits per cluster and pericarp thickness which were significant at (p < 0.05), indicating the presence of sufficient genetic variability in the genotypes and considerable scope for their improvement table 1. according to asati, et al. [17] analysis of variance revealed that significant differences among the genotypes for all the traits; aysh, et al. [18] reported that the highly significant differences observed among the genotypes evaluated indicates existence of a good deal of variability with respect of the characteristics assessed and offers ample chances for the genetic improvement of the tomato germplasm; meena agriculture and food sciences research, 2020, 7(1): 22-27 25 © 2020 by the authors; licensee asian online journal publishing group and bahadur [19] reported that there were significant differences among germplasm for all the traits studies indicating the presence of significant variability in the materials which can be exploited through selection; shankar shankar, et al. [20]; haydar, et al. [21] and singh, et al. [22] revealed that highly significant differences among the genotypes for all the characters indicating sufficient variability existed in the present material selected for the study and indicating the scope for selection of suitable initial breeding material for crop improvement; kumar, et al. [23] reported that significant differences among germplasm for all the traits studies indicating the presence of significant variability in the materials which can be exploited through selection. 4.2. estimation of genotypic and phenotypic coefficient variation the present result revealed that, higher gcv were recorded for number fruits per plant (25.29607%), fruit yield per plant (21.7934%), average fruit weight (21.93702%), fruit shape index (25.65%) and juice volume (23.9115%). moderate gcv value were recorded for days to first flowering date (15.76%), plant height (13.21%), number branches per plant (13.34%), number flowers per cluster (16.44), number fruits per cluster (13.35%), number clusters per plant (16.68%), fruit length (14.18%) and fruit diameter (12.67%). however total soluble solid (8.93%), pericarp thickness (8.91%), ph (7.58%), days to 50% flowering date (7.37%) and days to first fruit set (5.74%) had the lowest values. high pcv values were observed in fruit shape index (26.791%), number of fruits per plant (26.408%), juice volume (25.732%), average fruit weight (24.408%), fruit yield per plant (22.991%) and number of clusters per plant (20.207%). however days to first flowering (18.177%), number of flowers per plant(17.333%),number of branches per plant(17.148%), plant height (15.287%), fruit length(15.071%),pericarp thickness(14.085%),number of fruits per plant(14.978%), fruit diameter(13.586%) and total soluble solid(12.092), had moderate values. this indicated the presence of environmental influence on these characters [24]. 4.3. heritability and genetic advance heritability ranged from (55.22%) for total soluble solid to (98.08 %) for ph and characters such with very high heritability values were obtained for ph (98.08%), fruit shape index (95.65%), number of fruit per plant (91.75%), number of flowers per cluster (89.38%), fruit yield per plant (89%), fruit length (88.25%), fruit diameter (87.69%), juice volume (86.35%) and average fruit weight (80.05%); moderately high heritability values were obtained from number of fruits per cluster (79.13%), days to first flowering (75.15%), plant height (74.71%), days to first fruit set (71.95%), number of clusters per plant (68.09%) and number of branches per plant (60.53%); whereas pericarp thickness (55.14%) and total soluble solid (55.22%) had moderate heritability values table 2. table-1. the analysis of variance for the 18 characters tested at melkassa in 2017/2018. mean squares source of variation df dff d50%f dffs plh nbpl nflc nfc ncpl nfpt replications 1 107.56 14.22 24.5 16.43 28.8 0.294 28.34 1.28 2105.46 block within reps(adj) 10 80.56 11.25 4.083 31.37 3.7 0.176 0.2383 195.85 2643.04 genotypes un adjusted 35 50.5 22.01 23.3143 125.66 4.2521 0.6844 0.38 742.62 225.26 adjusted 35 46.540** 19.845* 22.55** 120.46* 3.986* 0.63** 0.34* 21.8** 312.08** error -rcb design 35 10.6 22.01 4.4 30.33 2.068 0.1419 0.17 376.04 5131.14 -intra block 25 11.5556 6.32 4.5 29.8 1.4211 0.1283 0.1424 7.207 24.8811 cv % 12.84 6.27 5.07 10.87 15.24 8.01 9.69 16.15 10.725 r2 87.1 85 88.66 86.4 85.78 89.02 92.48 83.911 95.79 efficiency of lattice over rcbd 91.34 108.65 97.32 100.11 123.77 102.66 106.94 126.27 125.03 note: where ** = highly significant at 1%, * = significant at 5%, df=degree of freedom, dff = days to first flowering, d50%f = days 50% flowering, dffs = days to fruit set, plh = plant height, nbpl = number of branches per plant, nflc = number flowers/cluster, nfc = number fruits/cluster, ncpl = number of clusters/plant, nfplt = number fruits/plant, r2 = coefficient of determination. table-1. (continued). mean squares source of variation df fl fd afw fypt pth fsi ph tss jv replications 1 0.2069 0.28 715.52 67230 0.0003 0.001 0.009 2.3112 27417 block within reps(adj) 10 0.2777 0.1023 126.41 208112 0.0064 0.021 0.016 0.2849 6109.51 genotypes un adjusted 35 2.215 1.07 1672.57 465169 0.014 0.24 0.0188 0.4072 58021 adjusted 35 2.15** 1.16** 1609.99** 468652** 0.014* 0.24** 0.27** 0.42** 55723.4** error -rcb design 35 0.2563 0.1237 270.19 2024662 0.0058 0.013 0.01735 0.2204 11853 -inter block 25 0.2477 0.132 327.7 42303.7 0.006 0.01 0.0047 0.1946 6917.014 cv % 7.32 6.722 15.488 10.341 10.53 7.49 1.512 11.372 13.3 r2 92.85 92.13 88.07 93.84 80 97.34 99.26 79.96 91.98 efficiency of lattice over rcbd 100.36 93.5321 82.4505 103.53 100.6 116.47 98.082 103.86 83.7648 note: where = ** = highly significant at 1%, * = significant at 5%, df = degree of freedom, fl = fruit length, fd = fruit diameter, afw = average fruit weight, fyplt = fruit yield/ plant, pth = pericarp thickness, fsi = fruit shape index, ph = power of hydrogen, tss = total soluble solid, jv = juice volume, r2 = coefficient of determination. agriculture and food sciences research, 2020, 7(1): 22-27 26 © 2020 by the authors; licensee asian online journal publishing group high heritability along with high genetic advance as percent of the mean are exhibited by number of fruits per plant, number of flowers per cluster, fruit yield per plant, fruit length, fruit diameter, average fruit weight, number of fruits per cluster, days to first flowering, plant height, fruit shape index and juice volume; high heritability along with moderate genetic advance as percent of the mean are exhibited by days to 50% flowering, days to first fruit set and power of hydrogen; and medium heritability along with moderate genetic advance as percent of the mean are exhibited by total soluble solid and pericarp thickness. high estimate of heritability may be the result of the diverse nature of the genotypes included in the study shushay, et al. [25]. sunil, et al. [26] reported that high heritability coupled with high genetic advance were noticed for number of fruits per plant, average fruit weight and fruit yield per plant in tomato. table-2. estimation of variability for 18 traits of tomato genotypes tested at melkassa in 2017/2018. traits range mean ± std sg 2 sp 2 se 2 pcv% gcv% h2 ga gam% dff 40.5 – 17 26.5277±5.62 17.47 23.25 11.56 18.177 15.76 75.15 7.47 28.18 d50%f 47.5 – 35 40.083±3.86 8.73 11.89 6.32 8.603 7.37 73.42 5.22 13.03 dffs 49 – 36 41.833±3.74 5.77 8.02 4.50 6.770 5.74 71.95 4.20 10.05 plh 77.6 – 40 50.211±8.78 44.02 58.92 29.80 15.287 13.21 74.71 11.83 23.56 nbpl 12.83 – 5.7 7.824±1.88 1.09 1.80 1.42 17.148 13.34 60.54 1.68 21.42 nflc 6.4 – 3.4 4.469±0.64 0.54 0.60 0.13 17.333 16.44 89.38 1.43 32.07 nfc 4.831 – 2.93 3.893±0.82 0.27 0.34 0.14 14.978 13.35 79.13 0.95 24.49 ncpl 23.4 – 11 16.628±3.97 7.69 11.29 7.20 20.207 16.68 68.09 4.72 28.39 nfplt 69.8 – 28.1 46.510± 14.59 138.42 150.86 24.88 26.408 25.30 91.75 23.25 49.99 fl 9.10 – 4.9 6.799±1.11 0.93 1.05 0.25 15.071 14.18 88.25 1.87 27.49 fd 6.879 – 3.35 5.409±0.77 0.47 0.54 0.13 13.586 12.67 87.69 1.32 24.48 afw 193.5– 65.8 116.881± 31.11 657.42 821.27 327.70 24.519 21.94 80.05 47.33 40.49 fyplt 3086 – 1407.9 1988.880± 574.51 187942.10 209094.00 42303.70 22.991 21.80 89.88 847.92 42.63 pth 0.849 – 0.498 0.71±0.09 0.00571 0.01 0.01 14.085 8.91 57.14 0.10 13.89 fsi 2.7082 – 0.8592 1.293±0.35 0.11 0.12 0.01 26.791 25.65 95.65 0.67 51.75 ph 4.44 – 3.995 4.570±0.13 0.12 0.12 0.00 7.580 7.58 98.08 0.71 15.49 tss 5 – 2.6 3.879±0.58 0.12 0.22 0.19 12.092 8.93 55.22 0.53 13.69 jv 1030 – 265 618.681± 186.63 21885.00 25343.55 6917.01 25.732 23.91 86.35 283.60 45.84 note: where, std = standard deviation, dff = days to first flowering, d50%f = days to 50%flowering, dfs = days to fruit set, plh = plant height, nbpl = number of branches per plant, nflc = number flowers/cluster, nfc = number fruits/cluster, ncpl = number of clusters/plant, nfplt = number fruits/plant, fl = fruit length, fd = fruit diameter, afw = average fruit weight, fyplt = fruit yield/ plant, pth = pericarp thickness, fsi = fruit shape index, ph = power of hydrogen, tss = total soluble solid, jv= juice volume. 5. conclusion the present investigations revealed the presence of good variability among genotypes with respect to different traits considered. thus there is an opportunity to improve tomato through direct selection or hybridization that involves crossing of different genotypes. references [1] v. bergougnoux, "the history of tomato: from domestication to biopharming," biotechnology advances, vol. 32, pp. 170-189, 2014.available at: https://doi.org/10.1016/j.biotechadv.2013.11.003. 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[26] m. k. sunil, v. rathod, j. c. bommesh, s. vijeth, and k. k. muthaiah, "genetic variability in tomato (solanum lycopersicum l.)," journal of environmental biology-science, vol. 30, pp. 47-51, 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 51 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 51-57, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.51.57 © 2020 by the authors; licensee asian online journal publishing group evaluate technical standards of implemented soil bund in central rift valley of ethiopia: the case of adama, lume and dodota districts kalkidan fikirie1 abebe bezu2 melat eshetu3 daniel bekele4 mohamed rabo5 ( corresponding author) 1holeta agricultural research center, holeta, ethiopia. 2,3,4,5melkassa agricultural research center, melkassa, ethiopia. abstract land degradation in the form of soil erosion is a serious problem in ethiopia. to combat this, the government undertaken swc measures across the country since 1970’s. this study therefore, investigated technical standard of implemented soil bund practice in crv of ethiopia. a mixed methods approach was employed, combining qualitative and quantitative data collection tools; hh survey, ki interviews, fgd, observation and desk literature review. hhs were randomly chosen from the sampling frame exist at pa level and a total of 150hhs were selected for this study. further, soil bund structures were measured and evaluated. the data analysis was carried out by using descriptive statistics, one way anova and chi-square tests. results of the study showed that soil bund parameters implemented under the standard are bund spacing, width and height of embankment. while bund top width, bottom width, berm width and top width of embankment were better. soil and stone bunds are mostly preferred, whereas check dam is the least preferred technology to implement in the study area. major challenges farmers faced during swc practices implementation, shortage of labor, lack of technical skills, fear for animal trampling, lack of awareness and lack of sufficient farm tools. hence, the study has concluded that every year swc practices implemented by community mobilization are not standardized and consequently it is difficult to manage the watershed in different areas. therefore, the study suggests that well organized training should be given for experts at different levels to fill the technical gap on their skills and extension services like demonstration should be held for all stakeholders before implementing the swc structures. keywords: farmers’ perception, land degradation, soil erosion, technical standard. citation | kalkidan fikirie; abebe bezu; melat eshetu; daniel bekele; mohamed rabo (2020). evaluate technical standards of implemented soil bund in central rift valley of ethiopia: the case of adama, lume and dodota districts. agriculture and food sciences research, 7(1): 51-57. history: received: 17 january 2020 revised: 21 february 2020 accepted: 26 march 2020 published: 5 may 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors like to thank government officials of adama, lume and dodota districts, development agents for their support during data collection phase of the study. special thanks go to adama, lume and dodota districts’ natural resource coordinators for their guidance and provision of detailed information about the soil and water conservation practices in the study area. funding: authors would like to thank ethiopian institute of agricultural research (eiar)/ melkassa agricultural research centre (marc) for the financial support to us to pursue this research. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 52 2. methodology ..................................................................................................................................................................................... 52 3. results and discussions .................................................................................................................................................................. 53 4. conclusion and recommendations ............................................................................................................................................... 56 references .............................................................................................................................................................................................. 57 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.51.57&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.51.57&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1592 agriculture and food sciences research, 2020, 7(1): 51-57 52 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by investigating technical standard of implemented soil bund practice in crv of ethiopia. 1. introduction 1.1. background and justification land degradation in the form of soil erosion is a serious problem in ethiopia. most productive land has been exposed to degradation and threat both economic and survival of the people [1]. soil erosion is a cause of land degradation that affects the soil physical, chemical and biological properties, results in onsite soil nutrient loss, soil structure and texture damage and offsite water pollution, flooding and sedimentation of water resources in the country [2]. in ethiopia, alarming rate population increment, continuous and steep slope farming, low vegetation cover, deforestation and insufficient soil and water conservation practices cause annual soil loss of about 1.5billion metric tons and resulted, 1.2 ton/ha/yr cereal yield reduction on average [3, 4]. in addition, the continually increasing land use change is exasperating the rates of soil erosion, soil fertility reduction, crop yield decline and food insecurity [5, 6]. continuous soil erosion threatens communities’ livelihoods especially in drought prone areas, where arable land is a scarce resource in the country. according to belay, et al. [7] report, deforestation for the expansion of agricultural farms and rangelands has driven to increasing soil losses and the growth of rock outcrops, soil nutrient depletion, decreasing productivity and macro environmental degradation in ethiopia. to combat these problem, ethiopian government promotes soil and water conservation practices through community mobilization to increase production and productivity of the land, food security, improve livelihood of the community and mitigate environmental degradation [8, 9]. hence, since, 2010 different physical and biological soil and water conservation practices implemented by community campaign on privately owned and communal lands [10, 11]. these physical and biological soil and water conservation practices implemented in a coordinated effort by the government with local community members [12]. nevertheless, mass mobilization approaches lead to the implementation of soil and water conservation practices with inappropriate design and resulting the implemented practices had lesser impact on conserving soil and water than expected and consequently gully formation in many part of the country. these are due to technical gap and lack of skilled manpower in the community. technical gap caused failure of implemented physical and biological soil and water conservation practices which in turn serious soil erosion and land degradation in the country and also in the study area, failure of implemented soil and water conservation practices was a serious problem like other part of the country due to improper design. thus, the purpose of this study was to evaluate the technical standard of implemented soil bund by community campaign in adama, lume and dodota districts, crv of ethiopia. subsequently, the result of the study could be helpful for soil and water conservation researchers and other academicians. the objectives of this study were: • to evaluate the technical standard of implemented soil bund in the study area. • to assess farmers’ perception on soil erosion and conservation practices. • to assess and describe major challenges farmers faced during soil and water conservation practices implementation. 2. methodology 2.1. study area description the study was conducted in three selected districts of crv of ethiopia namely lume, adama and dodota districts. lume district is found in the east shewa zone of oromia region and its located 70km to the east of addis ababa the capital city of ethiopia. the district is geographically located between 8024’-8051’ n latitude and 3901’39017’e longitudes [13]. the total land area of the district is 75,220ha, which comprises a total of 35 peasant associations. lume is bordered in the south with bora district, in the east adama district, in the north amhara region, in the west ada’a chukala district and in north-west with gimbichu district. the administrative center of the district is modjo town. adama district is bordered on the south by the arsi zone, on the southwest by koka reservoir which separates it from bora district, on the west by lume, on the north by the afar region and on the east by boset district. the altitude of the district ranges from 1500-2300m.a.s.l. the major soil type of the district is andosol and about 74.3% of the district coved by this soil type. the remaining 25.7% of the districts’ soil is cambisols and luvisols. adama district has an estimated total population of 180,710 of which 91,859 are men and 88,851 are women [14]. the population density of the district is 207.4 people per square kilometer, which is greater than the zone average of 189.6 and the average farmland size per household was 2.5ha. dodota district is a part of arsi zone and bordered on the south by tena, on the southwest by hitosa, on the north by the east shewa zone, on the east by jelu and on the south east by sude district. the administrative center of the district is dera and other towns include awash melkassa. the altitude of dodota ranges from 1400 to 2500m.a.s.l. the total land of the district shows that 23.2% cultivable, 10.6% pasture, 4.3% forest and the remaining 42% is considered swampy, mountainous or otherwise unusable. agriculture and food sciences research, 2020, 7(1): 51-57 53 © 2020 by the authors; licensee asian online journal publishing group figure-1. map of the study area. 2.2. sampling procedure multistage sampling technique was employed to select the population for the study which involved purposive and random sampling techniques. first, districts were purposively picked, and secondly, kebeles were selected using lottery method. finally, households were randomly chosen from the sampling frame (list of farmers) exist at kebele level. a total of 150 households were selected for this study. 2.3. data collection and analysis data was collected through structured questionnaire administered to sampled farmers by trained enumerators. before the actual survey, the questionnaire was pretested in non-sampled villages. the pretest was not only used to test the appropriateness of the tool in collecting the required data but also to evaluate the trained enumerators on the capability of administering the questionnaire. in addition to these, soil bund structure implemented in the study area were measured and evaluated. information related to perception of farmers on soil and water conservation technologies, farmer’s preferences of swc practices and major challenges farmers faced by before and during soil and water conservation implementation were gathered from the respondents. households’ socio demographic, institution and economic features were also collected. data were cleaned, organized and analyzed using spss software. both descriptive and inferential statistics were used to analyze the gathered and cleaned data. 3. results and discussions 3.1. respondents’ socio demographic structures the study was conducted in three districts of crv of ethiopia. the total numbers of respondents interviewed were 150 and of which 84% respondents are male, the rest 16% were female respondents. the age of respondents showed that, 93% of respondents included the age of 18-64 and the remaining 7% are > 64 age table 1. out of the total respondents, education levels of 50.7% of the respondents were elementary school and 16.7% are uneducated. concomitantly, 17.3% of respondents existed in high school education level. in this study, adama (16%) and dodota (12%) districts have the least illiterate respondents, and lume have the highest uneducated (22%) respondents. the high number of family size was recorded under dodota district than adama and lume which is smaller than the national average family size of 6.4 people per household. hence, from this result and personal observation during field work it is possible to conclude and suggest that labor availability is key factor influencing households’ decision to participate in soil and water conservation practices. table-1. socio-demographic characteristics by districts. variables lume (n=50) adama (n=50) dodota (n=50) pooled (n=150) gender in % male 84.0 90.0 78.0 84.0 female 16.0 10.0 22.0 16.0 household head age in % 18-64 92.0 96.0 92.0 93.0 >64 8.0 4.0 8.0 7.0 household head education level in % non educated 22.0 16.0 12.0 16.7 read and write 14.0 18.0 8.0 13.3 elementary school 52.0 44.0 56.0 50.7 high school 12.0 20.0 20.0 17.3 diploma 0.0 2.0 4.0 2.1 family size 5.6 5.2 6.3 5.7 agriculture and food sciences research, 2020, 7(1): 51-57 54 © 2020 by the authors; licensee asian online journal publishing group 3.2. standard of implemented soil bund in the study area to evaluate the quality of implemented soil bund in the study area, parallel transect walk method was used to measure the dimensions of the implemented soil bund structures. the major physical soil and water structures were found in the study area, soil bund, waterway, cutoff drain, check dams and others. these structures have mainly constructed by community mobilization. bund spacing, bund gradient, field slopes, bund cross sectional areas like embankment height, top and bottom width and channel depth and width were measured and evaluated. accordingly, the implemented soil bund structures were above and below the standards. soil bund parameters implemented below the specified standards were bund spacing, bund depth, and height of embankment and bottom width of embankment. while, bund top width, bund bottom width, berm width and top width of embankment are better table 2. during field observation, there was no any similarity between the existing soil bund structures in the area. the main reasons for soil bund implemented below the standards are knowledge and skill gap on soil and water conservation structures. this was not only farmers but also development agents have knowledge and skill gap on designing the structures. in the study area, usually communities perceived that implementing soil bunds in narrow spacing may create difficulty in plowing activities and reduces farm size at the same time needs much labor forces to implement. traditionally, farmers were using traditional drainage ditches on their farmlands to disposed excess runoff during high rainfall. nonetheless, this type of conservation measure was out of the standard which had negative impact on soil erosion control since the drainage ditches have on average high slope gradient which leads to erosion along traditional drainage ditches and create excess siltation on the lower part. similar finding by gizaw [15] reported that, farmers construct drainage ditches every year and sediments eroded from the ditches and accumulated down slope were common indicators of erosion on farmer’s farmlands. table-2. comparison between standards and implemented soil bund structure. parameters national standards lume adama dodota slope for the sites (%) >3 4.3 4.3 4.3 slope of the level bund (%) 0 0.2 0.1 0.1 bund length (m) 30-80 55.8 57.4 60.2 bund spacing (m) 34.9 10.7 22.8 21.5 bund depth (cm) 50 30 40 37 bund top width (cm) 50 72.9 77.5 76.3 bund bottom width (cm) 50 70.3 60.2 72.6 berm width (cm) 5 20 12.5 11.1 height of embankment (cm) 60 28.3 44.2 45 top width of embankment (cm) 30 43.3 36.7 35.2 bottom width of embankment (m) 1.1 1 1 1 3.3. farmers’ perception on soil erosion finding of the survey revealed that, 82.8% of lume districts’, 78.3% of adama district and 77.6% of dodota districts’ respondents were implemented soil and water conservation practices to reduce the risk of soil erosion. while 17.2% of lume district, 21.7% of adama district and 22.4% of dodota district respondents were not implemented any of the soil and water conservation structures to control soil erosion in the study area table 3. table-3. farmers swc structures implementers and non-implementer. swc implementers lume adama dodota swc implementers in % 82.8 78.3 77.6 swc non implementer in % 17.2 21.7 22.4 farmers had different interest in soil and water conservation practices. majority of farmers showed high interest to conserve their soil while limited number of farmers were reluctant. most of the time farmers whose lands were seriously degraded by erosion were highly interested to conserve the soil. these farmers were those residing in the upper part of the watershed. in contrary, farmers those lands were less degraded by soil erosion showed less interest. in terms of soil and water conservation structures users, 77.4%, 69.5% and 68.9% of respondents perceived the degree of soil erosion as sever from lume, adama and dodota districts, respectively. in case of non-user groups (non-conservation implementers), majority of the respondents perceived the degree of soil erosion was moderate to minor on their croplands table 4. the result of chi-square test showed that, respondents’ perception on the degree of soil erosion on their farmlands were significantly different between conservation practices implementers and nonimplementers. the reason is that, due to different in respondents’ education level and the degree of soil erosion problem on their cultivated lands. fgd and key informant interviewer also responded that the main causes of soil erosion in the study area were easily erodible nature of soils, uncontrolled grazing, inappropriate farming system, deforestation and poor watershed management. this result is in line with the finding of kibemo [16] who stated that, educated farmers have better understanding the existence of soil erosion than uneducated farmers. table-4. farmers’ perception on soil erosion in the study area. districts lume adama dodota degree of erosion swc user (%) nonuser (%) swc user (%) non-user (%) swc user (%) non-user (%) χ2 sever 77.4 1.7 69.5 8.8 68.9 5.3 0.067ns moderate 12.3 1.5 7.5 6.4 15 4.7 0.078ns minor 6.4 0.7 3.5 4.3 3 3.1 0.032ns total 96.1 3.9 80.5 19.5 86.9 13.1 agriculture and food sciences research, 2020, 7(1): 51-57 55 © 2020 by the authors; licensee asian online journal publishing group 3.4. farmers’ preferences of soil and water conservation practices preferences about soil and water conservation technologies strongly affects farmers’ adoption decision. in the study area, most of the time farmers prefer soil bund (95%) and stone bund (80%) to implement on their own lands table 5. as indicated in the focus group discussion, check dam and gully rehabilitation technologies were less prefered by respondents’ since these conservation practices needs skilled manpower and intensive labor. moreover, farmers have limited information about the values of these physical and biological conservation practices and destruction of biological materials by free grazing in the area. during field observation, it was witnessed that the nature of livestock husbandary was open grazing style this created damaging of structures by animal trampling over the structures. this makes that the concentration of runoff forms more erosion han inconserved areas. the chi-square test revealed that, there is insignificant difference among respondents’ perception on the effectivness of physical soil and water conservation practices. majority of respondents perceived soil and stone bund were effective to control soil erosion while some proportions perceived check dam maked them more effective to control erosion. this is may happned, due to the difference in degree of erosion among respondents’ land, severity and minor eroded soils, farmers implement different structures to control runoff and sediment movement. table-5. farmers’ preferences of soil and water conservation practices. types of structure farmers' preference in % rank soil bund 95 1 stone bund 80 3 waterway 78 4 cut off drain 77 5 gully rehabilitation 63 6 check dam 58 7 regarding structure maintainance, only 23%, 12.5% and 9.8% respondents from lume, adama and dodota districts, respectively have maintained the structures on their farmlands table 6. the absence of structural maintainance are due to farmers perception on consquences of erosion education, land ownership and family size or labor availability were major determinant farctors listed by farmers to not maintain the implemented structures individually. in most cases, farmers were not voluntary to maintain the implemented structures since hetrogeneity of the problem over landscape, variability in the size of land holding per househld and difference in perception to soil erosion. table-6. structures maintained by districts in the study area. districts structures maintained lume 23.1% adama 12.5% dodota 9.8% 3.5. farmers provided extension services and training on soil and water conservation practices 3.5.1. access to extension services according to the household survey, 69% of lume, 72% of adama and 66% of dodota respondents got better extension service on soil and water conservation technologies in the study area figure 2.the chi-square test confirmed that there is no significant difference between districts with regard to extension services (p>0.05). as indicated in the focus group discussion, frequent contact of extension worker is also related to the socio-economic status of the farmers. frequently, extension workers might prefer to visit farmers with more farmland or those who have already adopted the soil and water conservation technologies in the area. the importance of extension service is to initiate change that bring about sound soil and water conservation especially on the part of smallholder farmers as it offers them technical advice on necessary services. therefore, extension service is fundamental for the natural resource conservation, providing training, technical advice, accessing the supply of inputs timely and giving various information that ranges from production to marketing. moreover, it represent local farmers’ frequency of contact with das and frequency of participation in extension planning, training, field day, on-farm trial and demonstration regarding land management, agriculture and livestock production. thus, extension service has positive impact on enhancing soil and water conservation technologies. 3.5.2. access to training training on soil and water conservation technologies is one of the key factor than influence the participation of farmers in soil and water conservation practices. accordingly, 37% of lume, 39% of adama and 29% of dodota districts’ respondents have access to training on swc technologies figure 3. the statistical test result confirmed that there was a negative and significant relationship between districts with regard to participation in soil and water conservation training (p<0.05) with dodota having low participation. this indicated that the number of farmers got training were better at lume and adama districts. hence, during focus group discussants responded that most of the time training was given by different ngos like meret and slm (sustainable land management) in lume and adama districts. on the other hand, in dodota district, farmers got training access often from government bodies including political leaders. the more the local farmers get soil and water conservation training, the more likely that they acquire the relevant information along with the technical; known the importance of soil and water conservation technologies. training creates awareness and helps the local farmers to perform bring innovation and invention in soil and water conservation technologies. hence, training as the fundamental element to change the perception of local farmers in order to adopt and expand soil and water conservation technologies. agriculture and food sciences research, 2020, 7(1): 51-57 56 © 2020 by the authors; licensee asian online journal publishing group figure-2. farmers got extension services. figure-3. farmers got training on swc practices. 3.6. major challenges farmers’ faced during soil and water conservation structures implementation main challenges such as lack of labor, lack of technical skills, lack of farm tools, fear of animal trampling and lack of awareness are the major challenges in the study area though farmers appear to face such problems figure 4. in the study area, usually soil and water conservation practices carried out in the dry season when no or little labor is needed for agricultural activities. because, labor for the establishment constitutes the largest part of the investments in soil and water conservation practices. the technological options are less available in the study area to relieve humans. thus, farm execution of most farm related works require human labor, among other. physical soil and water conservation structures implementation and maintenance require intensive labor for which machinery has not been introduced or developed in the country. the durability of swc structures mainly based on the slope of the area, rainfall intensity, soil stability and land use type and management, but it needs frequent maintenance such as de-silting and repairing broken parts. a guideline by the ethiopian ministry of agriculture estimates 150-250pd for the construction of the kilometer of commonly practices soil and water conservation structures, such as soil bunds, fanyajuu, stone bunds [17]. in this case, labor may remain a challenge for the undertaking human labor based soil and water conservation activities in the area. regarding lack of farm tools, challenge raised by farmers was no provision of spades and dig to excavate the soil and fear of animal trampling in the area, since structures implemented less quality and ineffective in the area. that is also what is explained by kebede [10] which indicated that labor demand appears much more than the requirement for most ordinary farming business. figure-4. major constraints farmers faced during soil and water conservation implementation. 4. conclusion and recommendations the participation of women are better at dodota district than lume and adama (mean=16%). regarding family size, the high number of family size recorded under dodota district compared to lume and adama. soil bund parameters implemented below the standard are bund spacing, bund depth and height of embankment. whereas, bund top width, bund bottom width, berm width and top width of embankment are better. due to this, runoff generated over tops causing rill and gully formation which is difficult to control. with regard to swc technology preferences; soil and stone bund are mostly preferred while, check dam is the least preferred technology to implement in the study area. the study result shown that, swc implementers challenged by constraints like shortage of labor, lack of technical skills, fear for animal trampling, lack of awareness and lack of sufficient farm tools. to sum up, every year swc practices implemented by community mobilization are not standardized and thus it is difficult to manage the watershed in different areas. therefore, based on the above findings the following recommendations can be drawn for further consideration and improvement of soil and water conservation practices in the study area in particular and in the country in general. • well organized training should be given for experts at different levels to fill the technical gap on their skills. • extension services like demonstration should be held for all stakeholders before implementing the soil and water conservation structures. • soil and water conservation structures should be planned by experts at kebeles and district levels rather than planning at zonal or regional levels for its quality and effectiveness. agriculture and food sciences research, 2020, 7(1): 51-57 57 © 2020 by the authors; licensee asian online journal publishing group references [1] t. genene and g. abby, "review on overall status of soil and water conservation system and its constraints in different agro ecology of southern ethiopia," journal of natural sciences research, vol. 4, pp. 59-69, 2014. 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[16] d. kibemo, "farmers' perception on soil erosion and their use of structural soil conservation measures in soro district, southern ethiopia," msc thesis, addis ababa university, ethiopia, 2011. [17] d. lakew, v. carucci, w. asrat, and a. yitayew, "community based participatory watershed development: a guideline, part 1, ministry of agriculture and rural development, federal democratic republic of ethiopia ministry of agriculture and rural development, addis ababa, ethiopia, january 2005," 2005. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 50 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 50-56, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.50.56 © 2019 by the authors; licensee asian online journal publishing group effect of storage conditions on quality characteristics of spicy and nonspicy varieties of pork sausages subhajit ray associate professor, department of food engineering &technology, central institute of technology kokrajhar, assam, india. abstract two different varieties of pork sausages viz. spicy and non-spicy were formulated and cooked at 700c for 3 minutes. both of these two formulated varieties were packaged with low-density polyethylene and wax paper respectively and stored under refrigeration. sensory, nutritional and microbiological characteristics of both types were analysed during different storage periods viz. 1day, 7days, 14 days and 21 days respectively. evaluation of organoleptic characteristics showed greater acceptance of spicy variety in comparison to non-spicy variety. there were no significant variations in the nutritional composition of pork sausage varieties during storage. microbiological quality of the spicy product was superior to its non-spicy counterpart. sausages packaged in lowdensity polyethylene have the greater shelf stability than that of in waxed paper after storage periods of 14 days and 7 days respectively under refrigerated condition. experimental results also revealed that spicy formulation has greater shelf stability than that of the nonspicy formulation. keywords: pork sausage, refrigeration, storage stability, low-density polyethylene, wax paper. citation | subhajit ray (2019). effect of storage conditions on quality characteristics of spicy and nonspicy varieties of pork sausages. agriculture and food sciences research, 4(1): 50-56. history: received: 26 january 2019 revised: 1 march 2019 accepted: 8 april 2019 published: 4 june 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: the author is grateful to the department of food engineering and technology, central institute of technology kokrajhar, assam, pin: 783370, india for providing necessary laboratory facilities to carry out this research work. funding: this study received no specific financial support. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 51 2. material and methods ..................................................................................................................................................................... 52 3. results and discussions .................................................................................................................................................................. 54 4. conclusions ....................................................................................................................................................................................... 56 references .............................................................................................................................................................................................. 56 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.50.56&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.50.56&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/1830 https://orcid.org/0000-0003-0081-9836 agriculture and food sciences research, 2019, 6(1): 50-56 51 © 2019 by the authors; licensee asian online journal publishing group 1. introduction slaughter of animal produces a considerable amount of edible by products (head, meat, heart, tongue, tripe etc.) with high biological value and low palatability attributes [1]. comminuted meat products offer an attractive avenue for the utilization of low-value cuts and edible offal’s by replacing a certain proportion of skeletal meat to reduce their cost of production [2]. the most important component of meat is the muscle in which the myofibriller proteins form about 60% of the total muscle protein [3]. proteins derived from raw meat and cooked pork rind; exhibit a wide range of functional properties. they are able to form networks and structures, interact with other ingredients and thus plays an important role in the texture, sensory and nutritional qualities of foods [4].sausage is a food that is prepared from comminuted and seasoned meat and is usually into a symmetrical shape [5].fresh comminuted meats and ingredients used as raw materials in the formulation can be modified to yield desirable organoleptic and keeping qualities of the product. in sausage preparation, raw meat batters are formed by chopping meats along with other ingredients to form a coarse dispersion of main water, fat and protein [6].in sausages and restructured meats, gel formation of myofibriller proteins is responsible for retaining and stabilizing water and fat in comminuted meats and for binding meat pieces [4]. collagen is the main component of skin. collagen and other connective tissue proteins play a negligible role in gel formation of sausage batter. however they are typically included in formulations for improving water binding, processing yield, juiciness and palatability or even blend cost in processed meat [7-9]. many researchers have reported the addition of starch in various sausages in forms such as corn and potato starch, including modified starch, for evaluation of the preparation of bologna type beef sausage [10] potato flour and tapioca starch for preparation of pork sausage during refrigerated storage [1] or potato starch in low fat frankfurters[11]. broadly speaking the basic preparation of sausage generally involves the mincing, grinding or cutting of lean meat and fat, the addition of spices, curing salts or other ingredients, followed by filling into containers [12]. preparation of sausages predates recorded history. sausage making developed slowly, beginning with the simple process of salting and drying meats. this was done to preserve fresh meat that could not be consumed immediately. as the human community expanded geographically, the availability of raw materials and differences in climate effected variations in manufacturing procedures. as a result, the typical flavors textures and shapes of many sausages described day today as frankfurters, braunschweiger, pork sausage and salami were produced [5].consumers nowadays eat sausages due to its convenience, variety, economy and nutritional value. sausages are economical since they were commonly manufactured from the cheaper cuts of meat and from by products. in fact, sausage products no doubt resulted from the attempts of primitive man to utilize for future use the less desirable parts and by products from slaughtering. the motto of the sausage industry wills might be ‘buy a pound; serve a pound’[13].pork sausage belongs to the category of fresh sausage whereas dry and semidry sausages, cooked sausages, cooked smoked sausages, uncooked smoked sausages and cooked meat specialties were other varieties [5]. they were made from uncooked or cooked raw materials [14]. pork sausages utilized head, neck, regular trimmings as meat sources and were a minced, cased product [15]. this type of meat products regarding emulsion type sausage was heat treated to between 750c and 800c to achieve desirable organoleptic properties and bacterial stability [14].they usually differ from other fresh meats in that it contains ground pork, salt and spices. commonly but not necessarily, it has a somewhat short refrigerated shelf life because of the relatively large microbial population it contains when it was prepared from the large part of pork trimmings. they have a bright red color and were usually wrapped in oxygen permeable films such as cellophane, polyvinylchloride and polyethylene [15]. pork sausages comprised of 30%fat [5] soy proteins, milk proteins, starch, wheat flour and yeast as meat extenders, (3.0-3.5)% salt [15] sucrose and dextrose as sugar [15] spices viz. ginger, onion, cinnamon, cloves, black pepper, coriander seed, etc.[5] and other ingredients as curing salt viz. sodium nitrate and nitrite or sodium nitrite alone [5] monosodiumglutamate [16] vinegar [17-19] and soysauce [20]. the majority of natural casings for sausages are obtained from the intestine of pigs, sheep and cattle. the length of gut obtained from an animal is influenced by the breed and degree of maturity [12]. intestines intended for use as casings for sausages must be immediately processed after evisceration. first, they are emptied and well flushed. pig, sheep and goat small intestines are scrapped thoroughly without inversion to remove the exterior (serous) and interior parts (mucous membrane), preserving the middle elastic muscle [15]. all salted natural casings should be thoroughly flushed inside with running water in the morning before use. they are then dipped into warm water to regenerate their elasticity (small intestines 10-20 minutes and large intestines 30-60 minutes) and drained for a short time before use [14]. the basic aim in cooked pork sausage manufacturing is to make a heat stable meat batter, a meat emulsion, consisting mainly of water and protein. heat treatment transforms it from a viscous form to a rigid and elastic solid structure which can be considered as a protein gel with entrapped fat particles. if the protein gel is strong enough, the fat will not be separated during heat treatment [15]. the nutritional value of all foods including meat and meat products is being seriously considered in view of consumer interest and demand. nutritional constituents of some sausages viz. mortadella polish style pork sausage, raw and cooked sausages, dry and cooked salami, scraple, souse, thuringer, canned vinna sausage etc. were reported according to pearson and tauber [13]. proximate analysis of the main food components such as moisture, ash, fat, protein and carbohydrate and small numbers of specialized analyses based upon classical wet chemical procedures. in the age of more conventional food materials and simple food technology, such proximate and wet chemical analyses were probably sufficient for most quality control purposes [21]. the nitrogen content of meat proteins is about 16%, which means the protein content of meat is 6.25 times the nitrogen content [13]. a balanced intake of fat is essential to ensure the dietary supply of essential fatty acids and the fat soluble vitamins a, d&e [21]. processed meats are considered to be high-fat foods. fresh pork sausages and patties may have fat as high as 50% although the industry average is 36% [3]. high-fat contents also lead to reduced moisture content and give less firm sausage [22]. moisture is the most predominant component of cooked sausages, accounting for approximately 45-55% of total sausage weight but the exact amount varies depending on the amount added during preparation as well as the lean to fat ratios of the sausages [5]. the carbohydrate content of meat and meat products is usually negligible unless it is added during processing either as sugar or as carbohydrate material. never the less, the glycogen present at the time of slaughter, although it comprises only about 1%, plays a major role in determining the physical properties of meat. studies of the mineral content of meat have been largely confirmed to calcium, phosphorus, sodium, potassium and iron. it was reported that the content of magnesium, copper and zinc were agriculture and food sciences research, 2019, 6(1): 50-56 52 © 2019 by the authors; licensee asian online journal publishing group present in baby foods and several other processed items [13]. microbiological quality of pork sausage greatly affected by salmonella spp, coli forms, staphylococcus aureus and some spore formers [20]. salmonella spp may produce three main types of diseases i.e the enteric fevers, septicaemias and gastroenteritis [23]. coliform bacteria may reach the bloodstream and cause sepsis. certain coliform bacteria produce a potent enterotoxin resembling cholera toxin capable of producing acute diarrhoea, without invading intestinal epithelium. all strains of staphylococcus aureus are potential pathogens. under suitable conditions, strains produce a variety of toxins, the most important of which is probably α-toxin which in the animal test is lethal, dermonecrotic, haemolytic, leucocidal and causes platelet damage [24]. sensory characteristics by hedonic rating test was used to measure the consumer acceptability of pork sausage. the panelist was asked to rate the acceptability of the product on a scale, usually of nine points, ranging from like extremely to dislike extremely scale with different ranges and other experiences phrases could also be used [25]. storage of pork sausage under refrigerated condition can enhance the shelf stability. storage of meat products through proper packaging arrangement is an important consideration. since the cooked sausage is perishable, the storage quality of the product can be maintained in the refrigerator for about two week [26]. freezing is used in the food industry to extend the shelf life of the products. however, during frozen storage the formation of ice crystals can occur and lead to lower binding properties in the products [27].the packaging requirement of meat products was further influenced by the types of processing and mechanizing that may be applied to them [28]. plastic as a packaging material encompass a whole family of natural and manmade substances [29]. the traditional package for meats was made from various types of paperboard. papers are still extensively used, but for most applications, they are coated or combined with other materials to improve their functional qualities [28]. wrapping materials may also be required to provide barriers to liquid water, oils and greases [30]. waxed paper is extensively used for meat packaging. polyethylene meets the requirement of being a tough moisture vapor proof film which retains its flexibility at temperatures as low as 510c. its high porosity for oxygen was preferred for many fresh meat packaging applications [28]. now in the present course of work two different varieties of pork sausages viz. spicy and nonspicy were developed and their storage stabilities were carried out during different periods of interval under refrigerated conditions. sensory characteristics, nutritional characteristics and microbiological characteristics were also studied. 2. material and methods to carry out the research pork meat purchased from kokrajhar market, assam adjacent to the institution. wheat flour, common salt, sucrose, soy sauce, ginger, onion, clove, cinnamon, pepper, coriander, low-density polyethylene, waxed paper and casing of goat were also purchased from a local market. monosodium glutamate, sodium nitrate, sodium nitrate and acetic acid were collected from the food technology department of the institute. the methods of manufacturing of both the spicy and nonspicy varieties were represented by the following figure 1. figure-1. manufacturing layout of spicy and nonspicy pork sausage. source: subhajit ray, 2019. mincing completely destroys the meat structure. meat proteins in presence of salt, phosphate and cold water are dissolved forming a system consisting of a solution of salt-soluble proteins, muscle and connective tissue agriculture and food sciences research, 2019, 6(1): 50-56 53 © 2019 by the authors; licensee asian online journal publishing group particles. when fatty tissue is added the fat particles will be emulsified with the salt-soluble proteins during comminution. the proteins will, therefore, be able to cover the total fat surface, surrounding each fat particle and so stabilize the emulsion. during comminution, the structural breakdown of meat and fatty tissues occurs and a new system was formed i.e minced meat suspension after addition of salt, phosphate, ice water emulsion and other additives [14]. here the meat and fat were minced separately in a mincer through 5mm diameter perforated plate. mixer simply mixes the product to incorporate all of the ingredients [15]. chopping was carried out by a chopper which was composed of a revolving metal bowl that contains the meat mass. a chopper was often used as a means of batching the sausage mix, the mixed batch being transferred to an emulsifier for acquiring the desired texture [13]. a table of the bench should be placed under the outlet of stuffing horn or stuffer to support the casings as it was filled and to facilitate subsequent linking [15]. in recent years final internal temperature reached in particularly all cooked sausages have gradually increased. higher internal temperatures contribute to increase product shelf life and improved color development and stability [5]. 2.1. screening of sausage formulations pork emulsions were prepared by eight formulations e.g. a1, a2, a3, a4 and b1, b2, b3, b4 of non spicy and spicy varieties respectively. quality characteristics were studied by sensory evaluation. 2.2. preparation of ingredients the ingredients used in the preparation of meat emulsion and quantities were given in table 1. table-1. formulation of nonspicy and spicy varieties of pork sausage (mg/100gm). i a1 a2 a3 a4 b1 b2 b3 b4 pork meat 72 65 56 50 63 59 52 46 fat 18.7 16.5 15 14 16.3 15.4 14.4 12.5 binder 4.43 5.8 7.1 7.2 5.5 6.00 7.21 7.92 salt 1.44 1.44 1.44 1.44 1.44 1.44 1.44 1.44 soy sauce 1.44 1.44 1.44 1.44 1.60 1.60 1.60 1.60 sucrose 1.44 1.44 1.44 1.44 1.60 1.60 1.60 1.60 ginger 0.52 0.52 0.55 0.52 0.40 0.40 0.40 0.40 onion 2.42 2.45 2.45 2.45 vinegar 0.80 0.80 0.80 0.80 spice mix 0.20 0.20 0.20 0.20 water 7.83 17.0 23.93 6.71 11.08 17.87 25.06 nano3 0.02 0.02 0.02 0.02 0,02 0.02 0.02 0.02 nano2 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 note: legend: i – ingredients, a1, a2, a3 and a4 – non spicy variety pork sausage, b1, b2, b3 and b4 – spicy variety pork sausage. 2.3 sensory evaluation technique the product was fried in mustard oil and then served as coded samples for sensory evaluation. there was ten panellist including faculty members and students of the food technology department. the parameters used were color, flavor, texture, palatability and overall acceptability through nine-point hedonic rating test as suggested by ranganna [25]. the best and appropriate two products i.e. spicy and nonspicy was selected by doing statistical analysis of sensory evaluation according to watts, et al. [31]. these samples were used to proceed for storage stability determination under refrigeration. both the proximate analysis and microbiological analysis were carried out. 2.4. nutritional analysis both the two varieties of pork sausages viz. spicy and non spicy were undergoing for nutritional analysis. the moisture content was determined by air drying method according to aoac [32]. the crude fat content was determined according to gerrard [12]. the moisture free sample was used to determine protein content. carbohydrate content of the sample was determined by taking the sum starch and total sugars. the total ash of the sample was determined. for mineral contents determination, dry ashing was done and it was then used for estimation of calcium, iron and phosphorous. the titratable acidity was determined as acetic acid [26].the dried sample was used to determine the crude fibre content according to osborne and voogt [21]. 2.5. microbiological analysis microbiological analysis was carried out for both varieties of pork sausages. total colony count in terms of total viable organisms was determined by pour plate method according to harrigan and mccance [33]. presence of coli form group of organisms was enumerated by mpn technique according to refai [34]. detection of salmonella sppwas done according to varadaraj [35]. detection of staphylococcus aureus, a food poisoning organism was carried out according to colee, et al. [36]. agriculture and food sciences research, 2019, 6(1): 50-56 54 © 2019 by the authors; licensee asian online journal publishing group 3. results and discussions in this study the sensory characteristics, nutritional characteristics and microbiological quality during storage of different varieties viz.non spicy (a1, a2, a3 and a4) and spicy (b1, b2, b3 and b4) were determined. 3.1. sensory characterization the quality of different formulations of pork sausage viz. nonspicy (a1, a2, a3 and a4) and spicy (b1, b2, b3 and b4) were evaluated according to its major sensory attributes such as color, texture, flavor, palatability and overall acceptability by nine-point hedonic rating test. the result of the experimental study was shown by the following figure 2 and figure 3 respectively figure-2. sensory evaluation of nonspicy pork sausage. source: subhajit ray, 2019. minced and seasoned meat encased in skin and cooked or preserved, mainly to be eaten cold in slices. screening of sausage formulations was carried out by statistical analysis by anova [31]. according to figure 1 formulation, a1 was mostly accepted by the panelists among nonspicy formulations and this may be due to the high meat content, while it was significantly different with a2 and a4 but not different with a3 at 5% confidence level for the parameter overall acceptability. the formulation b2 was superior among spicy formulations according to the panellists which may be due to mild addition of figure-3. sensory evaluation of spicy pork sausage. source: subhajit ray, 2019. minced and seasoned meat encased in skin and cooked or preserved, mainly to be eaten cold in slices figure 3. water and fine communication lead to developing in flavor and palatability. now for the overall acceptability, the formulation b2 was significantly different with b3 but not different with b1 and b4 on the basis of overall acceptability at 5% confidence level. according to figure 2. so the product a1 and b2 were significantly different in color and overall acceptability while not intexture, flavor and palatability at 5% level of probability. now the statistical analysis showed that the formulations a1 and b2 were the best among nonspicy and spicy variety respectively. 3.2. nutritional characteristics during storage the superior pork sausages viz. a1 of nonspicy category, b2 spicy category were stored using low-density polyethylene and waxed paper as packaging materials under refrigerated storage condition. after 1st day and 14th agriculture and food sciences research, 2019, 6(1): 50-56 55 © 2019 by the authors; licensee asian online journal publishing group days of storage periods, they were analyzed for their chemical characteristics viz. moisture content, total ash, protein content, carbohydrate content, fat content, crude fibre, salt content, acidity and calcium, phosphorous and iron as minerals. the experimental results were shown in table 2 and table 3. table-2. nutritional characteristics of pork sausage during 1st day storage period. c1 pr1 f1 tc1 ca1 p1 fe1 sa1 w1 a1 ac1 cf1 a1 in ldpe 12.87 11.7 8.3 178 118 135 1.72 63.409 270 0.05 1.3 a2 in wax paper 12.85 11.8 8.3 175 119 130 1.70 63.336 270 0.05 1 .32 b2 in ldpe 11.04 11.5 8.01 214 127 140 1.85 65.129 290 0.1 1.7 b2in wax paper 11.03 11.6 8.00 13.8 129 142 1.77 65 290 0.1 1.72 note: legend: 11st day storage, c1composition(gm/100gm sample), pr1protein(gm), f1 – fat(gm),tc1total carbohydrate(gm),ca1 – calcium(mg), p1 – phosphorous(mg), fe1 – iron(mg), sa1 – salt(gm), w1 – water(gm), a1 – ash(gm), ac1 – acidity(%), cf1 – crude fat(gm),ldpelow density polyethylene. table-3. nutritional characteristics of pork sausage during 14th day storage period. c14 pr14 f14 tc14 ca14 p14 fe14 sa14 w14 a14 ac14 cf14 a1 in ldpe 12.8 11.2 8.4 180 119 130 1.70 50.0 272 0.05 1.3 a2i n wax paper 12.2 11.9 7.9 174 121 135 1.75 64.217 273 0.05 1.33 b2 in ldpe 11.3 11.2 8.1 220 125 148 1.86 65.033 294 0.1 1.72 b2 in wax paper 11.1 11.8 7.6 227 123 145 1.82 65.15 295 0.1 1.74 note: legend: 1414th day storage, c14composition (gm/100gm sample), pr14protein(gm), f14 – fat(gm),tc14total carbohydrate(gm),ca14 – calcium(mg), p14 – phosphorous(mg), fe14 – iron(mg), sa14 – salt(gm), w14 – water(gm), a14 – ash(gm), ac14 – acidity(%), cf14 – crude fat(gm), ldpelow density polyethylene. table 2 and table 3 showed that the proximate composition of the different varieties of pork sausages after 1st day and 14 days of storage periods under refrigerated condition, a slight variation in protein, fat and moisture was observed in case of low-density polyethylene packaging for both a1 and b2 formulations. so there were no such significant variations in composition. the very low water vapor transmission rate may be responsible for this result [37]. moreover, a decrement of protein and fat content after 1st day and 14thdays storage period and simultaneous increment of moisture content after 14 days storage period were observed in both formulations. this was due to the fact that the water vapor transmission rate and gas transmission rate of waxed paper was significant. 3.3. microbiological quality during storage the microbiological analysis of the stored pork sausages after 1st, 7th, 14th and 21st days respectively of both nonspicy and spicy varieties viz. a1 and b2 were carried out for enumeration of the total number of microorganisms by plate count technique, coliform count and detection of salmonella spp, staphylococcus aureus. the experimental results were shown in table 4. table-4. microbiological quality of pork sausage during different storage periods. sv sp tpc cf s sa a1 in ldpe 1 160 7 410 14 1.6x103 21 4.9x104 a1in wax paper 1 210 7 3.2x103 4 14 6.9x105 >110 21 8.4x106 >300 b2 in ldpe 1 240 7 520 14 1.2x103 21 4.3x104 b2 in wax paper 1 310 7 2.8x103 7 14 5.6x105 >1100 21 6.4x106 >2000 note: legend: svsample variety, spstorage period (day), tpctotal plate count, cfcoliform, ssalmonella spp, sastaphylococcus aureus, ldpelow density polyethylene. table 4 showed that the bacteriological quality of the stored sausages. sample b2 showed a higher initial population of bacteria in terms of total plate count than sample a1. both a1 and b2 showed negative response during the test of coliform, salmonella spp and staphylococcus aureus when packed in low-density polyethylene. both the spicy and nonspicy varieties packed in the wax paper showed high total plate count than low-density polyethylene. during different storage periods, there was an increase in the coliform count but occurrence wax only in wax paper packaged materials and at 14th day coliform count was observed maximum in spicy variety b2. total plate count increasing rate was low in sample b2 than in sample a1 because of antibacterial properties of spices [5] and acidic nature of the product which may control the population of microbes [16]. both the two major food poisoning bacterias viz. salmonella spp, staphylococcus aureus was absent in both these two categories of sausages. both of them were non spore formers weiser, et al. [38] and frobisher and grambtree [39]. so it was evident from the result that the pork sausages packaged in the wax paper could be consumed up to one week but the same product packaged in lowdensitypolyethylene could be consumed up to two weeks when both kept under refrigerated condition. agriculture and food sciences research, 2019, 6(1): 50-56 56 © 2019 by the authors; licensee asian online journal publishing group 4. conclusions sensory characterization revealed that among nonspicy variety and spicy variety a1 and b2 were superior to other formulations due to two important attributes viz. color and overall acceptability. the shelf life of both categories of sausages viz. a1 and b2 were greater when packaged in low-density polyethylene than packaged in wax paper under refrigerated storage condition. moreover, it was also observed that spicy sausage viz. b2 showed greater shelf stability than nonspicy counterpart viz. a1 in context to bacteriological stability point of view. nutritional characteristics of both these two varieties of sausages were not so prominently varied during refrigerated storage condition and as a result, showed its health beneficial importance. references [1] w. s. ruban, a. kalaikannan, and v. apparao, "physico-chemical characteristics of pork sausage during refrigerated storage," veterinary world, vol. 2, pp. 95-97, 2009. 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[28] j. m. ramsbottom, packaging. the science of meat and meat products, 2nd ed. sanfransisco: j.f.w.h. freeman and company, 1971. [29] r. c. griffin and s. sacharow, principles of packaging development. westport connecticut: the avi publishing company inc, 1978. [30] r. p. goddard, wrapping and packaging papers, ed. by paine, f.a. the packaging media. london: blackie and sons ltd, 1977. [31] b. m. watts, g. l. ylimaki, and l. g. elias, basic sensory methods for food evaluation. canada: the international development research centre, 1989. [32] aoac, official methods of analysis of aoac international 20th edition, 2016, editor:dr. george w. latimer, jr, 20th ed. vol. 45. usa: published by aoac international, 2012. [33] w. f. harrigan and m. e. mccance, laboratory methods in food and dairy microbiology. new york: academic press inc, 1976. [34] m. k. refai, manuals of food quality control, microbiological analysis. rome: fao.the united nations, 1979 [35] m. varadaraj, "methods for detection and enumeration of foodborne bacterial pathogens: a critical evaluation," journal of food science and technology, vol. 30, pp. 1-13, 1993. [36] j. g. colee, j. p. duguid, and b. p. marmion, practical medical microbiology, 8th ed. baltimore, usa: the williams and wilkins company, 1989. [37] export, export packaging note no.16, international trade centre. switzerland: unctad/ gatt, 2011. [38] h. h. weiser, g. j. mountney, and w. a. gould, practical food microbiology and technology, 2nd ed. westport, connecticut: the avi publishing company inc, 1971. [39] h. frobisher and g. grambtree, fundamentals of microbiology, 9th ed. japan: toppan company limited, 1974. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 73 © 2018 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 5, no. 2, 73-78, 2018 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2018.52.73.78 © 2018 by the authors; licensee asian online journal publishing group influence of agro-ecological areas on the antioxidant capacity of onion varieties grown in burkina faso virginie marie dakéné1  abdoulaye sérémé2 moumouni koala3 marius k. somda4 hagrétou sawadogo – lingani5 alfred s. traoré6 ( corresponding author) virginie marie dakéné1 1national laboratory of public health; 09 bp 24 ouagadougou 09; burkina faso 2natural substances department / irsat / cnrst, 03 bp 7047 ouagadougou 03; burkina faso 3department of medicine and traditional pharmacopoeia / irss / cnrst, 03 bp 7047 ouagadougou 03; burkina faso 5food technology department / irsat / cnrst, 03 bp 7047 ouagadougou 03; burkina faso 4,6department of biochemistry-microbiology; university ouaga i pr joseph ki-zerbo; 03 bp 7021 burkina faso abstract colored onion bulb is an important source of antioxidants. some ecological factors can significantly influence the content and quality of these substances. the aim of the present study is to determine the antioxidant and total polyphenol content of fresh bulbs of five (05) varieties of onion (galmi violet, damani violet, prema, safari and a local variety-lv) grown in burkina faso, and to evaluate also the influence of agro-ecological parameters on the antioxidant content of the most cultivated variety (galmi violet) of them. samples of onion bulb for analysis were collected in the six agro-ecological areas of burkina faso under the same cultivation conditions. antioxidant activities were evaluated using the method of ferric reducing antioxidant power. the evaluation of the total polyphenol contents was carried out using the folin-ciocalteu method. the results showed that antioxidant activities for the 5 varieties tested vary between 0.125 ± 0.001 and 0.149 ± 0.004 mg te / g, and total polyphenol contents from 0.172 ± 0.011 to 0.272 ± 0.003 mg eag/g. results on the assessment of the effect of ecology on the antioxidant content of galmi violet variety revealed significant variations in antioxidant content from 0.144 ± 0.002 to 0.155 ± 0.001 mg te / g, and total polyphenols from 0.208 ± 0.014 to 0.292 ± 0.012 mg eag / g depending on the ecological cultivation area. keywords: agrarian, cultivation. citation | virginie marie dakéné; abdoulaye sérémé; moumouni koala; marius k. somda; hagrétou sawadogo – lingani; alfred s. traoré (2018). influence of agro-ecological areas on the antioxidant capacity of onion varieties grown in burkina faso. agriculture and food sciences research, 5(2): 73-78. history: received: 12 october 2018 revised: 21 november 2018 accepted: 17 december 2018 published: 7 january 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this research project was supported by a grant from the west africa agricultural productivity program/national center of specialization fruits and vegetables (waapp / ncs-fv). competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 74 2. material and methods ..................................................................................................................................................................... 74 3. results ................................................................................................................................................................................................ 75 4. discussion .......................................................................................................................................................................................... 76 5. conclusion ......................................................................................................................................................................................... 77 references .............................................................................................................................................................................................. 77 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.52.73.78&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.52.73.78&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/1621 agriculture and food sciences research, 2018, 5(2): 73-78 74 © 2018 by the authors; licensee asian online journal publishing group 1. introduction air pollution, ionizing radiation, radioactive radiation (uv, x or ɣ) consequences of industrialization and urbanization, as well as tobacco and alcohol, are among the main risk factors responsible for diseases, related to oxidative stress. these diseases pose an increasingly serious public health problem [1]; [2]. indeed, diseases related to oxidative stress are the increasingly important causes of disability and premature death in both developed and developing countries such as burkina faso. in 2001, they were responsible for about 60% of the 56.5 million deaths worldwide, and 46% of the global burden of morbidity [1]. the same source indicates that this morbidity rate will rise to 57% in 2020, and diseases that are linked to oxidative stress will be responsible for nearly 3/4 of deaths worldwide. in developing countries deaths from these diseases dominate in mortality statistics [3]. one of the main causes of these diseases is the action of free radicals, chemical molecules naturally produced in the body during different reactions of metabolism. antioxidants are chemical molecules that not only fight against free radicals, but also repair the damage they cause. they therefore play a leading role in the prevention of diseases caused by oxidative stress [4]; [5]; [6]. synthetic antioxidants exist but are inaccessible to consumers because of their cost. in addition, at high doses, these synthetic antioxidants could become pro-oxidants potentially dangerous for health defraigne and pincemail [7]; edeas [8]; favier [9]. pincemail, et al. [10] pointed out that many studies show that a high level of antioxidants obtained through a diet rich in fruits and vegetables can reduce safely the relative risk of mortality due to oxidative stress. food and nutrition are therefore important factors in determining the prevention of oxidative stress related illnesses [3]. several foods are used as natural sources of antioxidants, but their antioxidant abilities are not well known generally, hence the present investigation on onion (allium cepa l.). because of its composition, onion is a plant with many nutritional and antioxidant properties. indeed, many studies attribute it properties to fight against certain diseases related to oxidative stress such as cancer, cardiovascular disease, diabetes, aging-related diseases [11]; [12]; [13]; [14]; [15]. burkina faso is one of the largest producers of onion in west africa where it ranks fourth after nigeria, niger and senegal [16]. onion bulb ranks first among market gardening. it is a consumer product both in urban centers and in rural areas. in 2012 the production was 329319 tons, and this production continues to grow [16]. the onion bulb produced in burkina faso is exported to the countries of the sub-region, particularly in côte d'ivoire, ghana, togo and benin [16]; [17]. its use as a natural source of antioxidants could therefore allow populations, and particularly vulnerable ones such as children, pregnant women and the elderly, to prevent diseases related to oxidative stress. however, some factors such as culivation techniques, environmental conditions, have an impact on the biosynthesis of antioxidant compounds in plants [18]; [19]; [20]; [21]; [22]. the aim of the present study is to check antioxidant and total polyphenol content of five (05) varieties of onion bulb (galmi violet, damani violet, prema, safari and lv ) grown in burkina faso, and to determine the influence of agro-ecological factors on the antioxidant content of galmi violet, the most cultivated variety. 2. material and methods 2.1. sampling the study is focused on the following five varieties: galmi violet, damani violet, prema, safari and a lv. samples of each variety of onion bulb were collected in six regions of burkina faso: mouhoun, high basins, north, central, west central and east, for laboratory analyzes. sampling was done randomly for each variety. since the cultivation techniques are identical for the six areas, two (02) batches each consisting of a 50 kg bag of bulb were collected by variety. varieties collected by region are presented in table 1. a total of twenty (20) lots of 05 onion varieties were collected for this study (table 1). the batches were kept at room temperature, in a ventilated place, away from solar, heat and light. table-1. varieties collected and number of lots by region region number of lots varieties collected mouhoun 02 galmi violet central 02 galmi violet west central 06 galmi violet, prema, lv east 04 damani violet, safari high basins 02 galmi violet north 04 galmi violet, safari source: field work data. 2.2. methods of analysis the determination of the antioxidant activities and the total polyphenol contents on fresh extracts of onion bulb concerned all the five varieties. the evaluation of the effect of the agro-ecological areas on the concentration of antioxidants and total polyphenols is done on the galmi violet variety only. 2.2.1. sample preparation and extraction of antioxidants for each variety, three (3) bulbs from each batch are taken randomly. the outer peel (dried flake) were removed and then cut into pieces using a stainless steel knife and crushed using a porcelain mortar. then, 2 grams of the fresh onion raw material was immediately weighed in a glass beaker and mixed with 3 ml of solvent consisting of a mixture: acetone, water, acetic acid (awa) in the proportions of 70: 29.5: 0.5 (v / v). after homogenization with the vortex mixer, the mixture was macerated for 24 hours under cold conditions (4 °c.). after filtration using whatman no 1 filter paper, the filtrate was stored at 4 °c protected from light in amber flasks for further analysis. this extraction operation was repeated three times in succession in order to ensure the total extraction of the sample. agriculture and food sciences research, 2018, 5(2): 73-78 75 © 2018 by the authors; licensee asian online journal publishing group 2.2.2. evaluation of the antioxidant activity of the varieties collected the frap method (ferric ion reducing antioxidant power) [23] was used to evaluate the antioxidant activity of the collected samples. the principle is to evaluate the effectiveness of an antioxidant by its ability to reduce iron (transition from ferric form, fe3 + to ferrous form, fe2+). the oxidant used for this purpose is a ferric salt, fe (iii) (tptz)2cl3 (tptz = 2,4,6-tripyridyl-s-triazine) which goes from colorless to intense blue when reduced. the absorbance of the solution is measured at 595 nm [24]. the procedure was as follows: a volume of 20 µl of extract was introduced into 30 µl of distilled water and then 200 µl of frap solution which is prepared by mixing 1 ml of tptz, 10 ml of a buffer solution of acetate of sodium (ph = 3.6) and 1ml of fe (iii), h2o solution. a blank consisting solely of 250 μl of the extraction solvent was also prepared. the whole was incubated at 37oc for 10 minutes after homogenization. absorbance was then read at 595 nm with a spectrophotometer (mp96, safas) for each sample. a calibration curve was established using trolox as a reference (antioxidant). the results were determined from the equation (y = 28.67 x + 0.066, r2 = 0.999) of the standard curve and were expressed in mg of trolox equivalent (te) per gram of fresh plant material. an average of the contents for each variety was calculated. 2.2.3. evaluation of the phenolic compounds content of the varieties collected the evaluation of the content of phenolic compounds was made using the method of folin-ciocalteu (r-fc) [25] whose principle is to react on the sample (the extract), the reagent of folin-ciocalteu (diluted 10 times) and absorbance measured at 765 nm. the procedure was as follows: a volume of 60 μl of extract was mixed with 60 μl of the folin-ciocalteu reagent diluted 10 times. the mixture was left at room temperature for 8 minutes to allow complete reaction of the reagent on the oxidizable compounds or on the phenolates. then, 120 μl of 7.5% (w / v) aqueous sodium carbonate solution (na2co3) was added to neutralize the residual reagent. after 30 minutes of incubation at 37 °c, the absorbance of the extracts was measured at 765 nm with the mp96 spectrophotometer, safas (96 well quartz microplates). a blank is made with 60 μl of r-fc and 120 μl of sodium carbonate solution. a calibration curve was established using gallic acid as a reference. the results were determined using the equation (y = 46.41x + 0.063, r2 = 0.998) of the calibration curve. they were expressed as mg gallic acid equivalent (gas) per gram of fresh plant material. all measurements were made in triplicate. 2.2.4. statistical analysis of the results the statistical analysis of the results was carried out using the genstat version 14 software. the comparison between the varieties was also performed by the multivariate analysis of genstat version 14. 3. results 3.1. antioxidant activities (aao) of onion bulb varieties collected in the 6 production areas the results of the antioxidant activity of the extracts of the 05 varieties of onion bulb collected in the mouhoun, high basins, north, central, west central and east regions are presented in the table 2. significant differences (p <0.05) between the antioxidant activities (aao) of different varieties of fresh bulb are observed. these contents range from 0.125 ± 0.001 to 0.149 ± 0.004 mg te / g fresh material. the varieties with the highest levels of antioxidants are lv, galmi violet and damani violet, with contents respectively 0.149 ± 0.004; 0.147 ± 0.004 and 0.139 ± 0.000 mg te / g fresh material. however, it is noted that the levels are substantially equivalent for the galmi violet (0.147 ± 0.004 mg te / g fresh material) and lv (0.149 ± 0.004 mg te / g fresh material) varieties. prema and safari have lower levels (0.125 ± 0.001 and 0.132 ± 0.002 mg te / g fresh material). table-2. antioxidant activity of the 5 varieties of onion bulb collected variety antioxidant activity (aao) (mg te / g mf) galmi violet 0,147 ± 0,004c lv 0,149 ± 0,004c damani violet 0,139 ± 0,000b safari 0,132 ± 0,002ab prema 0,125 ± 0,001a note: all assay results are expressed as mean contents ± standard deviation. the averages affected by the same letter are not significantly different. 3.2. total polyphenol (tpp) content of onion bulb collected in production areas the results of the total polyphenol contents of the extracts of the 05 varieties of onion bulb collected in the mouhoun, high basins, north, central, west central and east regions are shown in table 3. the results show a significant variation (p <0.05) from one variety to another. the contents range from 0.220 ± 0.013 to 0.272 ± 0.003 mg eag / g fresh material for the five varieties. the highest levels are obtained with galmi violet, prema and damani violet varieties, with respectively 0.272 ± 0.003; 0,270 ± 0,014 and 0,254 ± 0,017 mg of eag / g of fresh material. the safari variety has the lowest content (0.172 ± 0.011 mg eag / g fresh material). however, galmi violet and the prema have almost the same level of polyphenol. table-3. total polyphenol content of the 5 onion bulb collected variety total polyphenol (tpp) content (mg eag / g mf) galmi violet 0,272 ± 0,003c lv 0,220 ± 0,013b damani violet 0,254 ± 0,017bc safari 0,172 ± 0,011a prema 0,270 ± 0,014c note: all assay results are expressed as mean contents ± standard deviation. the averages affected by the same letter are not significantly different. agriculture and food sciences research, 2018, 5(2): 73-78 76 © 2018 by the authors; licensee asian online journal publishing group no significant correlation was observed between aao and tpp for the different varieties (r2 = 0.069) (figure 1) figure-1. correlation between aao and tpp of 5 varieties of onion collected in the production areas 3.3. effect of the agro-ecological areas of burkina faso on the antioxidant and polyphenol contents of galmi violet variety the results of the effect of the agro-ecological areas in burkina faso on the antioxidant and total polyphenol contents of the variety galmi violet are presented in table 4. two batches of fresh onion bulb were collected by region, the results are an average of the data obtained from the two lots. these results show that, there is a significant variation (p <0.05) of antioxidants and polyphenols contents from one ecological region to another. for aao and tpp these levels respectively range from 0.144 ± 0.002 to 0.155 ± 0.001 mg te / g fresh material, and from 0.208 ± 0.014 to 0.292 ± 0.012 mg eag / g fresh material (table 4). the highest levels of antioxidants are obtained with the mouhoun samples (0.155 ± 0.001 mg te / g fresh material). the north (0.152 ± 0.003 mg te / g mf), west central (0.152 ± 0.006 mg te / g mf) and high basins (0.147 ± 0.004 mg te / g mf) have equivalent levels. central samples show the lowest levels (0.144 ± 0.002 mg te / g fresh material). the highest levels of total polyphenols are obtained with north region (0.292 ± 0.012 mg eag / g fresh material), central region (0.289 mg eag / g fresh material), west central region (0.280 ± 0.002mg eag/g fresh matter) and mouhoun region (0.265±0.006 mg eag / g of fresh material). these 4 regions have substantially equal contents. high basins presented the lowest levels (0.208 ± 0.002 mg of eag / g of fresh material). table-4. antioxidant and total polyphenol content of galmi violet by locality region aao (mg of te / g of fresh material) tpp (mg of eag / g of fresh material) central 0,144 ± 0,002a 0,289 ± 0,000b west central 0,152 ± 0,006ab 0,280 ± 0,002b north 0,152 ± 0,003ab 0,292 ± 0,012b mouhoun 0,155 ± 0,001b 0,265 ± 0,006b high basins 0,147 ± 0,004ab 0,208 ± 0,014a note: all assay results are expressed as mean contents ± standard deviation. the averages affected by the same letter are not significantly different. 4. discussion 4.1. antioxidant activities (aao) and total polyphenol content (tpp) of onion varieties collected in production areas: mouhoun, high basins, north, central, west central and east the results obtained are in agreement with those of the literature concerning the variation of the concentrations of antioxidants and total polyphenols between varieties of the same species as pointed out by causse [26] whose researches have shown the existence of a wide range of variation of the concentrations of antioxidants and total polyphenols between varieties of the same species of fruits and vegetables, ranging from single to double. in fact, jeffery, et al. [20] showed, by comparing the β-carotene content of 50 varieties of broccoli, which could be 6 times greater from one variety to another. kurilich, et al. [27] showed a 5 times increase in total antioxidant capacity between eight (08) varieties of the same plant according to ecological areas. several factors would be responsible for the variation in antioxidant levels between varieties of the same species. indeed, as causse [26]. benbrook [28] these variations are physiological, genetical, agronomical (cultivation techniques, production method), or environmental (radiation, temperature, region of culture). to our knowledge very little work has been done on the antioxidants and polyphenols of onion bulb varieties grown in burkina faso. the variations observed in the case of onion bulb varieties of our research, could be explained by the combined effect of these factors. one could thus think that the equivalence of the levels observed between the varieties of galmi violet and the lv for the antioxidants, and between the varieties of galmi violet and prema for the polyphenols, would be the fact of genetic factors, since the cultivation conditions (agronomic and environmental) were the same for these varieties. r2 = 0.069 indicating that only 06.9% of the antioxidant capacity of the extracts is due to the contribution of the phenolic compounds. in other words, the richest extract of phenolic compounds is not necessarily the one which has the highest antioxidant activity. these results clearly show that polyphenols are not the only compounds responsible for the antioxidant activity of onion bulb extracts. similar results have been agriculture and food sciences research, 2018, 5(2): 73-78 77 © 2018 by the authors; licensee asian online journal publishing group previously reported by other authors. indeed, dabiré [29]; psarra, et al. [30] showed a weak correlation between antioxidant power and phenol content by working on extracts of tridax procumbens species and wine samples. carotenoids are also compounds with very high antioxidant activity contained in the onion. koala, et al. [31] showed a good correlation between total carotenoid levels and the antioxidant activities of eight (08) orange fleshed sweet potato varieties. we could therefore think that carotenoids would be the compounds that would contribute, to a large extent, to the antioxidant activities of the onion bulb varieties collected. 4.2. effect of the agro-ecological areas of burkina faso on the antioxidant and polyphenol contents of galmi violet variety the results of the effect of the agro-ecological areas of burkina faso on the levels of antioxidants and total polyphenols of galmi violet variety, show that the contents of these compounds vary according to the agroecological areas, especially between central, mouhoun, and the other 3 regions (west central, north and high basins) for antioxidants, and between the high basins and the other 4 regions (west central, north, central, mouhoun) for polyphenols. this shows that the areas have an influence on the concentration of these compounds in onion bulb grown in burkina faso. similar results have been observed by other authors. indeed, as pointed out by mauget [32] important differences in polyphenol and carotenoid contents are observed for the same variety of fruit or vegetable grown in different geographical areas. this has been confirmed by koala, et al. [31] which showed a 30% variation in polyphenol levels as a function of agro-ecological areas, working on orange-fleshed sweet potato (ofsp). as dragovic-uzelac, et al. [19] pointed out, that, without climatic characterization of the region, it is difficult to attribute these variations to any factor of the air or edaphic environment. the nature of the soils in the areas concerned is practically identical because the type of dominant soils in these areas is ferruginous soils, but the morphological characteristics (structure, texture, depth, thickness, color, porosity, etc.) and the analytical characteristics differ from an area to another bunasols (national office of soils) [33]; bunasols [34]; bunasols (national soil bureau) [35]; bunasols (national soil bureau) [36]. schreiner [22] has shown that in some vegetables, radiation and temperature play a more important role than water or fertilizer. variations in antioxidant and polyphenol levels from one area to another for the same variety of onion could therefore be explained by the effect of these factors, and the morphological and chemical composition differences observed at the soil level. the equivalence of concentrations noticed between the areas of west central, north and high basins, for the antioxidant content, on one hand, and west central, north, central and mouhoun for the content of polyphenols in other hand, could be explained by the existence in these areas of similar climatic and / or edaphic conditions. 5. conclusion the results of this work show that onion bulb varieties cultivated in burkina faso are rich in antioxidants, and concentrations vary from one agro-ecological area to another. the onion bulb could therefore be used as natural sources of antioxidants, and could thus contribute to the prevention of oxidative stress related diseases which today constitute a real public health problem in burkina faso and around the world. this onion bulb rich in antioxidant and others micronutrients makes it a vegetable that could help improve the nutritional status of vulnerable groups such as children, pregnant women and the elderly. however, for the future it is necessary that much further research studies based on the precise determination of the influence of the individual parameters of the ecological areas on the antioxidant content of the bulbous onion be carried out. this could lead to a better understanding of the impact of these parameters on the antioxidant concentration of onion bulbs and consequently to optimize the antioxidant biosynthesis of the bulbous onion produced in burkina faso. other studies could be focused on the determination of the carotenoids level in the bulb in order to determine the existence or not of a correlation between aao and total carotenoid content (tct). this will make it possible to know the contribution of these compounds to the antioxidant activity of the onion bulb grown in burkina faso, and consequently to optimize their biosynthesis within the plant. references [1] who (world health organization), "diet, nutrition and chronic disease prevention," technical report series no. 9162003. 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[36] bunasols (national soil bureau), "morpho-pedological study of provinces of yatenga, loroum and zondoma provinces. scale 1/100 000," technical report no. 122. burkina faso2004. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 1, no. 1, 1-4, 2014 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 1 production and nutritional studies of cookies and spiced millet drink nwachoko, ndidi 1* --jack, ibiba reuben 2 1,2 department of chemistry, rivers state university of science and technology, nkpolu-oroworukwo, port harcourt, rivers state, nigeria abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ............................................................................................................................................................. 2 2. materials and methods ........................................................................................................................................... 2 3. source materials ..................................................................................................................................................... 2 4. animal collection .................................................................................................................................................... 2 5. preparation of cookies ........................................................................................................................................... 2 6. preparation of spiced millet drink ....................................................................................................................... 2 8. proximate composition .......................................................................................................................................... 2 9. determination of glucose ....................................................................................................................................... 2 10. results .................................................................................................................................................................... 3 11. discussion............................................................................................................................................................... 4 12. acknowledgement ................................................................................................................................................. 4 references .................................................................................................................................................................... 4 nutritional studies of cookies produced from wheat (triticumaestivum) flour and spiced millet drink produced from millet (pinnesetumnigritarum) spiced with aframomumchrysanthum were investigated. the result of the proximate composition of cookies showed the moisture content to be 3.54%, crude protein 7.57%, fat 27.61%, ash 1.718% and total carbohydrate 59.562%. the spiced millet drink proximate values were moisture content 78.29%, crude protein 1.18%, fat 0.249%, ash 0.108% and total carbohydrate 20.173%. also the result of the nutritional studies carried out on the albino rat blood sample fed with these products at different percentages showed that the blood glucose values were (3.2, 2.90, 2.85, 3.33 and 3.63mmol/l), total protein (73.00, 69.67, 78.50, 86.00 and 83.33g/l) and total cholesterol (2.40, 3.03, 2.35, 3.20 and 3.53mmol/l) for group 1,2,3,4 and 5 respectively. comparing the result of the experimental groups with the control group, the obtained values for glucose, total protein and cholesterol were within the same range, whereas the blood triglyceride values (1.80, 2.50, 2.65, 2.27, and 2.07mmol/l) and high density lipoprotein (hdl) values (1.65, 2.07, 1.85, 2.67 and 1.83mmol/l) for group 1,2,3,4 and 5 respectively of the experimental groups were insignificantly above values of the control group. keywords: cookies, millet drink, aframomumchrysanthum, spiced, nutritional production. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2014, 1(1): 1-4 2 1. introduction in each passing day, humans either complain of hunger or food shortage. cookies are flat dry sweet biscuits that are made from wheat flour (composite form) and other natural food stuff such as yam, cocoyam and soybean, [1]. the word biscuit comes from the french word “biscuit”, meaning twice cooked and is a literal description of what happened in the early days of biscuit production. cookies are known to be of high nutritional value that could take care of almost all the classes of nutrients (carbohydrate, protein, fats and oil, minerals and vitamins). in most part of the world, baked foods based on wheat flour are popular food stuffs [1]. spiced millet drink had become one of the basic necessities of homes for sustenance of life. this is because of the enormous energy generated by food drink consumed at certain intervals of the day. it serves for carbohydrate, water and some other nutrients [2, 3]. the production of spiced millet drink assumes different forms in nigeria including kununzaki, kunungyada, kununakamu, kununjiko. however, kununzaki is the most commonly consumed type and has gained wider acceptability with some nigerian community. its preparation and processing is dependent on indigenous technology. this food drink is found to be one of the traditional non-alcoholic fermented beverage drink [4, 5]. cereal grains used for its preparation are sorghum maize, millet, guinea corn, or rice. the produced kunun is usually packed in plastic bottle under ambient temperature or cooled in refrigerator [6, 7]. 2. materials and methods 2.1. materials wheat flour, margarine, baking powder, coconut, egg, sugar, orange rind, sieve garri. millet, sweet potato, aframomumchrysanthum, water and sugar. 3. source materials wheat flour used for this study was purchased from rumuokuta market, port harcourt, and all other baking ingredients such as garri, baking powder, margarine, coconut, egg, sugar, orange rind. millet was purchased from a retail outlet in mile one market, port harcourt and the ingredient such as sweet potatoes, aframomunchrysanthum were also obtained from the same source. 4. animal collection fifteen (15) albino rats were collected from the department of biochemistry, university of port harcourt animal house. 5. preparation of cookies 200g of margarine and 125g of sugar were mixed until fluffy. 2 whole eggs, 1.5g coconut and 1.5g of orange rind were added while mixing continued for about 20 minutes. 300g of wheat flour, 5g of baking powder, and 50g of sieved garri were slowly introduced into the mixture. the dough was rolled and cut to various shapes with a cutter. place in a greased baking sheet and baked in the gas oven until golden brown and cooled. 6. preparation of spiced millet drink 200g of millet were steeped in water for 24hours, washed, ground with 0.5g of spices (aframomumchrysanthum) and 15g of sweet potato. hot water were pour into mixture then allowed to stand over night and strained the following morning, it was sweetened with 10g of sugar to taste and cooled in the refrigerator. 7. sensory evaluation the sensory properties of cookies and spiced millet drink were determined using a fifteen member panelists consisting of students from different departments of rivers state university of science and technology, cookies and spiced drink samples were subjected to sensory evaluation. the following attributes namely; taste, aroma, texture, appearance and over all acceptability were assessed on cookies and spiced millet drink samples. 8. proximate composition aoac [8] method was used for proximate analysis of cookies and spiced millet drink, and carbohydrate was determine by difference (carbohydrate = 100% (% moisture +% protein + fat/lipid + % ash). 8.1. sample preparation a total of (15) albino rats were divided into five (5) groups comprising of three (3) rats in each group. the groups were labeled 1-5. group 1 was used as the control and fed with their normal food and water throughout the period of the experiment. group 2 were fed with 20% of the product (cookies), 80% of the normal feed, group 3 were fed with 50% of the product, 50% of the normal feed, group 4 were fed with 70% of the product, 30% of the normal feed and group 5 were fed with 100% of the product (cookies). the experimental groups were given spiced millet drink only instead of water. on the fourteen day of feeding cookies and spiced millet drink to the experimental groups, the albino rats were sacrificed and blood sample were collected in lithium heparin bottles to avoid coagulation and they were labeled appropriately and sent for analysis. 9. determination of glucose the test tubes were labeled as test standard and blank, 1.0ml of glucose reagent was pipette into all the test tubes. 0.01ml of the albino rat blood samples were added into the appropriate tubes mixed and incubated at 37 o c for 10mins. the instrument was zeroed with the blank and the absorbance was read at 520nm. agriculture and food sciences research, 2014, 1(1): 1-4 3 lucose bsorbance of test bsorbance of tandard concentration of standard 9.1. protein the test tubes were labeled as test, standard and blank 1.0ml of the protein reagent was pipette into all the test tubes then 0.02ml of the sample was added into the appropriate tubes, mixed and incubated at 25 o c for 10mins, the instrument was zeroed with the blank and the absorbance was read. rotein bsorbance of test bsorbance of tandard concentration of standard 9.2. high density lipoprotein (hdl) the test tubes were labeled as test, standard and blank, 0.5ml of high density liquid (hdl) reagent was pipette into the appropriate tubes, the test tubes were labeled as test, standard and blank 0.1ml of the supernatant were added into the appropriate tubes and allowed to stand for 10mins at 25 o c. the instrument was zeroed with the blank and the absorbance were read at 540nm and calculated as: bsorbance of test bsorbance of tandard concentration of standard 9.3. total cholesterol the test tubes were labeled as test standard, and blank, 1.0ml of cholesterol reagent was pipette into all the tubes, then 0.01ml of the albino rat blood sample was added into the appropriate tubes, and water (h20) to the tube labeled blank and allowed to stand for 10mins at room temperature, then the absorbance was read, at 540nm and calculated as: total cholesterol bsorbance of test bsorbance of tandard concentration of standard 9.5. total triglyceride the test tubes were labeled as test, standard and blank 1.0ml of triglyceride reagent were pipetted into all the tubes, then 0.01ml of the albino rat blood sample were added into the appropriate tubes and water (h20) to the tube labeled blank and allowed to stand for 10 mins at room temperature, then the absorbance were read at 540nm and calculated as: trigl ceride bsorbance of test bsorbance of standard concentration of standard 10. results the proximate composition of cookies and spiced millet drinks and nutritional studies of albino rats blood samples fed with these products are shown in the tables below. table-1.proximate composition of cookies and spiced millet drink sample %moisture content % ash % fat %crude protein %total carbohydrate cookies 3.54 1.718 27.61 7.57 59.562 spiced millet drink 78.29 0.108 0.249 1.18 20.173 the above table showed the result of the proximate composition of cookies and spiced millet drink. table-2.nutritional studies of cookies and spiced millet drink using albino rat group fed ratio glucose (mmol/l) total protein (g/l) total cholesterol (mmol/l) triglyceride (mmol/l) high density lipoprotein (mmol/l) 1. control 3.200.57 73.004.24 2.400.28 1.800.14 1.650.21 2. 20% of the product 80% of normal feed 2.900.70 69.674.62 3.030.06 2.500.20 2.070.21 3. 50% of the product 50% of the normal feed 2.850.07 78.502.12 2.350.07 2.650.07 1.850.07 4. 70% of the product 30% of the normal feed 3.330.51 86.002.65 3.200.10 2.270.38 2.670.68 5. 100% of the product 3.630.68 83.333.51 3.530.15 2.070.06 1.830.06 the above table showed the result of the nutritional studies of cookies and spiced millet drink. values are expressed as mean sd. agriculture and food sciences research, 2014, 1(1): 1-4 4 table-3.sensory evaluation of cookies appearance flavour aroma mouth feel taste excellent 1 3 3 v. good 5 4 3 4 5 good 10 12 13 9 8 fair poor the above table showed the result of sensory evaluation of cookies done by 16 panelist. table-4.sensory evaluation of spiced millet drink appearance flavour aroma mouth feel taste excellent v. good 1 1 good 15 6 5 4 4 fair 10 11 9 10 poor 3 1 the above table showed the result of sensory evaluation of spiced millet drink done by 16 panelist. 11. discussion the result of the proximate composition of cookies in table1 showed that cookies are rich in carbohydrate (59.562%) and fat (27.6%). this percentage is quite moderate for consumption. the moisture content of the cookies was 3.54%. this low moisture content implies long shelf life. the ash and crude proteins content of the cookies are 1.718% and 7.57% respectively. the result of spiced millet drink revealed that spiced millet drink is high in moisture content (78.29%). this is good for all class of persons as water is essential for life. this high moisture content implies that the drink require a means of preservation such as refrigeration. table 2 showed the result of the nutritional study of albino rat blood sample fed with cookies and spiced millet drink. the result showed that the blood glucose values, total protein values and total cholesterol values of the experimental groups and the values obtained from the control group were within the same range. the total triglyceride values and high density lipoprotein (hdl) values were insignificantly above the values of the control group. table 3 and 4, showed the sensory evaluation of the cookies and spiced millet drink. the panelist noted that the products are quite good in appearance, flavour and aroma. 12. acknowledgement we are grateful to membere, martha d. and management of lively stones medical diagnostic laboratory. 1 rumuokparali road, off s.d.a. junction choba, for their contributions. references [1] h. barbara, from hardtack to home fries: an uncommon history of american cooks and meals. uk: free press, 2002. [2] s. a. odunfa, "microbiological quality of yoghurt and milk drink samples in ibadan, a nigerian city," journal of agriculture, vol. 2, pp. 43-46, 1988. [3] j. jango-cohen, "the history of food. twenty-first century books," pp. 4-8, cdb9267fr3z6, 2005. [4] r. e. lichtenwalner, g. i. glover, and c. c. shaw, "protease activity of water and acid, reconstituted grain sorghum," journal of agricultural food chemistry, vol. 27, pp. 352-362, 1979. [5] t. adenyemi and s. umar, "effect of method of manufacturing on quality characteristics of kunun zaki, a millet based beverage," nigerian food journal, vol. 12, pp. 34-40, 1994. [6] n. a. amusa and o. a. odumbuka, "microbiological and nutritional quality of hawked kunun (a sorghum based non-alcoholic beverage) widely consumed in nigeria," pakistan journal of nutrition, vol. 8, pp. 20-25, 2009. [7] a. t. ikekoronye and p. o. ngoddy, integrated food science and technology for the tropics. london: mcmillian publisher limited, 1985. [8] aoac, association of official analytical chemist, 14th ed. washington dc: association of official chemist, 1990. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. agriculture and food sciences research issn: 2411-6653 vol. 2, no. 2, 56-61, 2015 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 56 influence of processing methods on the tannin content and quality characteristics of cashew by-products emelike n.j.t 1 --ebere c.o 2* 1,2 department of food science and technology, rivers state university of science and technology, port harcourt, nigeria abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ............................................................................................................................................................................... 57 2. materials and methods ............................................................................................................................................................. 57 3. results and discussions ............................................................................................................................................................ 58 4. conclusion .................................................................................................................................................................................. 60 references ...................................................................................................................................................................................... 60 cashew (anacardium occidentale l.) kernel was processed using sand and gari roasting methods to study the chemical compositions, tannin reduction and sensory properties of the kernel in comparism with the reference sample (imported cashew kernel). oil was extracted from part of the gari roasted sample. soybean flour was processed, the oil equally extracted and the physiochemical characteristics of the oils studied. from the result, there was no significant difference (p>0.05) in all the chemical compositions studied irrespective the processing method from the reference sample. tannic acid of the kernel was observed to reduce to 0.1% in the gari roasted kernel and showed no significant difference to the imported kernel (0.1%). however, sand roasted cashew kernel had the highest tannic acid value of 0.2%. sensory attributes of colour, flavour and general acceptability of the gari roasted kernel (4.50, 4.70 and 4.50, respectively) compared well with the imported kernel. all the sensory attributes of the sand roasted kernel was significantly lower compared to the reference sample. the physiochemical characteristics of the golden yellow colour cashew kernel oil showed that the oil is high in acid value (1.57mg koh/g), saponification value (218.6mg koh/g), specific gravity at 25 o c of water (0.91%) and iodine value of 144.021gl2/100gm. this is an indication that gari roasting method is the best processing method in terms of the tannin reduction and the sensory characteristics. keywords: processing methods, tannin content, chemical compositions, sensory properties, physiochemical characteristics, cashew kernel, cashew oil. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(2): 56-61 57 1. introduction fruits such as cashew (anacardium occidentale l.) are rich in tannin and polyphenols [1] which gives it antioxidant properties and makes it an effective remedy against chronic dysentery in cuba and brazil [2, 3]. cashew trees are all over nigeria and second to almond in commercial importance [4]. cashew is like a table starting from lagos to kogi, kwara, nasarawa, plateau down to enugu, abia and cross river states [5]. large scale cultivation started in 1953 [6]. the total estimated output of raw cashew nut in nigeria is 50,000 tonnes while about 5,000 – 7,000 tonnes are produced annually mainly as an export crop [7] as a result of lack of adequate processing techniques to reduce its tannin content. cashew contains 0.64mg/100g of tannin in the case of total tannins but in condensed tannins, it is 0.18mg/100g. the main product from cashew tree is cashew kernel (true fruit), which is rich in fat and protein. aroyeun [8] reported 18.60% and 39.90% of protein in cashew kernel and cashew kernel meal, respectively. undefatted and defatted cashew kernels had protein values of 19.8% and 34.0%, respectively as reported by emelike, et al. [9]. in these reports, the protein values of cashew increased after processing which leads to the removal of tannin from the resultant products. tannin (schinopsis spp.) is water soluble polyphenolic compounds ranging in molecular weight from 500 – 3000 daltons that have the ability to precipitate proteins. butter [10] reviewed that tannin is any phenolic compound of sufficiently high molecular weight containing sufficient hydroxyls and carboxyls to form effectively strong complexes with protein and other macro-molecules under the particular environmental conditions being studied. apart from the ability to precipitate protein, tannin also decreases digestibility and palatability [11]. according to fao [12] tannin binds protein; fights digestion by inhibiting key enzymes involved in digestion and can also render iron and vitamin b12 unavailable. other nutritional effects which have been attributed to tannin include damage to the intestinal tract, toxicity of tannin absorbed from the gut, interference with the absorption of iron and possible carcinogenic effect [10]. if tannin is served in excess or food high in tannin is ingested over a long period, the liver could be damaged. it has been reported that tannins have liver and kidney toxicity, some evidence has shown human oesophageal cancer on long consumption of beverage containing tannins. scalbert [13] reviewed that tannic acids inhibits the absorption of glucose and methionine in the mouse intestine. recently, many researches have been carried out to reduce the annual wastage of cashew and its by-products in nigeria. compositional studies and physicochemical characteristics of cashew nut flour [7] nutritional enrichment of biscuits with cashew kernel meal [8] roasted and defatted cashew nut flour [14] cassava flour biscuit supplemented with cashew apple powder [15] effect of packaging materials, storage time and temperature on the colour and sensory characteristics of cashew apple juice [16] and wheat flour/cashew apple residue in cookie preparation [17] were all carried out for this purpose. this research work is therefore embarked to analyse the influence of processing methods such as sand roasting and gari roasting on the tannin content of cashew kernel. analyse the quality characteristics which included the chemical and sensory compositions and compare it with the imported cashew kernel. finally, mill part of the gari roasted sample, extract the oil and compare the physiochemical properties of the oil with soybean oil. 2. materials and methods 2.1. materials mature, ripe cashew (anacardium occidentale l.) apples, red and yellow varieties were harvested in an orchard at uturu, abia state, nigeria. the nuts were detached from the apples and a total of twenty kilograms of cashew nuts were used for the analysis. soybean (gycine max) was purchased from mile 3 market in port harcourt, rivers state, nigeria. five hundred grams of soybean was used. all chemicals used were of analytical grade. 2.2. preparation of cashew kernel cashew kernel was processed using the method earlier reported by emelike, et al. [9] for defatted and undefatted cashew kernel flour with some modifications. the nuts were sun dried for three days to prevent deterioration during storage. they were conditioned (mild spraying with water in a sieve) to increase flexibility and prevent scorching. the conditioned cashew nuts were divided into portions of 500g each for easy processing, placed in a metal basket and immersed in a pot of hot vegetable oil (corn oil) for 1 min to make the shell brittle for shelling. the nuts were stirred at intervals of 10 sec to prevent burning while in the hot oil. the cashew nut shell liquid (cnsl) of the nut extracted into the pot as the volume of the oil increases in the pot. the cashew nuts were poured out after 1 min and allowed to cool for about 1h. the brittle shell was broken with wooden mallet and the kernel extracted. the kernels were pulled together which yielded 13.14kg. 2.3. processing methods of the cashew kernel and the extraction of cashew kernel oil the cashew kernel was divided into two equal samples of 6.57kg each. one sample was roasted in gari for 30 min, cooled for a few min. the testa removed manually, sorted and packaged to get the gari roasted sample (grs). the other sample was roasted in sand for 30 min, cooled slightly and the testa removed manually using tooth pick. it was packaged after the removal of testa to get the sand roasted sample (srs). gari roasted sample was divided into two, one part was milled, the oil was screw pressed from the sample and used in comparison with soybean oil to study the physiochemical properties of the oils. agriculture and food sciences research, 2015, 2(2): 56-61 58 figure-1. flow chart for various processing methods of cashew kernel. source: author’s computation. 2.4. extraction of soybean oil soybean oil was extracted using the traditional method outlined by aremu, et al. [18] for oilseeds with some modifications. the bean was crushed after washing and milled into paste by using philips hr2000 blender. water was added to the paste; the mixture was stirred and manually kneaded by hand until the oil separates to the top and sides of the utensils being used for the kneading. it was suspended in boiling water and boiled for 30 min with liberated oil floating on the surface. further quantities of water were added after boiling to replace the evaporated water and to facilitate the floatation of the oil to the surface. the oil was carefully scooped from the surface of the water and reboiled for 5 min. it was allowed to cool in room temperature (28±2 o c), filtered and stored in the bottle for analysis. 2.5. physiochemical analysis of cashew kernel, cashew kernel oil and soybean oil moisture content of the kernel and the oils, protein, fat, crude fibre, total ash and tannin of cashew kernel were determined using the latimer [19] method. free fatty acid, peroxide value, saponification value, unsaponification matter, iodine value, refractive index and specific gravity were also carried out by the method of latimer [19]. clegg anthrone method [20] was used in the determination of the available carbohydrate. 2.6. sensory evaluation sensory analysis of the differently processed cashew kernel was carried out using a twenty member panelist consisting of staff and students of food science and technology department, rivers state university of science and technology, port harcourt, nigeria. criteria for selection were that panelist were 16 years of age, regular consumers of cashew kernel and were neither sick nor allergic to any nut/kernel. panelists were trained in the use of sensory evaluation procedures. at each session, samples were served on white disposable plates, properly coded with 5-digit random numbers to prevent bias. the sensory qualities evaluated were: colour, taste, flavour, attractiveness and general acceptability. a descriptive five point hedonic scale as described by iwe [21] was used, 1 and 5 representing the least score (dislike extremely) and the highest score (like extremely), respectively. necessary precautions were taken to prevent carryover flavour during the tasting by ensuring that panelists rinsed their mouth with water after each session of sensory evaluation. 2.7. statistical analysis results were expressed as mean values and standard deviation of five (5) determinations. data were analysed using a one-way analysis of variance (anova) using statistical packaging for social science (spss) version 20.0 software 2011 to test the level of significance (p<0.05). duncan new multiple range test was used to separate the means where significant differences existed. 3. results and discussions 3.1. chemical compositions chemical properties of locally processed cashew kernel such as sand roasted sample (srs) and gari roasted sample (grs) including the imported cashew kernel are shown in table 1. from the result, srs and grs had agriculture and food sciences research, 2015, 2(2): 56-61 59 protein content of 20.7 and 20.5%, respectively while the imported sample recorded protein value of 21.0%, which are not significantly different (p>0.05). this observation is in agreement with the report of emelike, et al. [9] for the protein value of undefatted cashew kernel flour (19.8%) while that of the defatted cashew kernel flour (34.0%) was relatively high compared to the result in this study. this is because during the process of defatting, the anti-nutrient (tannin) responsible for protein binding capacity is removed thereby making protein more assessable. the fat content of srs and grs ranged from 50.0 – 51.0%, respectively and showed no significant difference. fat value for the imported sample (48.4%) was significantly low compared to the locally processed samples. fat values reported in this study for both the imported and locally processed cashew kernel are in agreement with those of undefatted cashew kernel, melon seed, groundnut, non-defatted flour of brebra seed, undefatted cashew kernel flour with the values of 53.3%, 55.2%, 48.5% and 47.1%, respectively [9, 22]. this is an indication that cashew kernel is an oil seed which can be extracted and used as an economically important material for the production of soaps and oily creams. there was no significant difference (p>0.05) between the ash content of locally processed cashew kernel with the imported sample. pomeranz and clifton [23] recommended that ash contents of nuts/kernels, seed and tubers should fall within the ranges of 1.5 – 2.2% to be suitable for animal feeds. thus, ash value for sand and gari roasted cashew kernel including the imported kernel (5.2, 5.4 and 5.0, respectively) does not fit into this purpose. the sand and gari roasted kernel including the imported cashew kernel had crude fibre values of 1.4, 1.5 and 1.2 %, respectively and shown no significant difference. these values agreed with those reported by aremu, et al. [7]; vincent, et al. [14]; emelike, et al. [9] for cashew nut flour, roasted/defatted cashew nut flour and sand roasted/defatted cashew kernel flour (1.2, 1.42 and1.2%), respectively. moisture mean values of the three studied samples ranged from 4.7 – 5.0% and showed no significant difference. these values agreed with those reported by berhanu and amare [22] for non-defatted flour of brebra seed (4.24%). there was equally no significant difference between the sand and gari roasted cashew kernel to the imported sample with the values of 19.5, 20.0 and 20.1%, respectively. among the two processing methods studied, the reduction of tannin content in the grs (0.1%) compared very well with the tannin content in the imported sample (0.1%) and they were significantly high (p<0.05) when compared to the srs (0.2%). this is an indication that cashew kernels can be locally processed using gari roasting method in other to reduce the amount of tannin acid in the kernel which has been a limiting factor. table-1. comparative chemical compositions of differently processed cashew kernel parameters (%) sand roasted gari roasted imported protein 20.7±0.10 a 20.5±0.32 a 21.0±0.20 a fat 50.0±0.04 a 51.0±0.04 a 48.4±0.10 a ash 5.2±0.10 a 5.1±0.30 a 5.0±0.01 a crude fibre 1.4±0.12 a 1.5±0.10 a 1.2±0.20 a moisture 5.0±0.04 a 4.9±0.31 a 4.7±0.10 a carbohydrate 19.5±0.70 a 20.0±0.04 a 20.1±0.02 a tannin 0.2±0.01 a 0.1±0.02 b 0.1±0.01 b results represent means ± standard deviation of five determinations. values not followed by the same superscript along the rows are significantly different (p≥0.05). 3.2. sensory properties from the result, there was no significant difference (p>0.05) in the sensory attributes of taste and attractiveness of the both processing methods studied as presented in table 2. sensory attributes of colour, flavour and general acceptability between the imported and gari roasted cashew kernels also showed no significant difference. this is an indication that gari roasted cashew kernel was accepted by the panelist just as the reference sample (imported kernel). this is in agreement with the reported of aroyeun [8] who observed no significant difference in the sensory attributes of colour, taste, texture and general acceptability of biscuit produced with 10% substitution of wheat flour with cashew kernel meal to the digestive biscuit already in the market. ebere, et al. [17] also reported no significant difference (p>0.05) between the general acceptability of cookies prepared with 100% wheat flour and those substituted with 20% cashew apple residue. unlike flaxseed flour which significantly affected all the sensory attributes of chapattis produced from different levels of substitution as reported by hussain, et al. [24]. this indicates that cashew kernel are useful source of raw materials, it could be milled and used in formulation of products with no sensory difference from the already existing products in the market. gari roasting method could be suggested as the best method to be adopted in the processing of cashew kernel. table-2. sensory scores for processed cashew kernel parameters sand roasted gari roasted imported lsd colour 2.80 b 4.50 a 4.70 a 0.56 taste 3.80 b 3.60 b 4.40 a 0.70 flavour 3.90 b 4.70 a 5.00 a 0.51 attractiveness 3.80 b 4.00 b 4.80 a 0.32 general acceptability 3.70 b 4.50 a 4.70 a 0.38 means not followed by the same superscript in the rows are significantly different at 5% level of probability (p≥0.05). 3.3. physiochemical properties of the oils the utilization of oils from different sources depends mainly on their compositions and no oil from a single source can be suitable for all purposes, thus the study of their constituents is important. the comparison of the physiochemical properties of cashew kernel oil and soybean oil is shown in table 3. from the result, the golden yellow oils have moisture values of 0.075% and 0.060% for cashew kernel and soybean oils, respectively. the moisture values of these oils are very low when compared with those of legumes ranging from 5 – 11% as reported by aremu, et al. [18]. this is an indication that these oils can be stored for a longer time without going rancid (offagriculture and food sciences research, 2015, 2(2): 56-61 60 flavour) than legumes though soybean oil has the tendency of lasting longer compared to cashew kernel oil. the free fatty acids in this study are 0.06% and 0.03% for cashew kernel oil and soybean oil, respectively and they compared very well with the report of aboki, et al. [25] for sunflower seed oil (0.042%). the quality of the oils studied is high though soybean oil is better in terms of the smoke point that will be produced during refining or cooking. peroxide value of cashew kernel oil is 0.001meq/g while that of soybean oil is 0.010meq/g. these values are less than the peroxide values of 0.50meq/g for african bush mango seed oil [26]. high peroxide value is an indication of high levels of oxidative rancidity of the oil and also suggests low levels of antioxidant [27]. acid value is an indication of the quality of fatty acids in the oil and the edibility of an oil and suitability for use in the paint and soap industries [18]. the acid values are 1.57mgkoh/g and 0.60mgkoh/g for cashew kernel oil and soybean oil, respectively. acid value of cashew kernel oil falls within the ranges of jatroph curcas seed oil (1.50mg koh/g) and groundnut seed (1.79mg koh/g) as reported by akintayo [28]; ayoola and adeyeye [29] respectively. high acid value in oil showed that it may not be suitable for use in the food industries rather useful raw material for the production of paints, liquid soap and shampoos [18]. the saponification value of cashew kernel oil is 218.6mgkoh/g while soybean oil is 189.0mgkoh/g. this result is similar to that reported by aremu and akinwumi [30] for cashew seed oil (212.00mgkoh/g) and the value for soybean oil closely agreed with the result reported by akanni, et al. [31] for soybean seed oil (192.3mgkoh/g). the high saponification value of cashew kernel oil is therefore a good indication that the oil is suitable for soap making, oil-based cream and shampoos but non-edible. the value for unsaponifiable matter is 1.79meq/g for cashew kernel oil and 1.41meq/g for soybean oil. the iodine value of cashew kernel oil is 144.021 gl2/100gm and soybean oil is 121.698 gl2/100gm. these values are high than those of refined castor oil and brebra seed oil (84.8 gl2/100gm and 104.48 gl2/100gm), respectively [22, 32]. good drying oils should have iodine value above 130 [18]. cashew kernel oil is therefore classified as drying oil and could be useful raw material in the manufacture of vegetable oil-based cream [33]. refractive index of 1.446 and 1.466 was observed in this study for cashew kernel oil and soybean oil, respectively. the refractive index value of oils are within the acceptable range of 1.4677 – 1.4707 for virgin, refined and refined-pomace oils according to codex standards for fats/oils from vegetable and plant sources codex alimentarius commissionm [34]. yahaya, et al. [35] reported that specific gravity is commonly used in conjunction with other figures (refractive index) in assessing the purity of oil. the specific gravity at 25 o c of cashew kernel oil is 0.91 and 0.92 for soybean oil. this result is in agreement with the specific gravity of 0.906, 0.910 and 0.920 for soybean oil, moringa seed oil and cotton seed oil, respectively [31, 36, 37]. this is an indication that cashew kernel and soybean oils are denser than water and will make the oils to flow and spread easily on the skin and therefore, they are good sources of raw material for cream production [38]. table-3. physiochemical properties of cashew kernel oil and soybean oil parameters cashew kernel oil soybean oil moisture (%) 0.075 0.060 free fatty acid (%) 0.06 0.03 peroxide value (meq/g) 0.001 0.010 acid value (mgkoh/g) 1.57 0.60 saponification (mgkoh/g) 218.6 189.0 unsaponification (meq/g) 1.79 1.41 iodine value (gl2/100gm) 144.021 121.698 refractive index at 40 o c 1.446 1.466 specific gravity at 25 o c of water 0.91 0.92 smoke point 165 o c -- flash point 135 o c -- colour golden yellow golden yellow results are mean values of five determinations 4. conclusion processing methods studied did not utter significant changes in all the chemical compositions of the locally processed cashew kernel from the reference sample (imported cashew kernel). grs was observed to reduce tannic acid in the kernel to 0.1% which compared very well with the tannin content in the imported kernel (0.1%). there was no significant difference between the sensory attributes of colour, flavour and general acceptability of the grs and the imported kernel while there was significant difference in all the sensory parameters of the srs to the imported cashew kernel. from the result of the physiochemical properties, cashew kernel oil is a good source of raw material for the production of paints, liquid soaps and shampoos while soybean oil is a good raw material for food industries. references [1] m. adou, d. a. kouassi, f. a. tetchi, and n. g. amani, "phenolic profile of cashew (anacardium occidentale l.) of yamoussoukro, cote d’ivoire," pakistan journal of nutrition, vol. 10, pp. 1109–1114, 2012. 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[38] g. o. oyeleke, e. o. olagunju, and a. ojo, "functional and physicochemical properties of watermelon (citrullus lanatus) seed and seed-oil," journal of applied chemistry, vol. 2, pp. 29–31, 2012. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. 22 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 22-29, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.22.29 © 2019 by the authors; licensee asian online journal publishing group impact of osmodrying on qualitative properties of fruits viz. pineapple cube, apple cube and banana slice subhajit ray associate professor, department of fet, cit kokrajhar, assam, india. abstract nowadays osmodrying is proved to be an efficient technique of processing and preservation of fruits and vegetable varieties. the process of osmodehydration can promote the suitability of processing of various categories of fruits and vegetable produce with retention of organoleptic and nutritional characteristics. the aim of this project work is to prepare osmodried fruit cubes with satisfactory health benefits and good organoleptic acceptance. to carry out the project work, apples, banana and pineapple of fresh quality was purchased from local market adjacent to the institution. now these fruits were blanched, peeled and were formulated using different concentrations of sugar solutions viz. 60%, 65% and 70% respectively. the dehydrated fruits were then subjected to physico-chemical and microbiological analysis. the chemical quality indexes viz.moisture, ash, fat and sugar were analyzed for the osmodried products. for osmotically dehydrated apple, pineapple and banana cubes, the estimated moisture ,ash, fat and sugar content were found as 77%, 77.64%, 79.24% ;0.12%, 0.06%, 0.08%; 0.1%, 0.07%, 0.08%; 24.2%, 25%, 26.5% and 10.71%, 17.25%, 10.76%; 0.1%, 0.12%, 0.01%; 0.001%, 0.002%, 0.02%; 28%, 29.60%, 30% and 16%, 13.37%, 13.97%; 2%, 2.2%, 2.6%; 0.005%, 0.01%, 0.01%; 18%, 20.30%, 21.70% with respect to preservation in 60%,65%,70% sugar syrup respectively. microbiological analysis of osmodried apple, pineapple and banana revealed that negligible count of yeast and mold occurred at an incubation temperature and time of 37 degree centigrade for 72 hours with an optimum sugar concentration of 70%.sensory evaluation in terms of appearance, color, flavor, body, mouthfeelness and overall acceptability of pineapple cube, apple cube and banana slices in different sugar concentrations viz. 60%, 65%, 70% were studied. therefore in this study 70% sugar solution was proved efficient for osmodehydration in terms of its nutritional and organoleptic significance. keywords: osmodried fruits, preservation, nutritional quality, sensory characteristics, utilization. citation | subhajit ray (2019). impact of osmodrying on qualitative properties of fruits viz. pineapple cube, apple cube and banana slice. agriculture and food sciences research, 6(1): 2229. history: received: 6 december 2018 revised: 16 january 2019 accepted: 28 february 2019 published: 19 april 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group funding: the author gratefully acknowledged the infrastructural support of department of fet, cit kokrajhar, assam, india, pin: 783370 for completion of the work. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 23 2. materials and methods ................................................................................................................................................................... 24 3. result and discussions ................................................................................................................................................................... 26 4. conclusions ....................................................................................................................................................................................... 28 references .............................................................................................................................................................................................. 28 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.22.29&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.22.29&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://asianonlinejournals.com/index.php/aesr/article/view/1786 https://orcid.org/0000-0003-0081-9836 agriculture and food sciences research, 2019, 6(1): 22-29 23 © 2019 by the authors; licensee asian online journal publishing group 1. introduction osmodrying as a food preservation technique involves the addition of highly concentrated solution of osmotic agents e.g. sugar, salt into the food materials. fruits and vegetables dehydrated slightly can be added to various kinds of dairy products as well as bakery and confectionary items. finally the food materials are dried completely and can be considered as suitable components of cereal based products [1]. various process parameters involving in drying process as well as constituents responsible for osmosis can affect greatly on the nutritional and other quality attributes of osmotically dehydrated fruits and vegetables [2, 3]. though there is a significant loss of various nutrients during dehydration but at the same time the dried food materials enriched with concentrated solutions caused for osmosis can greatly increase its dietary value apart from its organoleptic properties. human health is highly benefited by the consumption of processed dehydrated fruits due to the potential availability of various microelements as well as antioxidants, soluble dietary fibers and also provides significant amount of energy where it is produced by osmosis by the application of concentrated sugar solution followed by dehydration as a result of convective air current. the process restricts the appearance potential of osmodried fruit items where consumers like weighty one. suitable alternatives of sucrose e.g. glucose, trehalose, polyols, fruit juice concentrate and different carbohydrate mixtures as osmotic agent has been tested and proved as potential osmotic substances though the knowledge about the influence of these substances on taste as well as organoleptic attractivity is unknown [4-8]. the process of osmosis results the elimination of water along with little amount of acid from fruits and vegetables where the semipermeable cell membrane can only permit water in comparison to sucrose which penetrates rapidly with respect to time. various factors e.g. temperature, concentration of sugar syrup as well as solution of osmotic agent, time etc. greatly influence the osmodried fruits and vegetable based product characteristics [9].these osmodried fruits have varied applications in various sweet dish preparations such as icecream, custard, pudding etc. one such application which was performed during this research work was the preparation of custard with osmo-dried fruits. the spoilage opportunity of fruit is more due to occurrence of more than 70% water. therefore proper storage as well as dehydration is the suitable way of keeping such kind of food materials for longer period. the mechanism of osmosis in fruit pieces involves the water migration because fruit pieces consisting of sucrose and other constituents where the structural entity of cells functions like effective semipermeable membrane. according to pointing, et al. [10] the influence of slight drying of fruit slices in sucrose solution reveals that the maximum extent of water removal will be 50% of the initial weight of different fruits like banana, papaya, mangoes and apples. 1.1. qualitative upgradation of food product osmosis followed by convective drying is always preferred to obtain the good quality of food product. according to several studies it has been observed that the sensory qualities of e.g. color, flavor and texture of food product is significantly enhanced by osmodrying [10, 11]. 1.2. energy efficicacy osmotic drying process is economical in use in juice and beverage industries because of less consumption of energy in comparison to other conventional drying process. the process of osmosis reduces water activity and extend the shelf stability thereby preserve the food products in a way where only drying process needs more amount of energy and can be avoided. the resultant osmotic solution can be used in juice or beverage industries as a product, improving process economy, or it may be re-concentrated for further drying. 1.3. reduction of cost due to ease of handling and transport to market the packaging and distribution cost of osmodried food products is significantly reduced. according to biswal [12] it has been observed that successive osmodrying and freezing of fruits and vegetables can save packaging and distribution costs effectively. 1.4. preservation without chemical treatment canned fresh apple slices can be well preserved by the addition of texture modifier e.g. calcium chloride [13]. osmo-canning process employed for apple slices results firmer texture and qualitative improvement. osmotic process discourages the reduction of enzymatic browning by using chemical [10]. 1.5. storage stability of osmodried product osmosis results better storage stability of fruits and vegetables than its untreated counterpart due to reduced level of water activity which lowers the extent of chemical reaction and the growth of toxin-producing microorganisms in the food. canning of fresh fruit and vegetable consisting of high moisture causes flavor reduction by dilution of sugar syrup with product water. osmo-canning process can suitably improve the storage stability of product and can prevent the flavor loss. 1.6. limitations of osmotic dehydration process addition of acid extracted from fruit in solution causes osmosis may overcome the problem of lowering of acidity [10].according to lenart and flink [1] osmodrying has been proved as a suitable alternative of other processes like sulphitation of fruits or blanching in comparison to enzyme activation as well as textural defects. syrup management strategy must be improved to overcome the problem in industrial application 1.7. selection of raw materials for osmodrying 1.7.1. internal properties of raw material the process of osmosis is governed by several controlling parameters including the variety, maturity, solube and insoluble solid content, intercellular spaces and most importantly enzymatic activity but solute concentration as well as process temperature has no effect on the gain of solid rate kinetics. agriculture and food sciences research, 2019, 6(1): 22-29 24 © 2019 by the authors; licensee asian online journal publishing group 1.7.2. morphology and dimensions of fruit slices loss of water during process of osmosis has a significant impact mainly adverse effect on the product morphology related to surface area [14]. it has been observed [11] that the partition coefficient of water into solute particles decreased with the increment of surface area, temperature, syrup concentration and thickness as morphological parameter. generally the dimension ranges from (3-10) mm was suggested of different shapes e.g. rectangular, circular or cubical for osmodrying treatment. 1.7.3. pretreatment for osmosis pretreatment techniques like blanching or freezing has an adverse effect i.e. qualitative deterioration before the loss of water during osmosis. different types of color degradation phenomena including enzymatic browning, discoloration etc. can be restricted by the addition of 1% citric acid,sulphur dioxide and acidic or alkaline solution of esters made of oleate before drying as well as osmodrying [15, 16]. a high quality osmodried food products based on papaya, mango slices etc. can be prepared by pretreatment with 0.4% ascorbic acid solution or 0.4 % ascorbic acid + 0.1% kms solution for 30 min time exposure. 1.7.4. dipping time a significant increase in elimination of water occurs due to rise in deeping time where solution concentration remains constant. it has been observed generally the maximum major solute migration took place within two hours of osmosis. according to tiwari and jalali [17] the rate decrement of osmodrying in mango and pineapple affects the increase in weight loss. according to the investigation [18, 19] it has been reported that osmodrying occurs at 500c for 3 hrs deeping time shown optimum level of water elimination and gain of sucrose as compared with 700c for same duration. 1.7.5. materials required for osmosis it has been observed from several reports that osmodrying process for fruits chiefly and vegetables can be suitably governed by materials viz. sugar, glucose, calcium chloride, monohydroxy ethanol and polyhydroxy compounds such as lactose, malt dextrin, corn syrup and mixtures and sodium chloride respectively. 1.7.6. stirring/mixing the rate of process of osmosis will be rapid during well stirred as well as mixed fruits with sugar syrup due to reduced mass transfer resistance (kl) which causes defect. it has also been observed [14, 19] that there is a significant effect of mixing speed on elimination of water. 1.7.7. fruit slices to osmotic agent ratio an optimum ratio is always maintained to achieve a good result. therefore conventionally in practice according to tiwari [19] a ratio of 1:2 or 1:3 is optimum. 1.7.8. mass transfer phenomena during osmotic dehydration in osmodehydration process followed by concentration development three major types of counter current mass transfer is suggested [19, 20]. a series of phenomena including water out flow from product to solution, a solute migration e.g. preservative, nutrients etc., improvement of organoleptic quality as well as leaching out of various solutes e.g. sucrose, organic acids, minerals, vitamins etc occurs during osmodehydration process. 2. materials and methods to carry out the project work, apples, banana and pineapple of fresh quality was purchased from kokrajhar market, assam adjacent to the institution. the manufacturing flow diagram is suitably indicated in figure 1. collection and sorting, grading of fruits. 2.1. utilization of osmodried fruits one of the suitable applications of osmodried fruits was done by the preparation of custard. the manufacturing flow diagram is indicated figure 2. agriculture and food sciences research, 2019, 6(1): 22-29 25 © 2019 by the authors; licensee asian online journal publishing group figure-1.general manufacturing flow diagram of osmodried fruits. source: subhajit ray,2019. agriculture and food sciences research, 2019, 6(1): 22-29 26 © 2019 by the authors; licensee asian online journal publishing group figure-2. general manufacturing diagram of custard. source: krishikosh.egranth.ac.in. refrigerated milk based product mixed with custard powder and finally osmodried fruit cubes are placed in the surface as topping material for display as well as sensory acceptability. 2.2. sensory evaluation sensory characteristics of osmodehydrated fruits (apple, pineapple, banana) were assessed by a group of trained panelists by using nine point hedonic rating test. the nine point hedonic rating scale is considered as standardas followed by sensory panel in any sensory evaluation study [21]. 2.3. determination of chemical quality parameters moisture content, fat content, ash content and carbohydrate content of osmodried fruits was measured using aoac [22] procedure. 2.4. microbiological examinations of osmodried fruits microbiological analysis of the osmodried fruits were carried out using salle [23]. 3. result and discussions 3.1. proximate composition of osmodried fruit table-1.evaluation of chemical quality of osmodried apple cube. % sugar concentration moisture (%) ash (%) fat (%) carbohydrate (%) 60 77 0.12 0.1 24.2 65 77.64 0.06 0.07 25 70 79.24 0.08 0.08 26.5 source: subhajit ray,2019. from table 1 it was found that after osmo dehydration of apple cube with sugar solutions of 60%,65%,70% concentration, the estimated moisture ,ash, fat and sugar content were found as 77%,77.64%,79.24%; 0.12%,0.06%,0.08%; 0.1%, 0.07%, 0.08%; 24.2%,25%,26.5. table-2.evaluation of chemical quality of osmodried pineapple cube. % sugar concentration moisture (%) ash (%) fat (%) carbohydrate (%) 60 10.71 0.1 0.001 28 65 17.25 0.12 0.002 29.6 70 10.76 0.01 0.02 30 source: subhajit ray,2019. agriculture and food sciences research, 2019, 6(1): 22-29 27 © 2019 by the authors; licensee asian online journal publishing group from table 2 it was found that after osmo dehydration of pineapple cube with sugar solutions of 60%,65%,70% concentration, the estimated moisture ,ash, fat and sugar content were found as 10.71%,17.25%,10.76%; 0.1%,0.12%,0.01%; 0.001%,0.002%,0.02% ;28%,29.60%,30%. table-3.evaluation of chemical quality of osmodried banana slice. % sugar concentration moisture (%) ash (%) fat (%) carbohydrate (%) 60 16 2 0.005 18 65 13.37 2.2 0.01 20.3 70 13.97 2.6 0.01 21.7 source: subhajit ray,2019. from table 3 it was found that after osmo dehydration of banana slice with sugar solutions of 60%,65%,70% concentration, the estimated moisture ,ash, fat and sugar content were found as16%,13.37%,13.97%; 2%,2.2%,2.6%; 0.005%,0.01%,0.01%; 18%,20.30%,21.70%. table-4.microbiological analysis of osmodried fruits. products concentrations(%) microbial count of yeast and mold at 37 degree centigrade (cfu/gm) finished product pineapple 60 57-63 65 negligible 70 negligible apple 60 34-36 65 10-12 70 negligible banana 60 78-87 65 72 70 24-29 source: subhajit ray,2019. it is observed from table 4that after osmo dehydration of first fruit viz.pineapple cubes with sugar solutions of three different concentrations, pineapples dehydrated using 65% and 70% sugar concentration shows negligible growth of yeast and moulds when incubated at 37 degree centigrade for 72 hours. similarly apple cubes dehydrated using70% sugar concentration shows negligible growth of yeast and mold. in case of banana slice 70% sugar concentration was found comparatively better than the other two concentrations as compared with growth of yeast and mold. 3.2. comparison of sensory evaluation datas using bar diagram figure-3. evaluation of sensory characteristics of osmodried apple by semi-trained panel of judges. source: subhajit ray,2019. from figure 3 it is evident that the appearance of osmotically dehydrated apple cube with 60% sugar concentration is a little low as compared to the other concentrations. apple dehydrated using 65% sugar concentration was found to be acceptable organoleptically with all respect as compared to the other two samples of apple dehydrated osmotically using 60% and 70% sugar concentration respectively. figure-4. evaluation of sensory characteristics of osmodried pineapple by semi-trained panel of judges. source: subhajit ray,2019. agriculture and food sciences research, 2019, 6(1): 22-29 28 © 2019 by the authors; licensee asian online journal publishing group from figure 4 it can be observed that pineapple cube dehydrated osmotically using 60% and 70% sugar concentration was found to have a good overall acceptability. but in case of the pineapple cube dehydrated using 65% sugar concentration the body and texture was not satisfactory and it also result a poor mouthfeelness. figure-5. evaluation of sensory characteristics of osmodried banana by semi-trained panel of judges. source: subhajit ray,2019. from figure 5 it was seen that the results on sensory analysis for banana slice after osmotic treatment viz.60%, 65% and 70% sugar solution respectively showed a marked decrease in the body and texture of the product compare to other evaluated datas. 4. conclusions osmodrying was performed to extend the shelf stability of fruit slices by using three different concentrations of sugar solutions viz.60%, 65%, and 70% respectively on three fruits viz. apple, pineapple and banana. the moisture, ash, fat and sugar analysis were performed for the final dehydrated products and the values were reported. for osmotically dehydrated apple, pineapple and banana cubes, the estimated moisture ,ash, fat and sugar content were found as 77%,77.64%,79.24%; 0.12%,0.06%,0.08%; 0.1%, 0.07%, 0.08%; 24.2%,25%,26.5% and 10.71%,17.25%,10.76%; 0.1%,0.12%,0.01%; 0.001%,0.002%,0.02% ;28%,29.60%,30% and 16%,13.37%,13.97%; 2%,2.2%,2.6%; 0.005%,0.01%,0.01%; 18%,20.30%,21.70% with respect to preservation in 60%,65%,70% sugar syrup after performing the microbiological analysis of dehydrated fruits it can be seen that after osmo dehydration of pineapple with sugar solutions of three different concentrations, pineapples dehydrated using 65% sugar syrup and 70% sugar syrup shows negligible growth of yeast and moulds when incubated at 37 degree centigrade for 72 hours. similarly apples dehydrated using70% sugar syrup shows negligible growth of yeast and mould. in case of banana 70% sugar concentration was found comparatively better than the other two concentrations as compared with the count of yeast and mould. therefore it can be concluded that for all the three fruits 70% sugar solution was found optimum for osmodehydration .experimental results also revealed that osmotically dehydrated fruit prepared by finite concentration of sugar syrup not only improve it’s shelf stability but also enhances nutritional value and sensory characteristics. references [1] a. lenart and j. flink, "osmotic concentration of potato. i. criteria for the end-point of the osmosis process," international journal of food science & technology, vol. 19, pp. 45-63, 1984. [2] i. mandala, e. anagnostaras, and c. oikonomou, "influence of osmotic dehydration conditions on apple air-drying kinetics and their quality characteristics," journal of food engineering, vol. 69, pp. 307-316, 2005.available at: https://doi.org/10.1016/j.jfoodeng.2004.08.021. [3] a. chiralt and p. talens, "physical and chemical changes induced by osmotic dehydration in plant tissues," journal of food engineering, vol. 67, pp. 167–177, 2005.available at: https://doi.org/10.1016/j.jfoodeng.2004.05.055. [4] m. fernando and w. e. l. spiess, "cellular response ofplant tissue during the osmotic treatment with sucrose, maltoseand trehalose solutions," journal of food engineering, vol. 49, pp. 115–127, 2001.available at: https://doi.org/10.1016/s02608774(00)00218-1. [5] n. rastogi, k. raghavarao, k. niranjan, and d. knorr, "recent developments in osmotic dehydration: methods to enhance mass transfer," trends in food science & technology, vol. 13, pp. 48-59, 2002.available at: https://doi.org/10.1016/s09242244(02)00032-8. [6] a. a. el-aouar, p. m. azoubel, j. r. barbosa, and f. e. x. murr, "influence of the osmotic agent on theosmotic dehydration of papaya (carica papaya l.)," journal of food engineering, vol. 75, pp. 267–274, 2006.available at: https://doi.org/10.1016/j.jfoodeng.2005.04.016. [7] r. moreira, f. chenlo, m. d. torres, and g. vazquez, "effect of stirring in the osmotic dehydration of chestnut using glycerol solutions," lwt – food science and technology, vol. 40, pp. 1507–1514, 2008. [8] d. konopacka, k. jesionkowska, m. mieszczakowska, and w. płocharski, "the usefulness of natural concentrated fruit juice as osmotic agent for osmo-dehydrated dried fruit production," journal of fruit and ornamental plant research, vol. 16, pp. 275-284, 2008. [9] m. le maguer, "osmotic dehydration: review and future directions," production symposium on progress in food preservation processes, vol. 1, pp. 183-309, 1988. [10] j. d. pointing, g. g. watterss, r. r. forrey, w. l. stangly, and r. jackson, "osmotic dehydration of fruits," journal of food science and technology, vol. 20, pp. 125-128, 1966. [11] m. s. rahman, "osmotic dehydration kinetics of foods," indian food industry, vol. 11, pp. 20-24, 1992. [12] a. biswal, "mass transfers in plant material in contact with aqueous solution of ethanol and sodium chloride equilibrium data," journal of food process engineering, vol. 11, pp. 159-176,, 1991. [13] r. l. dang, r. p. singh, a. k. bhatia, and k. verma, "studies on kashmir apples-canning as rings," indian food packer, vol. 30, pp. 9-16, 1976. [14] n. m. panagiotou, v. t. karathanos, and z. b. maroulis, "mass transfer modeling of the osmotic dehydration of some fruits," international journal of food science & technology, vol. 33, pp. 267-284, 1998.available at: https://doi.org/10.1046/j.13652621.1998.00167.x. agriculture and food sciences research, 2019, 6(1): 22-29 29 © 2019 by the authors; licensee asian online journal publishing group [15] i. hussain, m. iqbal, i. shakir, and n. ayub, "effect of sucrose and glucose mixture on the quality characteristics of osmotically dehydrated banana slices," pakistan journal of nutrition, vol. 3, pp. 282-284, 2004.available at: https://doi.org/10.3923/pjn.2004.282.284. [16] p. s. sunkja and g. s. v. ragharan, "assessment of pre treatment methods and osmotic dehydration of cranberries," journal of canadian bio systems, vol. 46, pp. 52-56, 2004. [17] r. b. tiwari and s. jalali, "studies on osmotic dehydration of different varieties of mango," in proceedings of first indian horticulture congress-2004 (6-9 november, 2004), new delhi, 2004. [18] a. m. mauro, s. m. mounnrat, and a. e. rodrogues, "vacuum drying of osmotic dehydrated apple slices drying," in proceedings of the 14th international drying symposium, brazil, 2004. [19] r. b. tiwari, "application of osmo air dehydration for processing of tropical fruits in rural areas," indian foodindustry, vol. 24, pp. 62-69, 2005. [20] a. karthiayani, "osmotic dehydration of fruits and vegetables with special reference to vacuum treatment," food and pack, vol. 39, pp. 82-84, 2004. [21] j. lim, "hedonic scaling: a review of methods and theory," food quality and preference, vol. 22, pp. 733-747, 2011. [22] aoac, official methods of analysis of aoac international 20th edition, 2016, editor:dr. george w. latimer, jr, 20th ed. vol. 45. usa: published by aoac international, 2012 [23] a. salle, laboratory manual on fundamental principles of bacteriology, 5th ed.: mcgraw-hill, 1961. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 145 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 1, 145-154, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.61.145.154 © 2019 by the authors; licensee asian online journal publishing group effect of pretreatments on the drying characteristics and quality of african star apple (chrysophyllum albidum) komolafe, o.m.1 hussein, j. b.2 adebayo, q.3 abiona, o.o.4 oke, m.o.5 ( corresponding author) 1department of food science technology, federal polytechnics, ilaro, nigeria. 2department of food science and technology, modibbo adama university of technology, yola, adamawa state, nigeria. 3department of food science and technology, federal university dutsinma, katsina state, nigeria. 4department of chemical sciences, osun state university, osogbo, osun state, nigeria. 5department of food science and engineering, ladoke akintola university of technology, ogbomoso, oyo state, nigeria. abstract fruits, due to high moisture content, deteriorate quickly when not adequately preserved. drying, a common preservation method, will affect quality of final product if not properly controlled. pretreatments prior to drying have been established as a way of retaining product qualities. this study investigated the effect of pretreatments on the qualities of dried african star apple flesh. african star apple fleshes were sliced (2, 4, 6 and 8 mm) and subjected to pretreatments [blanching (80 ºc for 3 min), lime juice (100%), ascorbic acid and salt solution (1:25 w/v)]. untreated samples served as control. samples were dried in cabinet dryer (50, 55 and 60 ºc) at 2 m/s constant air flow-rate, monitored at intervals, until constant weight was obtained. ascorbic acid and colour measurement of the fresh and dried sample were determined using standard method. the moisture contents of the african star apple flesh were observed to reduce from a mean value of 70.44%, 71.55% and 73.24% to 2.38%, 1.91% and 3.23% at temperatures of 50, 55 and 60 °c, respectively. the total drying time ranged between 7 to 8 h. colour of the dried african star apple was significantly preserved by the pretreatments used, and low overall color change (δe) was obtained at the lowest drying temperature (50oc). the ascorbic acid and lime pretreatments however had better colour overall. lime pretreatment for 2 mm thick at 50ºc gave the best result in terms of ascorbic acid retention. hence, lime juice pre-treatment has potentials of retaining quality of dried fruits. keywords: pretreatments, drying characteristics, african star apple, drying qualities. citation | komolafe, o.m.; hussein, j. b.; adebayo, q.; abiona, o.o.; oke, m.o. (2019). effect of pretreatments on the drying characteristics and quality of african star apple (chrysophyllum albidum). agriculture and food sciences research, 6(1): 145-154. history: received: 21 march 2019 revised: 30 april 2019 accepted: 4 june 2019 published: 15 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 146 2. materials and methods ................................................................................................................................................................. 146 3. results and discussion ................................................................................................................................................................. 147 4. conclusion ....................................................................................................................................................................................... 153 references ............................................................................................................................................................................................ 153 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.145.154&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.61.145.154&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/807 https://orcid.org/0000-0002-3537-1878 https://orcid.org/0000-0002-9029-5468 https://orcid.org/0000-0001-6490-1797 https://orcid.org/0000-0003-4928-8546 https://orcid.org/0000-0003-1175-0301 agriculture and food sciences research, 2019, 6(1): 145-154 146 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by investigating the effect of pretreatments on the qualities of dried african star apple flesh. 1. introduction the preservation of fruits and vegetables are essential for extending the shelf life and quality of the product. preservation technologies include drying, canning, dehydration and others. among these, drying (dehydration) is especially suited for developing countries with inadequate facilities. it offers a highly effective and practical means of preservation which reduces post-harvest losses and off-set shortages in supply [1]. african star apple (chrysophyllum albidum) is a forest fruit tree commonly found throughout tropical africa. it has a fleshy flesh of the fruits which is popularly consumed. the flesh can taste either very sweet or sour. the variation of the fruit colour is said to be correlated with the flesh taste. the covering of the sweet fruits are seen to be yellow while that of the sour ones are usually a mix of yellow and green colours when mature [2]. it is of great economic value due to its diverse industrial, medicinal and food uses. the fruits are not only consumed fresh but also used to produce jam, jellies, stewed fruit, marmalade, syrup and several types of soft drinks. it is also used for medical purposes due to properties of stalk and fruits. the leaves and seed of some of these fruits and vegetables are used in pharmaceuticals [3]. pretreatment of fruits prior to drying is highly recommended because it helps to keep light-coloured fruits from enzymatic browning during drying and storage. it also speeds the drying of fruits with tough skins, such as grapes and cherries [4]. different pretreatment methods have been developed for fruit drying, amongst which are lemon juice, salt solution, honey dip, ascorbic acid, sulfuring, osmotic pre-treatment and blanching [5]. the objectives of this study were to investigate the effects of pretreatments on the qualities (colour and ascorbic acid contents) of dried african star apple flesh. 2. materials and methods 2.1. experimental procedure the african star apples were sorted to remove over-ripe and damaged ones. the fruits were thoroughly washed and peeled before cutting into slices for drying. the flesh of the fruits was cut, using a simple improvised grater, into thin slices of 2, 4, 6 and 8 mm thicknesses. the average initial moisture content of the tropical almonds was determined using aoac [6] methods as 71.74 %, wet basis. 2.2. pretreatment of the samples the samples were pre-treated according to the methods of tunde-akintunde [7] and doymaz [8]. the completely randomized design (crd) was used for the drying experiments with the four pretreatments and one control replicated three times to ensure the consistency of the results. the pretreatments used on the fruit samples were on a weight per volume basis with 1 part of the solute in 25 parts of distilled water (1:25 w/v for 10 min) except for hot water blanching where no solute was added to the distilled water. the pretreatments were coded as shown below: • con: no pre-treatment solution. • bla: blanched in water bath (80 ºc for 3 min), removed and cooled immediately by placing in a large quantity of tap water for another 3 min to remove excess heat. it is drained in a metal sieve and the excess water blotted off before further processing. • asc: 1 part of ascorbic acid in 25 parts of distilled water (1:25 w/v for 10 min). • lim: 1 part of lime juice in 25 parts of distilled water (1:25 w/v for 10 min). • sal: 1 part of salt in 25 parts of distilled water (1:25 w/v for 10 min). 2.3. drying procedures for the drying experiments, 0.2 kg of fresh pretreated and untreated african star apple fleshes was spared on a single layer inside a perforated tray and dried at temperatures of 50 ºc, 55 ºc and 60 ºc respectively. the drying was accomplished using an air-convective cabinet dryer of rhong machinery manufacturing limited series cabinet dryer (model rcd-5) designed to dry food materials under controlled conditions. the temperature of the air in the drying chamber was controllable to suit the drying temperature desired. the air velocity in the drying chamber was controlled by a fan speed controller (adjustable air valve at air inlet and exhaust) constant at 2 m/s flow-rate. also, there was a time controller to monitor drying time. the moisture loss of the flesh during drying was measured at 5 minutes intervals for the first hour, 10 minutes for second hour and at 15 minutes intervals thereafter during the drying process. this was measured using a gallenkamp electronic balance (model mp 10001), which has 0 1,000 g measurement range with a reading sensitivity of ±0.01 g. the drying processes were stopped when the weight loss became negligible. the experiments were replicated three times and the average results were recorded. after drying, samples were packed and sealed in black polythene to prevent exposure to light and stored until further analyses. 2.4. determination of ascorbic acid ascorbic acid was determined using the aoac [6] method. an aliquot (10 g) of the sample was diluted to a fixed volume (100 ml) with 3% hpo3 and then titrated with 2, 6-dichlorophenolindophenol. a standard ascorbic acid solution of 5 ml was added to 5 ml of 3% hpo3 and titrated with dye solution to a pink colour, which persisted for 15 s. ascorbic acid (mg/100g) of reconstituted juice was calculated using the formula: 𝐴𝑠𝑐𝑜𝑟𝑏𝑖𝑐 𝑎𝑐𝑖𝑑 (𝑚𝑔 100𝑚𝑙) = 𝑇×𝐷𝐹×𝑉1 𝑉2×𝑉3 ⁄ (1) where, t = titre; df = dye factor; v1 = volume made up (100 ml); v2 = aliquot of extract taken for estimation (10 g) and v3= volume of sample taken for estimation (10 ml). agriculture and food sciences research, 2019, 6(1): 145-154 147 © 2019 by the authors; licensee asian online journal publishing group 2.5. colour measurements the fresh and dried fruit colour was determined by direct reading using a colorimeter (minolta model cr200; minolta camera co., ltd., osaka, japan) to obtain the colour values: l* (brightness/darkness), a* (redness/greenness) and b* (yellowness/blueness) as described by ashebir, et al. [9]. the instrument was calibrated using standard white and black plates before each color measurement. the measurements were taken from randomly selected fresh fruits at three different parts (around halfway from the blossom end) of a fruit and the mean of three readings at these random locations on the sample was used. it should be noted that these same fruit parts were dried and subject to colour measurement after drying. the fresh african star apple was taken as the ideal sample. the total colour change (∆e) parameter is calculated as follows: ∆𝐸 = [(𝐿∗ − 𝐿∗∗)2 + (𝑎∗ − 𝑎∗∗)2 + (𝑏∗ − 𝑏∗∗)2]0.5 (2) where; ∆e = indicates the total colour change of a sample in comparison to colour values of an ideal sample having colour values of l*, a* and b*. l* = the lightness of fresh samples; l** = the lightness of dried samples a* = the redness of fresh samples; a** = the redness of dried samples b* = the yellowness of fresh samples and b** = the yellowness of dried samples. 2.6. statistical analysis all experiments were performed in triplicate, and the results were expressed as means ± standard error (se). analysis of variance (anova) was carried out to determine any significant differences in measurements using the spss statistical software (spss 20.0 for windows; spss inc., chicago, il, usa) and considering the confidence level of 95%. the significance of the difference between the means was determined using the duncan multiple range test, and the differences were considered to be significant at p< 0.05. 3. results and discussion 3.1. drying characteristics the drying experiments gave different values for the moisture contents of the african star apple fruits at different temperatures and thicknesses. figures 1 – 3 show the drying curves for drying the flesh of african star apple of 2, 4, 6 and 8 mm thicknesses at temperatures of 50, 55 and 60 ºc. for drying carried out at 50 ºc, the equilibrium moisture content was attained in 8 h. also, for drying carried out at 55 ºc, the equilibrium moisture content was also attained in 8 h. on the other hand, for drying carried out at 60 ºc, the equilibrium moisture content was attained in 7 h. the moisture contents of the african star apple flesh were observed to reduce from a mean value of 70.44%, 71.55% and 73.24% to 2.38%, 1.91% and 3.23% for temperatures of 50, 55 and 60 °c, respectively. in this drying operation, a relative reduction in moisture content with increase in drying time was observed. this also resulted in the drying curve obtained showing a downward curve. figure-1. drying curves for african star apple flesh of 2 mm, 4 mm, 6 mm and 8 mm thicknesses dried at 50 °c. source: authors’ findings. agriculture and food sciences research, 2019, 6(1): 145-154 148 © 2019 by the authors; licensee asian online journal publishing group figure-2. drying curves for african star apple flesh of 2 mm, 4 mm, 6 mm and 8 mm thicknesses dried at 55 °c. source: authors’ findings. figure-3. drying curves for african star apple flesh of 2 mm, 4 mm, 6 mm and 8 mm thicknesses dried at 60 °c. source: authors’ findings. the downward trends of the drying curves show that moisture content reduces with increasing drying time. the highest moisture loss was observed at the early period of drying due to the removal of free water present at the surface of the fruits. over the course of the drying period, the moisture removal was observed to reduce; the agriculture and food sciences research, 2019, 6(1): 145-154 149 © 2019 by the authors; licensee asian online journal publishing group temperature of drying not commensurate to the amount of moisture being removed. at no point in the drying operations was the moisture removal at a constant rate. this confirms that the whole drying operation of the fruit samples largely took place in the falling rate period. this is in line with the results of taiwo and adeyemi [10] and tunde-akintunde [11] on banana slices and chilli peppers, respectively. the slice thicknesses of the samples and higher drying temperatures were also observed to have effects on the rate of moisture removal; which invariably affected the overall drying process. faster drying rates were observed at 2 mm and 4 mm thicknesses compared to the 6 mm and 8 mm thicknesses for drying of the african star apple. this result corroborated with hussein, et al. [12] findings in which the drying time reduced significantly as the thickness of slices decreases, because the resistance to moisture movement is relatively higher in thicker slices than in thinner ones. also, the rate of drying at 60 ºc was faster as shown in figure 3. this was due to high temperature gradients created by higher temperature which ease the migration of water from the inner part to the surface to get evaporated. the ease of this migration depends on the porosity of the samples, drying temperatures and the surface area available as reported by hussein, et al. [12]. the study also confirmed the reports of tunde-akintunde and oke [13] and sobukola [14] on plantain and apple thin layer drying, respectively. this was explained as higher temperatures leading to faster removal of moisture from food materials. 3.2. effects of temperature and pretreatments on drying rate the moisture contents of the fruits were observed to reduce with increase in the drying time. the drying rates of the pretreated samples were higher than the drying rates of the control sample in most of the drying conditions. however, these were not in all cases as the pretreatments were not significant in some drying conditions. figure 4 – 6 show the drying rate curves for the african star apple flesh. from the figures, the drying rates of the pretreated african star apple flesh were observed to consistently having higher drying rates; with the blanching pretreatment being more predominant. this may be attributed to the softer and looser structure of the flesh of the fruit, an observation reported by doymaz [15] on carrot pomace. the effect of blanching increasing moisture loss and drying rates of agricultural materials has also been reported by taiwo and adeyemi [10]. kaymak-ertekin [16] reported that blanching improves the drying rates of carrots because blanching increases the porosity of cell walls, leading to faster movement of water away from the food. this observation is also in agreement with the work of akintunde, et al. [17] on the blanching of green pepper; that blanching operation generally increases the drying rate. figure-4. drying rate curves for 2 mm, 4 mm, 6 mm and 8 mm african star apple flesh dried at 50 °c. source: authors’ findings. agriculture and food sciences research, 2019, 6(1): 145-154 150 © 2019 by the authors; licensee asian online journal publishing group figure-5. drying rate curves for 2 mm, 4 mm, 6 mm and 8 mm african star apple flesh dried at 55 °c. source: authors’ findings. figure-6. drying rate curves for 2 mm, 4 mm, 6 mm and 8 mm african star apple flesh dried at 60 °c. source: authors’ findings. the other pretreatment methods had slightly higher rates than the control samples, but not as predominant as the blanched samples. a similar observation was also made by abano [4] on drying of pineapples with different pretreatments. it was reported that there was an initial faster rate of drying with ascorbic acid pretreatment which was later reduced due to the leaching effects of ascorbic acid. salt pretreatment was also found to have a high drying rate in the study. the high drying rate was observed to be as a result of common salt made of sodium chloride in an acid-base reaction thus, heating caused the loss of moisture. the high drying rate of the salt pretreatment was attributed to this reaction. agriculture and food sciences research, 2019, 6(1): 145-154 151 © 2019 by the authors; licensee asian online journal publishing group 3.3. effect of temperature and pretreatment on ascorbic acid retention in the drying experiments, the fruits were dried by the reduction of their moisture contents; however, this was in tandem with the loss of valuable nutrients such as ascorbic acid content of the fruits. an important indicator of the efficient drying of the fruits is the ascorbic acid retention of the drying process. the results for the ascorbic acid retention of the drying experiments are shown in table 1. the total ascorbic acid content of fresh flesh was found to be 24.37 mg/100 g. the total ascorbic acid retention showed a consistent trend of samples pretreated with lime having highest values, followed by ascorbic acid, control and salt solution respectively. the values ranged from 14.75 mg/100 g to 19.17 mg/100 g at 50 ºc, 14.02 mg/100 g to 18.48 mg/100 g at 55 ºc and 13.72 mg/100 g to 18.15 mg/100 g at 60 ºc. the results also showed that there was higher ascorbic acid retention at lower temperatures and smaller thicknesses compared to higher temperatures and larger thicknesses. this was highlighted by the highest ascorbic acid retention at 50 ºc and 2 mm thickness compared to the lowest at 60 ºc and 8 mm thickness. table-1. ascorbic acid content of dried african star apple flesh. ascorbic acid content (mg/100 g) temperature pretreatment 2 mm 4 mm 6 mm 8 mm 50 °c control 17.29 ± 0.11c 16.89 ± 0.08c 16.61 ± 0.08c 15.89 ± 0.07b blanched 16.79 ± 0.24b 16.38 ± 0.32b 16.39 ± 0.08b 15.49 ± 0.29b ascorbic 18.10 ± 0.07d 17.68 ± 0.08d 17.12 ± 0.08d 16.59 ± 0.07c lime 19.17 ± 0.17e 18.83 ± 0.08e 17.58 ± 0.07e 17.17 ± 0.08d salt 16.21 ± 0.08a 15.81 ± 0.12a 14.74 ± 0.08a 14.75 ± 0.78a 55 °c control 16.94 ± 0.04c 16.51 ± 0.03c 16.18 ± 0.25c 15.58 ± 0.08c blanched 16.76 ± 0.03b 16.33 ± 0.06b 15.49 ± 0.07b 15.31 ± 0.16b ascorbic 17.83 ± 0.09d 17.44 ± 0.07d 16.84 ± 0.08d 16.46 ± 0.07d lime 18.48 ± 0.08e 18.07 ± 0.09e 17.32 ± 0.06e 16.81 ± 0.09e salt 15.84 ± 0.21a 15.49 ± 0.08a 14.37 ± 0.08a 14.02 ± 0.11a 60 °c control 17.64 ± 0.02c 16.39 ± 0.09c 15.89 ± 0.63c 15.48 ± 0.07c blanched 16.67 ± 0.08b 16.19 ± 0.05b 15.25 ± 0.09b 15.25 ± 0.09b ascorbic 17.64 ± 0.02c 17.19 ± 0.07d 16.74 ± 0.07d 16.21 ± 0.07d lime 18.15 ± 0.07d 17.73 ± 0.08d 17.19 ± 0.07e 16.63 ± 0.07e salt 15.25 ± 0.03a 14.79 ± 0.06a 14.15 ± 0.09a 13.72 ± 0.73a note: means within the same column at the same temperature with different alphabet(s) are significantly different at p ˂ 0.05. source: authors’ findings.’ the results showed a significant reduction in the ascorbic acid content of the dried samples, with increasing temperature and thickness also having great significance. the heating foods at higher temperatures cause losses of ascorbic acid because it is heat labile. the findings of gupta, et al. [18] corroborates this claim, where drying at temperatures above 60 ºc resulted in substantial ascorbic acid losses in the drying of cauliflower. it was also reported that prolonged exposure to higher temperatures destroys compounds in fruits. an explanation on the variations in ascorbic acid was given by garba and kaur [19] where the variations were attributed to the different pre-treatments used. joshi, et al. [20] also proposed that the ascorbic acid losses during most drying methods could be as a result of oxidation of ascorbic acid at high temperatures, which also supports the results of these drying operations. the ascorbic acid reduction of the fruit samples was consistent across the samples with pretreatments and control samples. this showed that hot air drying resulted in ascorbic acid degradation in all the samples. the higher values of ascorbic acid retention recorded for the lime and ascorbic acid pretreatment could be due to them being high sources of ascorbic acid. this could reduce the degradation kinetics of the ascorbic acid in the fruits during the drying operation. the control samples showed higher ascorbic acid retention than the blanched and salt pretreatments. this may be due to enzymatic and chemical degradation as well as leaching and heating operations, a similar observation noted by wawire, et al. [21] and toor and savage [22]. the results of these experiments were higher than the results reported by ruvini, et al. [23] on similar drying operations with similar pretreatments. the observations made by demiray and tulek [24] on ascorbic acid retention in fruits at temperatures below 70 ºc also support the ascorbic acid retention values for these experiments. the lower values for the salt pretreated samples across the fruits may be due to the inhibiting effect salt has on ascorbic acid [25, 26]. 3.4. effect of temperature and pretreatment on colour retention the final colour of the dried fruits is of great importance to the consumers. colour is one of the subjective quality characteristics which indicate the level of effects of the drying methods, the pretreatment effects or conditions. hence, colour is of great importance in deciding the preference of consumers [27]. tables 2 – 5 show the colour changes in the dried african star apple. the tables display the values for l*, a*, b* and δe which indicate the lightness, redness, yellowness and change in colour values respectively. the l*, a*, b* and δe values for african star apple were 40.29, 15.69, 18.54 and 37.78. the dried fruits all showed considerably lower values for all the parameters compared to the fresh samples. this is an indication that the slice thickness and pre-treatments were not significant because the colour changes were random and no specific sequence could be ascertained. the best colour retention for the african star apple were salt pre-treatment at 50 ºc for 2 mm, salt at 50 ºc for 4 mm, ascorbic at 50 ºc for 6 mm and salt at 50 ºc for 8 mm. agriculture and food sciences research, 2019, 6(1): 145-154 152 © 2019 by the authors; licensee asian online journal publishing group table-2. colour changes for dried african star apple flesh of 2 mm thickness. temperature pre-treatment l* a* b* δe 50 °c control 28.29 ± 0.62b 11.44 ± 0.90c 5.95 ± 0.87b 18.52 ± 1.34bc blanched 24.93 ± 0.97a 9.63 ± 0.64b 4.17 ± 1.38b 16.33 ± 1.77ab ascorbic 28.28 ± 3.60b 7.12 ± 0.17a 1.90 ± 1.43a 16.22 ± 3.56ab lime 24.33 ± 0.66a 6.64 ± 1.01a 1.90 ± 1.84a 13.14 ± 1.98a salt 32.31 ± 1.61c 7.70 ± 0.52a 5.74 ± 0.07b 21.57 ± 1.03c 55 °c control 27.66 ± 2.77b 11.29 ± 0.76b 5.32 ± 1.94d 17.71 ± 3.16b blanched 24.37 ± 0.60a 7.32 ± 0.53a 2.04 ± 0.48ab 13.54 ± 0.58a ascorbic 26.62 ± 0.98ab 6.59 ± 0.47a 1.45 ± 0.77a 14.48 ± 1.04a lime 26.61 ± 1.65ab 6.96 ± 0.44a 3.23 ± 0.49bc 15.65 ± 1.28ab salt 28.36 ± 0.81b 6.99 ± 1.04a 3.65 ± 0.27c 17.17 ± 0.49b 60 °c control 28.62 ± 1.73c 7.96 ± 0.61b 4.31 ± 0.70c 16.45 ± 13.59d blanched 26.54 ± 0.26ab 6.79 ± 0.06a 2.84 ± 0.60b 13.57 ± 0.53b ascorbic 24.97 ± 0.62a 6.77 ± 0.15a 1.88 ± 0.30a 11.84 ± 0.66a lime 26.82 ± 1.76abc 8.27 ± 0.52b 3.86 ± 0.13c 14.95 ± 1.11c salt 28.39 ± 1.13bc 7.90 ± 0.44b 4.51 ± 0.09c 16.30 ± 0.82d note: means within the same column at the same temperature with different alphabet(s) are significantly different at p ˂ 0.05. source: authors’ findings. table-3. colour changes for dried african star apple flesh of 4 mm thickness. temperature pre-treatment l* a* b* δe 50 °c control 27.01 ± 0.62ab 10.16 ± 0.90b 4.67 ± 0.87ab 17.24 ± 1.34a blanched 26.21 ± 0.97a 10.91 ± 0.64b 5.45 ± 1.38ab 17.61 ± 1.77a ascorbic 29.56 ± 3.60b 8.40 ± 0.17a 3.18 ± 1.43a 17.50 ± 3.56a lime 25.61 ± 0.66a 7.92 ± 1.01a 3.18 ± 1.84a 14.42 ± 1.98a salt 33.59 ± 1.61c 8.98 ± 0.52a 7.02 ± 0.07c 22.85 ± 1.03b 55 °c control 26.38 ± 2.77a 10.01 ± 0.76b 4.04 ± 1.94ab 16.43 ± 3.16ab blanched 25.65 ± 0.60a 8.60 ± 0.53a 3.32 ± 0.48ab 14.82 ± 0.58a ascorbic 27.90 ± 0.98ab 7.87 ± 0.47a 2.73 ± 0.79a 15.76 ± 1.04a lime 27.89 ± 1.65ab 8.24 ± 0.44a 4.51 ± 0.49b 16.93 ± 1.28ab salt 29.64 ± 0.81b 8.27 ± 1.04a 4.93 ± 0.27b 18.45 ± 0.49b 60 °c control 27.34 ± 1.73c 6.68 ± 0.61b 3.03 ± 0.70c 15.17 ± 0.97d blanched 25.26 ± 0.26ab 5.51 ± 0.06a 1.56 ± 0.60b 12.30 ± 0.53b ascorbic 23.69 ± 0.62a 5.50 ± 0.15a 0.59 ± 0.30a 10.56 ± 0.66a lime 25.54 ± 1.76abc 6.99 ± 0.52b 2.58 ± 0.13c 13.67 ± 1.11c salt 27.11 ± 1.13bc 6.62 ± 0.44b 3.23 ± 0.09c 15.02 ± 0.82d note: means within the same column at the same temperature with different alphabet(s) are significantly different at p ˂ 0.05. source: authors’ findings. table-4. colour changes for dried african star apple flesh of 6 mm thickness. temperature pre-treatment l* a* b* δe 50 °c control 28.73 ± 1.24a 8.94 ± 0.40b 5.74 ± 1.97a 18.51 ± 2.18a blanched 30.00 ± 3.69a 8.16 ± 0.18a 4.66 ± 1.74a 18.55 ± 3.75a ascorbic 29.53 ± 2.08a 7.80 ± 0.79a 5.52 ± 0.13a 18.60 ± 1.32a lime 26.52 ± 0.23a 8.15 ± 0.25a 3.59 ± 0.51a 15.36 ± 0.30a salt 26.88 ± 1.82a 7.58 ± 0.19a 3.33 ± 2.21a 15.25 ± 2.65a 55 °c control 24.66 ± 0.76a 6.49 ± 0.68a 1.68 ± 1.46a 12.28 ± 1.65a blanched 27.75 ± 1.06b 5.72 ± 0.44a 2.24 ± 0.71ab 14.66 ± 1.34ab ascorbic 24.22 ± 0.17a 7.22 ± 0.21ab 2.95 ± 0.27ab 13.07 ± 0.27ab lime 27.79 ± 1.86b 8.62 ± 0.27b 4.73 ± 0.78b 17.14 ± 1.71b salt 27.68 ± 3.32b 6.86 ± 1.91a 4.02 ± 3.48ab 15.98 ± 5.12ab 60 °c control 27.97 ± 0.54c 4.72 ± 0.69a 0.64 ± 0.49a 13.73 ± 0.36a blanched 24.90 ± 0.71a 6.08 ± 1.78ab 1.76 ± 1.81a 12.44 ± 2.16a ascorbic 26.39 ± 0.37ab 5.09 ± 0.38ab 1.66 ± 0.43a 13.11 ± 0.52a lime 26.68 ± 1.70bc 4.88 ± 0.31ab 4.09 ± 4.62a 13.26 ± 1.54a salt 25.30 ± 1.07ab 6.28 ± 0.57b 1.82 ± 0.47a 12.59 ± 1.17a note: means within the same column at the same temperature with different alphabet(s) are significantly different at p ˂ 0.05. source: authors’ findings. table-5. colour changes for dried african star apple flesh of 8 mm thickness. temperature pre-treatment l* a* b* δe 50 °c control 24.42 ± 0.37a 7.82 ± 0.60b 1.71 ± 0.81a 12.70 ± 0.82a blanched 29.16 ± 4.47b 7.82 ± 0.24b 4.41 ± 1.99b 17.70 ± 4.42b ascorbic 24.88 ± 0.54a 7.79 ± 0.60b 1.79 ± 0.60a 13.02 ± 0.35a lime 28.58 ± 1.82b 5.48 ± 0.51a 2.66 ± 0.59a 15.53 ± 1.67ab salt 29.24 ± 1.95b 7.70 ± 1.37b 5.15 ± 0.69b 18.14 ± 1.54b 55 °c control 28.50 ± 0.36a 7.54 ± 0.30a 3.54 ± 0.75b 16.53 ± 0.46a blanched 27.15 ± 1.28a 9.21 ± 0.59b 4.92 ± 1.58b 17.04 ± 2.08a ascorbic 27.12 ± 1.44a 6.82 ± 0.64a 1.87 ± 0.22a 14.34 ± 1.02a lime 27.12 ± 1.45a 7.06 ± 0.68a 4.22 ± 1.48b 16.23 ± 3.13a salt 27.12 ± 1.46a 7.36 ± 0.29a 4.53 ± 0.58b 16.32 ± 1.93a 60 °c control 27.74 ± 2.09a 7.34 ± 1.16b 4.26 ± 1.98b 16.31 ± 3.03b blanched 25.66 ± 3.34a 6.42 ± 0.86ab 1.08 ± 0.40a 12.84 ± 2.13a ascorbic 25.04 ± 0.89a 5.30 ± 0.14a 0.80 ± 0.86a 11.59 ± 1.18a lime 25.46 ± 0.35a 5.45 ± 0.66a 1.81 ± 0.90a 12.59 ± 0.87a salt 26.39 ± 1.34a 6.17 ± 0.71ab 2.49 ± 1.91ab 13.90 ± 2.34ab note: means within the same column at the same temperature with different alphabet(s) are significantly different at p ˂ 0.05. source: authors’ findings. the values recorded were lower compared to values available in literature for drying of similar foods, although their values for fresh samples were higher. also, comparable data on the drying of african star apple were not agriculture and food sciences research, 2019, 6(1): 145-154 153 © 2019 by the authors; licensee asian online journal publishing group available. however, doymaz, et al. [28] reported that higher l* and lower a*/b* values are required in dried food materials. the results obtained for the drying in this study are in line with that observation. an explanation for the colour changes experienced is given by fellows [29] where it was reported that colour changes in fruits and vegetables are caused by heat, oxidation during drying and residual enzyme activity which may cause browning during storage. a similar observation was reported by de-corcuera, et al. [30] on how blanching pretreatments made samples darker with loss of yellowness which gave lower l* and b* values. this was associated to the destruction of pigments by blanching. ali, et al. [31] reported a similar observation in the drying of guava slices. it was reported that the slices became darker during the drying process and it was attributed to prolonged drying time. it was also reported that temperature increase destroys colour quality; therefore, it was suggested that drying operations should be carried out at lower temperatures to better retain the colour of dried fruits. 4. conclusion the tropical african star apple fleshes were dried in the cabinet dryer. the drying rate of the african star apple flesh samples decreased as drying rate progressed while it increased with increase in drying temperature. the slice thickness and pretreatments were observed to affect the drying rates, with smaller thicknesses drying faster and blanching pretreatment having the highest drying rate of all. the increase in slice thickness was observed to reduce the drying rate and vice-versa. the drying time of the fruits ranged between 7 to 8 h. the structures of the samples were however observed to be differ compared to the control samples, despite significant ascorbic acid retention as a result of the pretreatments; especially the lime and ascorbic acid. the pretreatments were observed to help reduce the effect of ascorbic acid degradation by heat. the colour changes of the fruits were also significant as there were significant difference between the values of the fresh samples and the dried products. the ascorbic acid and lime pretreatments however had better colour overall. drying at 50 ºc with lime pretreatment gave the best result in terms of ascorbic acid retention. references [1] i. pardeshi, s. arora, and p. borker, "thin-layer drying of green peas and selection of a suitable thin-layer drying model," drying technology, vol. 27, pp. 288-295, 2009. available at: https://doi.org/10.1080/07373930802606451. 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[30] j. i. de-corcuera, j. r. cavalieri, and j. r. powers, blanching of foods. in d.r. heldman (ed.), encyclopedia of agricultural, food, and biological engineering. new york: marcel dekker, inc, 2004. [31] a. ali, y. a. yusof, n. chin, and m. n. ibrahim, "effect of different drying treatments on colour, quality and ascorbic acid concentration of guava fruit," presented at the 2nd international conference on agricultural and food engineering, cafe 2014, kuala lumpur, malaysia, 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 211 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 2, 211-220, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.62.211.220 © 2019 by the authors; licensee asian online journal publishing group heavy metals in soil and vegetables irrigated with extin mining ponds water in barkin ladi local government area plateau state, nigeria mafuyai g. m.1 eneji i. s.2 sha’ato r.3 nnamonu l. a.4 ( corresponding author) 1department of chemistry, faculty of natural sciences, university of jos, plateau state, nigeria. 2,3,4department of chemistry, college of science, university of agriculture, makurdi benue state, nigeria. abstract tin mining pond water irrigated fields can cause potential contamination with heavy metals to soil and vegetables, thus pose a threat to human beings. the current study was designed to investigate the contamination of the soil with toxic heavy metals and their accumulation in edible vegetable crops. the heavy metals pb, cu, cd, zn, cr, fe, mn and as were analyzed for their bioaccumulation factors to provide baseline data regarding environmental safety and the suitability of tin mining pond water for irrigation in the future. the contamination factor (cf) of these metals in the soil were calculated and indicated levels of metal contamination in the order of cd > zn > pb > cr > cu > as ˃ fe ˃ mn. the concentrations of pb, cd, cr, as, and mn in the edible vegetables were above the safe limit prescribe by fao/who, 2007 and eu, 2002 in all studied vegetables. the results indicated a potential pathway of human exposure to slow poisoning by heavy metals due to the utilization of vegetables grown on heavy metal contaminated soil that was irrigated with tin mining pond water sources. amongst the studied vegetables, as and cr were observed to exceeds tolerable limit. the irrigation source was identified as the source of the soil pollution in this study. thus, their consumption might pose substantial health risk to consumers and therefore need for proper remediation to reduce health risk and the extent of heavy metals contamination. keywords: heavy metals, contamination, vegetables, tin mining. citation | mafuyai g. m.; eneji i. s.; sha’ato r.; nnamonu l. a. (2019). heavy metals in soil and vegetables irrigated with extin mining ponds water in barkin ladi local government area plateau state, nigeria. agriculture and food sciences research, 6(2): 211-220. history: received: 15 july 2019 revised: 19 august 2019 accepted: 24 september 2019 published: 31 october 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: i give god almighty the glory for sustaining my soul with nourishment to end this research work. there are assured people whom without their support we would have great difficulties accomplishing this study. first, it has been a great benefit working with our supervisor professor i. s. eneji, whom without his guidance, support and fruitful comments and discussions, this research would never have been completed. special thanks are also extended to my wife for her support and prayers during the period. the technical staff at the chemistry department, university of jos, nigeria, thank you all for your kind support and understanding may lord bless you all. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 212 2. materials and methods ................................................................................................................................................................. 212 3. results and discussion ................................................................................................................................................................. 214 references ............................................................................................................................................................................................ 218 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.211.220&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.211.220&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1084 agriculture and food sciences research, 2019, 6(2): 211-220 212 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by providing a baseline research on heavy metals in ex-mining pond water accumulation in vegetables and the need for quotient. 1. introduction mining of solid minerals has been identified as a major entry point of heavy metals into the environment consequently polluting various components of the environmental media [1]. mining, which involves the extraction of naturally occurring minerals from the earth’s crust, is considered the world’s second oldest and most important industry after agriculture [2]. in the process of mining a particular metal the entire soil mass is excavated, laid bare and expose to the environmental agencies of weathering, degradation and transportation, this result in soil erosion and extensive contamination of the surrounding areas. the abandoned ponds and dumps tailing scattered still impact the environment by contaminating vegetables as well as pollution of underground water by discharged leachate [3]. heavy metals are considered as one among the most important environmental concerns because of their toxicity and accumulation in body [4],[5],[6]. heavy metals are those metals which have a specific density of more than 5 g/cm3 and harmfully affect the living organisms as well as environment and living organisms [7]. these metals are essential to maintain the normal body physiology and functioning when present in very low concentrations. however, they become lethal when certain threshold levels exceed. many studies have been conducted throughout the world in relation to plants and soil pollution with heavy metals through irrigation by urban and industrial effluents [8]. vegetables are important components of a healthy and perfect diet of human beings [9]. in a developing like country like nigeria, the fight against poverty, hunger, malnutrition and undernourishment continues to be a basic goal of development and a variety of strategies are being applied based on micronutrient rich food like vegetables considered essential. these vegetables are the fresh and edible portions of herbaceous plants. they are very popular in nigeria, though, its production is low in rainy season compared to dry season due to differences in environmental conditions, non-availability of high yielding varieties and cultural practices in the crop production [10]. they are important class of food substances highly beneficial for the maintenance of health and prevention of diseases [11]. the inclusion of vegetables in human diets has been identified as a major means of promoting balanced diets across populations of various income brackets [12]. this is because green vegetables have been recognized as one of the richest natural sources of essential minerals, protein and vitamins. in addition to being cheap sources of macro and micronutrients, vegetables could also be efficiently produced with limited resources. the regular consumption of vegetables, specifically the dark green leafy vegetables is highly recommended because of their potential in reducing the risks of chronic diseases [13],[14]. mining generally enriched heavy metals in soil leading to elevated levels in both plants and water samples [15]. sometimes the metals are very well preserved for hundreds of years and can remain a potential hazard for the environment. heavy metals are currently of environmental concern due to their bioaccumulation in soil and vegetables [16]. the soil, water and vegetable resulting from mining operations contain potentially carcinogenic levels of heavy metals and these elements could be related to the high prevalence of upper gastrointestinal cancer rates in the in the country [13]. the determination of heavy metal distribution in plants and environmental media is one of the most accurate criteria related to environmental pollution [17]. the accumulation of heavy metals in agricultural soils is of increasing concern because of food safety, potential health risks, and its detrimental effects on soil ecosystems [18],[19]. the accumulation of heavy metals in cultivated plants due to environmental media, fertilizers, and pesticides have a negative effect on plant growth and human health. the concentrations in plants can affect humans directly either through ingestion or through the food web by ingestion of crops or animals or indirectly-damaging environmental health [20]. the environmental exposure to heavy metals is a well-known risk factor for cancer [13]. some metal ions like cadmium, lead and, chromium have toxic effects on biochemical reactions in our body. heavy metals can be harmful due to their potential accumulation in different tissues of humans. even in low concentrations, heavy metals have detrimental health effects, because they are nonbiodegradable and persistent in nature [21],[17]. the health risks of heavy metals through consumption of vegetables from these mining soils are of great concern in the study area. irrigation has contributed significantly to poverty alleviation, food security, and improving the quality of life for rural populations. however, the sustainability of irrigated agriculture is being questioned, both economically and environmentally. the increased dependence on irrigation has not been without its negative environmental effects. thus, the need to investigate the levels of accumulation of heavy metal concentrations in the edible parts of some cultivated vegetables irrigated with ex – tin mining water in the area. 2. materials and methods 2.1. study area the study was carried out in barkin ldai local government areas of plateau state figure 1. the study areas lie between latitude 9°30’ 40’’n to 9°33’ 20” and longitude 8°53’20’’e to 8°54’40’’e. the study area played host to a lot of mining activities by foreign companies which rendered the area derelict with numerous waste dumps and ponds. soil and vegetables samples were collected through stratified random sampling method from four different farms each irrigated with tin mining ponds pond water.1kg of each of the vegetable samples including carrot, cabbage, garden egg, spinach, pepper and tomato from each stratum was collected. three replications of each of these samples were analyzed for their heavy metals concentration using flame atomic absorption spectroscopy (faas). agriculture and food sciences research, 2019, 6(2): 211-220 213 © 2019 by the authors; licensee asian online journal publishing group figure-1. map of barkin – ladi local government area showing study ponds. 2.2. soil sampling and preparation soil samples were obtained from four farms each irrigated with tin mining pond water in the study farms viz: b1 = opp. bethany acad. b2 = hosp. raod, b3 = gwol, b4 = behind highland bottling company. the soil samples was collected at depth ranging between 0 20 cm using a steel soil auger and kept in tagged polythene bags. the soil samples were air-dried and sieved to <0.25 mm, then stored in desiccators prior to analysis of heavy metals. 5g of dried and sieved subsoil sample was taken into 100 ml of conical flask. 20 ml of 1:1 hno3 was added to conical flask and covered with a watch glass. then the sample was evaporated to 5 to 8 ml on a hot plate. after cooling, 5 to 10 ml of hclo4 and 20 ml of metal-free distilled water added. then the sample was again evaporated to 10 to 12 ml on the hot plate. after cooling, the sample was filtered through whatman no. 42 filter paper and the filtrate transferred to a 100 ml volumetric flask and make up to mark with metal-free distilled water [22]. the concentrations of pb, cr, cu, cd, zn, mn and as were be determined by atomic absorption spectrophotometer (faas. pg990). 2.3. contamination factor (cf) the contamination factor was determined to express the level of metal contamination in surface soil and vegetables. the contaminant factor is calculated using the following formula: 𝐶𝐹 = 𝐶𝑚𝑒𝑡𝑎𝑙 𝐶𝑏𝑎𝑐𝑘𝑔𝑟𝑜𝑢𝑛𝑑 (1) where: cmetal is the concentration of a given metal in the soil and cbackground is a metal concentration of a control soil sample. the cf value for describing the contamination levels is summarized in table 1. table-1. level of contaminant based on (cf) values. cf values contamination remark < 1 no contamination 1 – 2 suspected contamination 2 – 3.5 slight contamination 3.5 – 8.0 moderate contamination 8.0 – 27 severe contamination > 27 extreme contamination source: sutherland [23]. 2.4. collection and preparation of vegetables for analysis the following vegetables were collected and analysed during dry and rainy season: carrot (daucus carota subsp. sativus), family: apiaceae; spinach (spinacia oleracea l.), family: amaranthaceae and tomato (lycopersicon esculatum l.), family: solanaceae, cabbage (brassica oleracea), red pepper (capsicum anum) and garden egg (solanum melongena), respectively. the vegetables (mature) samples were collected randomly from the both farms treated. healthy vegetables were collected carefully and washed thoroughly with tap water followed by distilled water and thereafter, kept on refrigerated under suitable condition analysis. the vegetables were cut into pieces with a stainless knife and dried at laboratory temperature and then powdered with mortar pestle before digestion to determine the heavy metals concentrations. 2.5. plant samples digestion for metal analyses one gram of each milled homogenized sample were weighed using a digital weighing balance into a conical flask. exactly 5 ml of 60% hydrochloric acid (hcl) and 10 ml of 70% nitric acid (hno3) was added into the weighed samples. the sample mixture was digested with moderate heat (50˚c) on a hot-plate until white fumes evolved, making it brownish solution. the heat was intensified further for few minutes to expel off most of the hydrochloric acid (hcl). exactly 50 ml of distilled water was added into the solution and heated for a few minutes, agriculture and food sciences research, 2019, 6(2): 211-220 214 © 2019 by the authors; licensee asian online journal publishing group allowed to cool before being filtered through whatman no. 42 paper (11 μm) into a dispensed transparent plastic container that is clean with detergent and treated successively with hcl and rinsed with deionized water. the filtered sample was left to stand for few minutes for the aspiration of the element. the digested samples was then analyzed for the concentrations of; lead (pb), manganese (mn), chromium (cr), cadmium (cd), zinc (zn), cupper (cu), iron (fe) and arsenic (as) using atomic absorption spectrophotometer (aas). 2.6. statistical analysis descriptive statistics viz., mean, standard error, standard deviation, minimum and maximum values of parameters are measured. 2.7. karl pearson’s coefficient of correlation coefficient of correlation (r) is a quantitative measure of the correlation between two variables. the correlation coefficient is measure of correlation is based on arithmetic mean and standard deviation. this method can be used to measure correlation for individual series as well as for grouped data. the following equation is used for getting pearson’s coefficient (r) is: ( )( )  −− = nsxsy yyxx r (2) where, r = coefficient of correlation, x = variable x, x̅ = mean of variable x. y = variable y, y̅ = mean of variable y, n = number of pairs of variables. sx= sd of variable x, and sy = sd of variable y. 3. results and discussion 3.1. heavy metals in soil heavy metals in the soil are derived from natural components or geological sources as well as from human activities or anthropogenic sources. normally heavy metals in soil are found in several forms. these forms are involved in their movement from soil to plant. the conversion of immobile or non-bioavailable forms of heavy metals to mobile or bioavailable forms is dynamic phenomenon in the soil and occurring continually is regulated by physical, chemical and biological processes and interactions between them. as a result it is found that any form is not stable for long time. in this study total content of soil pb, cu, cd, zn, cr, fe, mn and as are considered which are depicted in lead (pb): the mean concentration of pb was 67.9±8.2 mg/kg for the four different farm soil irrigated with tin mining pond. the highest concentration of 80.6 mg/kg was observed in farm b2 with the lowest 59.2 mg/kg in farm b1. the trend of the concentrations in the four different farms was b2 > b3 > b2 > b4 > b1. this result showed similar trend of increased in pb concentration in the soil irrigated with tin mining pond water. it was observed that pb is positively correlated with zn in dry season and negatively with cu, cr and fe while in rainy season pb is positively correlated with fe. the reported values are within he prescribed safe limit of irrigation soil [24] standard limits. the value of pb obtained in this work is higher compared with that reported in kassa ropp barkin – ladi [25]. similar work done revealed pb concentration in soil to range from 1 – 58 mg/kg mahmood and malik [26]. ratul, et al. [27] reported 28.13 mg/kg in agricultural soil irrigated with contaminated river water lower than the concentration reported in this study. pb in soil treated with mining pond water is positively correlated with mn and as in dry season and rainy season but a significant positive correlation is showed between pb and available phosphorus in the treated soil collected during rainy season. the study showed the significant difference (p < 0.05) in the pb content of the treated soils collected in both seasons. the pb enrichment in the mining pond water irrigated soils as compared to ground water irrigated soils are collaborated with the findings of 15.4 ±6.6 mg/kg and 9.31 ±2.2 mg/kg [26],[28]. copper (cu): the mean value of cu in mining pond water treated soil collected was 20.3mg/kg and the highest value of 27.2 mg/kg was obtained in farm b1 with the lowest 15.6 mg/kg at farm b2. this result is lower compared to the findings 42.0 – 111.6 mg/kg reported [3] which recorded extremely high concentrations of copper at industrial sites, and the also the reported 69.01 mg/kg in agricultural soil irrigated with contaminated river water [27]. the cu concentration of the soil irrigated with tin mining pond water is within the safe limit for cultivation [24]. the trend of cu in the soil sample farms is in the order of b1 > b4 > b3 > b2, respectively. the metals contents in both soil irrigation with mining pond water also corroborated with the findings of henry, et al. [29],singh, et al. [5]. cadmium (cd): the concentration of cd in the mining pond water treated soil with tin mining pond water in both sites studied shows a mean of 0.94 ± 0.4 mg/kg. the highest concentration was obtained in farm b1 1.17 mg/kg and the lowest 0.28 mg/kg at b4. the concentration obtained in this work for soil treated with tin mining pond water agrees with the 0.965 mg/kg reported [27] in agricultural soil irrigated with contaminated river water. comparing with the safe limit of cd in the soil it is found that both farm soils are within the prescribed standards of 3.0 mg/kg set [30],[31]. cadmium concentration in soil was relatively high, this may be attributed to applications of fertilizers and other farming practices including used of pesticides. generally, the high concentrations of metals in this area could also be as a result of the tin mine activities, wastes dumped and metals availability in the earth crust. the treated soil does not show any significant correlation with other parameter at the significant difference (p < 0.05). in this study cd concentration in both tin mining pond water irrigated farm soils are lower compared to the literature values reports of 15.38±6.6 mg/kg, 3.54±0.6 mg/kg and 9.91±1.1 mg/kg [26],[25],[32] respectively. zinc (zn): the mean concentration of zn in the mining pond water treated farm was 68.87 ± 18.4 mg/kg. the highest concentration obtained was in farm b2 soil 94.8 mg/kg and lowest at b3 soil 54.3 mg/kg. from the agriculture and food sciences research, 2019, 6(2): 211-220 215 © 2019 by the authors; licensee asian online journal publishing group concentration of zn obtained in the soil irrigated with tin mining pond water, it is clear that the water has potential for the development of zn enrichment. both the soil samples analyzed has been found to be enriched with more zn. the zn concentration in both studied farm soils is within the safe limit of 200mg/kg prescribed [31]. the zn concentration in the wastewater irrigated soil reported in this study is in agreement with wastewater irrigated soil concentrations of 50.80 ±28.6 mg/kg published [26] and reported concentration of 45.73 mg/kg [27] in agricultural soil irrigated with contaminated river water; but however, higher compared to the result of 3.9 ±0.1 and 6.03±1.7 mg/kg reported [28]. chromium (cr): the soil samples that are collected has a mean cr concentration in mining pond water treated soil as 12.5 ±4.1 mg /kg. it was observed that cr in both farm soils, the highest concentration was found in farm b1 soil 14.5 mg/kg and lowest in b2 soil 11.1 mg/kg. the result reported in this study is lower compared with 22.9 mg/kg reported by and 20.01±11.3mg/kg [15], [26] and 54.2 mg/kg reported in kassa ropp for similar tin mining soil and 69.75mg/kg reported by daniel, et al. [25], ratul, et al. [27] in agricultural soil irrigated with contaminated river water. however, the low value obtained may be as attributed to leaching of metals beneath the soil. the cr concentration in soil of both plots irrespective are within the 150 mg/kg safe limit standard [30]. the study also showed the significant difference (p < 0.05) in the cr content of treated soils collected in both seasons. iron (fe): the soil sample irrigated with tin mining pond water during at the various sampling farms has mean concentration of 491.48 ±28.5 mg/kg. higher concentration of fe were observed in the soil around farm b1 526 mg/kg and the lowest farm b2 478 mg/kg mining ponds, this may be as a result of the washing of mining piled dumps by runoff water during rainfalls. in the same vain fe correlated negatively with mn and as. the results show that iron (fe) is the most abundant essential metal in both soil and vegetable samples, samples. the trend of concentration of fe in the farm soils analyzed is in the order of b1 > b4 > b3 > b2. the variations in the absorption of fe from the soil by the plant’s tissues are evident in the low fe contents in the vegetable samples. the high concentrations of fe in the soil samples may suggest a very anthropogenic source of fe, which allows the percolation of fe to the soil depths rather the surfaces. boamponisem reported 14.08 mg/k lower values of fe as compared to this work [32]. manganese (mn): the soil irrigated with tin mining pond water in both farms showed a of 18.1 ±1.3 mg /kg, with the highest concentration obtained in farm b4 soil 19.8 mg/kg and the lowest at b1 soil 16.3 mg/kg. the trend in concentration in the farms soil is in the order of b4 > b3 > b2 > b1. this variation in concentration of mn may be attributed to accumulation washing from different places such as roads, ashes from burn vegetation washed down by rainfall. the concentration of mn reported in the soils studied is lower compared to limit of 80.0 mg/kg prescribed by usepa [31]. increased mn content in the mining pond water irrigated soils as compared with ground water irrigated soils agrees with findings of mahmood and malik [26]. arsenic (as): the mean concentration of as in tin mining pond water treated soil collected from both farms was 146 ±23 mg/kg. the highest concentration was recorded in b3 soil 174 mg/kg with the lowest obtained in b1 soil 114 mg/kg. the level of concentration of as in the farms is in the order of b3 > b4 > b2 > b1. the concentration of as recorded in this work is higher compared to the mean total arsenic concentrations of 50 – 60 mg/kg recorded for agricultural soils treated with arsenical pesticides [33],[34]. the result shows that there is accumulation of as in the soil as a result industrial wastes and pesticide applications which might increase concentrations. the as level in the irrigated soils is within the safe limit for cultivation. as, show positive correlation with mn cr and pb. naturally elevated levels of arsenic in soils may be associated with geological substrata such as sulfide ores therefore, anthropogenic contaminated soils can have several concentrations of arsenic. arsenic concentrations of up to 27 000 mg/kg were reported in soils contaminated with mine or smelter wastes [35]. soil on agricultural land treated with arsenical pesticides may retain substantial amounts of arsenic [34]. figure-2. mean heavy metal concentration in soil irrigated with tin mining pond water (mg/kg). table-2. contamination of heavy metals for soil at different irrigated soil. metals bethany academy (b1) hospital road (b2) gwol (b3) highland bottling company (b4) pb 1.26 2.25 2.37 1.32 cu 0.69 1.67 1.32 1.27 cd 3.53 3.39 4.15 3.84 zn 1.08 2.14 2.18 1.37 cr 1.52 2.93 2.35 1.09 fe 1.39 1.26 1.38 1.14 mn 1.33 0.30 1.36 0.99 as 1.02 1.31 1.22 1.33 agriculture and food sciences research, 2019, 6(2): 211-220 216 © 2019 by the authors; licensee asian online journal publishing group 3.2. contamination factor (cf) of heavy metals in soil contamination factor (ef) of heavy metals in the soil was calculated with help of the formula provided [36]. soils collected from the cultivated soils’ shows variation in metal concentration. soils collected from the various farms of the study sites shows sequence of contamination in descending order as: cd > zn > pb > cr > cu > as ˃ fe ˃ mn. based on the contamination categories as proposed [23] it is found that soils irrigated with tin mining pond water were suspected to contamination with cr, pb, and zn. the rest of the metals exhibit the minimal deficiency of mineral contamination in the soil and slight contamination was observed in the soil with cd in all farms studied. according [37] contamination values between 0.5 and 1.5 indicate the metal is entirely from the crust materials or natural processes, whereas contamination values greater than 1.5 suggest that the sources are more likely to be anthropogenic. higher transfer coefficients reflect high soil contents or greater potentials of plants to absorb metals and bio-accumulate into tissues [38]. however, low transfer coefficients have been reported to indicate strong sorption of the metals to soil colloids [39]. contamination factor (cf) of heavy metals in the irrigated soils is also highlighted as cd (2.46) ˃ cr (2.43) ˃ zn (1.67) ˃ cu (1.36) ˃ pb (1.31) [40]. table-3. mean ±sd concentration of heavy metals in vegetables (mg/kg). metals vegetables tomatoes garden egg pepper cabbage carrot spinach pb 0.491±0.11 0.396±0.05 0.307±0.26 0.164±0.03 0.465±0.02 0.455±0.04 cu 0.071±0.01 0.076±0.01 0.065±0.00 0.065±0.01 0.090±0.01 0.062±0.01 cd 0.009±0.00 0.006±0.01 0.006±0.00 0.005±0.00 0.022±0.01 0.014±0.01 zn 0.376±0.04 1.104±0.08 0.395±0.02 0.184±0.02 0.913±0.06 0.448±0.03 cr 0.154±0.03 0.116±0.00 0.106±0.01 0.109±0.01 0.148±0.02 0.147±0.03 fe 1.551±0.21 1.170±0.02 1.489±0.03 1.489±0.07 1.647±0.14 1.554±0.21 mn 0.454±0.39 0.825±0.11 0.177±0.01 0.166±0.01 0.791±0.15 0.115±0.01 as 0.076±0.01 0.043±0.01 0.033±0.02 0.028±0.01 0.466±0.39 0.046±0.00 3.3. heavy metals accumulation in cultivated vegetables tomato: the result shows that pb concentration in tomatoes is 0.49±0.1 mg/kg. the values obtained are similar to the value 0.26 – 0.70 mg/ kg mahmood and malik [26] and 1.15±0.29 mg/k reported values [41]; in a research on the level of as, cd, cr, hg and pb in soil and some vegetables which showed concentrations above maximum permissible limit and standard value. it also observed high levels of pb (2.40±0.99) mg/kg and cd (0.25±0.11) mg/kg in wastewater irrigated tomatoes in egypt [42]. higher concentration of 10. 75 mg/kg are also reported in wastewater irrigated tomatoes [43]. the mean concentration of arsenic was 0.076mg/kg. this concentration is high compared to 0.06±0.02 mg/kg reported [44] and 0.62±0.19 mg/kg [41]. the high value of arsenic in might be due the usage of arsenic containing chemicals such as pesticides and herbicides (calcium arsenate, cupper acetoarsenate and lead hydrogen arsenate). it was reported that these were used in pig and poultry feeds which in turn introduce arsenic into the soil and plants as chicken dug is used as fertilizers [45]. the cr concentrations were also high in the study areas, with a mean of 0.15 mg/kg. the concentration of cr obtained in this study is lower compared to 0.66 mg/kg reported in similar studies [46],[47]. mn has a mean concentration of 0.45±0.39 mg/kg in the tomatoes cultivated with tin mining pond water in the various farms studied. however, the might have been due washing by runoff water from the immediate vicinity since several anthropogenic processes took place and tin mining is still occurring in small scale by illegal miners. this is agree with the 0.12 mg/l reported on the study of vegetables in southern turkey [48]. the high concentrations of some of these metal in the areas could be as a result of the closeness of the mining ponds to major traffic highways. other metals such cu, zn, and fe, has concentrations of 0.071±0.01, 0.376±0.04 and 1.55±0.21, respectively. table-4. pearson's correlation coefficient matrix (mean) concentration of heavy metals in tomatoes. pb cu cd zn cr fe mn as pb 1.000 cu -0.750 1.000 cd 0.743 -1.000 1.000 zn 0.906 -0.399 0.389 1.000 cr -0.667 0.993 -0.994 -0.288 1.000 fe -0.410 0.911 -0.915 0.015 0.953 1.000 mn 0.869 -0.325 0.315 0.997 -0.211 0.095 1.000 as 0.365 -0.890 0.894 -0.064 -0.937 -0.999 -0.143 1.000 statistically (p-value < 0.05). pb is positively correlated with cd, zn and mn while zn with mn. a positively correlation was also notice between cd and as, and also cr is negatively correlated with cd and as. the order of metal concentration in tomatoes in this work is; fe ˃ mn ˃ zn ˃ pb ˃ cu ˃ cr ˃ as ˃ cd. heavy metal accumulation in tomato due to long-term wastewater irrigation was also highlighted [29], [43],[49]. garden egg: the mean concentrations of each heavy metals in the three farms shows that pb 0.40 ±0.05, cr (0.12 ±0.00) and mn (1.17±0.02) as (0.83 ±0.11) mg/kg were higher than the standard permissible limit [50] respectively. the concentration of cu 0.076±0.01 below the permissible standard value of 0.7mg/kg in both garden egg irrigated with tin mining pond water while the concentration fe and cd are lower compared to other metals standards. the order of heavy metals accumulation in garden egg was fe > mn > pb > zn > cu > cr ˃ as ˃ cd. pearson’s correlation shows that in garden egg pb is positively correlated with zn and mn, while cu is positively correlated with cr, and cd negatively correlated with cr. the results obtained in this study were agriculture and food sciences research, 2019, 6(2): 211-220 217 © 2019 by the authors; licensee asian online journal publishing group similar to the reported studies many scholars [43],[51],[52] who also reported high levels of zn (3.91) and cd (1.56) mg/kg in wastewater irrigated garden egg in kaduna, nigeria. table-5. pearson's correlation coefficient matrix (mean) concentration of heavy metals in garden egg. pb cu cd zn cr fe mn as pb 1.000 cu -0.750 1.000 cd 0.743 -1.000 1.000 zn 0.906 -0.399 0.389 1.000 cr -0.667 0.993 -0.994 -0.288 1.000 fe -0.410 0.911 -0.915 0.015 0.953 1.000 mn 0.869 -0.325 0.315 0.997 -0.211 0.095 1.000 as 0.365 -0.890 0.894 -0.064 -0.937 -0.999 -0.143 1.000 pepper: the mean concentration of the various heavy metals in pepper irrigated with tin mining pond water was pb (0.307±0.26), cu (0.065±0.00), cd (0.006±0.00), zn (0.395±0.02) cr (0.106±0.01), fe (1.489±0.03), mn (0.177±0.01) and as (0.033±0.02). the concentration of cd was slightly lower in all the study areas. the order of concentration of the heavy metals in pepper cultivated with tin mining pond water was fe > mn > pb > cu > zn > cr ˃ as ˃ cd. the concentrations of mn, cr, pb and cd in pepper in the studied areas have crossed the prescribed safe value [30],[50] respectively. the high concentration of this metals in pepper might be attributed to high level of pesticides and fertilizer on farmland for better yield of crops. wastewater induced heavy metal accumulation in vegetables is also reported as mn (19.1±1.7) mg/kg accumulation in vegetables grown on farmlands irrigated with treated sewage water in ghana [53]. 1.15±0.29 mg/kg accumulation of cd in pepper is also being reported [41]. pb and cu correlated positively cr, mn and as, while cd correlated positively with mn and as and cr negatively correlated with as. heavy metals concentration in both sites vegetables showed significant level (p < 0.05) accumulation except cd (p = 0.05). dorcas high levels of heavy metals are also reported vegetables irrigated with wastewater from mines and industrial discharges to soils [54],[55],[56],[57]. table-6. pearson's correlation coefficient matrix (mean) concentration of heavy metal in pepper (mg/kg). pb cu cd zn cr fe mn as pb 1.000 cu -0.751 1.000 cd 0.944 0.998 1.000 zn -1.000 -0.969 -0.952 1.000 cr -0.873 -0.707 -0.664 0.861 1.000 fe 0.763 0.910 0.933 -0.779 -0.351 1.000 mn 0.907 0.987 0.995 -0.917 -0.587 0.964 1.000 as 0.907 0.980 0.966 -0.999 -0.834 0.809 0.936 1.000 cabbage: the mean concentration of heavy metals in the studied cultivated cabbage irrigated with tin mining pond water was found to be pb (0.164±0.03), cu (0.065±0.01), cd (0.01±0.00), zn (0.18 ±0.02), cr (0.11±0.01), fe (1.49±0.07), mn (0.17±0.01) and as (0.03±0.01) mg/kg. the concentration of metals in cabbage irrigated with tin mining pond water is lower compared to the reported literature of 0.22±0.2 mg/kg and 6.25±1.2 mg/kg 1.61±0.2 [56],[58], [59] mg/kg). similar studies conducted show order of concentration of the heavy metals in is: fe > mn > pb > zn > cr > cu ˃ as ˃ cd [29]. it was therefore, noted that some of metals that are high in cabbage cross the standard permissible limits fao/who. the high levels of these metals in this vegetable might be as a result of the use of pesticides and herbicides and irrigated with tin mine water [60]. this also agrees with the findings that, the application of large volumes of partially treated or untreated wastewater in some parts of africa has adversely affected both surface water bodies and the urban and peri-urban farmers using these water bodies as sources of irrigation [61]. the pearson’s correlation shows that pb correlated positively with cu, mn, as, and negatively correlated cr. cd is positively correlated with mn, as and negatively correlated with cr at (p > 0.05). table-7. pearson's correlation coefficient matrix (mean) concentration of heavy metals in cabbage. pb cu cd zn cr fe mn as pb 1.000 cu 0.993 1.000 cd -0.148 -0.034 1.000 zn 0.958 0.984 0.143 1.000 cr -0.754 -0.824 -0.538 -0.911 1.000 fe -0.994 -0.974 0.260 -0.919 0.674 1.000 mn 0.932 0.884 -0.497 0.788 -0.464 -0.967 1.000 as 0.999 0.992 -0.163 0.953 -0.744 -0.995 0.937 1.000 carrot: the heavy metals in the carrot analyzed from the sampled farms shows that the concentrations are higher in all studied metals. the metals studied crossed the safe limit of european union (eu) [30]; fao/who [50] limit with the exception of cu, zn, and fe others were not safe for human consumption as their concentrations were above the maximum permissible limit, 0.30mg/kg. the mean concentrations of pb (0.46 mg/kg) and as (0.47 mg/kg) were higher compared to other toxic metals in carrot the studied metals are in order of their descending trend: fe ˃ zn ˃ mn ˃ as˃ pb ˃ cr ˃ cu ˃cd. studies showed that cadmium (cd) had mean concentration of 0.24 mg/kg in carrots in barkin ladi haipang compared [54] with the result of 0.02mg/kg recorded in this work. several results have been reported of cd in carrot due to irrigation with agriculture and food sciences research, 2019, 6(2): 211-220 218 © 2019 by the authors; licensee asian online journal publishing group wastewater [26],[62],[63]. pb correlated positively with cr and negatively with mn. cd and cr showed negative correlation in significant level of (p < 0.05). table-8. pearson's correlation coefficient matrix (mean) concentration of heavy metals in carrot. pb cu cd zn cr fe mn as pb 1.000 cu 0.993 1.000 cd -0.768 -0.687 1.000 zn 0.662 0.569 -0.989 1.000 cr 0.923 0.872 -0.955 0.899 1.000 fe 0.399 0.289 -0.894 0.951 0.721 1.000 mn -0.703 -0.615 0.995 -0.998 -0.922 -0.933 1.000 as -0.285 -0.395 -0.396 0.530 0.105 0.765 -0.481 1.000 spinach: the concentration of all the metal studied in all sites were found to have high metals in spinach but others show lower values. the mean concentration of spinach was pb (0.46 ±0.04) cu (0.06±0.01), cr (0.014±0.01), zn (0.45±0.03), cr (0.15±0.03), fe (1.55±0.21). comparing the concentration heavy metals studied in spinach, it is found that values obtained for pb are lower than the findings of 2.9±1.2 mg/kg 1.50±0.1 mg/kg,15.1 mg/kg 2.78 mg/kg, 3.31mg/kg [26],[32],[56],[64],[65]. the concentration of cr and cd in agricultural soil irrigated with contaminated river water are far higher than the value reported in this work [4],[27],[39],[66]. comparing with the prescribed standards it is found that pb, cr, as, mn and cd content in spinach have crossed safe limit [24],[30]. however, the major sources of vegetable contamination with heavy metals might be due to the waste water irrigation, solid waste disposal, sludge applications, vehicular exhaust. excessive accumulation of heavy metals in agricultural soils through the use of agrochemicals and by other sources may lead to elevated heavy metal up-take by vegetables and thus affect food quality and safety [66]. statistically heavy metals content in spinach in shows significant (p < 0.05). pb is positively correlated with cr and negatively correlated with mn and as. wastewater induced heavy metals enrichment in spinach was also studied [4],[39]. the variations in heavy metal concentrations in vegetables of the same site may be ascribed to the differences in their morphology and physiology for heavy metal uptake, exclusion, accumulation and retention [7]. it is worthy of note that low concentrations of fe in the vegetable samples relative to its abundant availability in the soils, can be attributed to; low absorption of fe by the tissues of the vegetable samples, possible leaching of fe from the soil surface and runoff during rainfall. table-9. pearson's correlation coefficient matrix (mean) concentration of heavy metals in spinach. pb cu cd zn cr fe mn as pb 1.000 cu -0.202 1.000 cd 0.597 -0.906 1.000 zn -0.771 0.780 -0.971 1.000 cr 0.988 -0.182 0.581 -0.758 1.000 fe 0.807 -0.741 0.955 -0.998 0.795 1.000 mn -0.796 -0.431 0.009 0.229 -0.809 -0.286 1.000 as -0.882 -0.283 -0.149 0.380 -0.891 -0.434 0.987 1.000 3.4. conclusion from the present study it is noted that tin mining pond water irrigation practice is not suitable for cultivating edible vegetables especially those that are consumed regularly. because excess heavy metals are considered as toxic elements due to accumulation in the biological systems 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[66] m. muchuweti, j. birkett, e. chinyanga, r. zvauya, m. d. scrimshaw, and j. lester, "heavy metal content of vegetables irrigated with mixtures of wastewater and sewage sludge in zimbabwe: implications for human health," agriculture, ecosystems & environment, vol. 112, pp. 41-48, 2006. available at: https://doi.org/10.1016/j.agee.2005.04.028. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 6 agriculture and food sciences research vol. 5, no. 1, 6-22, 2018 issn(e) 2411-6653 / issn(p) 2518-0193 doi: 10.20448/journal.512.2018.51.6.22 genetic differentiation of arc soybean [glycine max (l.) merrill] accessions based on agronomic and nutritional quality traits mofokeng, maletsema alina1  mashingaidze, kingstone2 1,2agricultural research council-grain crops, potchefstroom, south africa abstract soybean is one of the most important leguminous crops grown globally for food and feed. the study of genetic diversity is invaluable for efficient utilization, conservation and management of germplasm collections. the study aims at assessing genetic diversity present among the soybean genotypes using phenotypic markers. the restriction maximum likelihood revealed highly significant differences among the genotypes for eight quantitative traits. the principal component analysis revealed three most important pcs contributing 63.19%, 25.43% and 8.88% to the total variation of 97.5%, respectively. seed yield was highly significant and highly correlated with seed number per plant, pod weight per plant, pod number per plant, and hundred seed weight but negatively correlated with seed number per pod. the hierarchical clustering revealed three major clusters with further sub-clusters. the accessions 2015/06/12, 69 s 10, pr 154-14, r 5-4-2 m, hawkeye (ussr), and pr 145-2 were the most diverse. there were significant differences among the accessions based on nutritional quality traits such as oil, protein and stearic acid across the locations. the protein content varied from 29.1% to 35.6%, oil content varied from 10.6% to 20.7% whereas oleic acid and ash varied between 6.8% and 30.8%, and 4.3% and 8.2%, respectively. there was vast genetic diversity among the soybean genotypes. the presence of genetic diversity will aid breeders in selections and hybridization programmes for crop improvement. keywords: agro-morphology, genetic diversity, nutritional quality, soybean. citation | mofokeng, maletsema alina; mashingaidze, kingstone, (2018). genetic differentiation of arc soybean [glycine max (l.) merrill] accessions based on agronomic and nutritional quality traits. agriculture and food sciences research, 5(1): 6-22. history: received: 10 october 2017 revised: 17 january 2018 accepted: 22 january 2018 published: 25 january 2018 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: the authors would like to thank the department of science and technology for funding, mr paul rantso, and soybean breeding team for technical help. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ......................................................................................................................................................................................... 7 2. materials and methods ...................................................................................................................................................................... 7 3. results ................................................................................................................................................................................................... 9 4. discussion .......................................................................................................................................................................................... 15 5. conclusion ......................................................................................................................................................................................... 16 references .............................................................................................................................................................................................. 21 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2018.51.6.22&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://orcid.org/orcid-search/quick-search?searchquery=mofokeng,%20maletsema%20alina https://orcid.org/orcid-search/quick-search?searchquery=mashingaidze,%20kingstone agriculture and food sciences research, 2018, 5(1): 6-22 7 1. introduction soybean [glycine max (l.) merrill] (2n=2x=40) is one of the most important legumes produced worldwide. according to food and agriculture organisation of the united nations [1] the three major world-producing countries are u.s.a (90.6 million metric tonnes), brazil (68.5 million metric tonnes) and argentina (52.6 million metric tonnes). the total production in africa was 1.5 million tonnes with west africa producing 437,115 metric tonnes. nigeria is the leading producer in west africa with 393,860 metric tonnes [2]. in south africa soybean is produced in almost all provinces with the free state being the major producer. in 2014, south africa produced 8851 metric tons in an area of 502900 hectares. soybean is grown primarily for the production of seed and has several uses in the food and industrial sectors. it is the most important crop provider of proteins and oil used in animal nutrition and for human consumption. it contains 40 to 42% good quality protein and 18 to 22% oil comprising 85% unsaturated fatty acids and is free from cholesterol, it is highly desirable in the human diet [3]. besides fixing the atmospheric nitrogen, this crop has the ability to grow in a wide range of environments, to reduce soil erosion, to suppress weeds and to suit inter and sequential cropping patterns. it is valued finding to understand genetic diversity and relationship for facilitating the transfer of useful genes among cultivated species and maximizing the use of available germplasm resources. the extent of genetic diversity in germplasm can be assessed through morphological characterization. the characterized material then helps the plant breeders to select the accessions to be utilized in hybridization programme [4]. an investigation of genetic relatedness at a broad level may provide important information about the historical relationship among different genotypes. it reveals genetic backgrounds and relationships of germplasm and also provides strategies to establish unitize and manage crop core collections [5]. therefore the knowledge of the genetic variation within accessions from germplasm collections is essential to the choice of strategies to incorporate useful diversity into the program to facilitate the introgression of genes of interest into commercial cultivars, to understand the evolutionary relations among accessions, to better sample germplasm diversity and to increase conservation efficiency [6]. morphological characters, both quantitative and qualitative have long been used to identify species, genera, to evaluate systematic relationships, and to discriminate between varieties [7]. in breeding practice and seed production, the role of morphological descriptor is very important, since the distinguishing between varieties can be done quickly and precisely. qualitative traits are usually controlled by a few genes, thus easily observable and suitable for cultivar differentiation and identification. on the other hand, quantitative traits have more limitations in cultivar description, since they are affected by environmental effects, developmental stage of the plant and the generation of selfing of breeding material. according to khalid, et al. [8] scientific classification of the plant still relies on morphological traits. they are easier to work with, cost effective and easy to score and requires less time and finally they do not need any technical knowledge. kumar, et al. [9] evaluated genetic diversity and interrelationship of agro-morphological traits in soybean genotypes. vesna, et al. [10] determined genetic relatedness of soybean genotypes based on agro-morphological traits and dna markers and they found genetic differences among the genotypes. malek, et al. [11] also assessed genetic variability and association of characters among the soybean mutants and reported a vast genetic variability. khatab, et al. [12] reported the presence of genetic diversity among soybean genotypes assessed through agro-morphological descriptors. soybean has nutritional quality attributes such as protein and oil that makes it an important food crop. it has 40–42% high quality protein and 18–22% oil comprising up of 85% unsaturated fatty acid as well as 12% carbohydrates [13]. soybean not only contains high quality protein, but the protein content is also much higher than that of other plant foods. soy protein is valued as a healthy protein due to containing a balanced proportion of all of the important and essential amino acids required by the human body [14]. it can provide two fold more proteins as compared to any other vegetable crop or grain [15]. soy oil can serve as a good source of oleic and linoleic acid, even the partially hydrogenated soy oil contains 25% linoleic and 3% linolenic acid [16]. soybean oil is also a good source of vitamin e [17]. some recent studies revealed variations among the soybean accessions based on quality traits such as oil [18]. however, the nutritional quality traits vary depending on the varieties grown. there are limited studies on the analysis of genetic diversity among the soybean accessions based on nutritional quality traits in south africa. there is still need to understand and record variations due to other nutritional quality traits that soybean cultivars possess. this will aid breeders to improve the quality of soybean cultivars and for selection of best accessions for both quality agronomic attributes. the objective of the study was to assess genetic diversity using agromorphological and nutritional quality traits among soybean genotypes grown in south africa. 2. materials and methods 2.1. plant material. experimental layout and management ninety-eight soybean genotypes maintained at the agricultural research council-grain crops institute were planted in potchefstroom (26.7145° s. 27.0970° e) and brits (25.6100° s. 27.7960° e) in alpha lattice design replicated two times. each plot consisted of two 4 m length rows with a spacing of 75 cm between the rows and 10 cm between the plants. fertilizer 2:3:4 was applied before planting. the plants were irrigated using sprinkler irrigation system. the pre-emergent weeds were controlled by herbicide bateleur gold and post emergent weeds were controlled by both basagran ec and manually. lime ammonium nitrate was top dressed 45 days after germination i.e. before the plants flower. the cultural practices were applied as per soybean planting recommendations. 2.2. data collection and analysis the agro-morphological data recorded is indicated in table 2. at harvest, five plants were randomly taken from each plot to measure days to 50% flowering; number of branches per plant; hundred seed weight (g); pod weight (g); seed number per pod; seed number per plant; pod agriculture and food sciences research, 2018, 5(1): 6-22 8 number per plant; and seed yield (g). analysis of variance was performed for all traits in order to test the significance of variation among the genotypes. the data were further subjected to principal component analysis and correlations. the dendrogram was constructed to study the genetic relatedness among the tested accessions using hierarchical clustering in genstat 18th version. table-1. list of names of the soybean accessions used in the study. serial number gm number accession name serial number gm number accession name 1 220 69 s 10 50 673 1935/06/15 2 223 69 s 13 seleksie 51 675 1971/06/06 3 231 69 s 19 52 678 105/5 4 266 nim 53 679 2012/01/05 5 267 yubelejuaja 54 681 2015/06/12 6 268 hawkeye (ussr) 55 688 85/05/ 7 278 shelby 56 691 1964/06/17 8 284 gx gous 57 699 essex 9 290 chippewa 63 58 707 db 1601 10 291 r 5-4-2 m 59 715 crawford 11 292 r 2-11-3m 60 724 d64-4636 12 297 b 66 s 365 61 725 d66-8666 13 304 b 66s 385 62 861 oribi 14 544 columbia m 8 a 63 862 impala 15 568 grant 64 864 sss 3 16 571 hampton 266 a 65 873 nd 85 17 575 hawkeye 66 1120 ags 239 18 578 hernon 67 1363 ipb 212-81 19 582 jackson 68 1371 coc chum 20 592 mack 6 69 1380 f 82-7824 21 593 mandarin 70 1386 mtd 63 22 595 maksura 71 1390 jupiter 23 597 mojiana 72 1403 f 82-7145 24 598 n69-2774 73 1409 f 82-7656 25 604 pi 170889/(r56-49) 74 1449 ufv-1 26 607 pikett 75 1552 pr 118 (278) 27 617 s4-a.p.4 76 1554 pr 133 (484) 28 621 santa rosa 77 1555 pr 144-4 29 623 soja (pautena) 78 1556 pr 144-9 30 624 solar 12 79 1558 pr 145-2 31 626 sss 2 80 1572 pr 154-13 32 630 vaschadaka 81 1573 pr 154-14 33 631 vicoja 82 1575 pr 154-22 34 637 yeluanda 83 1578 pr 154-47 35 646 54 s 116 84 1590 pr 161-40 36 648 54 s 219 85 1594 pr 162-18 37 649 54 s 95 86 1595 pr 164-20 38 650 14/6/32 87 1596 pr 164-22 39 651 2017/06/11 88 1597 pr 165-3 40 652 2020/06/18 89 1598 pr 165-31 41 653 21/6/23/2 90 1599 pr 165-50 42 654 28/6/35 91 1600 pr 165-52 43 655 28/6/54 92 1659 tn 81-46 44 659 89/05/ 93 ibis ibis 45 660 104/5 94 egret egret 46 662 165/5 95 heron heron 47 664 rhosa ligte hilum 96 nguthu nguthu 48 666 2017/06/26 97 dundee dundee 49 668 2020/06/25 98 jimmy jimmy note: agricultural research council-grain crops in south africa table-2. list of abbreviations full name abbreviation hundred seed weight (g) hsw number of branches per plant bnp days to 50% flowering dfw pod number per plant pnp pod length (mm) pdl pod weight/plant (g) pdw seed number per pod snp seed number per plant sdp seed yield (g) sdy agriculture and food sciences research, 2018, 5(1): 6-22 9 3. results 3.1. agro-morphological diversity 3.1.1. analysis of variance of nine agro-morphological traits there were significant differences (p ≤ 0.05) observed among the accessions grown in potchefstroom based on seed number per plant and seed yield; and highly significant differences based on days to 50% flowering, pod weight per plant, and seed number per pod (p ≤ 0.001) (table 3). seed number per plant ranged between 46 and 47, and seed yield ranged between 5.58 and 156.3 g. days to 50% flowering ranged between 55 and 121,5. pod weight per plant ranged from 13.33 to 4.21 g. seed number per pod ranged between 1 and 7 pods. in brits, significant differences (p ≤ 0.05) were observed among the accessions based on number of branches per plant and pod number per plant (table 4). number of branches ranged between 3.8 and 8.3, and pod number per plant also ranged between 31.7 and 47.0. the genotypic effects were highly significantly different across the two locations based on number of branches per plant and seed number per plant; and there were significant differences on pod length, seed number per pod, and seed yield (table 5). hundred seed weight, days to flowering, pod number per plant and pod weight per plant were non-significant. number of branches per plant ranged between 3.6 and 8.0. seed number per plant ranged between 71.9 and 418.2, whereas pod length, seed number per pod, and seed yield ranged from 27.8 to 44.3 mm, 1 to 3, and 11.6 to 96.8 g, respectively. the two sites showed highly significant differences (p ≤ 0.001) in quantitative traits such as number of branches per plant, pod weight per plant, seed number per pod, seed number per plant, and seed yield and significant differences on days to 50% flowering. on the other hand, highly significant differences were observed on the genotype x site interaction for days to 50% flowering, seed number per plant and seed yield. the significant differences on genotype x site interaction could be attributed to the different reactions of the accessions to sites or due to differences between the sites. 3.1.2. principal component analysis the agronomic data were subjected to principal component analysis (pca), which revealed that the three most important pcs contributed 63.2%, 25.4% and 8.9% of the total variation, respectively (table 6). seed number per plant, pod number per plant, and pod weight were the traits that contributed the most variation in the first pc. seed number per plant and pod number per plant were the traits that contributed the most variation in the second pc, whereas pod number per plant, and seed number per plant were the largest contributors to the variation observed in the third pc. the principal component biplot (figure 1) grouped the tested soybean accessions into two major groups. the accessions exhibiting early flowering, high seed number per plant, high pod weight and high seed yield were grouped together. 3.1.3. correlation analysis among phenotypic traits the phenotypic traits were analysed using pair-wise rank correlations coefficients. the results and association of the traits are reported based on the significance levels of 5% (p < 0.05) and 1% (p < 0.001). seed yield was highly significant and highly correlated with seed number per plant, pod weight per plant, pod number per plant, and hundred seed weight but negatively correlated with seed number per pod. however, was also positively and significantly associated with pod length and days to 50% flowering. seed number per plant was highly significant and positively correlated with pod weight per plant, pod number per plant, days to 50% flowering but negatively correlated with seed number per pod. it was also significant and positively correlated with pod length. seed number per pod was highly significant and negatively associated with pod weight per plant, days to 50% flowering, but positively correlated with number of branches. pod weight per plant was highly and positively associated with pod length, pod number per plant, days to 50% flowering. pod length was significant and positively correlated with pod number per plant, and number of branches. pod number per plant was highly and significantly associated with branch number and days to 50% flowering, respectively. 3.1.4. cluster analysis the agro-morphological traits were analysed using agglomerative hierarchical clustering to construct a dendrogram (figure 2). three major clusters were formed among the soybean accessions. cluster i was composed of two accessions, s4-a.p.4 (54) and columbia m 8 a (41) which were genetically closely related. cluster ii consists of two accessions, 04-apr-2000 (16) and 03-apr-2000 (15) which were closely related. the third cluster was composed of 94 accessions, which were sub-clustered into three groups. the accessions within each sub cluster were closely related whereas the accessions between the clusters were unrelated. the accessions that were mainly distantly related from the other accessions were 2015/06/12 (54), 69 s 10 (1), pr 154-14 (81), r 5-4-2 m (10), hawkeye (ussr) (6), and pr 145-2 (79). table-3. means of agro-morphological traits of 96 soybean accessions planted in potchefstroom. gm hsw bnp dfw pnp pdl pdw sdp snp sdy 1120 16.57 14.00 70.50 114.8 40.01 102.00 260.1 7.000 49.58 1363 12.07 19.00 109.50 164.0 35.01 89.50 231.5 6.500 33.08 1371 13.23 13.00 77.51 100.3 38.33 75.00 223.1 6.500 28.19 1380 12.57 16.50 97.00 239.7 37.51 67.00 435.6 7.000 61.25 1386 13.57 10.50 55.00 108.2 39.51 92.00 300.5 7.000 41.08 1390 14.45 20.50 81.50 145.5 35.23 86.00 232.0 5.500 32.29 1403 15.95 13.00 76.00 188.0 39.23 137.00 399.5 7.000 64.79 1409 12.57 9.50 98.00 107.8 26.67 65.50 138.8 4.500 17.91 1449 16.57 9.50 94.50 117.2 31.67 63.00 178.3 4.000 21.58 1552 9.00 20.50 105.51 131.7 0.00 28.50 176.3 4.000 20.00 1554 19.57 13.50 74.00 134.2 40.84 43.00 309.3 7.000 59.41 agriculture and food sciences research, 2018, 5(1): 6-22 10 1555 17.45 15.00 84.00 174.4 34.23 48.50 310.6 5.500 54.46 1556 18.23 17.00 83.01 140.1 43.00 122.50 243.6 7.000 45.52 1558 16.41 17.00 99.50 158.1 34.99 26.00 230.8 5.500 35.67 1572 13.00 23.00 116.00 47.9 39.99 39.50 68.5 7.000 7.17 1573 11.00 14.00 121.45 89.7 40.00 28.00 102.3 4.000 12.33 1575 12.00 18.00 97.04 74.3 34.00 23.00 91.3 4.000 10.67 1578 16.48 10.00 76.04 113.1 26.20 69.00 194.7 4.500 29.74 1590 10.00 16.50 96.54 38.6 37.87 71.00 46.4 7.000 5.58 1594 10.00 14.50 90.58 141.3 41.67 41.50 223.3 4.000 22.67 1595 14.33 10.50 103.00 157.6 39.41 69.50 181.5 6.000 24.40 1596 13.83 17.50 101.00 220.6 39.75 113.00 334.8 5.500 45.56 1597 12.98 23.00 104.54 174.7 40.37 66.00 331.1 7.000 51.08 1598 17.48 17.00 76.04 97.7 43.70 71.50 191.7 7.000 30.74 1599 14.26 11.00 77.45 172.1 44.08 101.00 274.8 7.000 43.09 1600 15.83 9.50 74.00 119.6 45.08 105.00 277.0 7.000 44.40 1659 13.66 19.00 107.58 176.4 39.23 124.00 360.1 7.000 52.56 220 16.00 15.00 73.48 136.4 26.99 55.00 294.4 3.500 48.61 223 22.03 13.50 69.48 154.4 39.99 60.00 323.2 7.000 67.61 231 23.45 13.50 67.97 110.8 42.54 61.50 202.8 7.000 54.54 266 17.00 17.00 68.97 165.7 40.33 4.50 374.7 4.000 74.00 267 20.03 13.50 79.98 189.3 39.82 122.50 210.7 7.000 54.77 268 18.03 16.00 74.48 241.6 39.32 35.00 570.4 7.000 126.44 278 19.90 18.50 63.49 159.0 40.83 49.00 343.5 7.000 59.54 284 18.53 21.50 75.48 207.1 39.99 17.00 436.4 7.000 81.77 290 18.45 15.00 74.47 169.2 42.54 71.50 375.7 7.000 72.87 291 20.40 17.00 70.49 141.7 41.67 34.50 309.7 7.000 63.04 292 19.45 14.00 77.47 180.5 38.38 57.00 318.3 6.500 71.04 297 21.45 12.50 72.97 131.5 39.38 111.00 272.0 7.000 55.20 304 18.95 21.00 78.47 221.7 41.71 85.50 434.3 7.000 83.04 544 15.95 19.50 77.47 216.0 43.38 35.00 517.2 7.000 79.04 568 23.53 19.50 70.98 141.4 40.65 130.50 258.5 7.000 61.11 571 24.95 11.50 66.47 79.2 36.71 82.50 153.5 7.000 37.87 575 17.90 18.50 107.99 81.3 40.00 64.50 135.0 7.000 20.04 578 14.95 8.50 89.47 90.0 45.04 103.00 135.0 7.000 20.20 582 18.53 21.00 107.48 125.3 43.32 89.00 191.0 7.000 29.27 592 13.53 14.00 61.98 179.1 41.65 135.00 457.0 7.000 65.77 593 18.29 19.50 75.97 82.9 36.64 94.50 150.8 6.000 24.28 595 20.59 13.00 76.96 130.6 31.70 109.00 239.0 5.500 48.94 597 23.09 16.50 75.96 166.6 39.20 21.50 328.7 7.000 75.78 598 19.79 17.50 73.97 113.7 40.81 94.00 216.3 6.500 43.78 604 20.05 13.50 72.98 141.6 39.17 51.50 330.8 7.000 60.94 607 78.55 13.00 94.48 107.1 37.00 48.50 158.5 6.000 84.27 617 22.05 23.50 74.48 309.6 40.00 40.00 688.0 7.000 156.27 621 17.55 14.50 70.98 92.2 36.67 99.50 217.3 7.000 35.77 623 59.09 17.50 76.46 134.4 31.03 79.00 205.0 4.000 60.28 624 15.00 23.50 83.47 157.0 36.67 52.50 248.0 2.500 37.33 626 17.09 15.00 91.96 127.4 43.36 87.00 201.3 6.500 35.11 630 22.00 9.00 85.48 201.7 31.67 3.50 310.7 2.500 65.67 631 16.59 14.00 90.46 90.7 45.86 69.00 170.3 7.000 26.94 637 16.55 12.50 72.98 179.2 40.83 136.00 373.0 7.000 60.77 646 18.59 12.00 71.96 106.1 30.03 78.50 185.3 4.000 33.44 648 18.05 11.00 73.98 114.1 36.67 91.50 226.5 5.000 59.61 649 16.59 10.00 71.46 104.4 41.86 84.00 190.2 6.500 34.61 650 16.79 15.00 78.47 234.1 36.64 71.00 432.0 5.500 72.78 651 20.42 13.00 76.98 123.5 49.21 44.50 246.3 7.000 54.58 652 14.92 9.00 78.98 89.2 27.55 69.00 202.0 4.500 30.08 653 19.23 14.50 73.98 143.7 36.56 57.50 241.2 7.000 50.08 654 14.04 14.00 68.49 117.5 37.51 87.50 251.5 6.500 39.05 655 15.04 15.00 71.99 255.2 41.34 73.00 608.5 6.500 101.21 659 17.00 12.50 79.98 72.7 13.33 30.00 148.7 1.500 25.33 660 18.98 18.50 77.47 126.4 39.09 121.50 247.1 7.000 51.40 662 61.23 13.50 74.48 144.4 37.40 54.00 311.7 6.000 73.25 664 17.04 20.50 72.99 165.2 36.67 129.50 291.3 6.500 60.38 666 17.54 15.50 65.49 134.4 40.01 139.50 341.1 7.000 64.71 668 20.54 15.50 72.99 158.9 40.01 82.50 350.0 7.000 73.38 673 20.06 11.00 67.98 90.0 37.37 94.00 191.8 5.500 40.33 675 19.56 14.50 76.48 160.3 37.54 53.00 313.9 7.000 55.83 678 0.00 15.00 75.49 0.0 0.00 1.00 0.0 1.000 0.00 679 18.04 18.00 77.99 137.2 41.67 43.50 302.5 7.000 53.88 681 22.23 15.50 77.48 137.9 27.40 65.50 333.6 4.500 72.08 688 21.23 16.00 71.98 114.2 39.06 66.50 240.6 7.000 49.42 691 15.56 16.00 75.98 192.8 42.37 28.50 391.3 7.000 89.16 699 6.98 21.50 104.97 87.4 25.75 39.00 136.6 4.000 8.30 707 18.93 16.00 90.99 225.8 39.11 19.50 464.0 6.500 90.14 715 18.54 18.00 70.99 141.5 41.67 52.50 291.0 7.000 59.38 724 15.23 14.00 83.48 124.2 42.40 93.00 265.2 7.000 37.75 agriculture and food sciences research, 2018, 5(1): 6-22 11 725 16.23 14.50 76.51 139.6 40.00 37.50 283.9 7.000 47.85 861 20.23 13.50 74.01 600.9 42.67 34.00 236.1 7.000 66.85 862 16.95 16.50 72.50 119.4 41.73 97.50 255.8 6.500 43.29 864 12.07 15.00 75.50 179.8 41.51 62.00 363.3 7.000 55.25 873 16.57 14.00 70.50 179.3 36.67 101.50 351.1 7.000 54.25 dundee 17.98 18.33 65.01 149.4 39.87 33.33 344.7 5.000 62.58 dundee* 20.95 16.00 66.02 152.5 43.84 128.00 314.9 7.000 57.89 egret 12.94 13.72 67.01 111.2 39.89 75.12 247.5 6.328 35.31 egret* 19.95 15.00 68.02 127.7 43.17 102.00 233.9 6.500 45.56 heron 15.33 11.50 66.00 130.6 36.75 88.50 282.1 6.500 40.06 ibis 16.83 17.50 66.50 153.1 40.58 122.50 328.1 7.000 56.90 f-prob 0.490 0.193 <.001 0.487 0.090 0.012 0.002 <.001 0.003 lsd 25.78 4.870 11.239 191.6 11.820 77.84 230.5 2.870 48.96 cv 66.61 31.83 7.19 63.77 15.25 55.11 40.70 23.69 47.05 se 12.57 4.843 5.745 94.70 5.876 38.97 113.9 1.437 24.20 table-4. means of agro-morphological traits of 98 soybean accessions planted in brits. genotype hsw bnp dfw pdl pnp pdw sdp sdy 1120 24.02 4.135 58.49 39.51 80.2 62.00 114.8 31.30 1363 16.00 5.302 56.99 37.00 138.3 55.33 213.3 31.67 1371 16.61 6.838 68.98 38.32 156.5 61.57 210.3 34.66 1380 15.02 6.968 57.99 37.18 84.0 27.66 113.8 15.30 1386 21.02 3.635 66.99 37.51 82.8 49.33 118.8 31.97 1390 14.55 6.654 46.98 34.30 82.4 29.02 114.6 16.42 1403 19.00 5.654 57.48 42.13 111.6 47.52 142.1 31.25 1409 16.00 6.135 58.49 40.00 55.3 21.67 82.7 12.67 1449 17.52 7.635 67.99 34.18 114.0 39.50 152.3 22.80 1552 12.61 7.338 60.48 35.15 148.0 50.57 239.0 32.16 1554 17.52 5.333 60.99 47.01 94.0 57.50 172.8 37.30 1555 18.00 6.154 49.48 36.97 82.4 31.68 108.7 19.42 1556 18.11 8.338 69.98 41.65 161.0 67.24 219.8 39.83 1558 17.14 8.165 61.08 38.32 187.4 65.57 267.2 47.55 1572 11.64 6.832 76.08 41.82 81.8 34.40 147.7 31.39 1573 0.00 6.373 56.28 0.00 0.0 0.00 0.0 0.00 1575 15.00 4.844 62.26 41.67 128.0 51.67 192.7 31.67 1578 16.00 7.344 70.26 31.67 111.7 42.00 156.0 25.33 1590 16.00 6.510 60.26 37.33 91.3 43.33 124.3 21.67 1594 14.00 5.038 55.76 35.00 158.7 60.00 253.3 34.33 1595 14.10 7.647 64.91 39.28 158.9 51.92 223.3 31.88 1596 17.10 7.647 73.41 37.11 162.9 70.92 249.0 42.55 1597 13.61 6.177 47.26 35.09 131.3 55.69 209.1 34.10 1598 20.11 6.677 47.76 36.92 147.1 72.03 200.8 44.60 1599 19.65 5.206 49.78 39.20 125.8 64.72 211.3 41.26 1600 22.10 4.647 68.91 39.11 97.6 52.09 128.0 33.88 1659 14.29 6.705 44.76 39.25 102.3 44.05 174.7 26.10 220 21.43 5.012 46.49 39.33 107.2 51.35 171.2 33.50 223 22.43 3.845 63.99 41.00 119.2 65.52 180.5 39.50 231 25.00 3.848 47.50 40.11 38.8 23.40 53.0 12.89 266 25.00 5.181 55.50 43.28 64.4 32.74 76.9 19.89 267 26.00 7.012 67.49 36.67 67.2 48.68 101.4 23.50 268 21.93 6.512 44.99 38.67 105.5 60.18 168.5 36.67 278 21.87 5.348 59.50 41.55 86.6 44.88 124.9 28.42 284 25.00 5.345 60.99 36.67 46.5 27.52 68.7 16.50 290 19.00 7.348 51.50 41.45 75.1 42.07 119.9 70.89 291 17.37 5.181 53.00 39.30 85.0 36.88 116.9 23.50 292 22.00 7.015 59.00 41.33 55.3 33.00 90.3 20.33 297 25.00 6.681 63.50 40.78 59.6 40.24 91.5 26.39 304 13.41 5.515 57.50 42.95 65.6 28.74 104.2 16.89 544 13.91 8.015 47.00 41.61 163.4 71.74 240.4 42.39 568 0.00 4.512 48.99 38.00 49.7 22.02 58.7 11.50 571 19.91 5.515 68.00 39.61 135.3 71.90 205.5 47.39 575 20.37 5.514 63.50 41.97 129.2 74.72 197.7 36.17 578 15.41 6.681 56.50 39.95 198.8 85.40 272.2 47.39 582 11.43 7.345 65.49 39.17 172.0 79.85 233.4 42.84 592 17.43 4.345 69.99 40.50 56.0 26.02 86.9 15.00 593 20.00 5.163 50.01 40.67 77.3 34.67 98.7 19.33 595 23.01 6.666 60.50 42.42 102.1 48.93 140.7 31.83 597 20.51 6.166 56.50 38.42 117.8 65.76 186.3 36.83 598 24.10 6.496 55.01 41.34 113.1 60.10 161.8 38.12 604 23.55 6.499 53.99 40.81 111.1 62.33 185.8 41.89 607 21.00 5.332 54.99 32.00 106.7 47.33 146.3 30.00 617 23.55 6.832 53.49 37.47 247.6 50.50 141.8 35.23 621 20.55 5.832 50.49 41.14 101.2 60.00 180.8 39.56 623 20.00 6.333 59.50 35.08 72.0 32.43 89.3 18.33 624 17.60 6.996 55.51 37.51 137.3 55.60 177.8 31.12 agriculture and food sciences research, 2018, 5(1): 6-22 12 626 18.00 7.666 57.50 31.67 69.0 26.00 75.0 13.33 630 22.00 6.999 53.49 34.97 91.7 44.17 131.8 29.56 631 24.01 4.999 48.00 39.58 102.8 47.76 135.0 29.66 637 16.55 5.832 53.49 37.47 116.9 51.50 208.5 33.23 646 20.51 5.166 58.00 36.58 70.5 31.10 104.3 20.66 648 20.05 4.665 46.49 37.81 79.7 32.17 99.0 19.56 649 19.01 4.833 53.00 41.08 64.0 29.76 107.5 20.66 650 17.10 6.996 56.01 38.17 105.9 53.10 178.9 32.45 651 19.00 5.324 60.49 39.32 71.9 35.05 114.0 19.48 652 16.00 4.491 45.49 36.32 40.4 17.05 56.5 10.15 653 17.00 4.143 46.00 37.49 74.9 31.48 106.3 18.41 654 17.00 5.661 45.99 36.67 187.7 83.67 329.0 56.00 655 15.88 6.495 47.49 33.82 76.2 30.49 118.5 18.25 659 24.77 7.840 61.05 40.84 109.3 57.87 159.6 54.55 660 23.00 7.180 48.49 40.00 70.3 31.67 94.3 21.00 662 22.01 4.643 46.50 33.82 74.5 39.64 118.9 26.24 664 20.38 5.328 60.49 31.65 104.3 49.66 166.2 34.75 666 21.38 5.828 43.99 38.82 82.2 38.82 120.7 23.75 668 21.38 5.328 64.99 38.82 93.8 53.49 140.0 28.75 673 23.00 4.000 56.05 40.00 56.2 31.21 88.8 19.38 675 17.00 6.006 47.55 39.67 83.2 45.71 94.8 17.22 678 23.00 5.661 66.49 37.48 53.7 27.32 78.5 15.58 679 121.88 4.328 53.99 40.15 87.2 56.82 157.3 36.42 681 26.00 5.309 56.00 40.00 100.3 53.00 141.0 35.33 688 23.00 6.809 45.00 45.00 116.3 51.00 148.3 43.67 691 19.27 6.340 57.55 40.00 161.5 73.54 200.1 44.38 699 8.66 7.680 46.99 37.26 133.5 28.26 200.4 17.00 707 9.75 5.512 61.49 39.98 68.8 33.71 108.1 22.44 715 22.88 4.495 50.99 39.15 90.2 61.82 153.8 37.58 724 16.51 7.309 57.50 43.49 163.2 74.98 246.9 45.41 725 19.00 7.838 64.98 44.00 134.3 60.67 222.3 37.00 861 24.00 6.338 55.98 38.32 79.7 44.07 126.2 27.99 862 20.55 5.320 45.98 33.63 107.9 53.18 165.6 34.58 864 23.02 4.302 47.99 39.85 86.7 57.16 163.0 38.64 873 17.02 4.635 48.49 37.85 148.3 66.83 211.6 43.97 dundee 21.65 5.552 47.50 39.62 111.5 57.70 172.9 38.74 dundee* 18.87 5.029 45.38 37.90 95.0 49.06 122.1 26.39 egret 19.91 3.865 62.12 40.03 85.0 45.06 149.5 30.06 egret* 23.37 5.695 44.00 44.90 142.6 76.56 195.3 45.72 heron 20.10 3.981 43.00 38.28 91.3 46.59 167.1 29.72 ibis 21.60 4.647 60.41 40.61 130.3 69.09 218.1 43.22 f-prob 0.863 0.006 0.163 0.064 0.367 0.382 0.214 0.599 lsd 39.65 2.401 19.59 6.221 104.52 43.14 134.8 32.60 cv 86.77 20.60 17.32 7.67 47.97 42.02 42.07 50.54 se 17.72 1.216 9.700 2.985 50.15 20.70 64.67 15.64 table-5. means and mean squares of combined analysis of variance of agro-morphological traits of 96 soybean accessions planted across two sites. potchefstroom and brits. 2016/17 genotype hsw bnp dfw pnp pdl pdw snp sdp sdy 1120 19.94 4.415 64.50 98.6 39.75 65.97 3,00 189.8 40.80 1363 13.22 5.833 83.31 156.0 35.67 58.40 2,83 226.0 32.67 1371 14.84 5.574 73.25 130.9 38.34 56.24 2,83 218.9 31.70 1380 13.60 6.243 77.54 163.9 37.34 66.61 3,00 278.5 38.86 1386 16.94 3.580 60.97 96.5 38.51 55.67 3,00 212.3 36.81 1390 14.67 6.747 64.28 114.9 34.76 42.21 2,50 174.8 24.51 1403 16.90 4.994 66.76 150.9 40.65 70.96 3,00 273.4 48.32 1409 13.48 4.823 78.29 91.5 31.00 35.75 2,00 121.3 16.25 1449 16.90 7.580 81.27 116.4 32.91 40.08 2,00 166.7 22.39 1552 11.15 7.079 83.05 142.7 35.17 44.84 3,00 218.1 28.07 1554 18.50 4.777 67.51 115.2 43.89 73.07 3,00 243.3 48.75 1555 17.60 5.577 66.78 129.6 35.57 59.28 2,50 211.9 37.25 1556 18.20 6.990 76.51 153.3 42.34 75.54 3,00 234.0 43.05 1558 16.67 6.907 80.29 173.4 36.65 66.01 2,50 250.0 41.53 1572 12.02 7.249 96.04 65.5 40.91 26.79 3,00 108.7 19.09 1573 11.00 8.005 89.07 89.7 40.00 32.67 3,00 102.3 12.33 1575 13.40 5.417 79.78 100.7 37.79 36.76 3,00 141.2 21.00 1578 16.30 5.320 73.25 111.7 27.77 43.39 2,00 181.6 28.28 1590 12.95 5.994 78.53 55.1 37.45 22.53 3,00 71.9 11.59 1594 11.87 4.913 73.28 149.9 38.37 57.29 3,00 238.1 28.40 1595 13.94 7.747 84.04 155.8 39.34 47.18 2,67 199.1 28.03 1596 15.07 6.742 87.27 189.7 38.43 73.13 2,50 289.7 44.03 1597 13.87 6.917 76.06 151.5 37.69 75.84 3,00 268.8 42.70 1598 19.30 6.161 62.02 120.2 40.29 59.31 3,00 193.9 37.53 1599 16.77 4.410 63.77 149.5 41.53 66.02 3,00 244.7 42.24 1600 18.47 3.910 71.49 106.3 42.12 61.24 3,00 200.8 39.17 agriculture and food sciences research, 2018, 5(1): 6-22 13 1659 13.94 6.496 76.32 142.4 39.17 63.49 3,00 266.8 39.57 220 19.40 4.997 60.02 121.8 33.10 60.05 1,67 233.1 41.12 223 22.17 4.165 66.75 136.8 40.50 85.40 3,00 252.3 53.73 231 23.90 4.165 57.76 75.9 41.35 59.31 3,00 130.0 33.99 266 20.73 5.415 62.26 100.7 42.31 59.74 3,00 182.4 38.33 267 21.85 5.741 73.75 129.0 38.27 66.36 3,00 156.2 39.34 268 19.80 5.911 59.78 174.4 39.01 115.21 3,00 372.0 82.23 278 20.87 5.749 61.49 123.9 41.21 66.75 3,00 234.9 44.19 284 20.50 6.252 68.26 127.9 38.35 76.62 3,00 254.8 49.62 290 18.62 6.158 63.02 123.4 42.01 74.92 3,00 250.7 71.84 291 19.04 5.415 61.76 114.4 40.51 56.30 3,00 213.8 43.55 292 20.19 5.824 68.26 139.6 39.31 77.06 2,83 243.5 53.99 297 22.56 5.407 68.25 96.6 40.08 57.85 3,00 184.1 40.97 304 16.37 6.251 68.02 145.4 42.33 79.46 3,00 272.8 50.44 544 15.00 7.243 62.28 190.6 42.51 117.80 3,00 381.9 60.95 568 23.44 5.502 60.02 96.0 39.35 55.54 3,00 159.5 36.66 571 22.60 4.659 67.24 107.2 38.15 64.27 3,00 179.9 42.49 575 19.10 5.832 85.79 105.4 40.99 57.97 3,00 164.8 27.93 578 15.17 6.915 73.03 144.0 42.53 78.11 3,00 203.3 33.51 582 15.17 7.163 86.54 148.0 41.27 68.40 3,00 211.1 35.89 592 15.30 4.498 65.98 118.3 41.09 59.62 3,00 274.2 40.75 593 18.93 5.834 63.02 80.6 38.00 53.01 2,67 132.9 22.57 595 21.87 5.491 68.76 116.2 37.03 61.27 2,50 190.9 40.54 597 22.04 5.829 66.26 142.2 38.84 102.84 3,00 258.9 56.64 598 21.80 6.162 64.51 113.2 41.08 62.20 2,83 189.3 40.99 604 21.84 5.492 63.51 126.7 39.99 76.98 3,00 258.4 51.59 607 61.92 4.828 74.79 107.2 35.38 79.17 2,67 152.7 66.72 617 22.90 7.333 64.02 279.2 38.77 122.06 3,00 418.2 96.75 621 19.10 5.328 60.76 96.8 38.89 63.30 3,00 198.3 37.65 623 47.03 6.079 68.01 103.3 33.06 42.91 2,00 148.3 39.66 624 16.58 7.416 69.52 144.1 37.22 60.09 2,00 201.8 33.24 626 17.60 6.323 74.78 107.8 39.54 48.11 2,83 159.9 27.90 630 22.00 5.154 69.53 129.7 33.86 65.11 2,00 194.0 42.17 631 20.20 4.829 69.29 96.3 42.79 46.05 3,00 153.2 28.30 637 16.70 4.993 63.26 148.7 39.19 72.58 3,00 291.0 47.24 646 19.64 4.577 65.01 88.2 33.30 42.00 2,00 145.7 27.18 648 19.14 4.161 60.27 97.3 37.25 47.49 2,33 162.7 39.93 649 17.87 4.076 62.26 84.1 41.51 42.36 2,83 149.7 27.77 650 16.94 5.992 67.27 170.8 37.41 85.35 2,50 307.4 52.94 651 19.99 4.828 68.76 97.5 44.31 59.39 3,00 180.4 37.32 652 15.27 3.911 62.28 64.5 31.87 31.57 2,00 129.6 20.28 653 18.37 4.499 60.02 109.4 37.08 60.46 3,00 175.0 34.52 654 14.90 5.161 57.27 141.0 37.22 67.44 2,83 277.7 44.84 655 15.64 5.743 59.77 167.2 37.63 68.78 2,83 367.8 60.45 659 21.96 6.155 70.51 96.6 31.43 51.49 1,00 156.1 44.22 660 20.13 6.661 63.03 110.0 39.44 66.11 3,00 200.8 41.93 662 42.74 4.580 60.52 109.6 35.69 62.87 2,67 217.0 50.14 664 18.80 6.084 66.75 135.3 34.20 69.15 2,83 230.1 47.81 666 19.54 5.495 54.77 108.7 39.43 67.71 3,00 233.0 44.60 668 21.14 5.246 69.00 126.9 39.43 91.36 3,00 247.0 51.47 673 20.84 3.776 62.00 74.2 38.74 47.68 2,50 142.4 30.04 675 18.67 5.411 62.03 123.2 38.66 67.26 3,00 207.5 36.85 678 23.00 5.329 71.00 53.4 37.50 27.25 78.3 15.52 679 66.70 5.168 66.02 112.6 40.93 70.81 3,00 231.4 45.33 681 23.13 5.246 66.77 125.1 31.56 88.01 2,00 271.3 60.11 688 21.54 6.076 58.52 114.2 41.06 86.23 3,00 210.5 47.50 691 17.30 5.828 66.76 178.2 41.27 116.26 3,00 298.6 67.15 699 7.64 7.412 76.06 111.9 31.52 25.02 2,00 171.3 12.86 707 14.74 5.413 76.28 151.0 39.58 83.71 2,83 290.2 57.03 715 20.77 5.251 61.01 116.3 40.43 74.54 3,00 223.9 48.70 724 15.64 5.990 70.52 143.0 43.00 68.90 3,00 256.4 41.53 725 17.19 6.323 70.75 141.5 41.30 72.60 3,00 266.2 44.69 861 21.56 5.409 65.01 347.5 39.78 60.76 3,00 184.2 48.07 862 18.80 5.414 59.27 114.2 37.71 59.01 2,83 212.0 39.03 864 17.07 4.665 61.77 134.8 40.68 76.72 3,00 266.0 47.29 873 16.67 4.664 59.52 164.9 37.25 68.18 3,00 283.7 49.40 dundee 19.22 5.899 59.48 128.9 39.62 73.86 3,00 256.2 48.30 dundee* 19.77 5.164 55.76 124.6 40.95 68.89 3,00 221.3 42.41 egret 16.44 4.232 64.66 95.6 40.01 48.16 2,88 197.1 32.73 egret* 21.34 5.329 60.06 135.4 44.08 73.43 2,83 216.1 45.65 heron 17.27 3.913 58.31 108.8 37.49 52.94 2,83 222.7 34.92 ibis 18.77 5.250 63.50 139.4 40.58 75.87 3,00 271.1 50.12 hsw bnp dfw pnp pdl pdw 1,00 sdp sdy agriculture and food sciences research, 2018, 5(1): 6-22 14 genotype 236.7 3.851*** 278.09 5837 36.31** 1290.5 3,00 13538* ** 686.9* * site 334.6 32.707** * 56339.34 *** 182894* ** 9.27 81648.5 *** 0.4953 *** 149129 0*** 39431. 5*** genotype *site 177.8 1.935 189.56** * 5384 23.93 1092.0 0.0000 14512* ** 631.2* ** mean 19.10 5.49 67.68 123.79 37.82 62.68 0.0000 211.88 19.10 lsd 22.50 2.089 15.59 160.8 6.791 58.88 2,78 196.0 29.75 cv 75.53 27.04 11.62 62.26 12.04 44.93 0.7721 44.22 49.55 se 14.80 1.516 7.900 78.99 4.661 28.93 13.77 96.31 20.49 r2 table-6. factor loadings of the three pcs based on agronomic traits trait factor loadings 1 2 3 hsw -0.00436 0.00148 0.00187 bnp 0.00058 0.00631 -0.00191 dfw 0.03408 -0.10372 -0.05010 pnp 0.41799 0.26113 -0.86868 pdl 0.00519 0.00590 0.00401 pdw 0.20975 0.04508 0.07626 snp -0.22688 0.95126 0.17318 sdp 0.83968 0.11403 0.45054 sdy 0.15344 0.03245 0.06380 eigen values 6983426 2810062 981647 percentage variation 63.19 25.43 8.88 cumulative variation 63.19 88.62 97.50 table-7. correlations among nine agro-morphological traits of 96 soybean accessions sdy sdp snp pdw pdl pnp dfw bnp hsw sdy sdp 0.8805*** snp 0.1863*** 0.2436*** pdw 0.9186*** 0.8634*** 0.1831*** pdl 0.1564** 0.1547** 0.0500 0.1946*** pnp 0.6144*** 0.6451*** -0.0446 0.6164*** .1151** dfw 0.1219** 0.2074*** 0.6303*** 0.1749*** -0.0375 0.1422** bnp 0.0423 0.0690 0.3093*** 0.0756 .1668** .1799*** 0.0334 hsw 0.1808*** -0.0536 0.0212 0.0687 -0.0170 -0.0351 -0.0962 -0.0528 figure-1. a principal component biplot of nine agro-morphological traits of 96 soybean accessions. agriculture and food sciences research, 2018, 5(1): 6-22 15 figure-2. a dedrogram on agro-morphological traits of 96 soybean accessions. 3.2. nutritional quality diversity the nutritional quality traits were analysed using analysis of variance. there were significant differences (p< 0.05) among the soybean accessions planted in potchefstroom based on ash, moisture, and oil contents (table 8). the ash content varied between 4.3 and 10.9%. moisture content varied between 1.7 and 12.8% whereas oil content ranged between 7.1 and 23.9%. in brits, highly significant differences were observed in fiber, moisture, oil, palmitic acid, and protein contents, and significant differences were observed for linolenic acid (table 9). fiber content varied between 4.5 and 5.5%. moisture varied between 3.1 and 4.6%. oil content varied between 12.0 and 21.8%. palmitic acid ranged from 55.9 to 64.2% and protein content varied between 29.6 and 35.8% whereas linolenic acid ranged from 3.1 to 4.7%. when assessing the reaction of the soybean accessions across the two locations, highly significant differences were observed for ash, moisture, oil, oleic acid and stearic acid and significant differences were observed for protein content (table 10). ash content varied between 4.3% and 8.2%. moisture content ranged from 2.8% to 12.8%. oil content varied between 10.6% and 20.7%. oleic acid varied between 6.8% and 30.8% and stearic acid ranged between 31.6% and 263.5% whereas protein content was ranging between 29.1% and 35.6%. the two sites showed highly significant differences among the genotypes based on linolenic acid, moisture content, oleic acid, stearic acid and significant differences were observed for linoleic acid and oil content. the genotype x site interaction was significant for ash and non-significant for all other quality traits. 4. discussion the principal component analysis (pca) is a statistical method commonly used in population genetics to identify structure in the distribution of genetic variation across geographical location and ethnic background [19]. the goal is to evaluate the importance of each variable in relation to the total available variation among genotypes. the method provides an opportunity to exclude less important traits in the group studied [20] and simultaneously determine which traits are the most important. in this study, the traits that contributed to the most variation were seed number per pod, pod number per plant, seed number per plant and seed yield. the correlation information of the tested traits can be provided to the breeders in the direct and indirect selection programmes. the genotypes with significant correlations and desirable traits can be selected concurrently. moreover, understanding the relationship between yield and its component traits is of great importance to a breeder for making the best use of these relationships in selecting desirable genotypes for yield improvement programs [21, 22]. significant positive correlations of days to flowering, number of branches and pods per plant, pod length, seed number per pod with seed yield were reported by malek, et al. [11] which concur with the results in this study. this means that in selecting high yielding genotypes these characters should be given more emphasis as the best selection criteria. seed yield always showed a positive correlations with other desirable yield traits [23, 24] which indicates that the increase in one trait would result in the increase of the other; that is, simultaneous increase or decrease of both traits would be easy. the strong positive correlation of seed yield with other yield traits indicated that it would be very easy to identify a soybean genotype having higher seed yield simultaneously with higher number of pods per plant and but difficult with number of seeds per pod. similar results were reported in other studies [25-28]. traditional cluster analysis could provide an easy and effective way in determining the genetic diversity of germplasm collections [29]. it is commonly used to study genetic diversity and for forming core subset for agriculture and food sciences research, 2018, 5(1): 6-22 16 grouping accessions with similar characteristics into one homogenous category [30]. it is also used to summarize information on relationships between accessions by grouping similar units so that the relationship is easily understood. in this study, the accessions were clustered into three major groups of which two of them were further sub clustered into three groups based on the agro-morphological traits measured. the most diverse lines can be used as parents for hybridization for improvement of genes of novelty. genetic diversity evaluation among germplasm is a necessity and a prerequisite in any hybridization program and would promote the efficient use of genetic variations [31, 32]. to improve an efficient crop, it is essential to obtain the information on genetic diversity and relationships among breeding materials for a plant breeder. this is because the assessment of genetic diversity is important not only for crop improvement but also for efficient management and conservation of germplasm resources. in this study, the nutritional quality traits showed a vast diversity among the accessions. the oil and protein content were within the range recorded in other studies [33]. the oil content ranged from 13.8% to 22.5% whereas protein ranged between 37.0% and 50.1%) with a mean of 19% and 42.9%, respectively. the vast divergence of the accessions will assist breeders in selecting accessions with good quality for hybridization and conservation. the analysis of genetic diversity plays a fundamental role in identification of parents [34] and it can help to achieve long-term selection gain [35]. as a traditional method, morphological traits used to assess genetic divergence and classify existing germplasm materials. however, this technique is a low level but powerful taxonomic tool and has been utilized for the preliminary grouping of germplasm prior to their characterization using more precise marker technologies. although the genetic base of soybean cultivars is considered to be extremely narrow [36] studies of genetic diversity in soybean have been conducted using morphological characteristics [37, 38]. recently, de chavez, et al. [39] reported a vast phenotypic diversity among the soybean accessions in phillipines. khatab, et al. [12] reported the presence of genetic diversity among soybean genotypes assessed through agro-morphological descriptors. hamzekhanlu, et al. [40] studied 34 mutant lines including one control cultivar and detected variability for ten quantitative traits in soybean. iqbal, et al. [41] reported significant differences among all the assessed phenotypic traits. 5. conclusion most of the traits showed positive correlations between each other, which will assist in the combined improvement of these traits by selecting only highly and positively correlated and easily measurable phenotypic traits although most were highly significant and positively correlated with seed yield per plant. the accessions were clustered into three major groups with subgroups, which showed existence of a vast genetic diversity among the accessions. the most divergent accessions were 2015/06/12, 69 s 10, pr 154-14, r 5-4-2 m, hawkeye (ussr), and pr 145-2. the nutritional quality traits also varied significantly among the accessions. the presence of genetic diversity can be useful for breeding and selection of parents for transgressive segregation. table-8. means of nutritional quality traits of 96 soybean accessions planted in potchefstroom. gm ash fiber linol eic acid linole nic acid moistu re oil oleic acid palmi tic acid protei n steari c acid 1120 5.667 5.145 9.284 2.486 6.527 15.19 21.03 61.93 33.90 3.967 1363 5.952 5.215 9.489 3.886 6.272 16.10 23.59 60.36 33.70 4.232 1371 5.429 5.195 9.056 3.959 6.368 13.72 24.41 59.49 34.50 4.405 1380 5.552 5.000 9.974 4.111 8.677 13.90 22.87 60.02 33.77 4.007 1386 5.867 5.240 10.539 4.556 6.042 13.72 19.85 60.95 35.21 6.242 1390 4.983 4.932 10.179 4.318 7.569 14.11 25.29 58.55 33.76 4.595 1403 5.480 5.260 10.290 4.100 5.540 12.33 19.95 62.26 35.90 3.210 1409 5.040 4.870 8.360 3.750 12.450 17.26 30.02 57.60 31.86 5.590 1449 4.837 4.900 9.709 4.241 11.297 15.78 22.91 60.29 34.75 4.617 1552 6.190 5.220 10.110 4.410 7.130 17.15 23.15 60.14 31.80 4.920 1554 5.242 5.225 8.874 3.911 6.172 15.68 21.39 62.40 35.54 3.392 1555 5.123 5.207 9.129 4.098 6.484 14.87 24.51 61.01 35.49 2.225 1556 5.534 5.140 9.101 4.229 7.023 14.60 25.30 60.94 34.23 3.240 1558 5.051 5.030 8.378 3.566 9.278 17.17 29.86 56.36 32.98 5.793 1572 7.136 5.235 8.413 3.556 7.298 18.14 23.50 62.33 30.45 4.578 1573 5.456 4.819 8.846 3.892 12.784 17.46 26.54 59.14 31.34 7.295 1575 6.970 5.460 10.220 4.440 7.130 16.97 23.96 57.05 30.18 9.390 1578 5.299 5.348 9.399 4.193 5.395 14.88 25.29 61.33 35.70 3.672 1590 6.754 5.418 9.724 4.013 7.395 16.70 26.45 56.36 31.39 6.877 1594 10.938 2.350 29.234 15.461 -1.714 7.10 -33.03 55.74 28.55 10.460 1595 5.478 5.205 9.044 4.106 6.993 16.90 26.66 59.68 33.55 4.008 1596 5.108 5.335 10.424 3.966 6.468 16.37 21.65 60.71 33.61 3.693 1597 5.294 4.808 6.754 3.933 9.320 15.20 30.15 64.35 34.16 2.177 1598 4.469 5.153 9.529 4.093 8.230 16.61 23.57 60.18 34.48 4.442 1599 4.621 5.339 9.041 3.992 6.274 17.31 24.34 61.63 34.46 2.550 1600 5.008 5.365 9.689 4.036 6.133 16.79 22.29 61.73 34.66 2.493 1659 5.698 5.285 9.694 3.981 7.121 15.36 23.93 60.57 34.29 4.710 220 5.855 5.369 10.600 4.012 5.760 14.81 21.19 59.37 33.66 6.642 223 6.205 5.224 11.060 4.292 5.920 15.20 19.23 60.70 33.37 5.997 231 4.902 5.197 9.012 3.788 6.397 17.46 22.19 62.75 33.15 3.991 agriculture and food sciences research, 2018, 5(1): 6-22 17 266 5.072 5.152 9.687 4.258 5.942 14.11 23.09 59.86 34.70 4.866 267 4.950 5.234 9.710 3.922 5.965 14.28 19.94 61.29 35.58 4.992 268 5.140 5.164 9.680 4.252 6.340 14.61 23.31 59.86 35.46 4.317 278 5.342 5.152 9.396 4.084 5.757 13.56 30.40 59.59 34.93 1.981 284 4.770 5.204 9.405 3.912 6.320 16.76 20.86 61.25 33.94 4.932 290 4.782 5.107 9.047 4.133 6.087 14.95 21.95 60.73 34.10 5.741 291 4.662 5.212 9.016 4.089 6.207 15.83 22.22 60.86 34.77 5.021 292 5.192 5.187 10.017 4.073 5.857 14.07 21.79 59.79 34.88 4.331 297 4.857 5.142 8.892 3.938 6.092 15.51 21.81 60.86 33.99 5.366 304 4.587 5.162 8.782 3.973 5.932 16.74 21.69 61.92 33.99 5.291 544 5.107 5.132 10.362 4.448 5.937 13.78 22.94 59.23 34.65 4.891 568 5.000 5.164 10.230 3.917 5.965 13.43 23.38 55.52 35.76 3.497 571 5.842 4.302 16.497 9.988 5.912 22.58 15.15 63.07 26.23 5.891 575 5.137 5.272 9.836 4.239 7.212 17.13 23.89 60.13 33.36 2.721 578 5.482 5.177 8.937 3.783 6.012 18.28 26.01 60.20 32.48 4.026 582 5.000 5.229 8.920 4.102 7.115 17.11 25.44 60.06 33.53 1.987 592 5.345 5.159 9.425 4.132 6.095 12.84 23.94 59.64 35.59 4.352 593 4.304 5.104 7.879 3.956 8.242 16.44 27.93 59.13 34.27 2.980 595 5.177 5.201 9.463 4.026 6.669 15.76 23.28 60.71 35.51 3.263 597 5.102 5.221 9.033 3.881 6.354 15.54 23.78 60.49 35.40 2.818 598 5.159 5.224 8.529 3.916 6.597 16.17 23.63 61.31 34.98 2.590 604 5.153 5.314 9.553 3.918 5.920 15.50 22.87 60.50 35.05 3.008 607 5.088 5.169 8.213 3.718 6.370 16.44 24.43 62.18 34.76 2.273 617 4.878 5.234 9.528 4.003 5.860 14.48 20.01 61.45 35.75 5.613 621 4.743 5.164 8.638 4.143 6.020 15.38 24.89 59.69 34.62 5.428 623 5.112 5.181 8.743 4.056 6.274 17.93 22.89 62.27 32.44 4.563 624 5.054 5.159 9.219 4.221 6.402 16.49 23.73 60.77 33.90 3.795 626 5.087 5.011 9.943 4.036 9.414 17.59 24.72 59.85 34.66 4.443 630 4.370 5.060 8.380 3.830 8.650 18.39 28.17 58.57 32.17 3.360 631 5.542 3.941 13.968 9.856 6.344 23.90 16.71 46.62 24.46 2.373 637 5.103 5.169 9.518 4.103 6.845 15.07 22.32 60.43 34.12 4.918 646 5.157 5.131 9.113 4.151 6.394 17.02 22.83 59.95 33.41 7.483 648 5.938 5.369 7.493 6.868 6.475 16.08 13.38 41.59 33.73 33.723 649 4.387 5.126 5.118 4.916 5.399 18.87 20.04 61.29 33.25 3.568 650 5.134 5.069 8.774 4.246 5.682 14.45 22.78 62.33 34.89 5.160 651 4.831 5.153 9.445 4.110 6.015 17.04 23.79 59.84 33.18 4.519 652 5.941 5.128 9.805 4.445 5.875 14.44 21.66 59.92 32.98 6.599 653 5.293 5.229 10.036 4.036 6.031 15.45 12.09 61.07 34.62 4.009 654 5.184 5.195 9.086 4.249 5.785 15.57 22.68 61.96 33.80 4.522 655 4.869 5.180 9.071 4.139 5.395 14.32 22.28 62.96 34.82 3.577 659 5.720 4.920 9.260 4.050 5.240 10.83 22.18 60.25 35.27 8.500 660 4.619 5.228 8.759 3.764 6.234 17.25 22.03 62.23 33.75 4.525 662 5.083 5.184 8.866 4.006 6.431 14.58 23.94 60.78 34.19 2.099 664 5.054 5.145 9.086 3.914 6.285 15.43 23.44 59.93 34.11 3.772 666 4.809 5.150 9.021 3.979 6.220 16.37 21.66 61.51 34.33 4.497 668 5.004 5.045 9.356 3.994 6.195 14.28 22.88 59.44 33.90 4.572 673 5.532 5.276 9.296 3.895 5.642 15.31 22.64 61.62 34.37 5.398 675 5.177 5.251 9.551 3.985 6.012 15.29 22.99 60.59 34.77 3.373 678 4.894 5.195 9.626 3.884 6.080 16.71 22.18 60.25 33.61 4.907 679 5.474 5.130 10.371 4.384 5.575 13.30 23.11 59.60 34.39 3.247 681 4.748 5.064 7.856 4.106 6.421 17.04 27.41 59.14 33.78 4.844 688 4.898 5.104 9.986 4.086 6.036 14.81 21.40 60.03 34.15 4.814 691 5.327 5.226 10.631 3.970 6.707 15.88 21.00 60.60 34.35 3.913 699 6.404 5.318 8.919 4.309 6.794 20.25 26.43 60.59 30.60 4.705 707 5.418 5.244 9.576 3.934 6.617 14.95 23.45 60.24 34.72 3.763 715 5.289 5.185 9.136 3.809 5.945 13.93 21.78 61.84 35.65 3.602 724 5.158 5.019 8.686 4.151 5.326 14.32 24.83 61.34 32.45 4.229 725 5.190 5.080 9.120 4.650 6.180 15.18 26.75 58.94 31.94 5.470 861 5.144 5.265 9.466 3.964 6.253 16.93 21.56 61.14 33.62 4.635 862 5.103 5.102 9.704 4.308 5.599 14.66 21.66 60.26 34.39 5.625 864 5.002 5.185 9.464 3.866 5.927 15.94 20.59 60.99 33.72 5.617 873 5.077 5.130 10.029 4.041 6.187 13.79 19.55 61.08 34.45 4.967 dundee 5.764 5.112 9.966 4.083 5.968 11.99 22.34 59.17 34.70 4.383 dundee* 5.362 5.225 9.413 3.776 6.089 14.96 22.98 60.08 34.20 3.849 egret 5.521 5.053 10.072 4.244 6.261 15.13 20.19 59.91 33.16 4.892 egret* 5.137 5.235 9.133 3.796 6.114 14.28 21.85 61.57 35.17 3.814 heron 5.273 5.175 9.399 3.791 5.588 14.34 21.16 60.75 34.45 4.278 ibis 5.513 5.175 9.659 3.981 6.018 13.97 21.10 61.07 34.60 4.573 f-prob 0.02 0.42 0.622 0.492 0.038 0.047 0.802 0.771 0.396 0.512 agriculture and food sciences research, 2018, 5(1): 6-22 18 lsd 1.720 9626 7.039 4.214 3.524 4.509 20.47 8.438 4.649 9.342 cv 16.02 9.30 36.13 48.41 26.95 14.31 45.42 6.95 6.80 96.94 se 0.8506 0.4762 3.482 2.085 1.743 2.231 10.12 4.174 2.300 4.621 table-9. means of nutritional quality traits of 96 soybean accessions planted in brits genotype ash (%) fiber (%) linoleic acid (%) linolenic acid (%) moistu re (%) oil (%) oleic acid (%) palmitic acid (%) protein (%) stearic acid (%) 1120 5.511 5.063 9.472 4.178 7.726 15.25 23.15 60.16 33.51 3.413 1363 4.621 4.733 8.207 3.618 11.121 15.84 30.18 55.87 32.49 5.443 1371 5.726 5.084 9.129 3.877 6.014 12.46 22.49 60.08 34.53 4.279 1380 4.766 4.903 8.517 4.053 8.676 16.28 27.32 59.20 33.35 3.553 1386 5.746 5.283 9.627 3.898 5.241 15.00 21.38 61.78 35.26 3.618 1390 5.679 5.112 9.662 4.138 5.570 13.35 21.43 61.74 33.77 3.039 1403 5.859 5.277 9.687 4.298 5.415 14.72 24.31 61.46 34.90 1.619 1409 5.046 4.753 8.732 4.008 11.141 14.63 24.91 61.78 35.04 4.673 1449 5.331 5.068 7.622 3.088 5.951 15.15 21.49 64.23 33.69 3.593 1552 5.761 5.069 8.939 4.007 6.599 12.34 20.89 61.43 34.32 4.124 1554 5.921 5.243 9.012 3.728 5.861 14.85 22.02 61.98 34.85 3.083 1555 5.634 5.092 8.512 4.193 7.485 12.01 27.41 61.85 34.48 3.599 1556 6.011 5.114 8.959 3.982 5.849 14.39 22.04 62.60 32.98 3.694 1558 3.920 4.660 9.350 4.060 11.300 16.29 25.66 57.51 33.71 6.810 1572 4.724 4.511 8.359 3.809 13.055 16.60 24.42 60.51 32.23 7.096 1573 4.763 4.641 9.899 3.836 12.802 16.77 21.92 60.20 32.75 7.536 1575 4.865 4.739 8.732 3.892 10.789 13.52 26.00 58.65 33.96 5.822 1578 5.020 4.954 8.957 3.977 10.129 17.18 25.38 59.46 32.74 4.577 1590 4.730 4.844 8.937 4.097 10.764 15.98 24.09 59.59 32.87 4.192 1594 5.263 4.991 9.644 4.168 7.704 14.71 21.73 60.45 33.31 3.963 1595 5.283 5.167 8.376 4.174 8.012 16.31 23.11 62.55 33.13 5.225 1596 5.028 5.072 9.066 3.914 8.742 18.04 22.83 61.20 32.44 4.700 1597 5.660 5.169 7.837 3.842 6.614 14.85 26.16 62.01 33.83 3.127 1598 4.215 5.194 7.627 3.897 7.564 18.84 29.36 58.55 32.99 2.277 1599 4.738 5.276 9.069 4.001 5.597 17.77 24.59 61.12 33.97 2.191 1600 4.653 5.177 9.086 4.064 7.522 17.50 25.75 57.49 33.39 2.925 1659 5.508 5.111 8.919 4.048 7.034 13.85 22.07 63.04 35.10 3.188 220 4.621 4.980 8.438 4.435 5.853 15.84 25.74 61.12 33.95 2.471 223 5.186 5.065 8.848 4.395 5.168 17.82 23.88 62.27 32.87 2.421 231 5.255 5.078 8.329 3.955 7.664 17.96 28.31 59.36 31.26 2.216 266 5.185 5.188 8.994 3.111 5.934 15.87 22.93 62.60 34.25 2.571 267 4.466 4.950 8.738 4.210 10.298 18.04 23.03 59.75 33.07 4.056 268 6.310 5.230 8.750 3.340 5.810 16.66 23.73 62.38 33.66 2.450 278 4.770 4.999 8.859 4.591 6.883 15.31 30.95 58.00 34.78 0.953 284 4.260 4.930 8.850 4.100 10.320 17.71 24.35 60.10 32.85 4.400 290 4.825 5.258 9.069 4.040 5.854 16.98 22.98 62.98 34.76 3.776 291 5.395 5.224 8.659 3.581 5.138 17.29 25.83 62.11 33.81 0.928 292 4.640 4.680 9.830 4.240 11.910 16.97 26.03 56.44 32.59 6.420 297 4.340 5.078 7.804 3.875 7.104 17.40 27.07 58.74 34.09 3.166 304 4.900 5.030 7.680 3.860 8.040 18.83 24.67 61.52 31.42 4.400 544 5.400 5.250 8.420 3.880 5.360 16.38 24.39 62.49 34.49 1.780 568 5.540 5.180 8.870 4.000 5.320 16.77 25.02 61.43 33.08 1.910 571 5.415 5.518 8.619 4.095 5.134 15.50 24.35 63.03 34.47 0.991 575 5.290 5.229 9.004 4.606 5.358 17.35 21.30 63.40 32.68 3.538 578 5.085 5.243 8.254 3.880 6.449 17.87 25.23 60.86 32.89 3.716 582 5.081 5.215 7.783 3.935 5.478 19.92 23.28 63.27 31.39 3.376 592 4.796 4.915 9.248 4.095 8.703 16.99 30.87 57.01 33.58 3.326 593 4.809 4.965 7.562 3.866 8.112 18.12 28.42 59.12 31.99 3.220 595 5.201 5.186 8.956 4.398 6.523 16.88 24.88 60.73 34.11 1.035 597 5.201 5.116 8.706 4.033 6.813 13.53 24.79 60.36 35.77 2.425 598 5.044 5.055 8.607 3.901 6.782 13.63 23.84 61.46 35.40 2.740 604 5.370 5.260 9.600 4.220 5.780 14.49 23.76 60.74 35.65 1.740 607 5.310 5.190 9.110 4.020 5.650 15.53 22.08 62.03 34.04 3.090 617 4.320 5.020 8.530 3.840 8.730 16.89 24.84 58.43 33.21 5.340 621 4.520 4.970 8.570 4.320 8.780 13.53 27.00 57.83 35.10 4.540 623 4.516 4.956 7.176 3.708 8.358 19.85 28.21 60.96 31.07 3.085 624 4.799 5.005 7.532 3.891 8.162 17.36 25.71 60.04 32.22 4.365 626 5.291 5.036 9.201 4.028 8.833 16.80 25.46 60.46 33.75 4.480 630 5.081 5.305 7.952 3.687 5.888 18.86 23.46 63.57 31.41 3.763 631 4.971 4.981 7.211 4.148 7.793 13.54 33.05 56.50 33.98 1.360 637 5.456 5.235 9.242 3.772 5.483 16.52 21.18 63.03 33.57 4.063 646 5.306 5.171 8.776 3.803 6.663 15.94 23.16 60.11 33.59 4.210 648 4.961 5.055 8.402 3.937 7.608 17.85 14.17 61.54 31.93 3.428 649 4.466 5.121 8.301 4.028 5.898 16.47 25.97 61.16 34.12 2.720 650 4.969 5.115 8.122 4.451 6.837 18.22 24.76 63.32 32.86 2.420 651 5.194 5.231 13.829 4.645 5.617 17.10 25.38 60.94 33.25 2.148 652 5.714 5.126 8.424 3.955 5.682 16.22 26.71 60.96 32.97 1.638 653 5.609 5.254 8.592 3.871 5.610 17.03 22.92 63.58 32.52 1.248 agriculture and food sciences research, 2018, 5(1): 6-22 19 654 4.921 5.049 8.714 4.234 8.584 15.89 31.67 60.53 33.66 3.780 655 5.626 5.129 8.809 4.029 6.554 17.58 22.07 62.74 34.75 2.995 659 4.642 4.853 7.912 3.935 8.086 14.24 26.63 60.17 35.29 2.756 660 5.714 5.248 9.547 4.052 6.912 18.19 22.18 60.74 32.34 5.453 662 5.640 5.200 9.630 3.970 5.580 13.18 23.75 60.25 34.89 1.860 664 4.186 4.934 7.704 4.224 6.489 15.72 26.12 60.17 33.87 3.505 666 5.306 5.229 9.314 3.949 5.769 15.55 21.39 61.71 34.25 3.115 668 4.430 4.790 8.310 4.310 9.880 14.89 30.60 56.22 33.40 4.700 673 5.357 5.168 7.767 3.615 5.961 17.50 24.28 63.57 33.68 2.436 675 5.497 5.253 8.372 3.990 5.591 18.07 24.66 63.44 32.95 1.351 678 5.356 5.114 9.439 4.019 6.874 14.52 21.87 60.76 33.42 4.285 679 4.781 5.009 9.349 4.179 7.534 14.38 27.25 58.13 34.66 2.235 681 4.659 4.869 8.202 3.631 8.825 17.01 27.35 59.37 32.43 4.498 688 4.399 5.089 8.232 4.061 7.475 18.24 27.97 59.05 33.14 3.323 691 4.790 5.100 8.690 4.070 7.300 15.37 24.31 60.73 33.77 4.020 699 5.870 5.310 8.700 3.510 5.930 21.75 26.34 62.21 29.58 4.040 707 4.730 4.740 9.350 4.300 11.520 14.62 28.05 57.34 34.40 3.800 715 5.031 5.159 8.614 3.874 7.839 17.28 26.45 59.85 33.73 2.135 724 5.449 5.164 8.877 4.536 5.580 16.80 26.04 61.12 32.94 1.113 725 5.510 5.240 9.190 4.070 5.540 17.92 24.15 61.46 32.20 2.230 861 5.510 5.280 9.880 4.050 5.840 17.39 23.23 60.94 33.62 2.340 862 5.939 5.202 9.892 3.998 5.410 13.68 20.99 60.66 33.86 4.224 864 5.131 5.193 8.082 4.113 6.686 15.71 27.48 60.53 33.89 -0.610 873 5.631 5.228 9.792 4.058 5.721 14.84 25.23 60.90 34.46 2.433 dundee 5.763 5.160 9.701 4.137 5.509 14.29 24.18 59.75 34.30 2.896 dundee* 5.745 5.238 9.254 4.009 5.664 16.58 24.84 60.19 33.25 2.894 egret 4.911 5.031 8.949 4.072 8.081 16.14 25.65 58.18 33.02 3.510 egret* 5.245 4.968 8.694 3.724 8.944 17.75 28.28 59.01 32.43 4.264 heron 5.028 5.162 7.781 3.994 6.667 16.09 29.54 58.16 34.15 2.325 ibis 5.383 5.092 8.151 3.819 6.972 13.85 25.89 60.42 35.55 2.775 f-prob 0.224 0.004 0.079 0.030 0.012 0.001 0.217 0.018 0.002 0.069 lsd 1.1530 0.3602 2.105 0.6042 4.113 3.218 7.386 4.319 2.199 3.300 cv 10.55 3.33 11.30 7.10 26.81 9.35 13.94 3.34 3.09 47.16 se 0.5428 .1696 0.9908 0.2844 1.936 1.515 3.477 2.024 1.035 1.547 table-10. means of nutritional quality traits across the two sites. potchefstroom and brits in 2016/17. genotype ash (%) fiber (%) linoleic acid (%) linolenic acid (%) moisture (%) oil (%) oleic acid (%) palmitic acid (%) prote in (%) stearic acid (%) 1120 5.605 5.114 9.359 3.292 7.049 15.24 22.07 61.09 33.70 134.3 1363 5.319 4.990 8.851 3.741 8.565 15.99 26.79 58.21 33.11 184.0 1371 5.594 5.132 9.164 3.955 6.157 13.08 23.28 59.84 34.50 171.6 1380 5.184 4.962 9.251 4.068 8.617 15.07 25.04 59.64 33.56 142.7 1386 5.821 5.270 10.080 4.222 5.594 14.35 20.60 61.37 35.23 191.8 1390 5.319 5.023 9.894 4.220 6.583 13.78 23.54 60.08 33.75 147.3 1403 5.726 5.273 9.906 4.229 5.450 13.90 22.78 61.77 35.24 59.7 1409 5.058 4.803 8.606 3.921 11.537 15.58 26.69 60.31 33.90 181.9 1449 5.090 4.990 8.680 3.667 8.644 15.49 22.23 62.21 34.23 163.9 1552 5.915 5.124 9.353 4.153 6.783 14.05 21.70 60.99 33.42 175.9 1554 5.585 5.243 8.925 3.807 5.962 15.29 21.70 62.21 35.20 109.8 1555 5.370 5.155 8.796 4.133 6.953 13.52 26.04 61.41 34.98 97.6 1556 5.786 5.118 9.105 4.145 6.415 14.48 23.52 61.81 33.61 121.2 1558 4.702 4.908 8.715 3.739 9.917 16.93 28.40 56.73 33.18 250.1 1572 5.984 4.886 8.401 3.689 10.060 17.37 23.88 61.43 31.31 227.1 1573 5.128 4.723 9.362 3.881 12.784 17.07 24.14 59.59 32.03 263.5 1575 5.605 4.994 9.257 4.086 9.523 14.80 25.31 58.07 32.59 252.4 1578 5.147 5.144 9.197 4.117 7.730 16.01 25.34 60.34 34.23 154.7 1590 5.755 5.126 9.355 4.082 9.067 16.37 25.31 57.84 32.08 199.7 1594 8.197 3.611 19.842 10.049 2.843 10.75 -6.76 58.03 30.83 110.5 1595 5.378 5.178 8.749 4.162 7.493 16.61 24.83 61.02 33.33 178.4 1596 5.064 5.199 9.794 3.964 7.577 17.18 22.12 60.90 33.02 158.0 1597 5.455 4.970 7.292 3.917 8.045 15.05 28.22 63.14 33.97 108.7 1598 4.329 5.160 8.619 4.026 7.945 17.71 26.38 59.31 33.73 117.3 1599 4.686 5.299 9.060 4.013 5.936 17.48 24.30 61.32 34.24 67.2 1600 4.830 5.266 9.425 4.073 6.813 17.13 23.86 59.63 34.02 83.3 1659 5.618 5.180 9.430 4.081 7.066 14.59 22.73 61.78 34.66 150.2 220 5.249 5.176 9.559 4.216 5.926 15.29 23.53 60.00 33.80 148.6 223 5.703 5.143 9.995 4.341 5.676 16.45 21.62 61.24 33.12 159.1 231 5.087 5.140 8.621 3.860 7.036 17.80 25.23 61.10 32.16 105.9 266 5.140 5.170 9.291 3.692 5.963 15.06 23.09 61.17 34.40 134.2 267 4.707 5.092 9.247 4.061 8.188 16.09 21.58 60.33 34.35 189.0 268 5.528 5.187 9.365 3.950 6.169 15.29 23.47 60.68 34.86 137.9 278 5.068 5.073 9.141 4.338 6.384 14.45 30.78 58.76 34.82 35.7 284 4.603 5.115 9.214 3.973 7.644 17.08 22.04 60.86 33.57 190.6 290 4.816 5.181 8.998 4.079 5.999 16.03 22.52 61.81 34.34 187.9 291 5.021 5.213 8.848 3.850 5.769 16.58 24.08 61.41 34.26 115.7 292 5.010 5.022 9.936 4.113 7.864 15.00 23.24 58.65 34.14 200.6 agriculture and food sciences research, 2018, 5(1): 6-22 20 297 4.620 5.112 8.305 3.898 6.608 16.53 24.43 59.82 33.96 166.4 304 4.691 5.121 8.399 3.920 6.637 17.41 22.73 61.75 33.16 209.1 544 5.204 5.173 9.701 4.245 5.756 14.62 23.48 60.26 34.62 150.5 568 5.180 5.170 9.773 3.943 5.756 14.54 23.95 57.45 34.87 96.5 571 5.648 4.893 12.615 7.120 5.559 19.24 19.69 63.04 30.15 31.6 575 5.215 5.246 9.438 4.425 6.393 17.28 22.74 61.65 32.99 109.7 578 5.303 5.210 8.546 3.821 6.249 18.18 25.70 60.51 32.60 146.0 582 5.032 5.218 8.376 4.020 6.441 18.45 24.53 61.40 32.49 83.5 592 5.071 5.037 9.347 4.113 7.481 14.83 27.44 58.15 34.61 153.6 593 4.550 5.040 7.756 3.901 8.185 17.20 28.22 59.08 33.20 103.8 595 5.199 5.201 9.217 4.179 6.559 16.26 24.09 60.73 34.87 65.3 597 5.161 5.177 8.874 3.927 6.538 14.53 24.31 60.44 35.62 74.8 598 5.104 5.146 8.598 3.897 6.693 14.89 23.78 61.33 35.22 78.6 604 5.223 5.295 9.598 4.017 5.867 15.17 23.09 60.62 35.24 74.4 607 5.160 5.173 8.522 3.817 6.137 16.13 23.65 62.18 34.54 71.3 617 4.697 5.161 9.229 3.954 6.798 15.25 21.55 60.50 34.92 228.2 621 4.672 5.098 8.646 4.206 6.921 14.75 25.53 59.12 34.78 214.0 623 4.833 5.079 7.982 3.859 7.246 18.82 25.51 61.65 31.81 134.0 624 4.931 5.088 8.432 4.050 7.262 16.86 24.74 60.37 33.12 153.3 626 5.197 5.030 9.583 4.004 9.093 17.17 25.12 60.15 34.26 168.9 630 4.828 5.218 8.115 3.740 6.875 18.68 25.15 61.78 31.69 129.4 631 5.276 4.453 10.689 7.059 7.008 18.84 24.68 51.41 29.12 57.0 637 5.277 5.200 9.415 3.948 6.178 15.75 21.74 61.71 33.86 171.4 646 5.240 5.158 8.949 3.954 6.486 16.46 23.03 60.04 33.54 219.3 648 5.467 5.215 7.965 5.451 7.018 16.91 13.73 51.27 32.86 184.3 649 4.436 5.131 6.662 4.457 5.602 17.67 22.95 61.24 33.71 102.5 650 5.055 5.095 8.489 4.334 6.248 16.22 23.79 62.76 33.94 134.3 651 4.999 5.195 11.578 4.347 5.836 17.09 24.72 60.35 33.22 121.1 652 5.823 5.131 9.141 4.184 5.796 15.32 24.27 60.40 32.99 148.6 653 5.443 5.248 9.314 3.950 5.779 16.28 17.31 62.33 33.59 85.3 654 5.041 5.125 8.878 4.238 7.149 15.81 27.11 61.30 33.71 162.6 655 5.221 5.156 8.917 4.082 5.964 15.98 22.24 62.88 34.76 112.7 659 5.011 4.879 8.374 3.976 7.064 12.96 25.05 60.19 35.33 149.9 660 5.154 5.243 9.085 3.896 6.498 17.73 22.03 61.61 33.07 169.5 662 5.263 5.190 9.109 3.987 6.163 14.14 23.87 60.61 34.42 47.2 664 4.618 5.044 8.388 4.061 6.390 15.65 24.81 60.08 33.97 133.5 666 5.035 5.189 9.136 3.961 6.008 16.05 21.59 61.63 34.27 142.7 668 4.818 4.966 9.012 4.089 7.349 14.49 25.32 58.43 33.74 192.1 673 5.441 5.225 8.551 3.767 5.794 16.35 23.38 62.55 34.05 143.6 675 5.326 5.253 8.976 3.995 5.804 16.62 23.75 61.96 33.90 70.1 678 5.103 5.157 9.508 3.946 6.472 15.72 22.10 60.53 33.50 187.7 679 5.123 5.072 9.848 4.281 6.532 13.90 25.19 58.91 34.50 83.0 681 4.701 4.976 8.003 3.870 7.539 17.09 27.35 59.30 33.11 189.4 688 4.652 5.103 9.114 4.068 6.686 16.53 24.56 59.60 33.65 156.9 691 5.168 5.187 10.044 4.011 6.893 15.76 21.95 60.64 34.14 149.1 699 6.233 5.317 8.843 4.033 6.520 20.73 26.37 61.08 30.28 183.0 707 5.197 5.072 9.543 4.076 8.228 14.84 24.84 59.32 34.61 141.2 715 5.149 5.173 8.855 3.837 6.870 15.64 24.11 60.91 34.70 94.0 724 5.296 5.096 8.758 4.332 5.401 15.59 25.39 61.28 32.67 93.1 725 5.353 5.161 9.156 4.355 5.855 16.57 25.43 60.21 32.07 144.5 861 5.284 5.256 9.696 4.036 6.078 17.05 21.82 61.14 33.60 150.5 862 5.507 5.154 9.761 4.147 5.491 14.22 21.46 60.44 34.12 197.4 864 5.078 5.198 8.777 3.971 6.236 15.84 23.94 60.84 33.79 165.0 873 5.359 5.187 9.898 4.034 5.901 14.32 22.31 61.01 34.45 136.1 dundee 5.773 5.122 9.907 4.131 5.755 13.08 23.19 59.27 34.47 137.7 dundee* 5.550 5.218 9.397 3.938 5.906 15.71 23.70 60.04 33.72 121.8 egret 5.212 5.100 9.277 3.975 7.107 15.77 23.55 58.98 33.15 170.7 egret* 5.192 5.092 8.981 3.810 7.510 15.92 24.79 60.24 33.83 159.0 heron 5.149 5.160 8.643 3.913 6.118 15.19 25.12 59.44 34.28 118.0 ibis 5.444 5.126 8.956 3.926 6.487 13.91 23.32 60.71 35.04 138.5 genotype 0.8203** * .1472 0.93 2.282 6.577*** .766** * 55.09 12.59 4.906 * 8854** * site 3.0554 0.067 0 9.68** 9.210* 9.696*** 37.343 ** 666.57 *** 25.12 7.737 250321 *** genotype * site 0.6848* 0.136 3 0.79 2.150 3.906 5.117 41.15 8.64 3.219 4359 mean 5.12 5.00 9.03 4.07 6.70 15.56 23.11 59.15 33.02 139.23 lsd 1.480 0.780 4 5.479 3.208 3.865 4.281 15.93 7.018 3.798 129.4 cv 13.77 7.44 28.94 37.55 27.54 13.12 32.90 5.65 5.49 44.54 se 0.7204 .3799 2.667 1.562 1.882 2.084 7.756 3.409 1.849 62.24 r2 note: *** p<0.001, ** p< 0.025 and *p< 0.05 agriculture and food sciences research, 2018, 5(1): 6-22 21 references [1] food and agriculture organisation of the united nations, "fao statistics division, faostat 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journal of botany, vol. 40, pp. 2323-2328, 2008. view at google scholar asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://scholar.google.com/scholar?hl=en&q=phenotypic%20and%20molecular%20analysis%20of%20m7%20generation%20of%20soybean%20mutant%20lines%20through%20random%20amplified%20polymorphic%20dna%20(rapd)%20marker%20and%20some%20morphological%20traits https://scholar.google.com/scholar?hl=en&q=evaluation%20of%20soybean%20%5bglycine%20max%20(l.)%20merrill%5d%20germplasm%20for%20some%20important%20morphological%20traits%20using%20multivariate%20analysis https://scholar.google.com/scholar?hl=en&q=evaluation%20of%20soybean%20%5bglycine%20max%20(l.)%20merrill%5d%20germplasm%20for%20some%20important%20morphological%20traits%20using%20multivariate%20analysis 58 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 58-64, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.58.64 © 2020 by the authors; licensee asian online journal publishing group occurrence of fumonisins in some maize meal marketed to consumers in harare, zimbabwe princess mushonga1 wilson parawira2 loveness kuziwa nyanga3 ( corresponding author) 1,2department of biological sciences, faculty of science and engineering, bindura university of science education, chimurenga road, bindura, zimbabwe. 3institute of food, nutrition and family science, university of zimbabwe, mount pleasant, harare, zimbabwe. abstract mycotoxins are toxic secondary metabolites secreted by filamentous fungi which affect animals, plants and humanscausing a lot of diseases. fumonisins are the types that mostly affect cereal grains. the prevalence of fumonisins in harare marketed maize meal was investigated in 72 randomly selected samples. fumonisin b1 and fumonisin b2 were extracted from maize meal with methanol-water (3:1, v/v) using ultrasonic extraction. they were injected into an lc-ms system following centrifugation. a survey was also conducted to determine the measures taken by 5 major harare millers to mitigate fumonisin contamination in the maize meal. fumonisin b1 (fb1) was detected in all analysed samples at mean concentrations ranging between 61.45 and 265.79 µg/kg. fumonisin b2 (fb2) was detected in only 56.9 % of the samples analysed, with a range between 13.72 µg/kg and 76.93 µg/kg. the highest total fumonisin mean concentration (fb1+fb2) was detected in maize meal with maize bran added to it (342.72 µg/kg), while the least total fumonisin mean concentration was detected in maize meal with wheat bran added to it (61.45 µg/kg). all the samples analysed were within the maximum tolerable limit (mtl) of 1000 µg/kg. however, the average probable daily intake (apdi) of fumonisins ranged between 0.82 and 4.57 µg/kg body weight/day. maize meal with maize bran added and roller meal had pdmi above the regulatory limit of 2 µg/kg body weight/day recommended by the world health organisation. the high frequency of fumonisins consumed daily in these two types of maize meal could be posing some health risks to consumers. keywords: mycotoxins, maize meal, fumonisins, milling, risk assessment, world health organisation, zimbabwe. citation | princess mushonga; wilson parawira; loveness kuziwa nyanga (2020). occurrence of fumonisins in some maize meal marketed to consumers in harare, zimbabwe. agriculture and food sciences research, 7(1): 58-64. history: received: 22 january 2020 revised: 28 february 2020 accepted: 31 march 2020 published: 5 may 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: the authors would like to acknowledge the support they got from bindura university of science education and the pharmacy department of the university of zimbabwe for all the assistance throughout our work. authors also want to thank all the interviewed milling companies for the information they provided for the successful completion of their work. authors are indebted to mr. c. mutsimhu, dr. b. masamha and mr. k. basira who assisted them with the statistical analyses. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 59 2. materials and methods ................................................................................................................................................................... 59 3. results and discussion ................................................................................................................................................................... 60 4. conclusion ......................................................................................................................................................................................... 63 references .............................................................................................................................................................................................. 63 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.58.64&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.58.64&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1593 https://orcid.org/0000-0001-7735-2416 https://orcid.org/0000-0003-4576-6010 https://orcid.org/0000-0001-5585-4204 agriculture and food sciences research, 2020, 7(1): 58-64 59 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the levels of fumonisins in all the maize meal analyzed in this study were below the maximum tolerable limits of 1 000 µg/kg set by the european union. however, the apdi of fumonisins estimated for maize meal with maize bran added and roller meal were above the provisional apdi set by who. maize meal with maize bran added and roller meal showed high contamination levels probably because they contained the bran and some germ that are highly contaminated by fumonisins. the high frequency of fumonisins consumed daily in both types of maize meal pose some health risks to consumers. there is need for setting up measures to check the quality of maize-based foods in order to alleviate the dangers of developing esophageal cancer and other probable health complications that are a result of fumonisin contamination. 1. introduction maize (zea mays l) is an economically significant cereal that is grown in zimbabwe. it serves as the staple diet for the majority of the zimbabwean populace. maize is nutritionally important since it is an excellent source of proteins, starch, lipids, some bioactive compounds and some phytochemicals [1]. maize may contain a number of important b vitamins, folic acid, vitamin c, and provitamin a (i.e., precursor to vitamin a). maize is also rich in phosphorus, magnesium, manganese, zinc, copper, iron and selenium, and has small amounts of potassium and calcium. it can be consumed as raw grain or processed into maize meal. however, safety can be greatly compromised when the maize grains are contaminated by fumonisins, the secondary organic metabolites secreted by fusarium species [2]. fumonisin b1, b2 and b3 are the most predominant of all fumonisins and are often detected in maize and maize products [3]. a considerable volume of research on mycotoxins has been carried out in zimbabwe on maize and studies have shown that fumonisin b1 mainly contaminates the maize grains before and after harvest [3, 4]. fumonisin b1 which accounts for nearly 70 % of food contamination worldwide [5] is the most pernicious of the fumonisins. it has been associated with hepatocellular and oesophageal cancers, neural tube defects and impaired growth in children [6]. the b-series of fumonisins are generally thermo-stable at temperatures below 150 ºc but their concentrations get significantly lower at temperatures above this value [2]. insects, such as the maize stalk borer (busseola fusca), infest maize cobs in the field and subsequently cause contamination of the grains by spreading mycotoxigenic fungi [7]. when water activity is high (aw> 0.9), fumonisins are synthesised in maize before harvesting [8]. it is of paramount importance that maize kernels are adequately dried prior to milling in order to minimise the chances of contamination by the fumonisin-producingfungi. milling is a process that transforms grains into smaller consumable particles, such as maize meal, grits or starch. the milling process does not detoxify fumonisins, instead it re-distributes them, giving rise to higher or lower concentrations in the various milled fractions [9]. studies have shown that the germ and the hull of the maize kernels harbour very high levels of fumonisins compared to the endosperm [10, 11]. zimbabwean milling companies buy maize from the grain marketing board. they mill and package the maize meal which is then sold in the supermarkets. recently, there has been an improvement in the nutritional composition of maize meal produced by some companies. some are adding wheat bran or maize bran to the processed maize meal so as to enhance the fibre levels in their products. however, the brans might bring with them mycotoxins which contaminate the maize meal and pose some health risks to the consumers [12]. fumonisins have been found to be the most prevalent mycotoxin in zimbabwe. however, not much research has been done to determine levels of fumonisins in maize meal. the high frequencies of fumonisins observed in zimbabwean maize grain are a cause for concern to human health [4]. therefore, it is most appropriate to quantify the fumonisin levels in maize-meal since it is a direct product of maize consumed by many zimbabweans. the findings will help to raise awareness on the levels and dangers of fumonisins in the different types of maize meal being consumed in the country. 2. materials and methods 2.1. study area the study was conducted in harare, zimbabwe where maize meal was bought from three different supermarkets. these three supermarkets represented the major supermarkets selling maize meal to the population (2 123 132 people) in harare (zimstat. census 2012: provincial report). the supermarkets were selling maize meal from the zimbabwean millers. further, a survey was carried out on five major zimbabwean millers to establish the mitigation measures during the milling process to reduce fumonisin contamination. analysis of the maize meal samples was done at the laboratory of the pharmacy department of the university of zimbabwe, harare. 2.2. experimental design this study used a descriptive approach. quantitative observations were done by analysing and recording of quantifiable levels of fumonisin b1 and fumonisin b2 in the maize meal which was randomly purchased from harare supermarket between september and december, 2018 from 5 different grinders/millers. analysis of variance (anova) was used to statistically analyse the results. qualitative observation was done on the survey research. both the quality controllers and those who were milling (millers) answered to questionnaires with both open-ended and close-ended questions which mainly focussed on the measures taken by these 5 millers to mitigate the effects of fumonisin and other mycotoxins contamination in maize meal. 2.3. sampling of maize meal from each of the five major zimbabwean millers (m1, m2, m3, m4, m5), 2 kg maize-meal packets, 3 of each type (roller meal and super refined), were collected randomly from each of the three selected major supermarkets. in addition, 2 kg packets of each of super refined maize meal with wheat bran or maize bran were also collected from miller 2 using the same procedure as above. agriculture and food sciences research, 2020, 7(1): 58-64 60 © 2020 by the authors; licensee asian online journal publishing group one hundred grams of each type of maize meal were drawn from each of the 2 kg packets at 3 points (bottom, middle and top of the pack). similar types of maize meal samples collected from each individual miller were thoroughly mixed together at the laboratory of the pharmacy department, university of zimbabwe. the mixed samples were sealed in sterile plastic bags and stored at 20 ºc until further analysis. 2.4. fumonisin extraction from maize meal samples fumonisin extraction was done according to the protocol by li, et al. [13] with ultrasonic extraction which was modified. maize meal samples were blended and 5 g of homogenized maize meal were mixed with 25 ml of extraction solvent (methanol/water) in the ratio 3:1 (v/v). triphenyl phosphate (tpp) was used as the internal standard. samples were shaken by hand for 5 minutes and centrifuged at 4000 rpm for 5 minutes using a hettich zentrifugen (germany) centrifuge. the supernatant was placed into a 5 ml syringe. it was then filtered, through a 0.22 mm acrodisc syringe filter, into an hplc vial for lc-ms analysis. 2.5. calibration of standards five grams (5g) of blank (probrand) corn (corn without any fumonisin) were transferred into a 50 ml falcon tube. the calibration standards and quality control samples were matrix-matched by spiking known volumes of standards of fb1 and fb2 into blank samples to target concentrations 5 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 500 ng/ml, 1000 ng/ml, 2000 ng/ml, 3000 ng/ml and 5000 ng/ml. this was done using 25 ml (methanol/water) in the ratio 3:1 (v/v) as the diluent in which triphenylphosphate (tpp) was used as the internal standard at 500 ng/ml. the tubes containing the sample and extraction solvent were capped and shaken vigorously for 5 minutes. they were further centrifuged for 5 minutes at 4000 rpm. the supernatant was put into a 5 ml syringe and then filtered through 0.22 mm syringe filters into the vials for hplc-ms analysis. a calibration curve was made using 9 standards starting with 1000 ng/ml as stock. spiked samples were produced to validate the method using qc standards: hqc-4000 µg/kg; mqc-2000 µg/kg; lqc50 µg/kg. 2.6. hplc-ms conditions for fumonisin analysis the hplc used was agilent hplc 1260 system (california, usa) equiped with a binary pump, autosampler and thermostated column compartment. a phenomenex, luna column with dimmesions 50 x 2 mm, 3 micron was used for separation of the compounds. mobile phase a comprised 5 mm ammonium acetate; ph was adjusted with 16 ml formic acid to 3.1. mobile phase b consisted of 100 % methanol. the temperature of the column was maintained at 40 ℃ . the flow rate was 0.45 ml/minute while the injection volume was 80 µl. fifty percent methanol was used for needle wash. run time was set at 10 min. all the fumonisins eluted at 7.2 7.8 min. agilent q-tof 6530 mass spectrometer was used for detection of fumonisins. it was fitted with an electrospray ionisation (esi) probe and operated in the positive ionisation mode. the following parameters were optimised: capillary voltage, 4000v; drying gas temperature, 350 ºc and desolvation gas flow rate, 10 l/min. data acquisition and analysis was done using the mass hunter software version b.07.03 (509). 2.7. method validation an ‘in house’ validation was applied according to the internal procedure as well as the acceptance criteria for bio-analytical method development and validation. intra-day accuracy (97.54 %), linearity (0.995-0.999) and precision (2.78%) of the quality control (q c) samples for fumonisin b1 and fumonisin b2 were within the accepted range. inter-day quality control samples for both fumonisin b1 and fumonisin b2 also passed the acceptance criteria. the average recoveries for all quality control samples were between 85 % and 96 %. percent coefficient of variation was less than 2.78 % for all qc samples. calibration curves for both fumonisin b1 and fumonisin b2 showed that linearity were greater than 0.9900 according to the set regulations for method development and validation. limit of detection for fumonisin b1 and fumonisin b2 were 5µg/kg. in this regard, the lc-ms method developed passed all the acceptance criteria in accordance with fda principles for method validation and related regulations; therefore it was fit for routine sample analysis for fumonisins in maize meal. 2.8. estimation of the probable average daily intake of fumonisins in the 4 types of maize meal the estimation of the probable average daily intake (apdi) was done by calculating the average probable daily intake of fb1 and fb2 combined in the samples. the estimates of the average probable daily intake (apdi) of fb1 and fb2 combined in the samples were calculated using the formula: apdi=l×d/60kg, where: apdi is the average probable daily intake. l is the mean total concentration of fb1+fb2 in the maize meal samples. d is daily consumption of maize meal average of 800g [3, 4]. the proposed typical body weight of an adult was 60kg. 2.9. data analysis quantitative data were analyzed using unbalanced analysis of variance (anova) performed in genstat version 19. mean differences in the levels of fumonisin contamination were determined using least significant differences (lsd) at 5% level of significance. an unbalanced design anova was used because there was no equal number of observations since some fumonisin quantities were below the limit of detection. the qualitative data collected using the questionnaires were analyzed using content analysis. 3. results and discussion 3.1 survey results a questionnaire was administered to the millers and quality controllers of each milling company to establish the mitigation measures these milling companies were taking to reduce the levels of fumonisins in their maize meal and the responses are summarized in table 1. most of them knew very little about fumonisins, instead, during their quality checks they focused on aflatoxins, which are less abundant in zimbabwean maize than fumonisins [3, 4]. agriculture and food sciences research, 2020, 7(1): 58-64 61 © 2020 by the authors; licensee asian online journal publishing group their lack of knowledge about fumonisins may pre-dispose the maize meal consumers to health risks from fumonisin contamination. millers were not monitoring fumonisin levels in both the maize and the maize meal. this has potentially detrimental effects on the consumer’s health. mwalwayo and thole [14] stressed the co occurrence of aflatoxins with fumonisins in maize; therefore millers need to use cheaper and effective methods of testing for multiple mycotoxins in the maize meal, such as the liquid chromatography mass spectrometry technique (hplc/ms) [15]. all the millers were sorting and cleaning the maize kernels prior to milling. they did the sorting to remove physically damaged and infected grain based on physical damage and the presence of moulds. cleaning of the maize was done to remove cobs, chuff and stones. the sorting could explain the lower mean concentrations of fumonisins obtained in this study. research done by pietri, et al. [11] showed that fumonisin concentrations were lower in the cleaned maize than the unprocessed kernels. santini, et al. [16] also noted that cleaning, if done effectively, reduces fumonisins concentrations from the pericarp as well as from the damaged and broken grains by 50 %. matumba, et al. [17] and kimanya, et al. [18] also allude to the fact that sorting of maize prior to milling reduces contamination of maize by fumonisins since defective kernels are discarded. table-1. miller’s responses on mitigation measures to reduce fumonisin contamination in maize meal. parameter miller 1 miller 2 miller 3 miller 4 miller 5 vetting of grain for weevils ✓ ✓ ✓ ✓ ✓ monitoring moisture level ✓ ✓ ✓ ✓ ✓ treatment prior to storage with insecticides x ✓ x x ✓ covering of bags or silos x x ✓ ✓ ✓ cleaning of storehouse before storage ✓ ✓ ✓ ✓ ✓ removal of old grain before adding new grain ✓ ✓ ✓ ✓ ✓ use of pesticides during storage x ✓ ✓ ✓ ✓ sampling to monitor aflatoxin levels ✓ ✓ ✓ ✓ ✓ analysis for fumonisins before milling x x x x x sorting of grain ✓ ✓ ✓ ✓ ✓ testing for fumonisins in the maize meal x x x x x note: √ -being implemented xnot being implemented. however, despite the cleaning and sorting processes, some fumonisins were detected. this could be attributed to the cleaning method (screening and sifting), which did not completely eliminate fumonisins but only served to reduce them. nyangi [7] reported that the most noticeable sign of fusarium infection in maize grown in the tropical areas is kernel rot. however, fumonisins are often detected in maize kernels even if in the absence of invisible signs of infection. all the millers interviewed stressed that they checked the moisture content of their grain upon receipt from the grain marketing board (gmb). this mitigation measure helped in the prevention of moulds that cause fumonisin contamination. bacon and nelson [19] state that levels of kernel contamination by fumonisins under improper storage and the optimal growth condition for fumonisin-producing moulds are when moisture content of harvested maize is between 18-23 %. the monitoring of moisture levels within this range by these millers could be another reason for the low fumonisin levels detected in the maize meal analyzed in this study. millers stored their grain in silos and this made them more susceptible to cross-contamination by fumonisins. the use of hermetic technology for grain storage could be an effective alternative method in the reduction of fumonisins since the bags or silos suffocate the insects and fungi, preventing contamination by fumonisins. hove, et al. [4] noted that maize stored in polypropylene bags was less contaminated than un-bagged maize (mean fb1=263 µg/kg and 401 µg/kg respectively). most of the millers interviewed indicated that in the silos, tobacco weevil was the most challenging. weevils feed upon the kernels and at the same time spread the infection to other healthy kernels. in spite of vetting the grain for weevils upon its reception from the gmb, the weevils could mature in the grains stored in the silos, leading to contamination. miller 1 indicated that they did not apply any insecticides during storage of the grain. this could be the reason for the highest level of fumonisin contamination compared to other millers. other 4 millers were applying phosphin gas tablets at a rate of 10 tablets per silo so as to kill the weevils in order to reduce contamination by fumonisins. this data shows that fumonisin control was not intentional, rather indirect through direct practices destined to assure the control of food safety and quality. 3.2. fumonisin levels in maize meal analysis was done on the types of maize meal from different millers to determine fumonisin levels. there was a significant difference among the 5 millers in the concentration of fumonisins in the maize meal. there was also a significant difference between fumonisin b1 and fumonisin b2 levels in the 4 types of maize meal see table 2. again, a difference existed in the fumonisin concentration among the 4 types of maize meal (roller meal, super refined, super refined with maize bran and super refined with wheat bran). from the 72 samples analysed from the marketed maize meal produced by the large millers, all maize meal samples (roller meal, super refined, maize meal with wheat bran and maize meal with maize bran) had a fumonisin b1 level below the maximum tolerable limit (mtl) of 1 000µg/kg [20] see table 2. of the samples analysed for fb2 56.9 % had non-detectable levels. although half the samples of super refined maize meal had concentrations below the detection limit, all maize meal samples with wheat bran added did not present detectable concentrations of fumonisin b2. there was much variability in fb2 levels of super refined and this could be attributed to the low sample size. results from this study also showed mean fumonisin levels in the maize meal samples ranging from 61.64 µg/kg to 265.79 µg/kg for fb1 and 0.00µg/kg to 76.93 µg/kg for fb2 table 2. all analysed samples had fb1 but only 43.1 % were contaminated by fb2. the lower concentrations of fumonisins observed, compared to other studies [3, 4, 21] could reflect the effectiveness of the measures taken by millers to indirectly assure the control of safety and quality of foods. agriculture and food sciences research, 2020, 7(1): 58-64 62 © 2020 by the authors; licensee asian online journal publishing group a research conducted by doko, et al. [21] on zimbabwean maize meal showed that 83 % of the analysed samples had fumonisin b1, b2 and b3 ranging between 55 µg/kg and 2 735 µg/kg. murashiki, et al. [3] recorded fumonisin concentrations ranging between 10.43 µg/kg and 606.64 µg/kg in maize meal samples from shamva and makoni districts (p<0.05). these researchers analysed samples from small-holder farmers who took no precautionary measures against fumonisin contamination in the grains as large millers do, thus the high fumonisin levels detected. fumonisin b1 and b2 mean concentrations were significantly higher in maize meal with maize bran than other maize meal types (mean =265.79 µg/kg). roller meal had fumonisin mean concentrations which were slightly lower than those found in the maize meal with maize bran but higher than in super refined maize meal table 2. this is because the bran is the most colonized by the fusarium spp., followed by the germ fraction. these results are comparable to pietri, et al. [11], brera, et al. [22] and vanara, et al. [23] who reported that when the corn is milled, fumonisins are in elevated concentrations in the bran, followed by the germ fraction. broggi and others carried out a study in a commercial dry-mill in argentina and found a three-fold increase in fumonisin contamination in the germ and bran fractions than in the whole corn [7]. roller meal samples had fumonisin levels higher than super-refined maize meal samples because some pericarp and germ fragments probably found their way into the course meal during drymilling [24]. even though there was no bran or germ added to the super-refined maize meal, some fumonisins (80.49 µg/kg), were recovered from the samples. the main cause of contamination could be due to the floury endosperm that surrounds the germ where a higher formation of fumonisins is possible. in support of this, vanara, et al. [2] noted that the presence of amylopectin in the endosperm during kernel development instigates fb1 biogenesis. a study by gwirtz and garcia-casal [25] on processing of maize flour and corn meal products also showed a remarkable lowering of fumonisin concentrations in the endosperm during dry milling. levels of fumonisins obtained in maize meal where wheat bran was added to super refined were very low. at 5 % significance level, there was no difference between super -refined maize meal and super refined maize meal with wheat added table 2. this indicates that wheat was not a source of contamination by fusarium compared to maize. streit, et al. [26] state that fumonisin contamination commonly occurs in maize and maize products. however, although the results show that fumonisin concentration in the types of maize meal are within the maximum tolerable limit (mtl) of 1000 µg/kg by eu standard [12, 14] there is need for constant monitoring of fumonisin levels, especially in the maize meal with maize bran added and in the roller meal. fumonisin b1 was found to be more abundant in the maize meal samples than fumonisin b2 table 2. at a least significance difference of 5 %, the results from this research showed that there was a significant difference in the contamination of maize meal by the two types of fumonisins. fumonisin b2 was not detected in maize meal samples, such as maize meal with wheat bran added and some super refined maize meal yet fb1 was detected in the same samples at 2.5 µg/kg detection limit. this also confirmed that fumonisin b1 is the dominant type that contaminates zimbabwean maize. this is in line with the observations by marasas, et al. [27] and rheeder, et al. [28] who noted that fbi pre-dominates and contributes 70-80 % of all fumonisins whereas fb2 constitutes 15-25 % in maize grains. the differences in quantity between fb1 and fb2 could be because moulds produce higher quantities of the former. table-2. means of fumonisin concentration (µg/kg) for the different products from all millers combined. fumonisin type maize meal with maize bran roller meal super refined maize meal with wheat bran fb1 265.79±22.22a 229.85±10.28b 80.49±10.28c 61.45±22.22c fb2 76.93±31.43c 32.83±12.15d 13.72±14.57d 0.0e note: means in the same row and column with different superscripts are different at 5 % significance level. means with the same superscripts are the same at 5 % significance level. analysis was done to assess fumonisin mean concentration in the types of maize meal produced by the different millers. miller 1 had the highest mean fumonisin b1 concentration of 336.40 µg/kg table 3. the high levels in samples from miller 1, compared to other millers, may be attributed to the fact that they were not treating their maize grains prior to storage and as a result, weevils could have probably attacked the grains whilst in silos and caused further contamination by fumonisins. again, they did not cover the silos so it was easier for the insects to spread around and cause further contamination in, otherwise healthy, grains. hove, et al. [4] also noted that the damage caused by pests contributes a lot to mycotoxin contamination of the maize grains. that is why it is of paramount importance to apply chemicals that protect the grain from damage. miller 2 recorded the second highest mean levels of fumonisin b1 (158.39 µg/kg) because it was the only interviewed milling company that manufactures maize meal with maize bran added to it. as has been mentioned earlier on, the bran fraction contains the highest fumonisin levels in maize grains. furthermore, miller 2 was not covering the silos so this may have caused crosscontamination of grain by weevils. at 5 % level of significance, there was no outstanding difference in fumonisin b1 mean concentration among millers 3, 4 and 5. they all recorded the low mean concentration. miller 2 showed a marked difference from all other miller as it contained high levels of fumonisin b2 (81.87 µg/kg). this could also be attributed to the fact that they produced maize meal with maize bran, which normally contains a significant amount of fumonisins. table-3. overall fumonisin mean concentration (µg/kg) for different millers for all types of maize meal combined. fumonisin type miller 1 miller 2 miller 3 miller 4 miller 5 fb1 336.40±15.7a 158.39±13.8b 75.62±15.7c 94.96±15.8c 122.40±15.7 c fb2 10.05±18.6d 81.87±19.5ce 3.1419.1d 25.34±31.4d 19.60±19.1d note: means in the same row and column with different superscripts are different at 5% significance level. means with the same superscripts are the same at 5 % significance level. agriculture and food sciences research, 2020, 7(1): 58-64 63 © 2020 by the authors; licensee asian online journal publishing group 3.3. estimate of the average probable daily intake (apdi) for risk assessment the magnitude of fumonisin contamination obtained in the present study was below the tolerable regulatory level of 1000 µg/kg which was set by the european commission [20]. however, the joint fao/who expert committee on food additives (jecfa) set the health-based guidance value for fb1, fb2 and fb3 separately or combined at 2 µg/kg body weight/day [29]. in this study, risk assessment was done on the four types of maize meal to establish levels to which maize meal consumers are exposed to fumonisins daily. this study showed the average probable daily intake (apdi) of fumonisins by consumers table 4. the values for maize meal with maize bran added and roller meal were above the set value of 2 µg/kg body weight/day. it is an indication that consumers of these types of maize meal with maize bran in harare, zimbabwe, are at greater risk of vulnerability due to the detrimental accumulative levels of fumonisins unlike consumers of super refined maize meal and maize meal with wheat bran added. moreover, low level chronic intake of fumonisins in the maize and maize products can be more devastating than one-time high level intake a study carried out by williams, et al. [30] reported that 4. 5 billion maize consumers in the developing countries are exposed to chronic toxicity. the study conducted by murashiki, et al. [3] showed that populations in shamva and makoni districts, zimbabwe, had a probable daily intake of 5.76-8.09 µg/kg body weight/day. these figures are slightly higher than the values calculated in this study, most probably due to the mitigation measures taken by the registered millers and the type of milling done to reduce fumonisin contamination in the maize meal unlike with unregistered millers used by murashiki, et al. [3]. further, analysis done by hove, et al. [4] showed a maximum probable daily intake of 5.40 µg/kg body weight/day for children between 5-9 years, 4.40 µg/kg body weight/day for adolescents and 2.30 µg/kg body weight/day for adults in zimbabwe. high fumonisin intakes (141.97 µg/kg body weight/day) have also been reported in tanzania by kimanya, et al. [18]. other studies in the more economically developed countries, such as spain and brazil, have indicated fumonisin levels as low as 0.1 µg/kg body weight/day and 0.063 µg/kg body weight/day respectively [31]. these low values are perhaps due to the advanced and effective mitigation measure taken by these countries to mitigate fumonisin contamination in foods. this is coupled to the low intake of maize–based foods [31] unlike in zimbabwe where the staple diet is primarily maize-based. on grounds of the high average probable daily intake of fumonisins detected in some types of maize meal being sold in harare markets, it is recommended to adopt some measures to mitigate the deleterious effects of fumonisin consumption. these mitigation practices include; sorting and cleaning of grain to remove broken and moldy kernels, application of grain-protection chemicals to kill weevils and monitoring of moisture levels to reduce contamination of grain by fumonisins [29]. table-4. total mean fumonisin concentrations (fb1 + fb2) and probable intakes of marketed maize meal in harare, zimbabwe. type of maize meal total mean fumonisin concentration (µg/kg) average probable daily intake (µg/kg bw/day) maize meal with maize bran added 342.72 4.57 roller meal 262.68 3.50 super refined 94.21 1.26 maize meal with wheat bran added 61.45 0.82 4. conclusion the levels of fumonisins in all the maize meal analyzed in this study were below the maximum tolerable limits of 1 000 µg/kg set by the european union. however, the apdi of fumonisins estimated for maize meal with maize bran added and roller meal were above the provisional apdi set by who. maize meal with maize bran added and roller meal showed high contamination levels probably because they contained the bran and some germ that are highly contaminated by fumonisins. the high frequency of fumonisins consumed daily in both types of maize meal pose some health risks to consumers. there is need for setting up measures to check the quality of maize-based foods in order to alleviate the dangers of developing esophageal cancer and other probable health complications that are a result of fumonisin contamination. results from this study also suggested that some of the practices that are being taken by the millers are effective at reducing fumonisin contamination in the maize meal. the milling process itself, which includes dehulling and de-germination of maize kernels, seems to dilute and redistribute fumonisins into fractions that mostly become animal feed. it is of paramount importance to undertake further research on smallscale millers that will assist them to meet international quality standards and adopt those practices that minimize risks to fumonisin contamination in the maize meal they produce. references [1] s. siyuan, l. tong, and h. l. rui, "corn phytochemicals and their health benefits," journal of food science and human wellness, vol. 3, pp. 185-195, 2018. 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[31] g. cano-sancho, a. j. ramos, s. marín, and v. sanchis, "occurrence of fumonisins in catalonia (spain) and an exposure assessment of specific population groups," food additives & contaminants: part a, vol. 29, pp. 799-808, 2012. available at: https://doi.org/10.1080/19440049.2011.644813. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.who.int/foodsafety/foodsafetydigest/en/, 79 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 79-88, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.79.88 © 2020 by the authors; licensee asian online journal publishing group optimization of proximate and minerals compositions of sweet potato, soybean and rice bran composite flours for production of low glycemic index dough meal awolu, o.o.1 ajibola, c.f.2 adeloye, j.b.3 ogwu, f.4 ( corresponding author) 1,2,3,4department of food science and technology, federal university of technology, akure, nigeria. abstract dough meal with low glycemic index was produced from sweet potato, soy bean and rice bran composite flours. effect of the composite flour samples on the glycemic index, as well as proximate and minerals composition of the dough meal were optimised using optimal mixture design of response surface methodology. the dough meal had considerably high protein and fibre contents. the moisture and ash contents were low. the results indicated that the addition of sweet potato enhances the minerals composition, while soybean and rice bran had highest effect on the protein and fibre contents respectively. the results of the optimisation showed that the dough meal had glycemic index, meaning that the dough meal prepared from sweet potato (65.085.0 g/100g), soy bean (10.0-25.0 g/100g) and rice bran (2.5-10.0 g/100g) had acceptable glycemic index. the dough meal with 75.00 g/100g sweet potato flour, 15.00 g/100g soybean meal flour and 10.00 g/100g rice bran flour, however had the lowest glycemic index. keywords: dough meal, fibre, glycemic index, optimisation, protein. citation | awolu, o.o.; ajibola, c.f.; adeloye, j.b.; ogwu, f. (2018). optimization of proximate and minerals compositions of sweet potato, soybean and rice bran composite flours for production of low glycemic index dough meal. agriculture and food sciences research, 7(1): 79-88. history: received: 7 february 2020 revised: 16 march 2020 accepted: 20 april 2020 published: 18 may 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 80 2. materials and methods ................................................................................................................................................................... 80 3. results and discussion ................................................................................................................................................................... 81 4. conclusions ....................................................................................................................................................................................... 83 references .............................................................................................................................................................................................. 83 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.79.88&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.79.88&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1643 https://orcid.org/0000-0002-5499-1888 agriculture and food sciences research, 2020, 7(1): 79-88 80 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to the existing literature on low glycaemic index foods by utilizing locally, cheaply and widely available raw materials (sweet potato, soybean and rice bran) in the preparation of low glycaemic index foods, rich in protein and dietary fibre. the effects of the raw materials on the chemical composition of the food were optimized using response surface methodology in order statistically arrive at best blends rich in protein, dietary fibre and minerals composition. 1. introduction the understanding of the impact of foods on actual blood sugar is defined as the glycaemic index. it differs from measurement of the carbohydrate contents in food. in this wise, food are ranked as very low, low, medium or high in glycemic index. it has been observed that foods with very low or low glycemic index are very good for people with cardiovascular diseases (especially type 2 diabetes, metabolic syndrome, stroke and depression). sweet potato is a popular low-fat diet and with a low glycemic index (gi), which makes it very healthy for diabetes patients. the most important edible parts are the roots and immature leaves, which are used for human consumption, animal feed and to some extent, for industrial purposes. sweet potato (ipomoea batatas) is rich protein, carbohydrates, minerals (calcium, iron, and potassium), betacarotenoids, dietary fibre, vitamins (especially c, folate, and b6), very little fat, and sodium [1]. as an excellent source of vitamin a, it can play an important role in the fight against vitamin a deficiency (vad). rice bran is a by-product of rice processing and rich in nutrients. it has very high fibre content and contain proteins are of high nutritional value [2]. the proteins are also rich in essential amino acids, especially lysine, hence can be used as ingredients in food recipes [3]. it is rich in vitamins including vitamin e, thiamin, niacin and minerals like aluminum, calcium, chlorine, iron, magnesium, manganese, phosphorus, potassium, sodium and zinc [4, 5]. the soybean seeds contain high quality and quantity of protein. its amino acid composition is approximate to composition of animal proteins, and therefore it is often used as replacement component of meat protein. lysine, a limiting amino acid in cereal proteins is abundant in soybeans hence the use of soybeans as supplementary and complementary lysine source in cereal-based products. the application of composite flour has resulted in flours and food products with improved compositional and nutritional qualities [6, 7]. use of composite flour encourages the utilization of local crops and reduces the expenses on wheat importation. in addition, composite flour has been optimized using response surface methodology in order to evaluate the significant effect of the raw materials on some nutritional and rheological qualities of composite flour [7, 8]. the aim of this research was to investigate the proximate and minerals compositions of dough meal from composite flour comprising of sweet potato flour, rice bran flour, and soymeal flour. the compositions will be optimised using response surface methodology, in order to evaluate the significant effect of the raw materials on the minerals and proximate compositions. the glycemic index of the optimized meal will be evaluated. 2. materials and methods 2.1. materials sweet potato was obtained from a local market, akure, ondo state. rice bran was purchased from igbemo –ekiti, ekiti state. soybean meal was obtained at jof, owo, ondo state. all chemicals and reagents used was of analytical grade. 2.2. sweet potato flour preparation sweet potato tubers were sorted, washed, blanched at 80 oc for 5 min, peeled, sliced and dried at 60 oc for 24 h. thereafter, it was milled using the hammer mill and passed through mesh size of 250 µm to have a fine flour which was then packed in an airtight container for further use. 2.3. soybean meal flour preparation the soybean meal was milled and sieved to obtain a fine soybean meal flour according to iita procedure [9]. 2.4. rice bran flour preparation the rice bran was destoned, milled and sieved through a 250 µm mesh size to produce a fine rice bran flour [10]. 2.5. experimental design the experimental design was carried out using optimal mixture model of response surface methodology (design expert 8.0.3.1 software). the responses were proximate properties (protein, carbohydrates, ash, crude fibre, fat and moisture) and mineral (calcium, iron, potassium, magnesium, phosphorus, sodium and zinc) composition, while the variables were the raw materials (sweet potato flour, rice bran flour and soybean meal flour), as presented in table 1. agriculture and food sciences research, 2020, 7(1): 79-88 81 © 2020 by the authors; licensee asian online journal publishing group table-1. factor design and their level of mixture. run component 1 a: g/100g component 2 b: g/100g component 3 c: g/100g 1 65.000 25.000 10.000 2 85.000 12.500 2.500 3 85.000 10.000 5.000 4 77.500 16.500 6.000 5 72.500 25.000 2.500 6 76.667 20.833 2.500 7 85.000 12.500 2.500 8 75.000 15.000 10.000 9 85.000 10.000 5.000 10 70.000 20.000 10.000 11 68.750 25.000 6.250 12 72.500 25.000 2.500 13 65.000 25.000 10.000 14 80.000 10.000 10.000 15 80.000 10.000 10.000 16 73.125 20.750 6.125 note: where, a= sweet potato flour; b= soybean meal flour; c= rice bran flour. 2.6. dough preparation dough was made from the composite flours in the right proportion table 1 with hot water in a ratio of 100 g of flour to 25 g of water to form a thick paste and then was placed on a gas cooker at a temperature of 45 oc for about 10 min. a good stirring was done and dough obtained was allowed to cool. 2.7. determination of the proximate composition of dough meal the proximate composition including moisture, ash content, crude protein, crude fat and crude fibre of the dough meal were determined using aoac methods [11]. the carbohydrate was estimated by difference. 2.8. mineral analysis of the dough meal determination of calcium, magnesium, zinc and iron was by atomic absorption spectrophotometer (aas), while potassium, phosphorus and sodium was determined by flame photometry method following aoac procedures [11]. 2.9. glycemic index of the dough meal the glycemic index was determined by using wolever method [12]. the glycemic index of the food samples were determined by feeding the samples to rat for a period of 2 h. the bloods of the rats were then used to calculate for the glycemic index level in the food samples. 2.10. statistical data analysis all analysis was done in triplicate and results presented as the average of triplicate determinations, expressed as mean ± standard deviation (s.d). the data were subjected to the statistical analysis of variance (anova) using spss software version 17.0. 3. results and discussion 3.1. proximate compositions of dough meal the result of the proximate composition is presented in table 2, and the contour plots presented in figures 1a to 1f. the carbohydrate content had no significant model terms, while the r2 and adjusted r2 values were also low figure 1a, despite the fact that the values of the carbohydrate contents vary from 57.32 to 62.52%. the values could indicate that although, the dough meal would be a good source of energy, yet, it would not be a high glycemic index food. the fat content was moderately high (9.09-13.19 g/100g). soybean supports the fat content of the composite flour as shown in the contour plot figure 1b. the significant model terms (linear mixture, ab, a2bc) also showed that sample c (rice bran) did not significantly (p> 0.05) contribute to the fat content of the composite flour table 2. the fat content of the composite flour is pointer to the fact that it would be good as flavour retainer, and a carrier of fat-soluble vitamins. agriculture and food sciences research, 2020, 7(1): 79-88 82 © 2020 by the authors; licensee asian online journal publishing group table-2. anova results for proximate composition. parameter model (significant level) r2 adj r2 lack of fit (p  0.05) significant (p  0.05) terms carbohydrate cubic 0.8253 0.5633 0.3988 nil crude fat special quartic (< 0.0001) 0.9948 0.9889 < 0.0001 linear mixture, ab, a2bc crude protein cubic (< 0.0001) 0.9998 0.9995 0.4135 linear mixture, ab, ac, bc, abc, ab(ab), ac (a-c), bc (bc) crude fibre special quartic (< 0.0001) 0.9998 0.9995 0.0002 linear mixture, ac, bc, abc2 total ash linear mixture (0.3131) 0.1636 0.0349 0.9189 nil moisture special quartic (0.0003) 0.9607 0.9157 0.5897 linear mixture, ac, ab, ab2c, abc2 the dough meal showed significant (p <0.05) increase in crude protein content ranging from 8.75 10.87% with increase in soybean meal and rice bran flours. however, dough meal with high sweet potato flour and low soymeal flour had low protein content [13]. the optimisation results table 2 showed that the protein had high r2 (0.9998) and adjusted r2 (0.9995) values. the model (cubic) and model terms (linear mixture, ab, ac, bc, abc, ab(a-b), ac (a-c), bc (b-c)) were significant (p 0.05) with a nonsignificant lack of fit. this r2 and adjusted r2 values together with the significant model terms for protein contents indicated that the composite flour prepared in this study had reasonable and acceptable protein contents. high r2 and adjusted r2 values (close to one) indicates statistical models that are fit [10, 14]. the contour plot representing the effect of the raw materials on the protein content is shown in figure 1c. these results showed that the composite flours making the dough significantly (p 0.05) effect the protein content. the areas shaded red in the contour plot are the areas with highest protein content. it could be seen that the areas with high protein content were as a result of the soy bean and potato flours incorporation. the crude fibre ranged from 1.96-5.83%, with high r2 and adjusted r2 values table 2. rice bran flour was the best source of crude fibre as indicated by the r2 value, adjusted r2 value, and significant model terms table 2. the contour plot is shown in figure 1d. rice bran is rich in soluble fiber like beta-glucan, pectin, gums helpful in reduction of serum cholesterol, certain forms of cancer and constipation [15]. nutritional and functional properties of rice bran are well suited for baked products like cookies, muffins, breads, crackers, pastries and pancakes [16]. the ash contents too had no significant model terms together with very low r2 and adjusted r2 values figure 1e and table 2. although the total ash contents ranged from 1.49 to 3.00%. the moisture content decreased significantly (p 0.05) as in soybean meal and rice bran flour contents increased. the r2 and adjusted r2 values were 0.9607 and 0.5897 respectively. the drastic reduction in the adjusted r2 value is an indication that the composite flour does not strongly support water content, which indicates that the dough meal would be shelf stable. the contour plot for carbohydrate content is shown in figure 1f. 3.2. minerals compositions of dough meal the optimisation results of the minerals composition, consisting the model, r2 value, adjusted r2 value, lack of fit and significant model terms is presented in table 3. the contour plot showing the graphical illustration of the effect of the raw materials on the dough meal minerals composition is shown in figures 2a to 2g. from the optimisation results in table 2, only potassium (k), magnesium (mg) and calcium (ca) had considerable r2 and adjusted r2 values meaning that they were enhanced by the raw materials. in addition, the dough meal had considerable effect on iron (fe) content figure 2a. from the contour plot in figure 2a, sweet potato and soy bean flours were the main sources of iron in the dough meal. it has been reported that sweet potato is a reliable source of mineral iron [17]. the dough meal had low sodium content, as it had no significant (p0.05) model term table 3. the sodium content of the dough meal ranged from 23.62-58.72 mg/kg while the potassium content ranged from 180.77-266.79 mg/kg. the contour plot figure 2b also showed that the raw materials contributed very little to the dough meal. none of the raw material increased the sodium content. the considerable high potassium content in relation to the considerable low sodium content is an indication that the dough meal would be suitable for better cardiovascular functions. high k/na ratio is recommended for patients with high blood pressure (k/na < 1.0). sweet potato had positive effect on the potassium content figure 2c. the plot also indicated that sweet potato was the main source of potassium. agriculture and food sciences research, 2020, 7(1): 79-88 83 © 2020 by the authors; licensee asian online journal publishing group table-3. anova results for minerals composition. parameter model (significant level) r2 adj r2 lack of fit (p  0.05) significant (p  0.05) terms fe cubic 0.8241 0.5603 0.8907 nil k linear mixture (p< 0.0001) 0.7734 0.7385 < 0.0001 linear mixture mg linear mixture (p< 0.0001) 0.8150 0.7866 0.9625 linear mixture ca special quartic (p< 0.0001) 0.9968 0.9931 0.0028 linear mixture p linear mixture (p=0.0579) 0.3549 0.2557 0.9267 nil na quadratic (p=0.1890) 0.4817 0.2225 0.1485 nil zn mean 0.0000 0.0000 0.8917 nil the calcium content of the dough meal significantly increased (p<0.05) as the level of soybean meal flour increased figure 2d. the contour plot figure 2d clearly shown that soybean was the main source of calcium, followed by rice bran flour. the phosphorus content of the dough meal significantly (p<0.05) increased as the substitution levels of sweet potato in the dough meal increased figure 2e. the 3d plot showed revealed that sweet potato was the richest source of calcium among the raw materials, followed by rice bran. the zinc content ranged from 0.34-2.09 mg/kg. it was observed from the 3d plot figure 2f that the raw materials were not rich in mineral zinc. zinc is a micronutrient; hence, the content could be sufficient for daily nutritional needs. soybean flour was the best source of magnesium followed by rice bran flour figure 2g. the magnesium contents for the samples varied from 30.58 to 43.44 mg/kg. 3.3. glycemic index of the dough meal the results of the optimisation of the proximate and minerals composition has given pointers to the possibility of the dough meal to have cardiovascular benefits. the r2 and adjusted r2 values for the optimisation carbohydrate composition were 0.5633 and 0.3988 respectively, while it has no significant model terms. this showed that the raw materials may not support sugar (from carbohydrate metabolism) contents in the dough meal. it implies that the dough meal would be useful for consumers against incidences or prevention of diabetic mellitus. the glycemic index (gi) of the dough meal ranged from 22.66 to 45.37. they can however, be classified as low gi meal. low gi foods have been shown to release glucose slowly and steadily [18]. they are slowly digested, absorbed and metabolised, leading to a slower rise in blood glucose level. the sample with 75.00 g/100g sweet potato flour, 15.00 g/100g soybean meal flour and 10.00 g/100g rice bran had the least glycemic index. the profile of glycemic evaluation of the dough meal from composite flour blends of sweet potato, soybean meal and rice bran in their different mixing ratio is shown in figure 3. 4. conclusions the dough meal prepared from flour blends of sweet potato, soybean meal and rice bran flour exhibited good and acceptable glycemic index properties. in addition, the dough had high protein and fibre contents. the na/k ratio of the dough meals were lower than 1; 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[18] v. gourineni, m. l. stewart, r. skorge, and t. wolever, "glycemic index of slowly digestible carbohydrate alone and in powdered drink-mix," nutrients, vol. 11, p. 1228, 2019. available at: https://doi.org/10.3390/nu11061228. figure-1a. contour plot showing the effect of variables on carbohydrate content. figure-1c. contour plot showing the effect of variables on protein content. figure-1d. contour plot showing the effect of variables on fibre content. agriculture and food sciences research, 2020, 7(1): 79-88 85 © 2020 by the authors; licensee asian online journal publishing group figure-1e. contour plot showing the effect of variables on ash content. figure-1f. contour plot showing the effect of variables on carbohydrate. figure-2a. contour plot showing the influence of variables on iron (fe). agriculture and food sciences research, 2020, 7(1): 79-88 86 © 2020 by the authors; licensee asian online journal publishing group figure-2b. contour plot showing the influence of variables on sodium (na). figure-2c. contour plot showing the influence of variables on potassium (k). figure-2d. contour plot showing the influence of variables on calcium (ca). agriculture and food sciences research, 2020, 7(1): 79-88 87 © 2020 by the authors; licensee asian online journal publishing group figure-2e.contour plot showing the influence of variables on phosphorus (p). figure-2f. contour plot showing the influence of variables on zinc (zn). figure-2g. contour plot showing the influence of variables on magnesium (mg). agriculture and food sciences research, 2020, 7(1): 79-88 88 © 2020 by the authors; licensee asian online journal publishing group figure-3. the glycemic index of the dough meal. : asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 166 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 2, 166-171, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.62.166.171 © 2019 by the authors; licensee asian online journal publishing group nutrient content assessment of six citrus species parts and their feedstuff significance chinelo a. ezeabara1 okeke c.u.2 ( corresponding author) 1,2department of botany, nnamdi azikiwe university, p.m.b. 5025, awka, anambra state, nigeria. abstract insufficient and high cost of good quality feed is the primary limitation to livestock operation. hence, assessment of surplus atypical feed resources for their nutritive potential to support livestock productivity becomes topical. dry matter bases of the roots, stems, stem barks, leaves and peels of six citrus species that are commonly available in southeastern nigeria were investigated for nutrient composition. high value of crude protein ranging from 15 to 18 percent was found in the leaves of all the species. high carbohydrate values ranged from 70 to 81 percent, in the peels of c. grandis, c. reticulata, c. paradisii and c. sinensis. high percentage of fat was also present in the range of 12 to 13% in the peels of c. aurantifolia and c. sinensis. ash contents were very high in the leaves of c. aurantifolia, c. grandis, c. limon and c. paradisii, ranging from 10 to 14%. therefore, leaves and peels of these citrus species could be regarded as low cost and locally available alternative high-quality animal feeds. keywords: animal feed, animal protein, animal nutrition, citrus leaves, citrus peels, crude protein, feed quality, nutritive value. citation | chinelo a. ezeabara; okeke c.u. (2019). nutrient content assessment of six citrus species parts and their feedstuff significance. agriculture and food sciences research, 6(2): 166-171. history: received: 28 march 2019 revised: 8 may 2019 accepted: 10 june 2019 published: 22 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group contribution/acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 167 2. materials and methods ................................................................................................................................................................. 167 3. results .............................................................................................................................................................................................. 168 4. discussion ........................................................................................................................................................................................ 169 5. conclusion ....................................................................................................................................................................................... 169 references ............................................................................................................................................................................................ 170 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.166.171&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.166.171&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/866 agriculture and food sciences research, 2019, 6(2): 166-171 167 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study suggested that citrus aurantifolia (christm.) swingle, c. grandis osbeck, c. limon (l.) burm. f., c. paradisii macf., c. reticulata blanco and c. sinensis (l.) osbeck leaves and peels are potential alternative high-quality animal feeds. 1. introduction citrus belongs to the family rutaceae. six species of citrus are commonly available in southeastern nigeria. they include: citrus aurantifolia (christm.) swingle (lime), c. grandis osbeck (shaddock/pummelo), c. limon (l.) burm. f. (lemon), c. paradisii macf. (grapefruit), c. reticulata blanco (mandarin/tangerine), and c. sinensis (l.) osbeck (sweet orange).the members of this genus are small trees or large shrubs, reaching 5 to 15 meters tall, with spiny shoots and alternately arranged evergreen leaves with an entire margin [1, 2]. the coriaceous evergreen leaves are gland dotted and unifoliate-compound with a joint between leaf blade and petiole [1, 3]. citrus fruits are one of the world’s most crucial, tasty and juicy fruit of rutaceae [4, 5].they have long been regarded as important sources of vitamin c; medicinal plant[6]and in the manufacture of household products. however, only the juices and the essential oils, which the antimicrobial property [7] and anti-aflatoxigenic activity [8] have been investigated. the use of citrus pulp as a common animal feed had also been reported [9]. thus, it becomes absolutely imperative to investigate other citrus plants parts for nutritional composition with the view to support livestock productivity. good nutrition is an integral part of successful livestock operation. nutrients are chemical substances in food that nourish the body by providing energy, building materials, and factors to regulate needed chemical reactions in the body [10]. the groundwork of livestock production system is the accessibility of quality and quantity of animal feed, with feed costs taking up to 70% of the cost of production [11]. moreover, improper animal feeding and nutrition, either unbalanced diet, under or over feeding of livestock affect the general well-being of the animals, including lowering their body immune system. protein is one of the most expensive nutrients and an extremely important nutrient in all animal feed. consumption of small protein and food energy could develop protein-energy malnutrition [12]. the deficiency eventually results in body wasting, primarily of lean tissue, and increased susceptibility to infections. moreover, the deliberation for herdsmen to rear cattle in ranches in nigeria is ongoing. nigeria had over 14.73 million cattle consisting of 1.47 million milking cows and 13.26 million beef cattle in 2008 [13, 14]. the number must have undoubtedly increased by now. plant materials are the chief diets of livestock. high cost of animal feeds is among the major factors limiting animal production in nigeria [15, 16]. there is shortage of good quality feeds needed to sustain livestock growth especially during dry season, as a result of scarcity of forages. although many livestock farmers formulate their own animal feed, they have challenges of access to low cost, good quality and easily available raw materials. besides, feeding animals with crops could reduce the available food for humanity, considering the problem of food shortage as a result of human population explosion in developing countries. hence, other alternative sources such as agro-industrial by-products and crop residues are being assessed for their nutritive quality to support livestock productivity. the objective of this study therefore, is to determine the nutritive value of various citrus species parts that are predominant in southeastern, nigeria. 2. materials and methods 2.1. sources of materials the roots, stems, stem barks, leaves and fruits of citrus aurantifolia, c. grandis, c. limon, c. paradisii, c. reticulata and c. sinensis were collected in the months of november – december at optimum maturity, from agricultural and natural resources department market garden, amawbia, anambra state, nigeria. the voucher specimens were deposited at department of botany herbarium, nnamdi azikiwe university, awka, anambra state, nigeria. 2.2. preparation of plant materials the rinds of healthy ripe fruits of the six citrus species were peeled off with a knife. the roots, stems, stem barks and peels were sun dried for seven days whereas the leaves were air dried in the laboratory at room temperature for ten days. the dried samples (about 1.5kg) were then crushed with mortar and pestle before grinding into fine powder using a manual grinder (corona, usa). 2.3. proximate determination the nutrient content of the six citrus species parts were determined according to the methods described by onwuka [17]. 2.3.1. moisture content determination the dishes were washed thoroughly and dried in the oven. they were latter put inside the dessicator to cool. after which they were weighed. sample was put into the weighed dish and weight taken. the sample was dried in the oven at 70 ᴼc for 2 hours and at 105 ᴼc for the next 4 hours. the sample was cooled in the dessicator and the dry weight of sample plus dish taken. the moisture content was calculated as follows: percentage (%) moisture = w2− w3 × 100 w2− w1 1 where: w1 = initial weight of empty crucible w2 = weight of crucible + sample before drying w3 = final weight of crucible + sample after drying 2.3.2. ash content determination agriculture and food sciences research, 2019, 6(2): 166-171 168 © 2019 by the authors; licensee asian online journal publishing group five grams (5g) of finely ground dry sample was weighed into a tarred silica crucible. the sample was charred on a heater inside a fume cupboard, to dry off most of the smoke. the sample was transferred into a pre-heated muffle furnace at 550 ᴼc. it was left at this temperature for 2 hours. after which it was cooled in a dessicator and reweighed. percentage (%) ash = weight of ash × 100 weight of original sample 1 = w3 –w1 × 100 w2 –w1 1 where: w1 =weight of empty crucible w2 =weight of crucible +sample before drying w3 = weight of crucible + ash 2.3.3. crude fibre content determination water reflux was boiled for 30 minutes with 200 ml of a solution containing 1.25g of h2so4 per 100 ml of solution. the solution was filtered through two fold of cheese cloth on a fluted funnel. the residue was washed with boiling water until there was no longer acid. the residue was transferred to a beaker and boiled for 30 minute with 200 ml of a solution containing 1.25g of carbonate-free naoh per 100ml.the final residue was filtered through a thin but closed pad of washed and ignited asbestos in a gooch crucible. it was dried in an electric oven and weighed. it was finally incinerated, cooled and weighed. the loss in weight after incineration × 100 is the percentage of crude fibre. 2.3.4. crude protein content determination a measured weight (2g) of sample was weighed into a 250ml beaker. after which 75ml of hot water was added and brought to boil. it was stirred vigorously and added 25ml of 6% copper sulphate solution. it was again brought to boil, stirred vigorously and added 25ml of the 1.25% sodium hydroxide solution. the mixture was stirred vigorously, removed from the flame and allow to settle. it was filtered through a 15 cm, no. 4 whatman paper. the precipitate was cleaned from the sides of the beaker with a ‘policeman’. the paper was washed free from sulphate with very hot water 6 times. it was allowed to drain well, and then transferred to a kjeldahl flask containing about 10g anhydrous sodium hydroxide and a trace of selenium. thirty milliliters (30ml) of conc. h2so4 was added, nitrogen content determined and hence the protein content of the sample. percentage (%) protein = % n × f where: f = conversion factor (6.25) percentage (%) nitrogen = vs − vb × nacid × 0.028 × 100 w where: vs = volume of acid required to titrate sample in milliliters vb = volume of acid required to titrate blank in milliliters nacid = normality of acid (0.1n) w = weight of sample in grams therefore, percentage (%) protein = % nitrogen ×6.25. 2.3.5. fat content determination the sample was mixed with a mixture of methanol and chloroform in such proportions to give a single phase miscible with water. further, extra chloroform was added to give a separation of phases. the solvents were separated by centrifugation. the chloroform layer containing the dissolved fat was removed and fat residue left behind weighed. percentage (%) fat = weight of fat × 100 weight of sample 1 2.3.6. carbohydrate content determination carbohydrate content was determined by difference method: percentage (%) carbohydrate = 100 – (% moisture + % ash + % crude fibre + % crude protein + % fat) 2.4. statistical analysis the data obtained were statistically analyzed using one-way-anova and the significant difference was established at (p>0.05). the results were expressed as mean + sd. 3. results considerable high levels of the basic nutrients were present in parts of the six citrus species table 1. high levels of carbohydrate were detected in the stems of c. aurantifolia (67.53+0.08%) and c. limon (50.13+0.04%) as well as in the peels of c. grandis (81.2+0.06%), c. reticulata (75.4+0.08%), c. paradisii (80.6+0.4%) and c. sinensis (70.4+0.05%). high percentages of fat and crude fibre occurred in the peels of c. aurantifolia (12.0+0.4, 8.30+0.4), c. limon (9.01+0.04, 9.37+0.02) and c. sinensis (13.89+0.02, 7.86+0.02) respectively. fat content was also high in the leaf of c. reticulata (11.10+0.05%) and in the stem bark and root of c. paradisii (6.51+0.06, 6.28+0.4%) respectively. in addition, high levels of crude fibre were contained in the leaf of c. grandis (7.27+0.03%); peel of c. reticulata (9.75+0.03%) and leaf of c. paradisii (8.31+0.02%). ash contents were very high in the leaves of c. aurantifolia (14.11+0.02%), c. limon (14.15+0.02%) and c. paradisii (13.01+0.01%).it was also high in the roots of c. agriculture and food sciences research, 2019, 6(2): 166-171 169 © 2019 by the authors; licensee asian online journal publishing group grandis (12.72+0.04%) as well as in the stem bark of c. reticulata (15.18+0.02%) and c. sinensis (17.01+0.01%). appreciable high values of ash were also detected in the leaves of c. grandis (10.18+0.02%) and root of c. paradisii (10.14+0.01%) table1. 4. discussion high carbohydrate values were ranged from 70 to 81 percent in the peels of c. grandis, c. reticulata, c. paradisii and c. sinensis. carbohydrate and fat provide animal body with energy. energy (mainly from carbohydrates) and protein are the primary macro nutrients to take into account, for beef cattle nutrition [9]. high percentage of fat was also present in the range of 12 to 13 in the peels of c. aurantifolia and c. sinensis. fat is a solvent for fat-soluble vitamins and hormones. reasonable levels of crude fibre were found in the leaves (except c. sinensis) and peels (with the exception of c. grandis), of all the citrus species tested. fibre promotes digestion, cleanses the digestive tract, prevents absorption of excess cholesterol [18] and aids in bowel movement by adding mass to the intestinal content [19]. digestion of animal feed is therefore, determined by the crude fibre content [20]. ash contents were very high in the leaves of c. aurantifolia, c. grandis, c. limon and c. paradisii, ranging from 10 to 14%. the ash values were higher than the ones found in leaves 10.85% of ipomoea batatas (l.) lam. (sweet potatoes) [15]. ash of a food material contains mineral matter. mineral salts are needed for regulation of body metabolism. crude protein was found in high levels ranging from 15 to 18 percent in the leaves of all the citrus species investigated. the percentage crude protein values were higher than the levels present in the zea mays l. (corn) grain 9.5−11.2, sorghum bicolor (l.) moench (milo, sorghum) grain 10.3−11.0 and glycine max l. merr. (soybean) leaf 11.0−13.1 [21]. moreover, the values in the peels of c. aurantifolia, c. limon, c. reticulata and c. sinensis that ranged from 6.4−10.7% were higher than the levels found in leaf 6.2−7.8%, husk 3.0−4.0%, cob 2.1−3.8% and stalk 3.4−4.9% of corn; stem 3.6−4.5% and pod 4.5−9.0% of soybean, and stalk 3.3−3.9% of milo. beef is the major meat consumed in nigeria and the meat quality is primarily dependent on the feed quality. pastoralism, agro-pastoralism and ranching systems are three major beef cattle production systems in nigeria [22] with the pastoralism and agro-pastoralism being the principal systems. ranching system involves semi-intensive and intensive feeding systems, where the animals’ nutritional needs are supplemented with diets as additional feed. the nutrient requirements of animals differ because of a number of factors, including age, body condition, body size, pregnancy and lactation. these factors as well as forage quantity and forage quality are considered in determining the supplementation need of animals. for sustainable farming, easy availability of local feeds and supplements is essential. this can be achieved by sourcing for local plant species of high nutritional values. according to united states department of agriculture [23] the development of animal feeds entails mixing various readily obtainable and low cost feed materials into a ration that will provide nutrient requirements for animals. in addition, having the information on the chemical composition of the various available feed ingredients is a critical factor. the dietary crude protein used in diets formulated by some consultants ranged from 12.5 to 14.4% but most consultants use at least 13% crude protein [24] whilst at least 15 percent crude protein is required for creep feeds or forages for nursing calves [25]. this indicated that leaves of these citrus species could be used as protein supplement to enrich animal feed, because protein is needed by animal body in large amounts. sweet potato vine silage contained 136.1 g kg-1 dm crude protein and had been suggested for supplementing tropical cattle [26]. soybean contained 44% [27] and 49.0-52.0% crude protein value[21]. soybean meal is the most common protein source for all compound feeds for pigs, poultry and dairy cattle globally; however, it is seldom too costly [27].the finding therefore, suggested an inclusion of citrus plants leaves and peels in plant food protein, indicating that they can potentially be used as easily available protein sources for livestock production. providing adequate protein in beef cattle diets is essential for animal health and productivity as well as ranch profitability [25]. besides, young, growing cattle, in particular, need relatively high levels of crude protein in their diets to support muscle growth. moreover, when forage crude protein levels fall below 6 to 8%, the protein supplementation is highly needed[28]. this is as a result of low quality of the forage. citrus parts could therefore, be used to solve the problem of shortage of good quality feeds needed to sustain livestock growth especially during the dry season, which in turn, will support livestock productivity. insufficient good quality animal feeds result to serious effects on cattle such as lessened pregnancy rates; loss of body condition of the cow, and reduce milk production, hence lowers weaning weights[29]. in addition, other alternative feeds are mostly in short supply and costly. cost of feeding is the most expensive part of livestock production [30]. cost is also the primary factor for supplementing additional protein in finishing diets [24]. these indicated that the leaves and peels of these citrus species could potentially be used as inexpensive and readily accessible feedstuffs for livestock production. moreover, high level of crude protein in these citrus species leaves indicated that they can serve as cheap sources of protein. 5. conclusion the study revealed that the leaves and peels of these citrus species contained high nutritive values. they are also environmentally friendly, readily available and relatively cheap. hence, suggesting their potential use in development of animal feed for more sustainable feeding system. in addition, protein is the most expensive and critical nutrient needed by animal body in large amounts. therefore, incorporation of these parts as protein supplement in animal feeds to enhance animal nutrition is recommended. further research however, is necessary in order to assess the effect of the diet on animal nutrient digestibility and growth performance. agriculture and food sciences research, 2019, 6(2): 166-171 170 © 2019 by the authors; licensee asian online journal publishing group table-1. proximate composition of leaf, peel, stem, stem bark and root of six citrus species. citrus species nutrients plant parts (%) leaf peel stem stem bark root citrus aurantifolia crude protein 16.69+0.07 6.38+0.4 6.27+0.04 7.81+0.01 9.38+0.02 carbohydrate 41.76+0.04 53.89+0.4 67.53+0.08 56.86+0.12 55.17+0.01 fat 9.39+0.01 12.0+0.4 2.88+0.03 6.17+0.03 5.71+0.01 ash 14.11+0.02 5.82+0.04 6.47+0.01 11.27+0.03 13.61+0.01 crude fibre 6.77+0.02 8.30+0.4 5.74+0.03 5.68+0.03 5.23+0.4 moisture 12.86+0.04 11.29+0.06 11.13+0.01 12.07+0.02 10.87+0.02 c. grandis crude protein 16.27+0.04 1.0+0.02 6.52+0.01 6.32+0.4 8.77+0.3 carbohydrate 49.72+0.1 81.2+0.06 66.63+0.10 62.14+0.05 56.85+0.4 fat 5.24+0.4 0.7+0.01 2.77+0.03 5.85+0.4 5.25+1.01 ash 10.18+0.02 2.1+0.05 7.27+0.2 8.18+0.02 12.72+0.04 crude fibre 7.27+0.03 4.5+0.4 5.28+0.01 4.81+0.01 5.85+0.4 moisture 11.33+0.03 19.2+0.03 11.45+0.05 12.69+0.01 10.57+0.01 c. limon crude protein 15.51+0.04 7.30+0.01 4.80+0.01 7.02+0.03 10.25+0.01 carbohydrate 39.03+0.01 41.35+0.03 50.13+0.04 47.05+0.1 42.03+0.02 fat 7.23+0.03 9.01+0.04 1.13+0.03 5.25+1.01 4.35+0.01 ash 14.15+0.02 5.91+0.01 6.01+0.01 12.08+0.04 11.05+0.05 crude fibre 6.80+0.01 9.37+0.02 5.86+0.01 6.30+0.01 7.52+0.02 moisture 12.31+0.04 15.31+0.05 13.01+0.05 14.07+0.02 12.70+04 c. reticulata crude protein 15.86+0.04 8.95+0.03 7.29+0.01 6.21+0.02 8.79+0.04 carbohydrate 42.17+0.06 75.4+0.08 69.31+0.06 56.84+0.05 56.14+0.07 fat 11.10+0.05 9.02+0.4 3.07+0.01 6.29+0.01 6.04+0.02 ash 11.53+0.08 13.04+0.04 4.28+0.02 15.18+0.02 13.34+0.06 crude fibre 6.81+0.3 9.75+0.03 4.91+0.01 4.74+0.02 5.30+04 moisture 12.54+0.02 15.32+0.02 11.16+0.04 10.76+0.03 10.41+0.02 c. paradisii crude protein 18.13+0.05 0.4+0.01 6.43+0.4 6.01+0.03 8.41+0.03 carbohydrate 40.65+0.04 80.6+0.4 60.48+0.07 58.31+0.05 51.02+0.01 fat 5.03+0.4 0.3+0.01 2.83+0.03 6.51+0.06 6.28+0.4 ash 13.01+0.01 1.30+0.02 6.32+0.04 7.82+0.04 10.14+0.01 crude fibre 8.31+0.02 2.3+0.05 5.01+0.01 4.98+0.02 4.71+0.01 moisture 13.31+0.02 17.4+0.03 9.33+0.02 14.05+0.05 12.31+0.04 c. sinensis crude protein 16.03+0.02 10.74+0.01 8.09+0.02 6.35+0.01 9.31+0.04 carbohydrate 39.97+0.07 70.4+0.05 61.48+0.02 53.71+0.02 41.08+0.02 fat 11.53+0.08 13.89+0.02 5.01+0.03 7.31+0.03 8.01+0.02 ash 10.61+0.02 11.90+0.07 5.31+0.05 17.01+0.01 11.57+0.03 crude fibre 4.93+0.02 7.86+0.02 5.05+0.01 5.31+0.07 4.27+0.01 moisture 14.61+0.02 15.71+0.03 12.09+0.01 10.93+0.03 9.91+0.4 note: results are in mean ± se of triplicate determinations, (p>0.05). references [1] b. p. pandey, a text book of botany angiosperms, 2nd ed. new delhi: chand and company limited, 1981. 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[30] u. k. saha, s. s. sonon, d. hancock, n. s. hill, l. stewart, g. l. heusner, and d. e. kissel, common terms used in animal feeding and nutrition. university of georgia: uga cooperative extension bulletin 1367, 2017. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 17 © 2022 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 9, no. 1, 17-21, 2022 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v9i1.3793 © 2022 by the authors; licensee asian online journal publishing group microbial quality and sensory properties of ogiri produced from watermelon (citrullus lanatus) seed akele, e. o.1  akinyosoye, f.a.2 oyetayo, v. o.3 ( corresponding author) 1department of microbiology, adekunle ajasin university, akungba akoko, ondo state, nigeria. 1email: akele.olatunbosun@aaua.edu.ng tel: 08035422430 2,3department of microbiology, federal university of technology, akure, ondo state, nigeria. 2email: faakinyosoye@futa.edu.ng tel: 08035017616 3email: ovonew67@gmail.com tel: 08034278243 abstract the aim is to study the microorganisms involved in fermentation, their occurrence and the organoleptic properties of fermented watermelon (citrullus lanatus) seeds. the watermelon seeds were dehulled, sorted, washed and boiled for 4 hours and fermented naturally for nine days. microorganisms isolated from the fermenting watermelon seeds were bacillus sp, lactococcus sp, pediococcus sp, streptococcus sp, micrococcus sp and alcaligenes ps for bacterial, while the fungi obtained were mucor sp, rhizopus sp, fusarium sp, candida sp and saccharomyces sp. the bacterial count increased from 4 x101 cfu/ml to 1.21 x 1010 cfu/ml while fungal count also increased from 8 x 101sfu/ml to 1.2 x 109sfu/ml during the period of fermentation. the ph and temperature increased from 6.3 to 8.7 and 29oc to 40oc respectively. the total titratable acidity decreased from 0.176% to 0.027%. the mean sensory characteristics value of fermented sample as scored by the panelist shows that the texture of the fermenting sample become softer as fermentation progressed. the aroma became more pungent with a strong ammonia odour responsible for the characteristic flavour and a darker colour of product. the overall acceptability of fermented sample was adjudged like very much while the sample bought from the market was adjudged like moderately. keywords: fermentation, microorganisms, temperature, sensory, panelist, acceptability, organoleptic, titratable acidity. citation | akele, e. o.; akinyosoye, f.a.; oyetayo, v. o. (2022). microbial quality and sensory properties of ogiri produced from watermelon (citrullus lanatus) seed. agriculture and food sciences research, 9(1): 17-21. history: received: 14 january 2022 revised: 23 february 2022 accepted: 8 march 2022 published: 18 march 2022 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. authors’ contributions: all authors contributed equally to the conception and design of the study. competing interests: the authors declare that they have no conflict of interest. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study followed all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 18 2. materials and methods ................................................................................................................................................................... 18 3. results ................................................................................................................................................................................................ 19 4. results and discussion ................................................................................................................................................................... 19 5. conclusion ......................................................................................................................................................................................... 21 references .............................................................................................................................................................................................. 21 mailto:akele.olatunbosun@aaua.edu.ng mailto:faakinyosoye@futa.edu.ng mailto:ovonew67@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v9i1.3793 agriculture and food sciences research, 2022, 9(1): 17-21 18 © 2022 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by revealing that watermelon seed an unconventional substrate was fermented. organisms involved were microbiota from harvest and post-harvest environment, and they played a substantial role in the fermentation process. which resulted to the biochemical transformation which altered the appearance, texture and flavor of fermented watermelon seeds given it an overall acceptability adjudged like very much. which supports that fermented watermelon seeds could be additives in animal and baby weaning feed. 1. introduction ogiri is a fermented product of african oil bean. in nigeria castor oil, melon, fluted pumpkin, african yam and bambara seeds are used for the production of ogiri. such class of fermented product are known as condiment. condiment are fermented products usually produced from fermented vegetable proteins, they are craft-based [1]. the methods employed in the production of fermented condiments differ from one community to another in many areas. in nigeria, they are made in traditional ways, observance of good manufacturing practices and control of environmental conditions during the production phase gives a good product, consistency in product quality and stability is difficult to maintain because the fermentation is spontaneous and not controlled using starter culture [28]. the substrates used for the production of condiment are diverse as regions are diverse and more than one raw material can be used to produce condiments. most of the seeds used for condiment production are inedible in their raw, unfermented or cooked state [1]. watermelon (citrullus lanatus) is a horticultural crop known for its juicy sweet fruit [9-11]. it originated from namibia but has been domesticated in almost all parts of the world especially warmer climates [12]. it is an important but underutilized crop. the seeds are considered waste but research has reported it as a highly nutritive seed with large amounts of proteins and beneficial minerals [13]. watermelon seeds are one of the main underutilized fruits grown in warmer parts of the world [12]. therefore, knowledge on the nutritive and anti-nutritive content of watermelon seed will encourage their consumption in different ways and the re-utilization of the large quantity of seeds thrown away as waste even with their hidden nutrients [12]. presently, there are limited literatures on the microbiological effect on the nutritional properties. in this study, the microbial quality and sensory properties of fermented seeds of watermelon used to produce a condiment was evaluated. 2. materials and methods 2.1. sampling sites matured, fresh, healthy watermelon fruits were bought from sasa market in akure, ondo state nigeria. the fruits were cut open with a sterile knife, the seeds were collected then washed in distilled water until clean, dried then dehulled and kept in airtight container until used. 2.2. sample collection and confirmation the sample was taken to the horticulturist in the department of plant science and biotechnology, adekunle ajasin universiy, akungba akoko, ondo state, nigeria where the seeds were confirmed as watermelon (citrullus lanatus) seeds. 2.3. sample preparation and microbiological analysis the prepared water melon seeds (600 g) were pressure cooked in the autoclave for 4 hours inside sterilized aluminum pot, excess water was drained off. a sterilized spatula made of stainless steel was used to take part of the boiled, watermelon seeds in to the fermenter. the boiled watermelon seeds were wrapped with jute bags put into calabash and covered with sterile cocoa leaves for nine days. 2.4. isolation and characterization of organisms involved one gram (1g) fermenting water melon seeds were aseptically collected at 24 hours intervals for nine days, grind into paste with a sterile mortar and pestle and was aseptically transferred into the stock solution, a test tube containing 9.0 ml sterile distilled water labelled as 10-1. the dilution process was carried out serially until it got to the test tube labelled as 10-9. experiment was in triplicate. nutrient agar (na) and potato dextrose agar (pda) were used for the isolation of bacteria and fungi respectively. the 10-3 to 10-9 of serially diluted sample above was used. in each diluted sample, 0.5 ml was transferred to the center of a sterile petri dish. about 20 ml of molten na and pda agar cooled to 45oc were then added to each plate respectively and rocked gently to facilitate mixing the agar with the sample. the na plates were placed at 37 ± 2oc for 24 hours, while pda plates were kept at 25 ± 2oc for 48-72 hr. the distinct bacterial colonies that grew on the na were examined physically for their morphological characteristics such as colour, shape, size, elevation, surface and edges. they were then manually counted. fungi were also observed for their morphological characteristics such as pigmentation during growth, hyphae form, conidia form, growth rate and diameter size of each colony was measured using a meter rule during growth on the pda plates. colony counting was carried out visually by counting the number of visible colonies that appeared on the plates. calculation of colony forming unit (cfu) per gram for the bacteria and the spore forming unit (sfu) per gram for the fungi was based on the formula: cfu/gram or sfu/gram = number of colonies × dilution factor ml of sample suspension 2.5. physico-chemical analysis 2.5.1. determination of ph fermenting watermelon seed (2g) was weighed and grind into smooth paste then homogenized with 20ml of sterile distilled water in a 50 ml beaker. the ph electrode was immersed into the solution and the reading was noted. this was done for nine days. a ph 211 microprocessor, hanna instruments. agriculture and food sciences research, 2022, 9(1): 17-21 19 © 2022 by the authors; licensee asian online journal publishing group 2.6. temperature and total titratable acidity (tta) the temperature was taken by inserting a sterile thermometer into the batch of fermenting watermelon seeds while standard method of aoac [14] was used for tta. 2.7. sensory evaluation twenty panelists were selected which consist of venders, producer and individuals. these panelists were used to evaluate the ogiri produced from watermelon seeds for organoleptic characteristics using the nine-point hedonic scale. 3. results table 1. microbial counts of watermelon seeds fermented for the production of a condiment. days cfu/ml sfu/ml 1 4 x101 8 x101 2 3.6 x104 5.2 x103 3 7 x 107 7.3 x 106 4 1.48 x109 1.68 x 107 5 1.82 x 109 1.82 x 108 6 1.78 x 1011 1.98 x 109 7 1.74 x 1011 1.72 x 109 8 1.56 x 1011 1.41 x 109 9 1.21 x 1011 1.20 x 109 4. results and discussion result of the total viable count during fermentation of watermelon seeds as shown in table 1 reflects that the organisms were in their exponential phase of growth [15] and they increased all through the fermentation period which shows that the fermenting watermelon seed was a suitable substrate for microbial growth. table 2a. cultural, morphological and biochemical characteristics of bacterial isolates obtained during fermentation of watermelon to produce condiment. s / n o g ra m s ta in in g c e ll s h a p e m o ti li ty sp o re s in d o l p ro d u c ti o n c it ra te u ti li za ti o n n it ra te r e d u c ti o n m e th y l re d v o g e s p ro sk a u e r o x id a se t e st c a ta la se t e st g lu c o se m a lt o se m a n it o l fr u c to se su c ro se la c to se so rb it o l t e n ta ti v e i d e n ti ty is1 rod + + + + + + + alcaligenes sp is2 + rod + + + + + + + + + bacillus subtilis is3 + rod + + + + + + + + bacillus megaterium is4 + rod + + + + + + + + bacillus pumilis is5 + rod + + + + + + + bacillus firmus is6 + cocci + + + + + + + micro coccus sp is7 + cocci + + + + + + + + + streptococcus sp is7 + rod + + + + + + lactococcus sp is9 rod + + + + + + pseudomonas sp is10 + cocci + + + + + pediococcus sp note: key: + positive, – negative. table 2b. cultural and morphological characteristics of fungal isolates, obtained from watermelon seeds fermented to produce condiment. s/n growth rate diameter of colony after 7days on pda pigmentation during growth hyphae form conidium form name of organism 1 fastidious at 25oc on pda 8cm 3days white to yellow tater dark grey black sporangia upright phialide branched out from the phialide mucor hiemalis 2 fastidious 8cm in 4days white to brownish upright phialide branched out from the phialide rhizopus stolonifer 3 moderate 7cm white to pink short, broadly ending nonproliferating phialides conidiophores richly branched but not in spirodochia usually one celled macro-conidia fusarium verticilliodes 4 moderate 4cm creamy pseudohypha formed ovid shape produced laterally mycelium candida krusei 5 moderate 5cm creamy pseudohypha formed ovid shape produced laterally mycelium saccharomyces sp 6 moderate 4cm creamy pseudohypha formed ovid shape produced laterally mycelium candida tropicalis agriculture and food sciences research, 2022, 9(1): 17-21 20 © 2022 by the authors; licensee asian online journal publishing group at day one, the total viable bacterial count was 4.0 x 101 cfu/g and increased to 1.21 x 1010 cfu/g on the ninth day. fungal count was 8 x101 sfu/g it increased to 1.20 x 109 sfu/g table 1. the cultural and morphological characteristic showed a continuous and consistent succession of microorganisms as reflected in table 2(a and b). the most predominant bacteria were bacillus species, and alcaligenes sp while for fungi sacharomyces cerevisae, candida sp, and mucor sp. were the most predominant. table 3. the presences of some organisms maybe due to contamination or microbiota from harvest or post-harvest environment. from the list of organisms isolated, only few of them can be considered to play a substantial role in the fermentation process. bacillus species were prominent from the start to the end of the fermentation process, bacillus species can grow on ammonia or nitrogen source during melon fermentation to produce condiment [16, 17]. the combination of bacillus species, and alcaligenes species during fermentation of melon condiment are capable of producing the quality characteristic of good fermented melon condiment [11]. result of physico-chemical parameters is shown in table 4. temperature increased from 29oc to 40oc. this may be due to increase in cell population and the metabolic activities going on within the fermenting condiment. the ph increased from 6.3 to 8.7. this could be due to the high protein content of the fermented melon which was hydrolysed to amino acid and ammonia by the dominant microorganisms as reported by odunfa [2]; sanni, et al. [18]. the total titratable acid (tta) decreased from 0.127% to 0.027%. the observed increase in ph and decrease in titratable acidity of the fermented food could be due to presences of some lactic acid bacteria which degrades carbohydrates table 3. these observations agreed with earlier studies by chukwu, et al. [19]; ojokoh, et al. [20] and ojokoh and orekoya [15]. table 3. occurrence of bacterial and fungal isolates during the fermentation of watermelon seed for in the production of condiment. name of organism day 1 day 2 day3 day4 day5 day6 day7 day8 day 9 alcaligenes sp bacillus subtilis + + + + + + + + + + + + + + + + bacillus pumilis + + + + + + + + bacillus firmus bacillus megaterium + + + + + + + + + + + + candida krusei + + + + + + + + + candida tropicalis + + + + + + + + fusarium verticuliodes + + + lactococcus lactis + + + + micrococcus sp + + + + mucor hiamalis + + + + + pseudomonas sp + + + + + + + pediococcus sp + + + rhyzopus stolonifera + + + + + saccharomyces sp streptococcus lactis + + + + + + + + + + + + note: key: + present, absent. table 4. physio-chemical parameter of watermelon seeds fermented for the production of a condiment. days ph tta (%) temp. (oc) 1 6.3 0.176 29 2 6.6 0.149 32 3 7.2 0.104 33 4 7.6 0.099 36 5 7.8 0.081 35 6 8.1 0.068 36 7 8.2 0.059 37 8 8.4 0.045 39 9 8.7 0.027 40 the organoleptic characteristics of the fermented sample was compared to ogiri sample bought from the market figure 1 generally, the colour of the laboratory fermented sample was darker, with a stronger ammonia odour [2123] in all the panelist preferred the fermented sample to the sample bought from the market. therefore, the fermented sample was adjudged like very much while the sample bought from the market was adjudged like moderately. figure 1. sensory properties of fermented watermelon seed and commercial condiment. agriculture and food sciences research, 2022, 9(1): 17-21 21 © 2022 by the authors; licensee asian online journal publishing group 5. conclusion the results from this study revealed that fermented condiment produced from watermelon seed was more accepted by the test panel. this unconventional substrate could therefore serve as a good alternative for the production of ogiri, a local condiment. references [1] o. achi, "the potential for upgrading traditional fermented foods through biotechnology," african journal of biotechnology, vol. 4, pp. 375-380, 2005. 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[20] a. ojokoh, m. daramola, and o. oluoti, "effect of fermentation on nutrient and anti-nutrient composition of breadfruit (treculia africana) and cowpea (vigna unguiculata) blend flours," african journal of agricultural research, vol. 8, pp. 3566-3570, 2013. available at: https://doi.org/10.5897/ajar12.1944. [21] m. n. chukwu, n. o. kabuo, e. u. onyeka, t. c. odom, o. nwogu, n. j. nwokocha, and n. j. c., "production and organoleptic attributes of ogiri-ahuekere produced from groundnut (arachis hypogaea linn) seeds," research journal of food science and quality control, vol. 3, pp. 63-72, 2017. [22] m. chukwu, c. nwakodo, q. alozie, and j. ndulaka, "comparative studies on organoleptic properties of ogiri-ahuekere and ogiriegusi condiments," research journal of food science and quality control, vol. 4, pp. 11-19, 2018. available at: https://doi.org/10.2139/ssrn.3517482. [23] a. atere, v. oyetayo, and f. akinyosoye, "effects of starter culture on the proximate, antioxidant, antinutritional and mineral composition of fermented parkia biglobosa seeds to produce are," international journal of food sciences and nutrition, vol. 4, pp. 6165, 2019. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 30 © 2021 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 8, no. 2, 30-35, 2021 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2021.82.30.35 © 2021 by the authors; licensee asian online journal publishing group growth and yield components of some turmeric varieties (curcuma longa l.) as affected by inorganic fertilizer levels in umudike, south-eastern, nigeria akpan, a. u.1 orji, k. o.2 uhala, s. c.3 ( corresponding author) 1,2,3department of agronomy, college of crop and soil sciences, michael okpara university of agriculture, umudike, south eastern, nigeria. 1email: annahudo@yahoo.com 2email: regentko@yahoo.com tel: +2347065223591 3email: uhalamrs@yahoo.com tel: +2348139700708 abstract a research on growth and yield components of some turmeric varieties (curcuma longa l.) as affected by inorganic fertilizer levels in umudike, south eastern nigeria was carried out at the teaching and research farm of the university, during 2018/2019 cropping seasons. it was laid out in a split plot design with three replications. results, showed that plant height and number of tillers for both varieties and fertilizers levels increased at 2, 4, and 6 months after planting (map) during 2018 and 2019 cropping seasons. variety 021 and 400 npk (15:15:15) kg-1 produced tallest plants and highest number of tillers over other varieties and fertilizer levels. crop growth rate was positive at 2 – 4 map and negative at 4 – 6 map, with 021 and 400 npk (15:15:15) kgha-1 producing higher growth rate. heaviest rhizomes fresh weight (kg plot-1) of 3.74 and 3.75, rhizome dry matter content of 14.49 and 16.8%, rhizome dry weight of 23.57 and 23.90g and harvest index of 2.60 and 2.70% was yielded by umt 021 variety over other varieties. the level of 400 npk (15:15:15) kg ha-1 produced heaviest rhizome fresh weight of 4.47 and 4.57 kg plot-1; rhizome dry matter content of 19.42 and 25.8%; rhizome dry weight of 27.54 and 27.8g and harvest index of 2.81 and 3.20%, over other levels. consequently, variety 021 and 400 npk (15:15:15) kgha-1 are recommended for sole production of turmeric in umudike, south eastern nigeria. keywords: growth, yield, turmeric, varieties, inorganic fertilizers, levels. citation | akpan, a. u.; orji, k. o.; uhala, s. c. (2021). growth and yield components of some turmeric varieties (curcuma longa l.) as affected by inorganic fertilizer levels in umudike, southeastern, nigeria. agriculture and food sciences research, 8(2): 3035. history: received: 8 september 2021 revised: 12 october 2021 accepted: 17 november 2021 published: 28 december 2021 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 31 2. materials and methods ................................................................................................................................................................... 31 3. results and discussions .................................................................................................................................................................. 32 4. conclusion ......................................................................................................................................................................................... 35 references .............................................................................................................................................................................................. 35 mailto:annahudo@yahoo.com mailto:regentko@yahoo.com mailto:uhalamrs@yahoo.com http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.doi.org/10.20448/journal.512.2021.82.30.35 agriculture and food sciences research, 2021, 8(2): 30-35 31 © 2021 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the research has shown that turmeric requires lot of nutrients for growth and yield. however, excessive application of such nutrient might also be non-profitable to the farmers, due to the seemingly drop in both growth and yield attributes. a benchmark on inorganic fertilizer application for turmeric production has also been established. 1. introduction tumeric (curcumalonga l.) is a very important commercial crop grown for its aromatic rhizomes, which is used both as a spice and also in the preparation of different medicines. tumeric is a native of tropical south asia and its cultivation is mostly confined to south east asian countries. nigeria and bangladesh together contribute about 6% of the global turmeric production [1]. turmeric is a tropical perennial monocotyledonous herb belonging to the family zingiberaceae [2, 3]. tumeric can be propagated vegetatively, through mother rhizomes (primary rhizomes), finger rhizomes or split mother rhizomes. tumeric thrives best in well drained sandy or clay loamy soils, well rich in organic matter. the ideal ph is 4.5 – 7.5, while water logged and alkaline soils must be avoided. it also performs well in humid climatic conditions with temperature range of 20°c to 35°c and annual rainfall range of 80mm to 2500mm respectively. perhaps, this was the reason [4] reported that variation in turmeric yield could be due to variety used, the type of soil and the prevailing weather situations during the cropping seasons. turmeric rhizomes and the ground powder are used as spice or colouring agent. it helps in treating some diseases such as asthma, rheumatism, diabetes, wounds, cancers, urinary tract infections and liver ailments [5]. tumeric also helps in reducing the level of cholesterol in the human body. it helps to regulate the level of blood sugar as well as possess some antibacterial and antiseptic properties. the average composition of turmeric include: moisture (6.0%); protein (6.5%); ash (6.0%); crude fibre (3.0%), starch (5.0%); oil (3.5%); volatile oil (4.5%) and curcumin (3.1%) [6]. it is also used as a natural dye for cloth, leather, silk, fibre, wool and cotton. adeniji [7] and frances [8], stressed that the demands for turmeric is increasing because of its uses in the food and pharmaceutical industries. several studies have reported that nitrogen has significant effect on growth yield and yield components of turmeric [9, 10]. it has also been reported that nitrogen not only increases the yield of turmeric but also enhances the quality attributes [9]. application of nitrogen also enhances the efficacy of other nutrients like p and k, that improve the yield of turmeric [11]. n is involved in chlorophyll formation and influences stomatal conductance and photosynthetic efficiency and is responsible for 26 – 41% of crop yields ivonyi, et al. [12]. ojikpong [13], in his research on effect of planting dates and npk (15:15:15) fertilizer on the growth and yield of turmeric reported that 300 kgha-1 gave best values for all the measured growth and yield components. several studies have acknowledged that turmeric is a heavy feeder crop with high demand for n [10, 14, 15]. the high nutrient requirements of turmeric as reported by singh, et al. [14] is due to shallow rooting and potential to produce large amount of dry matter per unit area. also, the crop has a long growing period, extending up to 9 – 10 months, thereby making nutrient requirements period also prolonged. in view of the high nutrient demanding status of turmeric, there is a greater need to frequently boost the fertility status of soils, through prompt fertilizer application. this research therefore, evaluated the growth and yield components of some turmeric varieties (curcumalonga l.) as affected by inorganic fertilizer levels in umudike, south eastern, nigeria. 2. materials and methods the research was conducted at the teaching and research farm of michael okpara university of agriculture, umudike, south eastern, nigeria during 2018 and 2019 cropping seasons. umudike is located on latitude 05°29` north, longitude 07° 23` east at an altitude of 120.0m above sea level. it has a bimodal rainfall pattern with rainy season commencing from march to july and a dry spell in august followed by another rainfall session from september to november. annual rainfall pattern ranges from 2074.3mm to 2420mm mean, maximum and minimum temperatures and relative humidity are 31.4°c, 28°c and 63.4°% respectively [16]. the research was laid out in a split plot design with three replications. the main plot comprised npk (15:15:15) fertilizer levels of 0, 200, 400 and 600 kgha-1, while the subplot comprised the adopted turmeric varieties (umt 016, umt 019, umt 021). there varieties were on the field for 8 months while the rhizomes were sourced from the national root crops research institute (nrcri) umudike. the research layout was 29.5m x 8m or 0.0236ha. the main plot measured 7m x 2m, while the subplot measured 2m x 2m. the distance between plots and replicates were 0.5m and 1m respectively. healthy, pests and disease – free mother rhizomes (primary rhizomes) with healthy buds were used for planting. planting was conducted on 15th may for both cropping seasons on beds at the spacing of 20cm x 20cm, resulting in the population of 100 stands per plot and 250,000 stands per hectare. the plots were mulched after 3 days of planting with dry guinea grass (panicummaximum) at the rate of 20tha1[17]. fertilizer application was done by placement method at 3 weeks after planting, while weeding was conducted three times. mosquito nets were used to protect the entire research layout against rodents, squirrels and associated destructive pests [16]. harvesting of the rhizomes was manually conducted at 8 months after planting, when to plant becomes brownish and the leaves have fallen. digging forks and hand trowels were used to soften the soil, before the clumps were carefully pulled out of the soil. the mother (primary) and finger rhizomes were properly washed thereafter. plant height was conducted by measuring the height of four stands from the base to the terminal point at 2, 4 and 6 map, while the total was divided by four. the total number of tillers from four stands were divided by four to have number of tillers per plant. crop growth rate was obtained from the formula below: cgr = w2 – w1 sa (t2 – t1) (gm-2 day-1) [18] where: w1& w2: crop dry weight at the beginning and end of the interval. t1& t2: corresponding days. sa: soil area occupied by the plant. agriculture and food sciences research, 2021, 8(2): 30-35 32 © 2021 by the authors; licensee asian online journal publishing group rhizome fresh weight (kg plot-1) was by weighing total rhizomes harvested from four stands of turmeric per plot and subsequently converted through the formular: = weight of rhizomes per plot plot size × 1000 rhizomes dry matter content (%) was by the formular: rhizome dry weight rhizome fresh weight × 100 1 rhizome dry weight per plant was by weighing the total number of rhizomes from four stands, while the total weight was subsequently divided by four. harvest index was obtained through this formular: harvest index (hi) = economic yield biological yield or rhizomes fresh weight shoot dry weight before the commencement of the research, composite soil samples were collected randomly at the depth of 0 – 20cm from the research site for physico – chemical analyses. particle size distribution was determined by the bouyocous hydrometer method as described by benton [19]. soil ph in soil to water and soil to cacl2 at the ratio of 1:2:5, soil water and soil – cacl2 respectively, was by using glass electrode ph meter [20]. organic carbon was by wet oxidation method [21]. available p was determined by bray 11 method of bray and kurtz [22] as described by udo, et al. [20]. total nitrogen was done by the macro kjeildhal digestion method [23]. calcium and mg were determined by atomic absorption spectrometry, while potassium and sodium were by flame photometry [24]. both growth and yield data were subsequently subjected to analysis of variance (anova) method [25], while fisher’s least significant difference (f – lsd) was adopted to compare significant means at 5% probability level. 3. results and discussions 3.1. meteorological properties of the research site meteorological properties of the research site table 1, revealed that the months of january, february and march recorded the lowest rainfall situations of 75.4 and 0.0mm; 84.8mm and 43.7mm as well as 40.8mm and 78.7mm during 2018 and 2019 cropping seasons. monthly rainfall situations increases from may and dropped in november, preparatory for dry period (table 1). both maximum and minimum temperatures and relative humidity fluctuated in favour of rainfall patterns with dry months recording high temperatures and low relative humidity table 1. the relevance of weather to the productivity of turmeric was earlier reported by karthikeyan, et al. [4], that variation in turmeric yield could be caused by prevailing weather situations during cropping seasons. table-1.meteorological properties of the research site during 2018 and 2019 cropping seasons. months rainfall (mm) tempt °c r/h (%) max. min. 2018 2019 2018 2019 2018 2019 2018 2019 jan 75.4 0.0 32.0 33.4 22.0 21.5 31 49 feb 84.8 43.7 33.0 33.9 24.0 23.2 36 38 mar 40.8 78.7 33.0 33.2 22.0 23.4 66 54 apr 92.8 136.8 32.0 32.2 22.0 23.5 70 62 may 466.1 249.2 31.0 31.9 23.0 23.4 72 71 june 239.4 281.8 29.0 30.5 23.0 24.2 72 79 july 280.5 114.9 30.0 30.0 22.0 24.0 81 78 aug 237.1 436.5 34.0 29.6 23.0 23.3 81 78 sept 318.0 412.4 30.0 29.8 22.0 22.9 77 73 oct 184.8 169.1 30.0 31.0 23.0 23.6 69 74 nov 99.5 147.4 32.0 31.6 23.0 23.5 64 65 dec 90.8 0.0 32.0 32.7 22.0 21.8 51 48 total 2210.0 2068.5 table-2.soil physico-chemical properties of the research site, during 2018 and 2019 cropping seasons. soil properties values sand (%) 72.4 74.2 silt (%) 12.8 11.4 clay (%) 14.8 11.4 textural class sandy loam sandy loam chemical properties ph 5.09 5.04 phosphorus (mgkg-1) 11.06 12.4 nitrogen (%) 0.38 0.27 organic carbon (%) 0.3 0.62 calcium (cmolkg-1) 3.6 3.2 magnesium (cmolkg-1) 1.24 2.0 potassium (cmolkg-1) 0.65 0.52 3.2. soil physico-chemical properties of the research site the physico-chemical properties of the research site table 2, showed that the textural class of the site was sandy loamy soil with <80% sand fraction for both cropping seasons. some elements like phosphorus (11.06 and 12.4 mgkg-1) and organic carbon (0.3% and 0.62%) during 2018 and 2019 cropping seasons were low table 2. the agriculture and food sciences research, 2021, 8(2): 30-35 33 © 2021 by the authors; licensee asian online journal publishing group ph of 5.09 and 5.04 for both cropping seasons were moderate and capable of sustaining the production of turmeric. apart from nutrient availability, hossain and ishimine [26], attributed the performance of turmeric in the field to soil properties including: soil ph, moisture contents, bulk density, aeration and soil microbial activities. 3.3. effect of npk (15: 15: 15) fertilizer levels on the growth of turmeric plant height for variety and fertilizer levels increased at 2, 4 and 6 map during 2018 and 2019 cropping seasons table 3. variety umt 021 produced tallest plants of 11.8 and 12.3cm (2 map), 30.5 and 29.8cm (4 map), 69.6 and 70.2 cm (6 map) in 2018 and 2019 cropping seasons over other varieties. the 400 npk (15: 15: 15) kgha1 produced tallest plants of 12.4 and 12.6cm (2 map), 38.5 and 38.2m (4 map); 77.3 and 76.6cm (6 map), compared with control (0 level) table 3. plant height was significantly (p < 0.05) affected by variety at 2 and 4 (map), while 6(map) was not significantly (p > 0.05) affected for both years. plant height was significantly (p <0.05) affected by fertilizer levels at 2, 4 and 6 (map), while interaction (variety x fertilizer levels) was not significant. turmeric height will therefore vary with the cultivated variety and the level of inorganic fertilizer applied. the observed drop in height at 600 kgha-1, suggested that fertilizer addition beyond 400 kgha-1 will not be economical, and might lead to luxury consumption with attendant low yield. atugwu, et al. [27], reported increased turmeric height with age, while ojikpong [13], reported tallest turmeric at 300 npk (15: 15: 15) kgha-1 as well as drop at 450 npk (15:15:15) kgha-1. this research therefore corroborates the earlier report by ojikpong [13]. table-3. effect of npk (15:15:15) fertilizer level on plant height of turmeric varieties at 2, 4 and 6 map during 2018 and 2019 cropping seasons. tumeric varieties plant height (cm) 2 map 4 map 6map 2018 2019 2018 2019 2018 2019 umt 016 10.9 11.0 27.8 27.3 66.9 66.2 umt 019 9.9 9.9 26.2 26.2 66.9 64.7 umt 021 11.8 12.3 30.5 29.8 69.6 70.2 fertilizer levels (kgha-1) 0 9.0 9.1 16.7 16.5 52.1 51.0 200 10.9 11.1 29.0 28.1 68.4 66.6 400 12.4 12.6 38.5 38.2 77.3 76.6 600 11.2 11.5 28.3 28.1 73.4 74.8 map 2 4 6 2018 2019 2018 2019 2018 2019 variety 0.78 0.56* 1.78 1.64 ns ns fertilizer 1.00* 0.79* 2.75* 2.94* 10.88* 9.14* variety x fertilizer ns ns ns ns ns ns note: f-lsd (p< 0.05). table-4.effect of npk (15:15:15) fertilizer levels on number of tillers of turmeric varieties at 2, 4 and 6 map during 2018 and 2019 cropping seasons. tumeric varieties plant height (cm) 2 map 4 map 6map 2018 2019 2018 2019 2018 2019 umt 016 2.0 3.0 5.2 5.1 9.6 9.3 umt 019 1.7 2.7 4.8 4.9 9.8 9.2 umt 021 2.8 3.1 5.8 5.8 9.9 9.9 fertilizer levels (kgha-1) 0 1.6 2.1 3.8 3.7 7.0 7.4 200 2.1 3.0 4.9 4.8 9.8 9.8 400 2.9 3.9 7.0 7.2 11.7 11.2 600 2.1 2.7 5.3 5.4 10.6 9.6 map 2 4 6 2018 2019 2018 2019 2018 2019 variety 0.52 ns ns 0.68 ns ns fertilizer 0.50* 0.74 0.80* 1.03* 1.52* 1.88 variety x fertilizer ns ns ns ns ns ns note: f-lsd (p< 0.05). number of tillers per plant table 4, for both variety and fertilizer levels increased at 2, 4 and 6 map during 2018 and 2019 cropping seasons. variety umt 021 produced the highest number of tillers of 2.8 and 3.1 (2 map); 5.8 and 5.8 (4 map); 9.9 and 9.9 (6 map), over other varieties, while 400 npk (15:15:15) kgha-1 recorded highest number of tillers of 2.9 and 3.9 (2 map); 7.0 and 7.2 (4 map) as well as 11.7 and 11.2 (6 map) compared with other levels for both cropping seasons. number of tillers per plant was significantly (p < 0.05) affected by the variety at 2 map in 2018 cropping season, as well as 4 map in 2019 cropping season, while 6 map was not significantly (p > 0.05) affected by fertilizer levels at 2, 4 and 6 map. interaction (variety x fertilizer levels) did not significantly (p > 0.05) affect number of tillers for both cropping seasons. the observed reduction in number of tillers at 600 npk (15; 15:15) kgha-1, suggested that it was not economical to the farmer. number of tillers the research showed will equally vary with the variety planted and the level of fertilizer applied. the increased number of tillers with age, corroborates an earlier report by atugwu, et al. [27], who reported increase in number of tillers per plant as turmeric grew older. agriculture and food sciences research, 2021, 8(2): 30-35 34 © 2021 by the authors; licensee asian online journal publishing group crop growth rate table 5, for variety and fertilizer levels increased positively at 2 to 4 map and as well dropped negatively at 4 – 6 map for both cropping seasons. variety umt 021 recorded highest growth rate of 0.518 and 0.503 gm-2day-1 at 2 – 4 map as well as -0.123 and -0.0932 gm-2day-1 at 4 – 6 map. the 400 npk (15:15:15) kgha-1 level equally produced highest growth rate of 0.655 and 0.661 gm-2day-1 at 2 – 4 map and also 0.105 and -0.1043 gm-2day-1 at 4 – 6 map. crop growth rate was not significantly (p > 0.05) affected by variety at 2 – 4 and 4 – 6 map. fertilizer levels significantly (p < 0.05) affected crop growth rate at 2 – 4 map, but was however not significant (p > 0.05) at 4 – 6 map for both cropping seasons. interaction (variety x fertilizer levels) did not significantly (p > 0.05) affect crop growth rate. this result has shown that growth processes in turmeric are active from 1 to 4 map and that the formation and bulking of tender rhizomes may likely commence before 4 map. this may cause the vegetative and early reproductive phases of turmeric to occur simultaneously. the early commencement of the reproductive phase of turmeric as well as the effect of senescence during the steady growth phase may explain the negative growth rate and the crop grows older. crop growth rate is likely to be influenced by the age of turmeric and fertility status of the soil. table-5.effect of npk (15:15:15) fertilizer levels in crop growth rates of turmeric varieties at 2 – 4 and 4 – 6 map during 2018 and 2019 cropping seasons. crop growth rate (gm-2day-1) turmeric variety 2 – 4 map 4 – 6 map 2018 2019 2018 2019 umt 016 0.444 0.457 -0.089 -0.0751 umt 019 0.426 0.416 -0.069 -0.0921 umt 021 0.518 0.503 -0.123 -0.0932 fertilizer rates (kgha-1) 0 0.463 0.429 -0.103 -0.0872 200 0.446 0.454 -0.098 -0.0872 400 0.655 0.661 -0.105 0.1043 600 0.286 0.291 -0.068 -0.0706 map 2 – 4 4 – 6 2018 2019 2018 2019 variety ns ns ns ns fertilizer 0.146 0.1428 ns ns variety x fertilizer ns ns ns ns note: f-lsd (p< 0.05). table-6.effect of npk (15:15:15) fertilizer levels on rhizome fresh weight (kg plot-1), rhizome dry matter content, rhizome dry weight and harvest index of turmeric varieties during 2018 and 2019 cropping seasons. turmeric variety rhizome fresh weight (kgplot-1) rhizome dry matter content (%) rhizome dry weight (g) harvest index (%) 2018 2019 2018 2019 2018 2019 2018 2019 umt 016 3.19 2.85 13.00 15.4 17.25 17.4 2.56 2.2 umt 019 2.50 2.28 13.61 13.6 13.57 13.5 2.07 1.9 umt 021 3.74 3.75 14.49 16.8 23.57 23.90 2.60 2.70 fertilizer rates (kgha-1) 0 1.60 0.67 11.94 12.1 6.97 6.70 1.99 1.0 200 3.05 3.27 12.10 11.5 18.81 19.0 2.47 2.5 400 4.47 4.57 19.42 25.8 27.54 27.8 2.81 3.2 600 3.35 3.34 11.73 11.7 19.78 19.6 2.38 2.4 rhizome fresh weight (kgplot-1) rhizome dry matter content (%) rhizome dry weight (g) harvest index (%) 2018 2019 2018 2019 2018 2019 2018 2019 variety ns 0.672 ns ns 5.837 5.62 ns 0.52 fertilizer 1.395 0.911 ns 4.08 6.177* 6.32* ns 0.70* variety x fertilizer ns ns ns 7.17 ns ns ns ns note: f-lsd (p< 0.05). the evaluated yield parameters table 6, showed that the heaviest rhizome fresh weight per plot (kgha-1) of 3.74 and 3.75 kgha-1; highest rhizome dry matter content of 14.49 and 16.8%; heaviest rhizome dry weight per plant of 23.57 and 23.90g and harvest index of 2.60 and 2.70 were yielded by umt 021 variety compared with other varieties during 2018 and 2019 cropping seasons. the fertilizer level of 400 npk (15:15:15) kgha-1 equally yielded heaviest rhizome fresh weight of 4.47 and 4.57 kg plot-1; rhizome dry matter content of 19.42 and 25.8%; rhizome dry weight per plant of 27.54 and 27.8g as well as harvest index of 2.81 and 3.20 for both cropping seasons. a drop in yield components was observed at 600kgha-1 level, suggesting that fertilizer addition beyond 400 kgha-1 will be uneconomical and may as well engender luxury consumption. rhizome fresh weight (kg plot-1) was significantly (p < 0.05) affected by variety in 2019 cropping season, while fertilizer levels significantly (p < 0.05) affected rhizome fresh weight (kg plot-1) in both cropping seasons. rhizome dry matter content was not significantly (p > 0.05) affected by variety in both cropping seasons, while fertilizer levels significantly (p < 0.05) affected dry matter content only in 2019 cropping seasons. rhizome dry weight was significantly (p < 0.05) affected by the variety and fertilizer levels in both cropping seasons. harvest index on the other hand was significantly (p < 0.05) affected by both the variety and fertilizer levels only in 2019 cropping seasons. interaction (variety x fertilizer levels) significantly (p < 0.05) affected rhizome dry matter content in 2019 cropping season, while other yield parameters were not significantly (p > 0.05) affected. these results on yield agriculture and food sciences research, 2021, 8(2): 30-35 35 © 2021 by the authors; licensee asian online journal publishing group components slightly support [13] who reported increases in yield components of turmeric at 300 npk (15: 15: 15) kgha-1 as well as a drop in yield parameters at 450 npk (15: 15: 15) kgha-1 respectively. 4. conclusion the research has revealed that variety umt 021 and 400 npk (15:15:15) kgha-1 performed better that other varieties and fertilizer levels respectively. this further suggests that umt 021 variety was more adaptive to the research site while 400kgha-1 provided ample nutrients for the optimum growth and yield performances of turmeric. consequently, for a successful production of turmeric as a sole crop in umudike, south eastern nigeria, umt 021 variety and 400 npk (15:15:15) kgha-1are recommended. application of fertilizer up to 600 npk (15:15:15) kgha-1 was unproductive due to the observed drop in both growth and yield parameters. this could lead to luxury consumption, often characterized by luxuriant growth with attendant low yield. the growth and yield performances of turmeric, the research revealed will likely vary with cultivated variety and the level of inorganic fertilizer applied. references [1] k. r. kumar, s. n. rao, and n. r. kumar, "evaluation of turmeric (curcuma longa l.) cultivars at agency areas of north coastal andhra pradesh," progressive research journal, vol. 10, pp. 2417 – 2420, 2015. 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[26] m. a. hossain and y. ishimine, "growth, yield and quality of turmeric (curcuma longa l.) cultivated on dark-red soil, gray soil and red soil in okinawa, japan," plant production science, vol. 8, pp. 482-486, 2005. [27] a. i. atugwu, v. n. onyia, and r. u. eze, "evaluation of a turmeric variety for emergence and yield under four growth media. in: sustainable horticulture production system intensified. aba, s. c. & baiyeri, k. p. (editors)," in proceedings of horticultural society of nigeria conference held at the university of nigeria nsukka, between 25 – 31 october, 2020, 2020, pp. 353 – 361. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.worldcat.org.p.2,/ http://www.scribd.com,/ 131 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 2, 131-138, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.72.131.138 © 2020 by the authors; licensee asian online journal publishing group evaluation of starch components, pasting and functional properties of pleurotustuberregium flour from air dried sclerotia amaechi, n.c1 okoronkwo, c.u2 agbaeze, t3 ( corresponding author) 1,2,3department of food science and technology, abia state university, uturu-abia state, nigeria. abstract pleurotus tuberregium is one group of mushroom that produce edible sclerotia and is used for various culinary and medicinal purposes. flour was produced from the air dried sclerotia and starch components, pasting and functional properties were evaluated. it had a total and resistant starch contents of 73.63% and 12.37% respectively while its amylose content was 20.03%. its amylose and resistant starch contents classified it as regular and high resistant starch flour. its pasting properties revealed it had pasting temperature and peak time of 60.16ºc and 5.97min respectively. results on breakdown viscosity, set back viscosity and set back ratio revealed a high tendency of the flour to retrograde after gelatinization and when shear stress is applied. it had low swelling capacity, moderate dispersibility index and high gelatinization temperature with values of 4.17g/g, 53% and 94ºc respectively. this therefore suggests that flour from air dried pleurotus tuberregium sclerotia can be utilized in food formulations where high resistant starch is relevant for nutritional purposes and as thickeners in food products. however, there is need for technological modification to improve its pasting properties so as to generate flour with a high stability ratio and a low set back ratio and so withstand retrogradation and with improved resistance to shear stress. keywords: pleurotus tuberregium sclerotia, air-dried flour, resistant starch, pasting properties, dispersibilty index, amylose, amylopectin, set back ratio, stability ratio, gelatinization temperature. citation | amaechi, n.c; okoronkwo, c.u; agbaeze, t (2020). evaluation of starch components, pasting and functional properties of pleurotustuberregium flour from air dried sclerotia. agriculture and food sciences research, 7(2): 31-38. history: received: 15 september 2020 revised: 12 october 2020 accepted: 29 october 2020 published: 18 november 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 132 2. materials and methods ................................................................................................................................................................. 132 3. results and discussion ................................................................................................................................................................. 134 4. conclusion ....................................................................................................................................................................................... 136 references ............................................................................................................................................................................................ 136 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.72.131.138&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.72.131.138&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/2336 https://orcid.org/0000-0002-8378-1775 https://orcid.org/0000-0002-5943-8248 agriculture and food sciences research, 2020, 7(2): 131-138 132 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study has provided information on the nutritional, technological usefulness and functionality of flour from air dried pleurotus tuberregium sclerotia. the research has shown that flour from air dried sclerotia has resistant starch but there is need for technological modification to improve its pasting properties. 1. introduction edible mushrooms form part of diet in many human populations because of their unique flavour, texture or for their nutritional benefits such as having antioxidant activity. some mushroom species produce sclerotia which are edible and used for various purposes. the search for new food ingredients with good functionality and nutritional value has resulted to the exploitation of mushrooms and has been of interest to food technologists lately. the utilization of mushroom flours is limited due to absence of knowledge about their functional and compositional characteristics and their interactions [1]. pleurotus tuberregiumis a tropical edible mushroom that produces edible sclerotium/underground tuber as well as mushroom [2]. the sclerotium of pleurotus tuberregium is a compact mass of hardened mycelium containing food reserves which help the fungi survive environmental extremes [3]. it is highly nutritious containing good quality protein and carbohydrate with little fat [4]. major reserve carbohydrates in foods is starch comprised of polymer units namely amylose and amylopectin. the relative proportion of amylose and amylopectin and their organization within starch granules determine the functional properties of the starch and consequently its range of industrial applications in foods, pharmaceuticals etc [5]. starch contributes to textural properties of many foods and is commonly used in food and industrial applications as thickener, stabilizer and gelling agent [6]. pasting properties are functional properties associated with the ability of an item to act in paste-like manner [7]. dietary starches are important sources of energy for humans which during digestion, a portion goes undigested in the upper gastrointestinal tract and so is referred to as resistant starch. lately, there is a lot of interest in resistant starch for its potential health benefits as is to soluble fibre as well as its functional properties [8]. the physical properties of resistant starch makes it a functional ingredient that provides good handling and improve texture in the final product [9]. mushrooms have the ability to develop a morphological form known as sclerotium which is a compact mycelia structure under unfavorable conditions and can remain dormant until there is suitable environmental conditions required for fruiting bodies germination [10]. this mycelia structure known as the sclerotia serve as food reserve for fruiting bodies and is consumed as food by man. it is in view of this that starch components, pasting and functional properties of flour produced from air dried pleurotus tuberregium sclerotia was evaluated. this will provide information about its usefulness in food formulation and its functional characteristics as a healthy food ingredient. 2. materials and methods 2.1 sclerotia collection and preparation of flour sample pleurotus tuberregium sclerotia was purchased from a local market in umuahia, abia state, nigeria. the outer dirty brown back was peeled using stainless steel knife while the inner white mass of sclerotia was cut in small pieces of 5mm thickness and air dried on stainless tray in the laboratory (room temperature 25±2ºc). the air dried sclerotia was milled using a blender (master chef, india) at speed no2 into fine flour and was packaged in an airtight container and stored at room temperature prior to analysis. 2.2 analysis of starch components. 2.2.1 determination of total starch: total starch was determined by the aoac method 996.11 described by mccleary, et al. [11]. 50g of the flour was passed through a 0.5mm screen and 100mg of the sieved flour was weighed into a glass centrifuge. the tube was tapped to ensure all the samples falls to the bottom of the tube before 0.2mi ethanol (80% v/v) was added and the contents were stirred on a vortex mixer before 2ml dimethylsulphoxide was added and stirred continuously on the vortex mixer for 3min. subsequently the tube with its contents were placed in a vigorously boiling water bath for 5min. immediately3ml of thermostable α-amylase was added and the tube was incubated in a boiling water bath for 6min with intermediate vigorous stirring after 2,4, and 6min to ensure homogeneity. after which, the tubes were placed in a water bath at 50ºc and 0.1ml amyloglucosidase suspension was added to the mixture and the contents were stirred on a vortex mixer and incubated at 50ºc for 30min. after this step, the contents were transferred to a 100ml volumetric flask and distilled water was used to rinse tube contents thoroughly and the volume was adjusted to 10ml and mixed thoroughly before centrifuging at 3000rpm for 10min. 1ml of the supernatant was diluted to 10ml with distilled water. a reagent blank, glucose standard and test sample were subjected to endpoint analysis using god-pap reagent. for the test sample, 0.1ml of diluted supernatant was dispensed into a test tube and 3ml god-pap reagent was added, the blank contained 3ml god-pap reagent and 0.1,m water while the standard contained 3ml god-pap reagent plus 0.1ml glucose standard. all these were incubated at 50ºc for 20min and absorbance were read at 510nm against the reagent blank. total starch was calculated as shown in equation 1: starch (%) = ∆a x f w x fv x 0.9 (1) ∆a is sample god-pap absorbance read against reagent blank. f is the factor used to convert from absorbance to µg of glucose. w is weight of sample analysed in mg. fv is final volume of solution used. 2.2.2. determination of amylose and amylopectin amylose was determined by the iodine colorimetric method described by mohana, et al. [12]. the flour sample was defatted prior to analysis.100mg of the defatted flour was weighed into 100ml volumetric flask to which 1ml agriculture and food sciences research, 2020, 7(2): 131-138 133 © 2020 by the authors; licensee asian online journal publishing group 95% ethanol and 9ml 1n naoh were added and mixed thoroughly. after which it was heated on boiling water bath to gelatinize the starch and later on cooled to room temperature. 5ml of the gelatinized starch solution was dispensed into a 100ml volumetric flask to which 1ml of 1n acetic acid and 2ml of iodine solution were added and the volume made up to 100ml with distilled water. all the contents were thoroughlyvortexed mixed and allowed to stand for 20mins. the absorbance was measured at 620nm in a spectrophotometer using a blank containing 5ml 0.09n naoh, 1ml acetic acid and 2ml iodine solution and made up to 100ml volume using distilled water. the amylose content was determined based on the standard curve prepared using potato amylose. amylopectin was calculated by difference stated by juan, et al. [13] shown in equation 2: amylopectin (%) = 100 amylose (%) (2) 2.2.3. evaluation of resistant starch resistant starch was evaluated by the method described by goñi, et al. [14]. 100mg of the sample was dispensed into centrifuge tube and 10ml kcl-hcl buffer (ph1.5) was added and the mixture homogenized. 0.2ml pepsin was added to the homogenized sample mixture and incubated at 40ºc for 60min with constant shaking in a water bath. after which the sample was cooled to room temperature, 9ml tris -maleate buffer (ph 6.9) was added alongside 1ml α-amylase was added to the mixture, shaken well and incubated for 16h at 37ºc in a water bath with constant shaking. subsequently the sample was centrifuged for 15min at 3000rpm and the supernatant was discarded leaving the sediment. the sediment was washed once with 10ml distilled water, centrifuged and the supernatant discarded before adding 3ml distilled water and 3ml 4m koh. the contents were mixed thoroughly and left to stand at room temperature for 30min with constant shaking before 5ml buffer (ph 4.75) and 0.08ml amyloglucosidase were added and mixed. the sample mixture was incubated for 45min at 60ºc in a water bath with constant shaking. the mixture was clarified by centrifuging at 3000rpm for 15min and the supernatant siphoned into a volumetric flask. the residue was washed twice with 10ml distilled water and clarified by centrifuging each time and the supernatant recovered and combined with was put into the volumetric flask previously. the recovered supernatant solution was made up to 100ml using distilled water. a standard curve containing 10-60ppm glucose was generated. 0.5ml water, sample and standard glucose solutions were dispensed into test tubes. 1ml god-pap was added to each to each test tube and incubation was done for 30min at 37ºc in a water bath. absorbance of test and standards were read at 500nm against reagent blank. the standard curve was used to calculated glucose concentration of the sample. resistant starch was calculated as shown in equation 3: resistant starch (%) = mg of glucose x 0.9 (3) 2.3. pasting properties determination pasting properties of pleurotustuberregiumsclerotia flour was evaluated using rapid viscoanalyser (dingling rvu 232015, usa) by methods described by aacc [15]. a 3g sample was dispersed in an aluminium canister containing 25ml of distilled water. the sample mixture underwent a controlled heating and cooling cycle under constant shear where it was held at 50ºc for 1min, heated from 50 to 95ºc at 6 ºc/min and held at 95ºc for 5min. finally each sample was cooled to 50ºc and held for another 2min. the starch viscosity parameters measured were peak viscosity(pv), trough viscosity~viscosity at the end of holding time at 95ºc (hpv), breakdown viscosity (pvhpv), cool paste viscosity (cpu) ̴ final viscosity-viscosity at the end of the hold time at 50ºc; setback viscosity (sbv) ̴ (cpu-hpv), pasting time= time from onset of pasting to peak viscosity, pasting temperature= temperature from onset of pasting to peak viscosity. stability ratio (sr) = 𝐻𝑃𝑉 𝑃𝑉 and setback ratio (sbr) = 𝐶𝑃𝑉 𝐻𝑃𝑉 were calculated prescribed by julianti, et al. [16]. 2.4. functional properties determination 2.4.1 bulk density bulk density was determined by the method described by onabanjo and ighere [17]. a 50g weight of pleurotustuberregiumsclerotia flour was put into 100ml measuring cylinder. the cylinder was tapped several times on a laboratory bench to a constant volume. the volume of sample was calculated as shown in equation 4: bulk density (g/cm3)= 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑎𝑓𝑡𝑒𝑟 𝑡𝑎𝑝𝑝𝑖𝑛𝑔 (4) 2.4.2. water absorption capacity (wac) and oil absorption capacity (oac) these were determined by the methods described by onabanjo and ighere [17]. for wac, 1g of pleurotustuberregiumsclerotia flour was dispensed into 25ml centrifuge tube and 15ml distilled water was added to it and the tube was agitated on a vortex mixer for 2min. the suspension was centrifuged at 1000rpm for 20min and after which, the clear supernatant was decanted and discarded. the wet flour residue was reweighed and water absorption was expressed as weight of water bound by 100g dried flour. the same procedure was used for oil absorption capacity except that water was replaced with vegetable oil of specific gravity of 0.98g/ml. water absorption capacity and oil absorption capacity were expressed as shown in equation 5: wac/oac (g/g) = 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑒𝑑𝑖𝑚𝑒𝑛𝑡 𝐼𝑛𝑖𝑡𝑖𝑎𝑙 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑓𝑙𝑜𝑢𝑟 …………………………………………………….(5) 2.4.3. emulsion activity this was determined by the method described by yasumatsu, et al. [18]. a mixture of 1g flour sample, 10ml distilled water and 10ml soybean oil was prepared in a calibrated centrifuge tube. the emulsion was centrifuged at 2000rpm for 5min. the ratio of the height of emulsion layer to the total height of the mixture was calculated as emulsion activity in percentage shown in equation 6: emulsion activity (%)= 𝐻𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑒𝑚𝑢𝑙𝑠𝑖𝑜𝑛 𝑙𝑎𝑦𝑒𝑟 𝑏𝑒𝑓𝑜𝑟𝑒 𝑐𝑒𝑛𝑡𝑟𝑖𝑓𝑢𝑔𝑎𝑡𝑖𝑜𝑛 𝑇𝑜𝑡𝑎𝑙 ℎ𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑡ℎ𝑒 𝑚𝑖𝑥𝑡𝑢𝑟𝑒 𝑎𝑓𝑡𝑒𝑟 𝑐𝑒𝑛𝑡𝑟𝑖𝑓𝑢𝑔𝑎𝑡𝑖𝑜𝑛 x 100 1 ………………..(6) agriculture and food sciences research, 2020, 7(2): 131-138 134 © 2020 by the authors; licensee asian online journal publishing group 2.4.4. emulsion stability this was determined by the method of yasumatsu, et al. [18]. emulsion stability was estimated after heating the emulsion in the calibrated centrifuge tube which was obtained from the determination of emulsion activity at 80ºc for 30min in a water bath. this was followed up by cooling for 15mins under running tap water and centrifuged at 2000rpm for 15min. emulsion stability was expressed as shown in equation 7: emulsion stability (%)= 𝐻𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑒𝑚𝑢𝑙𝑠𝑖𝑓𝑖𝑒𝑑 𝑙𝑎𝑦𝑒𝑟 𝑇𝑜𝑡𝑠𝑙 ℎ𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑡ℎ𝑒 𝑚𝑖𝑥𝑡𝑢𝑟𝑒 x 100 1 ……………………………………(7) 2.4.5 foam capacity and foam stability foam capacity and foam stability were determined by the methods described by narayana and narasinga rao [19]. foam capacity was evaluated by dispensing 1g flour sample into a blender and 10ml deionized water (ph adjusted to 7.4 using 1n naoh and 1n hcl) was added. the mixture was blended for 5min before turning into a 250ml graduated cylinder and the foam volume was recorded immediately. foam capacity was calculated as shown in equation 8: foam capacity (%) = 𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑓𝑜𝑎𝑚 𝑎𝑓𝑡𝑒𝑟 𝑤ℎ𝑖𝑝𝑝𝑖𝑛𝑔 𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑓𝑜𝑎𝑚 𝑏𝑒𝑓𝑜𝑟𝑒 𝑤ℎ𝑖𝑝𝑝𝑖𝑛𝑔 x 100 1 ………………………………(8) foam stability: foam stability was evaluated by recording foam volume in the cylinder 1h after whipping as percent of initial foam volume, shown in equation 9. foam stability (%)= 𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑓𝑜𝑎𝑚 1ℎ 𝑎𝑓𝑡𝑒𝑟 𝑤ℎ𝑖𝑝𝑝𝑖𝑛𝑔 𝑉𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑓𝑜𝑎𝑚 𝑖𝑚𝑚𝑒𝑑𝑖𝑎𝑡𝑒𝑙𝑦 𝑎𝑓𝑡𝑒𝑟 𝑤ℎ𝑖𝑝𝑝𝑖𝑛𝑔 x 100 1 …………………….(9) 2.4.6.swelling index and swelling capacity these were analysed by the methods described by ukpabi and ndimele [20]. swelling index was determined by dispensing 25g of flour sample into 250ml measuring cylinder. 150ml deionized water was added and the mixture was shaken and allowed to stand for 4h before observing the extent of swelling. swelling index was calculated as shown in equation 10: swelling index (g/g)= 𝑉𝑜𝑙𝑢𝑚𝑒 𝑎𝑓𝑡𝑒𝑟 𝑠𝑜𝑎𝑘𝑖𝑛𝑔 − 𝑉𝑜𝑙𝑢𝑚𝑒 𝑏𝑒𝑓𝑜𝑟𝑒 𝑠𝑜𝑎𝑘𝑖𝑛𝑔 𝑊𝑒𝑖𝑔ℎ𝑡 𝑏𝑒𝑓𝑜𝑟𝑒 𝑠𝑜𝑎𝑘𝑖𝑛𝑔 ……………………...(10) swelling capacity: the gel obtained after determining swelling index was used in calculating swelling capacity as shown in equation 11: swelling capacity (g/g) = 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑔𝑒𝑙 𝑎𝑓𝑡𝑒𝑟 𝑠𝑜𝑎𝑘𝑖𝑛𝑔 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑏𝑒𝑓𝑜𝑟𝑒 𝑠𝑜𝑎𝑘𝑖𝑛𝑔 ………………………………(11) 2.4.7. dispersibility this was determined by the methods described by kulkarni, et al. [21]. 10g of flour sample was weighed into a graduated cylinder and 100ml distilled water was added to it. the mixture was shaken vigorously and allowed to stand for 3h before the volume of settled particles was recorded. 2.4.8. gelatinization temperature it was determined by the method described by shinde [22]. 1g flour sample was weighed into a 20ml screw capped tube and 10ml water was added to it and shaken vigorously before it was heated slowly in a water bath until a solid gel was formed. the temperature at which a gel was formed was recorded in ºc. 3. results and discussion 3.1. starch components resultson starch components of pleurotus tuberregium flour from air dried sclerotia is shown in table 1. its total starch content of 73.63% was comparable to total starch content of pleurotus ostreatus and had a value of 75% [23] but higher than starch from green cadaba banana flour subjected to various drying methods and had values between 29.24 and 40.77% [24]. its amylose and amylopectin contents were 20.03% and 53.60% respectively. the ratio of amylose and amylopectin in starch influences mechanical property of starch polymer [25]. its amylose content (20.03%) was comparable to amylose content of p. ostreatus(21.7%) [23] but lower than amylose content of native banana and plantain starches which had values of 42.07% and 38.79% respectively [6]. amylopectin content (52.62%) observed in this work was lower than amylopectin content of p. ostreatus(78.30%) [23]. amylose content is the underlying condition for categorizing starches into waxy, semi-waxy, regular and high amylose types when amylose content is 0-2%, 3-15%, 15-35% and >40% of the total starch respectively [26]. in the current study, starch from air dried pleurotustuberregium flour can be classified as regular starch. resistant starch content observed for p. tuberregiumwas 12.37%. this value was lower than resistant starch content of green cadaba banana flour subjected to different drying methods with values that ranged between 16.83 and 27.53% [22] but higher than resistant starch content of different cassava varieties which ranged between 1.12 and 4.14% reported by chisenga, et al. [27]. goñi, et al. [14] classified food materials with resistant starch content of 5-15% as high resistant starch foods. this suggests that flour from air dried sclerotia of pleurotustuberregiumcan be classified as high resistant starch flour. resistant starch functions physiologically as fibre in promoting a healthy gastrointestinal environment such as being a prebiotic compound [8]. therefore, incorporation of p. tuberregiumflour into food formulations will be of benefit in promoting a healthy gut environment. agriculture and food sciences research, 2020, 7(2): 131-138 135 © 2020 by the authors; licensee asian online journal publishing group table-1.starch fractions of pleurotus tuberregium flour from air dried sclerotia parameter value total starch (%) 73.63±0.00 amylose (%) 20.03±0.00 amylopecyin (%) 53.62±0.02 resistant starch (%) 12.37±0.01 note: values are means ± standard deviation. 3.2. pasting properties table 2 shows results on pasting properties of flour from air dried p. tuberregium sclerotia. its pasting temperature was 60.16ºc while its peak time was 5.97mins. pasting temperature obtained in this work was lower than pasting temperature (86.70 ºc) and peak time (7.90min) reported by kolawole, et al. [28] for p. tuberregium flour obtained from sclerotia oven dried at 40ºc. subroto, et al. [29] reported an increase in pasting temperature of potato starch oven dried at 40ºc when compared with pasting temperatures of starch obtained by heat moisture treatment. this suggests that pre-treatment such as oven drying affects pasting temperature which causes molecular rearrangement of starch molecules resulting to closed packed structure of amylose and amylopectin molecules hence influencing water absorption within the amorphous and crystalline regions. pasting temperature indicates the temperature at which viscosity begins to rise [30] and it ensures swelling, gelatinization and subsequent gel formation during processing [31]. peak time is a measure of cooking time [32]. low peak time is desirable to save energy required to form gels from starch. therefore p. tuberregium flour obtained from air dried sclerotia has a moderate pasting temperature and peak time hence, will not consume much energy to cook. peak viscosity (pv) was 350.55rvu. this was in contrast to peak viscosity reported by kolawole, et al. [28] for p. tuberregium flour which had a value of 101.33rvu. the high pv reported for p. tuberregium flour obtained in our work could be due to less interaction between water molecules and amylose-amylopectin granules through hydrogen bonds. charles, et al. [33] attributed a high peak viscosity to be caused by low levels of amylose and failure to re-associate with amylopectin and strengthen the molecular network. peak viscosity can be affected by amylose/amylopectin content and ratios, molecular weight, intermolecular conformation and the degree of polymerization of amylose and amylopectin, amount of amylopectin branching and the presence of minor components [34]. hot paste viscosity (hpv, i.e trough) was 101.50rvu. this was higher than 72.62rvu reported kolawole, et al. [28] for p. tuberregium flour. hpv is the minimum viscosity value which measures the ability of paste to withstand breakdown during cooking [16]. breakdown viscosity (bd) obtained in this work for p. tuberregium flour from air dried sclerotia was 149.51rvu while kolawole, et al. [28] reported a value of 24.52rvu for p. tuberregium flour obtained from sclerotia oven dried at 40ºc. breakdown viscosity is a crucial factor in determining paste stability during heating and mechanical shear stress encountered during processing arinola, et al. [35]. adebowale, et al. [32] reported that a higher breakdown viscosity results to a reduced ability of flour to withstand heating and shear stress during cooking. therefore, in this current study it can be deduced that air dried p. tuberregium flour has a low ability to withstand heating and shear stress. final viscosity (i.e cold paste viscosity) and set back viscosity were 391.56rvu and 290.06rvu respectively while kolawole, et al. [28] reported values of 68.64rvu and 41.10rvu for final viscosity and set back viscosity respectively for p. tuberregium flour obtained from sclerotia oven dried at 40ºc. the extent to which amylose molecules join to form a strong gel after cooling affects the final viscosity subroto, et al. [29]. final viscosity indicates the re-association of starch granules especially amylose during cooling time after gelatinization and the formation of gel network [36]. set back viscosity represents viscosity of starch after heating to 50ºc. the lower the set back viscosity, the higher the resistance to retrogradation [37]. high set back viscosity obtained in this study suggests that flour produced from air dried p. tuberregium sclerotia can easily retrograde while the final viscosity indicated a high re-association of starch granules during cooling time after gelatinization. stability ratio and set back ratio were 0.29 and 2.77 respectively. stability ratio obtained for p. tuberregium flour was lower than stability ratio of wheat flour (0.53) reported by julianti, et al. [16]. set back ratio of p. tuberregium flour was higher than set back ratio of wheat flour (2.02) [16]. stability ratio provides information on the resistance of a starch paste to viscosity breakdown as shear is applied while set back ratio is an index of starch retrogradation tendency after gelatinization [16]. this confirms that p. tuberregiumflour obtained from air dried sclerotia has low resistance to viscosity breakdown as shear is applied and a high tendency to retrograde. table-2. pasting properties of pluerotus tubberregium flour from air dried sclerotia parameter value pasting temperature (ºc) 60.16±1.51 pasting time (min) 5.97±0.0 peak viscosity (rvu) 350.55±0.77 hot paste viscosity (rvu) 101.50±2.09 breakdown viscosity (rvu) 149.51±0.69 final viscosity (rvu) 391.56±2.17 setback viscosity (rvu 290.06±0.08 stability rat 0.29±0.01 setback ratio 2.77±0.05 note: values are means ± standard deviation. 3.3. functional properties results on functional properties of pluerotus tubberregium flour from air dried sclerotia is shown in table 3. its bulk density (bd) was 0.4g/ml. this value was higher than bd of 0.29g/ml reported for pluerotus tubberregium flour by kolawole, et al. [28]agaricusbisporus (0.22g/ml) and pluerotus ostreatus (0.28g/ml) reported by ishara, et al. [1] but lower than bd of termitomyces heimii (0.74g/ml) reported by due, et al. [38]. bulk density is influenced by agriculture and food sciences research, 2020, 7(2): 131-138 136 © 2020 by the authors; licensee asian online journal publishing group the structure of starch polymers and loose structure of starch polymer could result to low bulk density [39]. a low bulk density is ideal for infant meal [40]. this suggests that the bulk density obtained for flour from air dried pluerotus tubberregium sclerotia could be suitable for the production of infant formula. its water absorption capacity (wac) and oil absorption capacity (oac) were 4.20g/g and 4.13g/g respectively. wac observed in this work was lower than wac reported for a. bisporus (5.43g/g) and p. ostreatus (7.19g/g) reported by ishara, et al. [1]. flour with a water absorption capacity greater than 125ml/100g (ie 1.25g/g) points out good bakery property [41]. therefore it is an indication that pluerotus tuberregium flour from air dried sclerotia can be good in bakery products. as regards oac, the value obtained in this research was higher than oac of p. tubberregium flour (0.22ml/g) reported by kolawole, et al. [28]t. hemii (125.25%=1.2525g/g) due, et al. [38] but lower than oac a. bisporus (548.3%=5.483g/g) and p. ostreatus (462.6%=4.626g/g)[1]. variation in oac of flour from different food items could be influenced by differences in nonpolar side chains of proteins which bind hydrocarbon side chain of the oil through hydrophobic interactions [42]. the fewer the non-polar amino acids, the less the hydrophobic interactions with hydrocarbon chain of oil, hence resulting to low oil absorption capacity. emulsion activity (ea) and emulsion stability (es) obtained in this study were 30.22% and 45.12% respectively. ea of p. tuberregium flour was lower than ea of wheat flour (43.88)[33] while its es (45.12%) was higher than es of wheat flour (38.38%) [33]. foam capacity (fc) obtained in this study (7.27%) was lower than fc of t. heimii flour (16.67%) due, et al. [38] pearl millet (11.30%), quinoa flour (9%)[43] as well as wheat flour (12.92%) [33]. the foam stability (fs) obtained in this study (31.55%) was lower than fs of species of p. ostreatus flour which ranged between 45.7% and 66.8%[44] but higher than fs reported for wheat flour (1.94%)[33]. an inverse relationship exists between foam capacity and foam stability[33]. flours with low foam capacity could form small air bubbles enclosed by a dense and more stiff protein film as such may not collapse easily hence, resulting to a higher foam stability. swelling capacity and swelling index for p. tuberregium flour in this study were 4.17% and 1.61% respectively. mushroom flour has been reported to have low swelling capacity [45]. similarly, ojo, et al. [46] reported a significant decrease in swelling properties of cassava-mushroom flour blends with increased substitution of mushroom flour. the ability of starch particles to retain water and swell is based on the extent of water retention through hydrogen bonding and is controlled by its amylose content as well as its amylopectin side chains [47]. dispersibilty index for p. tubberregium flour was 53%. dispersibilty is an index that measures how well a flour/flour blends can be rehydrated with water[21]. a high dispersibilty enhances better reconstitution of starch in water to give fine and functional paste[31]. this suggests that p. tuberregium flour from air dried sclerotia will rehydrate moderately in water to give a fine paste. gelatinization temperature obtained for p. tubberregium flour in this study was 94ºc. this was quite higher than gelatinization temperatures of s. citritum (70 ºc) and p. ostreatus (80 ºc) reported byaniekemabasi, et al. [23]. gelatinization temperature of starch is affected by botanical origin, amylose content and the structure of the amylopectin in the molecules aniekemabasi, et al. [23]. high gelatinization temperature is associated with low amylose content and suggests thermal stability aniekemabasi, et al. [23]. it was observed that p. tubberregium flour from air dried sclerotia had a regular amylose content which could be categorized as low table 1 and this suggests its high gelatinization temperature. table-3. functional properties of pluerotus tubberregium flour from air dried sclerotia. parameter value bulk density (g/cm3) 0.40±0.00 water absorption capacity (g/g) 4.20± 0.12 oil absorption capacity (g/g) 4.13±0.12 emulsion capacity (%) 30.22±1.62 emulsion stability (%) 45.12±1.82 foam capacity (%) 7.27±0.10 foam stability (%) 31.55±1.58 swelling capacity (g/g) 4.17±0.88 swelling index (g/g) 1.61±0.07 dispersibility index (%) 53.00±1.00 gelatinization temperature (ºc) 94.00±0.00 note: values are means ± standard deviation. 4. conclusion it is concluded that flour from air dried pluerotus tubberregium sclerotia has a regular (ie low) and high resistant starch based on values for its amylose and resistant starch contents. therefore, it can be incorporated in food formulations for nutritional purposes where its resistant starch can function as a prebiotic. the pasting properties indicated that the flour can retrograde based on its set back viscosity and set back ratios and a low tendency to withstand shear stress based on its breakdown viscosity and stability ratio. its functional properties indicated that it can be used in food formulations where low bulk density is desirable such as in weaning foods and will be useful as a good thickener and in low fat foods based on its water and oil absorption capacities. however, there is need for technological modification to improve its pasting properties so as to generate flour with a high stability ratio and a low set back ratio and so withstand retrogradation and with improved resistance to shear stress. references [1] j. r. ishara, d. n. sila, g. m. kenji, and a. k. buzera, "nutritional and functional properties of mushroom (agaricus bisporus & pleurotus ostreatus) and their blends with maize flour," american journal of food science and technology, vol. 6, pp. 33-41, available at: https://doi.org/10.12691/ajfst-6-1-6. 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[47] h. tang, t. mitsunaga, and y. kawamura, "relationship between functionality and structure in barley starches," carbohydrate polymers, vol. 57, pp. 145-152. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 9 © 2022 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 9, no. 1, 9-16, 2022 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v9i1.3768 © 2022 by the authors; licensee asian online journal publishing group effects of demographic factors on population dynamics in imo state, nigeria; implications for farm labor availability and supply anyanwu u.g.1 osuji e.e.2 nwaiwu i.u.o.3 tim-ashama a.c.4 ibekwe c.c.5 osuala m.o.6 eze e.u.7 praise c.n.8 ( corresponding author) 1,3,4,6department of agricultural economics, federal university of technology owerri imo state, nigeria. 1email: uchechigerarda@gmail.com tel: +2348036900536 3email: niuche2004@yahoo.com tel: +234703 133 6540 4email: timashama5@gmail.com tel: +2347034773356 6email: mercbright1@yahoo.com tel: +2348036900536 2department of agriculture, alex-ekwueme federal university ndufu-alike abakaliki, nigeria. 2email: osujiemeka2@yahoo.com tel: +2348037351597 5department of agricultural science, alvan ikoku federal college of education, owerri, nigeria. 5email: ibchigozie@yahoo.com tel: +2347035202787 7,8department of cooperative economics and management, institute of management and technology, enugu, nigeria. 7email: emy4jesus@gmail.com tel: +2348034809184 8email: praisechris4@gmail.com tel: +2348037132395 abstract the study assessed the effects of demographic factors on population dynamics in imo state, nigeria. multi-stage sampling technique was used select 60 respondents. data collected were analyzed using descriptive statistics and the ordinary least square regression technique. results showed that the area was dominated by female farmers 56.7%, and are married 66.7%. most of the farming lands used was inherited, with more of hired laborers 76.7% used due to rural-urban drift. results also showed that over 70% of the people migrated from rural to urban communities. result further showed that age, gender, educational status, income level and poverty index were important and significant factors affecting population dynamics (expressed as index of rural– urban migration). the study recommended the crop farmers to join cooperative societies to raise funds to support large-scale production while the government is to provide basic rural infrastructures to checkmate rural-urban drift in the area. keywords: population dynamics, farm labor supply, rural-urban drift, farm productivity, demographic factors, white collar jobs, efficient labor utilization, food shortage, cost of labor, imo state. citation | anyanwu u.g.; osuji e.e.; nwaiwu i.u.o.; tim-ashama a.c.; ibekwe c.c.; osuala m.o.; eze e.u.; praise c.n. (2022). effects of demographic factors on population dynamics in imo state, nigeria; implications for farm labor availability and supply. agriculture and food sciences research, 9(1): 9-16. history: received: 5 january 2022 revised: 17 february 2022 accepted: 3 march 2022 published: 11 march 2022 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. authors’ contributions: all authors contributed equally to the conception and design of the study. competing interests: the authors declare that they have no conflict of interest. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study followed all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 10 2. methodology ..................................................................................................................................................................................... 11 3. results and discussion ................................................................................................................................................................... 11 4. conclusion and recommendations ............................................................................................................................................... 15 references .............................................................................................................................................................................................. 16 mailto:uchechigerarda@gmail.com mailto:niuche2004@yahoo.com mailto:timashama5@gmail.com mailto:mercbright1@yahoo.com mailto:osujiemeka2@yahoo.com mailto:ibchigozie@yahoo.com mailto:emy4jesus@gmail.com mailto:praisechris4@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v9i1.3768 https://orcid.org/0000-0001-8605-834x agriculture and food sciences research, 2022, 9(1): 9-16 10 © 2022 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the paper posited novel implications of rural-urban drift in imo state, nigeria as it affects farm labor availability and supply. it equally averred the various socio-demographic factors influencing population dynamics, productivity and efficient utilization of labor supply in imo state, nigeria. 1. introduction in recent times, there has been concerted campaign to increase the food production level in nigeria to feed her teeming population of about 162 million [1]. efforts have been geared at indigenously encouraging food production, production of agricultural raw materials, agricultural value-chain addition and establishing a formidable platform for the overall growth of the agricultural sector [2]. these and many others are periodically affected by population dynamics which influences agricultural production cum farm labor availability. population dynamics refers to short and long term changes in the size and age composition of populations and the biological and environmental processes influencing those changes. it deals with the way populations are affected by birth rate, death rate, immigration and emigration [3]. population dynamics is the study of how and why populations change in size and structure over time. world bank [4], viewed “population dynamics” as a media used to convey changes in the numbers, age, class distribution, sex ratio, and behavior of a population through time and space, determined by inherent characteristics of the individuals and mediated by environmental conditions, food resources, and interacting biotic agents. population dynamics is centered on studying peculiar characteristics of a selected population; these characteristics include changes in population growth rates, age structures and distributions of people [5]. it is the responsive and long-term changes in the numbers, individual weights and age composition of individuals in one or several populations, and the associated physical discrepancies influencing the population at a time [4]. population dynamics in this study is expressed as an index of rural-urban migration, (internal migration) which is in contrast to international or intercontinental migration, and this refers to movement within a country. it refers to the movement of people from the countryside that is from rural areas to cities often metropolitan cities of the country [3]. this change of residence is often related to labor migration and career change, i.e. from the primary to the second or third sector. this movement of people usually results in two outcomes that is one side or the destination area gains population, while the other side loses people respectively. in order to ensure a sustainable growth of both rural and urban areas, cooperation, networking and coordination between both parties are completely important and must be taken into consideration [6]. in developing countries such as nigeria, numerous factors are known to cause rural-urban migration mainly the push and pull factors. push-factors drive migrants out of rural areas while the pull factors draw migrants to urban areas. migration factors and determinants are very complex and can be separated into economic and non-economic factors. economic drivers include rural unemployment or underemployment, low wages and no assets while non-economic drivers play an additional role and primarily involve poor rural infrastructures relating to housing, roads, educational opportunities, healthcare systems, thus, these causes’ people to move in search for new jobs and better economic opportunities [7]. again, natural disasters, drought, famine, war and conflicts are additional factors that cause people to migrate. the nexus between population dynamics and farm labor availability is very complex; intense debates and widespread discourse have continued over several decades on population growth as it affects agriculture and farm labor availability. for the past decades, agricultural growth has been declining rapidly due to the mass movement of people out of the rural areas to the urban cities/town partly due to unfavorable living conditions and patterns and this exodus had resulted in the shortage of farm labor which is not readily available again [8]. agricultural intensification in the state is ought to be boosted by farm labor supply arising from increasing population but this is not the case in the area as most of their able-bodied young men had migrated to cities/towns in search of greener pastures. the high population growth leads to intensified pressures on available farm lands, forest and other economic resources, thus leading to increasing poverty and low agricultural productivity [7]. furthermore, overpopulation has resulted in land resource scarcity, fragmentation of farm plots, and ecological imbalance and degradation such as increasing emissions, soil erosion, deforestation, and over-use of natural resources, thus producing adequate food for the rapidly growing population remains a prime challenge. despite series of agricultural development policies in the state, it had suffered multifaceted challenges ranging from land scarcity, deforestation, malnutrition, recurrent drought and lack of improved agro-technologies [9]. the existing agricultural land is unable to feed the over growing population and thus many of the crop farmers had remained trapped in vicious circle of poverty, disease and hunger. rising hikes in food prices, unemployment, lack of pasture for livestock, and intensive removal of natural vegetation further aggravates food shortages [10]. land holdings has remained small and subjected to more divisions and fragmentations due to increasing or rising population as more demand is made on available lands, and however, land re-distribution is not anticipated in the near future. consequently, the rising population is expected to provide farm labor both in the short and long run basis, of which when efficiently utilized would results in increased agricultural production and output [11]. on accounts, where farm labor supply is grossly unavailable and inadequate due to labor migrations, this evidently undermines agricultural production and further induces food importations. the issue of population dynamics is quite necessary to provide government; the adequate tools needed for fresh proper planning, implementation, monitoring and evaluation of any proposed developmental plan, without which the expected economic development will remain a mirage and will not be achieved. this is because the ability of a nation to function effectively as a social, cultural and economic entity is based on its population dynamics [12]. efforts need to be put in place to bring forth a formidable growth plan for agricultural development especially (the rural areas of the country being at the core front as a result of cultivable lands available to them with the requisite labor supply readily available relative to the urban areas). nigeria as a nation has over 71 million hectares of cultivable land of which half of it is barely used, with the nation relying solely on food importations which depletes our foreign reserves. it has reached at an appalling stage that importation of food has sustained an increase of 11% per annum [6]. this rising percentage has continued to affect both local and/ or indigenous production as well as increased unemployment in the nation. again, increase in population density has further exacerbated and degraded the vegetation’s of most cultivated arable lands, hence forcing farmers to cultivate on marginal lands, with its attendant consequences [12]. in addition, population pressure has remained the most potent force for increased poverty and soil infertility among agriculture and food sciences research, 2022, 9(1): 9-16 11 © 2022 by the authors; licensee asian online journal publishing group household farmers. studies had shown that efforts are being made to cushion soil infertility via fertilizer distribution based on soil maps so that farmers can apply a custom-based fertilizer to mitigate the depletion of soil nutrients. this operation is implemented by means of internally adopted and externally induced technologies. it is on this backdrop that the paper presents the implications of demographic factors on population dynamics and its effects on farm labor availability in imo state, nigeria based on the current evidences in knowledge on agriculture, population and farm labor supply or availability. however, the novelty of this study is consequent upon the implication of population dynamics on farm labor supply in the state which has not been documented. 2. methodology the study was undertaken in imo state, which is one of the 36 states in nigeria. it lies in the southern part of nigeria with owerri as its capital and its largest city. imo state has 27 (twenty-seven) local government areas and it is located between latitude 4° 45ˡn and 7°15ˡ and longitudes 6° 50ˡe and 7° 25ˡ. imo state is bounded on the east by abia state, north by anambra state, south by rivers state and on the west by delta state and river niger. the state has a total land mass of 5530 square kilometers (2,140 square miles) [13]. the national population census of 2006 indicated that imo state had a population of about 3,934,899 people which comprise of both males and females. imo state was chosen for this study due to the high level of rural-urban migration recorded in the area which urgently calls for concern. a multi stage sampling technique was used for the sample selection. recall that imo state is divided into 3 (three) agro ecological zones which are owerri, orlu and okigwe zones respectively. firstly, one local government area (lga) was randomly selected from each of the 3 agricultural zones of the state namely oforola west lga for owerri agricultural zone, njaba lga for orlu agricultural zone and onuimo lga for okigwe agricultural zone. secondly, one autonomous community was again randomly picked from the above selected local government areas, for owerri west lga; oforola autonomous community, for njaba lga; umuele amazano autonomous community and for onuimo lga, owerri okwe autonomous community. thirdly, 20 farmers (respondents) each were randomly selected across the various autonomous communities from the sampling frame provided by the agricultural development program (adp) coordinators in the respective zones, making a total of 60 respondents from whom information were sourced and used for data analysis. data collected from the research were analyzed using descriptive statistical techniques such as percentages, means, frequency distributions and ordinary least square regression technique. the ordinary least square regression technique is expressed implicitly as y = f (x1, x2, x3, x4, x5, e) (1) where: y = population dynamics (index of rural-urban migration in the area). x1 = age composition of the farmers. x2 = educational level. x3 = gender composition of farmers. x4 = income level of farmers. x5 = poverty level (estimated using poverty line index). e = error term. note that individual ith households indicated dependents/household heads who were involved in rural-urban migration in the area. the ordinary least square regression is explicitly stated thus: linear functional form: y = bo + b1x1 + b2x2 + b3x3 + ……….. bnxn (2) power functional form iny = bo + b1inx1 + b2inx2 +b3inx3 + ……..+ bninxn (3) semi-log functional form: y = bo + b1inx1 + b2inx2 + b3inx3 +………… + bninxn (4) exponential functional form: iny = bo + b1x1 + b2x2 + b3x3 + ………….. + bnxn (5) (note: x1, x2, x3, x4 x5 remains the same as stated earlier) 3. results and discussion 3.1. demo-graphic characteristics of the farmers table 1 showed the age distribution of the respondents in the area. majority of the respondents, 30.0% are between the ages of 51 and 60 years of age. furthermore, 20.0% of the farmers lie within the age bracket of 61 70 years with a mean age of 50 years. this implies that the respondents in the area are advancing in age and not likely to migrate but rather becomes conservative and non-receptive towards the use of improved technologies. moreover, they provide the needed farm labour used in agricultural production [14]. table 2 revealed the marital distribution of the respondents in the area. it showed that 66.7% of the respondents were married while 8.3% and 25.0% were single and divorced. the 66.7% of the married group implied that they are likely to have some dependents who may likely support their farming activities (farm labor supply) and vis a vis less likely to migrate. this also calls for diversification of economic activities in order to meet family obligations [15]. agriculture and food sciences research, 2022, 9(1): 9-16 12 © 2022 by the authors; licensee asian online journal publishing group table 1. distribution of respondents by age. age frequency percentage 21 – 30 6 10.0 31 – 40 11 18.3 41 – 50 11 18.3 51 – 60 18 30.0 61 – 70 12 20.0 71 – 80 2 3.3 total 60 100.0 source: field survey data, (2018). table 2. distribution of respondents by marital status. marital status frequency percentage single 5 8.3 married 40 66.7 divorced 15 25.0 total 60 100.0 source: field survey data, (2018). table 3 revealed gender distribution of the respondents in the area. from the table, 56.7% of the respondents were females while 43.3% were males. it therefore means that females dominated the farming activities in the area and this formed a major reason while the females stayed back in the area unlike their male counterparts who largely migrated and are involved in non-farming activities in the cities. more so, women are culturally restricted from migration due to native tradition, customs and believe systems which inhibit their movement and thus, provided the needed farm labor utilized in agricultural production [16]. table 3. distribution of respondents by gender. gender frequency percentage male 26 43.3 female 34 56.7 total 60 100.0 source: field survey data, (2018). table 4 showed the education distribution of the respondents in the area. according to the table, 43.3% of the respondents have no formal education, 28.3% of them had completed primary education, 21.7% had secondary education and only 6.7% had tertiary education. it was gathered that respondents who had tertiary education abandoned their farm work and migrated to cities/towns in search of white collar jobs due to poor returns from their farming enterprises. it could be noted that their exit affected farm labor supply and/or availability in the area. education is seen as a planned process of bringing desirable changes in the behaviors, skills, attitudes and knowledge of individuals. it is believed that educated farmers are more exposed to new ideas which enhance decision makings [17]. table 4. distribution of respondents by level of education. level of education frequency percentage no formal education 26 43.3 primary education 17 28.3 secondary education 13 21.7 tertiary education 4 6.7 total 60 100.0 source: field survey data, (2018). table 5 revealed occupation distribution of the respondents in the area. 60% of the respondent took farming as a major occupation, 16.7% took to trading and 16.7% engaged in other economic activities (hunting, okada riding, fishing, etc.) as their source of livelihood. farming in the area was characterized with small land holdings, low income due to small scale engagement and low investment, hence resulting to high poverty profile, rural-urban drift and shortage of farm labor [18]. table 5. distribution of respondents by occupation. major occupation frequency percentage farming 36 60.0 trading 10 16.7 artisan 4 6.6 others 10 16.7 total 40 100.0 source: field survey data, (2018). table 6 showed the farming experience distribution of the respondents in the area. 41.7% of the respondents had between 11–20 years of farming experience. the mean farming experience in the area was 18.13 years, and this indicated that majority of the farmers were relatively experienced to handle farm challenges as they arise. their years of experience could also help to increase profitability of the farm business. again, experience farmers are less likely to migrate since they are well exposed to handle and overcome farm pressures and challenges and thus enhance farm labor supply [19]. agriculture and food sciences research, 2022, 9(1): 9-16 13 © 2022 by the authors; licensee asian online journal publishing group table 6. distribution of respondents by farming experience. farming experience frequency percentage 1 – 10 19 31.7 11 – 20 25 41.7 21 – 30 5 8.3 31 – 40 8 13.3 41 – 50 3 5.0 total 60 100.0 note: mean farming experience = 18.13years. source: field survey data, (2018). table 7 indicated the household size distribution of the respondents in the area. most of the respondents, 40.0% had between 9–11 persons in their household; 36.7% had between 7–9 persons. the mean household size is 7 persons per household. this indicated that on the average respondent had relatively large household size which implies that household head could engage them in agricultural activities (family labor supply) and hence reduce expenses on hired labor. it invariably indicated that the household head has more people to cater for which could cause them to migrate in search of greener pastures [6]. table 7. distribution of respondents by household size. household size frequency percentage 3 – 5 3 5.0 6 – 8 22 36.7 9 – 11 24 40.0 12 – 14 9 15.0 15 – 17 2 3.3 total 60 100.0 note: mean household size = 7 persons. source: field survey data, (2018). table 8 revealed the farm size distribution of the respondents in the area. it showed that 20.0% of the respondents had between 1.5–1.9 ha of farmland; only 16.7% of the respondents had farm size which ranges between 2.6–3.0 ha. the mean farm size is 1.7 ha. this indicated that majority of them possess relatively small farmland and practiced farming on a small-scale level. this could be as a result of the issue of land fragmentations which inhibit large scale agriculture. it could also be that the small land holdings of the farmers triggered ruralurban drift in the area, hence shortage of farm labor supply [20]. table 8. distribution of respondents by farm size. farm size frequency percentage 0.1 – 0.4 12 20.0 0.5 – 0.9 13 21.7 1.0 – 1.4 8 13.3 1.5 – 1.9 12 20.0 2.0 – 2.5 5 8.3 2.6 – 3.0 10 16.7 total 60 100.0 table 9 showed the methods of land acquisition distribution of the respondents in the area. it revealed that 76.7% of the respondents acquired their land through inheritance. this means that such land is usually passed to offspring after owner’s death. this acquisition nature often leads to non-usage of such lands for agricultural purposes as the beneficiaries may not be interested in agriculture. it was gathered that most of the beneficiaries of inherited lands in the area sold off their lands and migrated to cities/towns to set up businesses with the money got from the sale off thus, creating a reduction in farm labor supply. however, 5.0% and 13.3% of the respondents hired and purchased their lands respectively [21]. table 9. distribution of respondents by method of land acquisition. land ownership frequency percentage inheritance 46 76.7 hired 3 5.0 purchased 8 13.3 gift 3 5.0 total 60 100.0 source: field survey data, (2018). table 10 revealed the distribution of the respondents based on the type of labor used in the area. from the table, 76.7% of the respondents used hired labor for their farming activities, while only 23.3% of the respondents used family labor. this indicated a grave consequence of rural–urban migration drift as most able-bodied young members of the households had migrated to the cities and towns in search of better economic activities and this also created a huge gap in farm labor supply [22]. table 10. distribution of respondents by types of labor used in farming. types of labor frequency percentages family 14 23.3 hired 46 76.7 total 60 100.0 source: field survey data, (2018). agriculture and food sciences research, 2022, 9(1): 9-16 14 © 2022 by the authors; licensee asian online journal publishing group table 11 revealed the distribution of the respondents according to age categories of labor used in the area. 33.3% of the respondents reported that they engaged people in age category of 14–60 years as hired laborers in farming activities, 40% used children under 14 years category while 26.7% employed people of above 60 years. it implied that 66.7% of the people that worked on the farms in the area were under aged children (less than 14 years) and advanced aged (above 60 years). this means that the active aged populace of the area had migrated to the cities and towns where economic activities are more favorable thus leaving the farming activities to the children and aged categories to handle [23]. table 11. distribution of respondents according to age categories of labor used. age categories frequency percentages 0 – 14 24 40.0 14 – 60 20 33.3 above 60 16 26.7 total 60 100.0 source: field survey data, (2018). 3.2. cost of labor in farming activities table 12 revealed distribution of the respondents according to the cost of labor in farming activities in the area. majority, 33.3% of the hired laborers earned wage between n1, 000 – n3, 999, 21.7% earned between n 4,000– n 6,999. the mean wage was n4, 682.92 which is less than $1 per day. it indicated the low income returns and status of the hired laborers in the area and hence formed the major reason for rural–urban migration drift in the area; as active aged category tend to move to cities and towns where they could attract higher wage rate per man day thereby leading to a shortage in farm labor supply in the area [24]. table 12. distribution of respondents by cost of labor in farming activities. cost of labor input frequency percentages 1,000 – 3,999 20 33.3 4,000 – 6,999 13 21.7 7,000 – 9,999 4 6.7 10,000 – 12,999 5 8.3 13,000 – 15,999 9 15.0 ≥ 16,000 9 15.0 total 60 100.0 source: field survey data, (2018). 3.3. factors affecting availability and efficient utilization of farm labor supply table 13 revealed the distribution of the respondents according to factors affecting availability and efficient utilization of labor supply in the area. it showed that 88.3% of the respondents cited scarcity of labor as a major constraints and impediment to availability and efficient utilization of farm labor. this is due to the migration of able-bodied men to cities and towns in search of better economic activities they considered less strenuous vis a vis farming. 76.7% were concerned about high cost of labor, this resulted from the huge migration of the active aged populace to the cities, thus the available remaining active category now places a high premium on labor cost 71.7% revealed inadequate fertile lands as a major issue impeding availability and efficient labor utilization as most available lands are infertile and less productive, hence requires little or no labor. furthermore, 63.3% complained about inadequate funds to hire laborers for farming activities, it could be deduced that this factor arose from high cost of labor force as a result of rural–urban migration drift in the area [20]. table 13. distribution of respondents according to factors affecting availability and efficient utilization of labor supply. factors frequency* percentage* high cost of labor 46 76.7 scarcity of labor 53 88.3 lack of adequate fund 38 63.3 inadequate fertile land 43 71.7 note: *multiple responses recorded. source: field survey data, (2018). 3.4. reasons for emigration of labor force categories in the area table 14 revealed the major reasons for emigration of labor force out of the area. it was evident that economic interest 70%, i.e. eagerness of the respondents to look for better economic opportunities outside their domain; social stratification 68.3% i.e. people desire to belong to a higher social strata, occupation difference 60%, i.e. changing from one higher occupation to another; standard of living 58.3%, i.e. improvement in living standard; social mobility 36.7%, i.e. innate desires to move from one place to another; environmental difference 13.3% i.e. changing to a new environment; and rural community difference 11.7% were major reasons for rural–urban migration drift in the area. hence, the quest to attain higher standard of living and social and economic strata largely contributed deeply to the movement of people from the rural areas to the cities and towns [20, 24]. table 2 revealed the marital distribution of the respondents in the area. it showed that 66.7% of the respondents were married while 8.3% and 25.0% were single and divorced. the 66.7% of the married group implied that they are likely to have some dependents who may likely support their farming activities (farm labor supply) and vis a vis less likely to migrate. this also calls for diversification of economic activities in order to meet family obligations [15]. agriculture and food sciences research, 2022, 9(1): 9-16 15 © 2022 by the authors; licensee asian online journal publishing group table 14. distribution of respondents according to reasons for emigration of labor force categories. reasons for emigration frequency* percentages* rural community difference 7 11.7 environmental difference 8 13.3 occupation difference 36 60.0 economic interest 42 70.0 social mobility 22 36.7 standard of living 35 58.3 social stratification 41 68.3 note: *multiple responses recorded. source: field survey data, (2018). x linear, semi-log, cobb-douglas and exponential functional forms were employed to determine significant factors affecting the population dynamics in the area. in order to choose the lead equation, the functional forms were evaluated in terms of the statistical significance of the explanatory variables, the coefficient of multiple determinations (r2), and highest f-value. among the four functional forms estimated; double log functional forms was chosen as the lead equation based on the above criterion. the table revealed a coefficient of multiple determination of 0.813, indicating that 81.3% of the total variations in the dependent variable were explained by the independent variables investigated. the significant fvalue indicated that the model has a good predictive ability and goodness of fit. the result shows that age, gender, educational status, income level and poverty index were important and significant factors affecting population dynamics (expressed as index of rural–urban migration). it was further revealed that gender, educational and poverty status have positive relationship with the dependent variable while age and income level had an inverse relationship with the dependent variable. gender which comprises of both males and females’ influences population dynamics to an extent, as the males being anxious to acquire wealth at all cost migrate to the cities/towns in search of better economic opportunities and good standard of living, thus leaving the female folks with no choice other than farming, thereby affecting farm labor supply and availability [21]. empirical literatures had consistently reported the dominance of women in agriculture more than their male counterparts as a result of rural-urban drift [20]. as expected, educational level is positive and significant meaning that education exposes and equip individuals for better opportunities as the more educated individuals in the area migrate to the urban areas (cities, town) looking for white collar jobs which often not readily available, hence abandoning farming to the less educated ones who cannot read nor write. it connote that their absence created a huge gap in farm labor supply and availability in the area. it is known fact that education is a veritable tool which influences rural-urban drift as it allows individuals to take major decisions as it affects their wellbeing and environment [22]. poverty status was equally positively significant as any percentage increase in the poverty level of the respondents will evidently induce rural–urban drift. the deeper the poverty level, the increase in rural–urban drifts. poverty pushes individuals to seek alternative means of survival and livelihood which often may take him/her out of their immediate environment in search of greener posture hence, creating a gap in farm labor supply in rural areas [12]. age of the respondents and income level had an inverse relationship with population dynamics; hence any increase in age and income level of the respondents will automatically reduce rural-urban drift. this means that as farmers advance in age the eagerness to migrate to cities/towns in search of better opportunities diminishes as they prefer to stay back home and engage in any meaningful economic activities for survival and in return providing the needed farm labor supply and availability without any hindrance. most times, the farm labor provided may not be really efficient and effective because advancement in age of the respondents [15]. this phenomenon literally explains why agriculture is still in the hands of ageing populace. whereas, as income level of an individual increases, he/she is more relaxed and much comfortable to live in his/her immediate environment and therefore less anxious to consider rural–urban migration, thus closing the huge gap in farm labor supply and availability [18]. table 15. multiple regression estimates of demographic factors on population dynamics. variables linear exponential *double-log semi-log age in years 0.325 (0.692) -0.061 (-25.935)*** -0.294 (-2.242)** 0.602 (0.808) gender -0.047 (-0.684) 0.034 (0.502) 0.065 (12.438)*** 0.047 (0.364) educational status 0.022 (1.971)* 0.023 (8.300)*** 0.005 (8.407)*** -0.32 (0.244) income level -0.268 (-0.569) 0.096 (0.208) -0.278 (-5.313)*** 0.382 (5.433)*** poverty index 0.431 (1.832)* -0.033 (-0.145) 0.558 (2.468)** 0.360 (0.838) r2 0.779 0.786 0.813 0.669 adj. r2 0.739 0.747 0.768 0.550 fvalue 18.556 20.391 28.319*** 5.618 note: n/b *** = significant @ 1%, ** = significant @ 5%. * = significant @ 10%, tvalues are figures in parentheses. 4. conclusion and recommendations population dynamics connote to short and long term changes in the size and age composition of populations and the structural and environmental processes influencing those changes. its impacts have really being felt in agriculture and as a result initiated rural-urban migration drift among the rural populace. findings of the study showed that majority of the respondents were females, married with a household size of 7 persons. respondents were majorly in the age brackets of 51 and 60 years. educational status showed that only 6.7% had tertiary education which initiated rural-urban drift in the area. it is noted that education equips one for better economic agriculture and food sciences research, 2022, 9(1): 9-16 16 © 2022 by the authors; licensee asian online journal publishing group opportunities which was the case of the educated respondents met in the area. due to the ceaseless migration of the populace, high cost of labor versus labor scarcity was highly rated among the factors influencing the availability and utilization of farm labor in the area. again, people migrated due to occupational difference, rural community difference, social mobility, economic interest, social stratification, etc. the result further showed that age, gender, educational status, income level and poverty index were important and significant factors affecting population dynamics (expressed as index of rural–urban migration). the study recommended the farmers to join cooperative societies as to raise funds to support large-scale production while the government is to provide basic rural infrastructures to checkmate rural-urban drift. references [1] j. anson, w. bartl, and a. kulczycki, roots and fruits of population growth and social structures: demographic and sociological vistas”, in studies in the sociology of population. cham: springer, 2019. [2] t. a. oluyemi and o. p. oluwaseyi, "population growth and human resource utilization nexus in nigeria," journal of humanities and applied social sciences, vol. 1, pp. 1-12, 2020. [3] w. mekuria, "the link between agricultural production and population dynamics in ethiopia: a review," advanvced plants agricultural resource, vol. 8, pp. 348-353, 2018.available at: https://doi.org/10.15406/apar.2018.08.00336. [4] world bank, "nigeria’s booming population requires more and better jobs," ed usa: world bank group, 2016, p. 126. [5] fma, "agricultural transformation agenda, a document on the developmental plan for agriculture in nigeria," ed abuja, nigeria: federal ministry of agriculture, 2019, pp. 134-140. [6] e. b. essien, "population growth and economic growth performance in nigeria (1981–2014)," turkish economic review, vol. 3, pp. 143-159, 2016. [7] world bank., world development indicators. usa: world bank group, 2019. [8] n. k. jha and r. saritha, "female labour force participation (flfp) and its significance in economic growth of india: an overview," research journal of humanities and social sciences, vol. 10, pp. 270-278, 2019.available at: https://doi.org/10.5958/23215828.2019.00049.4. [9] p. v. okoye and r. a. ezejiofor, "the effect of human resources development on organizational productivity," international journal of academic research in business and social sciences, vol. 3, pp. 250-268, 2013.available at: https://doi.org/10.6007/ijarbss/v3i10/295. [10] j. saghir and j. santoro, "urbanization in sub-saharan africa," in meeting challenges by bridging stakeholders. washington, dc, usa: center for strategic & international studies, 2018. [11] e. h. tartiyus, m. i. dauda, and a. peter, "impact of population growth on economic growth in nigeria (1980-2010)," iosr journal of humanities and social science, vol. 4, pp. 115-123, 2015. [12] m. m. yakubu, b. n. akanegbu, and g. jelilov, "labour force participation and economic growth in nigeria," advances in management and applied economics, vol. 10, pp. 1-14, 2020. [13] mlsupo, documented reports of imo state, nigeria. imo state, nigeria: ministry of land survey and urban planning owerri, 2017. [14] m. afzal and s. awais, "population and poverty nexus in pakistan," journal of independent studies and research, vol. 12, pp. 63-75, 2014.available at: https://doi.org/10.31384/jisrmsse/2014.12.2.5. [15] e. mbah, c. ezeano, and m. agada, "effects of rural-urban youth migration on farm families in benue state, nigeria," international journal of agricultural research, innovation and technology, vol. 6, pp. 14-20, 2016.available at: https://doi.org/10.3329/ijarit.v6i1.29207. [16] r. mistura, a. ogunniyi, k. k. salman, m. oyeyemi, and m. salawu, "migration, labor mobility and household poverty in nigeria: a gender analysis," economies, vol. 7, pp. 1-24, 2019.available at: https://doi.org/10.3390/economies7040101. [17] food and agriculture organization (fao), "a report of food and agriculture organization, rome." retrieved from: http://www.fao.org/faostat/en/#home, 2017. [18] m. bell, e. charles‐edwards, p. ueffing, j. stillwell, m. kupiszewski, and d. kupiszewska, "internal migration and development: comparing migration intensities around the world," population and development review, vol. 41, pp. 33-58, 2015.available at: https://doi.org/10.1111/j.1728-4457.2015.00025.x. [19] h. ghebru, a. mulubrhan, m. george, and o. adebayo, role of land access in youth migration and youth employment decisions: empirical evidence from rural nigeria (no. 58). washington, dc: international food policy research institute (ifpri), 2018. [20] ilo, global and continental policy frameworks guiding labour migration and labour mobility in africa. abidjan: international labour organisation (ilo), regional office for africa, 2019. [21] u. mukhtar, z. zhong, b. tian, a. razzaq, and t. hina, "does rural–urban migration improve employment quality and household welfare? evidence from pakistan," sustainability, vol. 10, pp. 42-81, 2018.available at: https://doi.org/10.3390/su10114281. [22] osuji, "impact of sustainable soil management techniques on land productivity and poverty levels of arable crop farmers in south east, nigeria," unpublished dissertation, department of agricultural economics, michael okpara university of agriculture, umudike nigeria, 2017. [23] a. ogunniyi, o. k. oluseyi, o. adeyemi, s. k. kabir, and f. philips, "scaling up agricultural innovation for inclusive livelihood and productivity outcomes in sub-saharan africa: the case of nigeria," african development review, vol. 29, pp. 121-134, 2017.available at: https://doi.org/10.1111/1467-8268.12267. [24] a. ogunniyi., m. george, o. kehinde, f. olusegun, and m. r. adedoyin, "the paradigm of governance quality, migration and its implication on food and nutritional security in sub-saharan africa: what does dynamic generalized method of moments estimation reveal?," presented at the 2018 conference international association of agricultural economists, (no. 275994), vancouver, bc, canada, july 28-august 2018, 2018. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.fao.org/faostat/en/#home 97 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 97-104, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.97.104 © 2020 by the authors; licensee asian online journal publishing group utilization of soursop (annona muricta) flour for the production of chin-chin n.j. deedam1 m.a. china2 h.i. wachukwu3 ( corresponding author) 1,2,3department of food science and technology, rivers state university, nkpolu oroworukwo, port harcourt, rivers state, nigeria. abstract the present study was aimed at utilizing soursop flour for the production of chin-chin. soursop (ss) was processed to flour. chin-chin was prepared from blends of wheat and soursop flours using 90:10, 80:20, 70:30, 60:40, 50:50 of wheat flour to soursop flour (ssf), and 100% wheat flour as control. proximate and sensory analysis of the chin-chin was determined using standard methods. the samples were also stored for 3 weeks and evaluated at weekly intervals for total bacterial and fungal counts. proximate composition of the chin-chin revealed a significant (p<0.05) increase in ash (0.42-0.96%), fat (33.31-39.29%), crude protein (5.32-7.94% protein), crude fibre (0.95-1.12%), and moisture content (4.85-7.65%) with a decrease in carbohydrate content (55.14-42.94%) as substitution of soursop flour increased. energy content decreased as substitution of soursop flour increased, but beyond 30%, level, the energy content was observed to increase significantly. substitution of soursop flour with wheat flour at the level of 10% compared favorably with the control sample suggesting that acceptable chin-chin could be produced at ssf substitution of up to 10%. the samples presented adequate microbiological conditions after storage of 3 weeks with counts ranging from 5.20×103-7.00×104cfu/g and 4.00×1046.00×104cfu/g, for total bacterial and fungal counts, respectively. the study therefore showed that soursop can be utilized for the development of chin-chin with improved nutritional value over 100% wheat flour thereby serving as a nutritious household food which will help address the problem of protein-energy malnutrition. keywords: soursop, chin-chin, wheat, proximate, household, utilization. citation | n.j. deedam; m.a. china; h.i. wachukwu (2020). utilization of soursop (annona muricta) flour for the production of chin-chin. agriculture and food sciences research, 7(1): 97-104. history: received: 20 april 2020 revised: 26 may 2020 accepted: 29 june 2020 published: 22 july 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 98 2. materials/methods .......................................................................................................................................................................... 98 3. results and discussion ................................................................................................................................................................. 100 4. conclusion ....................................................................................................................................................................................... 102 references ............................................................................................................................................................................................ 103 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.97.104&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.97.104&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1919 agriculture and food sciences research, 2020, 7(1): 97-104 98 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to literature by enhancing the utilization of soursop flour towards the production of chin-chin. the nutritional composition of chin-chin was enhanced by the incorporation of soursop flour. the acceptable level of soursop flour to wheat flour for chin-chin production was also established. 1. introduction in nigeria and other developing countries, the use of wheat flour for the production of baked goods has been on the increase. these products are highly consumed due to their cost, ready-to-eat nature and the availability of wheat flour as wheat flour is being imported consistently into the country [1]. most cereals are limited in essential vitamins and minerals while nuts and fruits are rich in them. thus a combination of such food products will improve the nutritional blend that will give it better nutrition value compared to wheat flour constituents alone. wheat flour which is a product of wheat grain is low in protein, vitamins and minerals. sengev, et al. [2]reported that wheat flour have a nutritional value inferior to flours produced from composite flours of cereals, fruits, tubers or nut. the fortification of wheat flour with fruits and nuts flour for the production of nutritious snacks would improve the nutritional value of wheat based products. with the current trend in nutrition, consumers are highly conscious of their health which has resulted to the consumption of foods with health-promoting effects [3]. this has also promoted research by food professionals and industries in the development of baked goods from non-wheat flour blends. this involves incorporation of food materials that are rich in nutrient such as fibre, protein, minerals and vitamins with wheat flour, a process called composite flour technology. several studies have been carried out on the chemical and physical properties of various flours blended with wheat flour and this showed that composite flours produced from cereals, tubers, fruits, legumes and nuts are preferable and have an advantage of enhancing the overall nutrition and sensorial properties of the finished product than products produced from single wheat flour [4-8]. however, the selection of components to be used in composite blends depends on the availability and nutritional potentials of the raw materials [2]. soursop (annonamuricata) belongs to the family annonaceae and a native of tropical north and south america [9]. they are either irregular, ovoid or heart shaped fruits which is 15-30 cm long and a width of 10-30 cm. it has a thick skin which is dark green with sparse curved spines [10]. the fruit mesocarp resembles white cotton and possesses a stingy and sweet-sour taste containing many dark seeds [11]. according to iombor, et al. [11] the oven dried soursop contains 8.10% moisture, 21.30% protein, 2.30% fat, 16.30% fibre, 11.40% ash, 40.70% carbohydrate and 2871.60kcal of energy. soursop is often consumed as a dessert fruit or utilized by food industries for the production of beverages, ice cream, wine, candy and syrup [9, 12]. the utilization of soursop flour for the production of bread has also been reported by zabidi and yunus [13]. iombor and banjo [1] also investigated the effect of soursop flour inclusion to wheat flour for bread production and reported that the flour from soursop has quality attributes which could be utilized for the production of bread and other baked goods thereby diversifying the utilization of soursop and improving the quality of food products produced from it. zabidi and yunus [13] also added that soursop flour could be utilized in enhancing the nutritional content (especially dietary fibre, minerals and protein contents) of various food products. akomolafe and ajayi [14] also reported that soursop possesses some therapeutic properties such as antioxidant and anticancer. they further added that the soursop fruits can be explored as a viable source of natural antioxidants for the production of functional foods. chin-chin is a fried or baked snack which is popular across nigeria and other parts of west africa [15]. it is a sweet, doughnut-like product prepared primarily from wheat flour, butter, eggs and milk. this is made into a stiff paste, rolled, cut and shaped into ¼ inches and then fried using vegetable oil or baked until it is golden brown and crispy [16]. chin-chin is usually consumed by children and adolescents. as a result of the general acceptance of chin-chin, there is a need to enrich it with nutritionally rich ingredients such as soursop flour. in nigeria, soursop fruit is highly underutilized. little industrial value is placed on the fruit due to high post harvest losses arising from poor storage and preservation technologies [13]. this poses a lot of concern for nutrients wasted which are not properly utilized. current estimates also have it that over 60% of fruits are lost to poor post harvest handling and storage [1]. soursop is also mainly consumed in the form of fruit juice or taken in its fresh form and no record of soursop flour used for the production of chin-chin has been carried out. in order to increase the demand and reduce wastage of this fruit, it can be incorporated into wheat flour for the production of baked products. the consumption of these products will contribute to ensuring food security at the household levels and also improve the nutritional status of households. the aim of this work therefore was to utilize soursop flour for the production of chin-chin and also to evaluate the proximate composition, sensory and microbiological quality of the product. this was with a view to reduce post harvest losses, encouraging utilization of soursop and also, increase overall nutrient of the products. 2. materials/methods 2.1. sources of materials soursop fruit was obtained locally from fruit garden market at d-line area port harcourt city, rivers state, nigeria. refined wheat flour and other ingredients such as margarine, eggs, brown sugar, salt, milk and vegetable oil were purchased from mile 3 market diobu, port harcourt. chemicals used for all analysis were of analytical grade. 2.2. processing of soursop flour soursop fruits were processed into flour as shown in figure 1. matured and ripedsoursop fruits (3 kg) were washed under running water and peeled gently with stainless knife. the peeled fruits were sliced into 5 mm thickness and the core and seeds removed, then the pulp was cut into small pieces and oven dried at 60ºc for 48 hr. the dried fruit was milled into flour using a nutri-blender (bl487q model) and was passed through a us70 (180µm diameter) sieve. the flour obtained was stored in an air tight plastic container at room temperature (37oc) to prevent spoilage of sample until used for further analysis. agriculture and food sciences research, 2020, 7(1): 97-104 99 © 2020 by the authors; licensee asian online journal publishing group figure-1. flow chart for the production of soursop flour. source:zabidi and yunus [13]. 2.3. preparation of chin-chin chin-chin was prepared using the method of akindele, et al. [17]. the ingredients except vegetable oil table 1 were mixed and rubbed together in a large bowl. the dough was placed on a flour surface and kneaded until smooth and elastic. the kneaded dough was then rolled out to appropriately 2 cm thickness and then cut into ¼ (2 cm by 2 cm) in size. vegetable oil (400 ml) was poured into a deep fryer mc 1800 model and allowed to be hot enough. thereafter, the cubes were poured into the hot oil and the chin-chin deep fried for 8 min until golden brown. the fried chin-chin was removed, drained off excess oil and allowed to cool then packaged for analysis. table-1.formulation of wheat and soursop composite flour for the production of chin-chin. ingredients chin-chin samples a b c d e f refined wheat flour (g) 100 90 80 70 60 50 soursop flour (g) 10 20 30 40 50 sugar (g) 40.00 40.00 40.00 40.00 40.00 40.00 margarine (g) 25.00 25.00 25.00 25.00 25.00 25.00 baking powder (g) 2.00 2.00 2.00 2.00 2.00 2.00 nutmeg (g) 1.00 1.00 1.00 1.00 1.00 1.00 eggs (whole) 1 1 1 1 1 1 milk (g) 15.00 15.00 15.00 15.00 15.00 15.00 water (ml) 15.00 15.00 15.00 15.00 15.00 15.00 salt (g) 0.5 0.5 0.5 0.5 0.5 0.5 vegetable oil (l) 2.00 2.00 2.00 2.00 2.00 2.00 source: akindele, et al. [17]. 2.4. proximate analysis and energy determination of chin-chin proximate analysis (moisture, ash, protein, fat and crude fibre) of the food products was determined using the method of aoac [18] while total available carbohydrate was calculated by difference using the formula: 100% (% moisture + % ash + % crude protein + % fat + % crude fibre) the energy content (e) was calculated using atwater factor method as described by adegunwa, et al. [15]. e = (9 x protein) + (4 x fat) + (4 x carbohydrate) 2.5. sensory evaluation of chin-chin the chin-chin samples were subjected to sensory evaluation 30 minutes after preparation using 9-pont hedonic scale. this was carried out using a 20 panelist made up of students of department of home science/hospitality management and tourism, michael okpara university of agriculture, umudike, abia state, nigeria. the following attributes namely appearance, crispness, texture, flavour and overall acceptability were assessed. the assessment ranged from 9 (like extremely) to 1 (dislike extremely) as described by iwe [19]. the criterion for selection of panelist was based on their knowledge of the products to be evaluated. the panelists were asked to sit on the laboratory stools with spaces apart and with coded paper given to each of them according to the samples to be evaluated. the panelists were instructed to rinse their mouth with water before and after tasting the samples. 2.6. microbial evaluation of chin-chin during storage chin-chin from different blends of soursop and wheat flour was crushed separately with the aid of a sterile ceramic mortar and pestle. the samples were packaged in an airtight container, stored at room temperature (37oc) agriculture and food sciences research, 2020, 7(1): 97-104 100 © 2020 by the authors; licensee asian online journal publishing group and analyzed for their total bacterial and fungal populations at weekly intervals for 3 weeks. one gram (1.0 g) of each crushed sample was transferred into a 10ml sterile normal saline, separately. the mixtures were shaken vigorously, and then 0.1ml each mixture was inoculated on nutrient agar (na) plate and sabouraud dextrose agar (sda) plate in duplicates using the spread plate method [20]. the inoculated na plates were incubated 370c for 24 h while the inoculated sda plates were incubated at ambient temperature for 5 days. after incubation, counts of the colonies on the na and sda plates were used to calculate the bacterial and fungal population respectively, with the aid of the equation below. population (cfu/g) = colonyx 10ml 0.1ml x 1g 2.7. statistical analysis results were analyzed statistically using analysis of variance (anova). significant differences in mean values was calculated by least significant difference (lsd) test and duncan’s multiple range test (dmrt) using the statistical package for the social sciences (spss) version 23.0 at the level of p<0.05. 3. results and discussion 3.1. proximate composition of chin-chin produced from wheat and soursop flour blends table 2 shows the proximate composition of chin-chin produced from wheat and soursop flour (ssf) blends. the range of moisture content for all the chin-chin samples was between 4.85% and 7.65% with control sample as least and sample with 50% ssf substitution as highest. there was a significant increase (p<0.05) in the moisture content as the substitution with soursop flour increased. samples substituted with 20, 30 and 40% ssf were significantly (p<0.05) similar. the increase could be attributed the increase to the hygroscopic nature of soursop flour due to the presence of reducing sugars. the trend is in concordance with the study of deedam and mbah [21] for granola produced from wheat and soursop flour blends. they reported the increase in moisture content of granola on substitution with soursop flour to be due to the high crude fibre content of soursop of 28.21% [22] the moisture values from the present study is in line with the findings of akindele, et al. [17] who reported values between 4.17-6.80% for chin-chin enriched with pumpkin and indian spinach vegetables. it was also comparable to the moisture content of chin-chin made from trifolate yam flour enriched with pumpkin seeds (5.38-7.30%) as reported by adelakan, et al. [23]. the range of moisture obtained from this study is at the minimum limit of moisture for baked goods [24]. the low moisture content of the chin-chin samples shows that the product will store better, ensuring high shelf stability [25]. the ash content of the chin-chin samples ranged between 0.420.96% with sample substituted with 50% ssf as highest and control sample as least. an increase in the ash content of chin-chin samples was observed as substitution with soursop flour increased. the control sample was significantly (p<0.05) different from others while samples substituted with ssf at all levels were significantly similar (p<0.05). the increase in ash content is attributed to the high ash content of 11.4% soursop flour [11]. this study agrees with the statement of wijewardana, et al. [22] that high ash content of soursop flour can be used to enrich the lost nutrients in wheat flour during refining. ibidapo, et al. [26], equally reported an increase in ash value of cookies enriched with carrot powder with values between 1.47-1.82%. ash content from this study is comparable with the findings of adebayooyetoro, et al. [27] with values between 0.470.75%. it was also similar to the study of eke-ejiofor and beleya [28] who reported ash content of 0.40-1.09% for chin-chin produced from high quality cassava flour and tiger nut residue flour blends. the result therefore indicates that the chin-chin samples would contribute mineral elements to the body. the substitution of wheat flour with soursop flour resulted to an increase in the fat content of chin-chin samples from 33.31-39.29%. chin-chin from 50% soursop flour substitution recorded the highest value while the control sample recorded the least. this increase in fat content did not differ significantly (p>0.05) among samples substituted with 30, 40 and 50% ssf while control sample was significantly (p<0.05) different from others. this increase in fat content of chin-chin from composite flours as compared to chin-chin from wheat flour could be due to substitution effect, as a result of higher fat content of soursop flour. this confirms the earlier reports of fasakin, et al. [9] that soursop fruit has high fat content of 22.57%. this trend was also observed by akindele, et al. [17]and sengev, et al. [2] for vegetable enriched chin-chin (11.67-17.34%) and wheat/sweet potato/mango mesocarp flour blends chin-chin (11.00-24.00%), respectively. fat content from the study was higher than the findings of adegunwa, et al. [15] and eke-ejiofor and beleya [28] for millet-wheat composite chin-chin and high quality cassava flour/tiger nut residue chin-chin, respectively. the high fat content of the chin-chin from this study is attributable to the fat from the frying oil and difference in recipes. the protein content of all the chin-chin samples ranged from 5.32% in control sample to 7.94% in sample substituted with 50% soursop flour. protein content of control sample and samples substituted with 10, 20 and 30% ssf were significantly (p<0.05) similar. a significant (p<0.05) increase in protein content was observed at 40 and 50% ssf substitution. this result shows that there was an increase in the protein content with corresponding increase in the substitution of soursop flour. this increase is attributed to the high protein content of soursop fruit (15.62%) as reported by emelike and akusu [29]; prescott, et al. [20]. similar result was also reported by deedam and mbah [21] for granola substituted with soursop flour. the protein content of wheat-soursop chin-chin samples were lower than those reported for wheat-tiger nut flour chin-chin (7.66-11.58%) and millet-wheat chinchin (12.63-19.50%) as reported by adebayo-oyetoro, et al. [27] and adegunwa, et al. [15] respectively. these differences could be attributed to the differences in recipe and also the use of soursop. crude fibre content of the chin-chin ranged from 0.95-01.12% with sample substituted with 50 ssf having the highest and sample with 10% ssf as lowest. there was an increase in the crude fibre of the chin-chin samples as substitution with soursop flour increased; however, this increase was not significant (p<0.05) between control sample and samples substituted with 10, 20, 30 and 40% ssf. the significant (p<0.05) increase in crude fibre content at 50% ssf substitution is due to the high crude fibre content of sourso (16.30%) as reported by iombor, et al. [11]. similar increase was also reported by deedam and mbah [21] for granola substituted with soursop flour. crude fibre of chin-chin from this study was lower when compared with millet-wheat composite chin-chin (4.84agriculture and food sciences research, 2020, 7(1): 97-104 101 © 2020 by the authors; licensee asian online journal publishing group 5.23%) and high quality cassava flour/tiger nut residue chin-chin (11.08-12.95%) as reported by adegunwa, et al. [15] and eke-ejiofor and beleya [28] respectively. the crude fibre value was however, comparable with wheat and tiger nut flour chin-chin (0.28-0.66%) as reported by adebayo-oyetoro, et al. [27]. these differences could be due to the recipe used in the formulation process and the levels of fibre contained in the flour being used. a significant (p<0.05) decrease was observed in the carbohydrate content, which ranged from 55.14% to 42.94% with the highest value in control sample and the lowest in sample substituted with 50% soursop flour. the increase in proportion of soursop flour brought about a decrease in the carbohydrate content of the granola samples. this result is in concordance with the findings of adebayo-oyetoro, et al. [27] who reported a decrease in carbohydrate content (62.76-52.95%) of wheat/tiger nut flour chin-chin as substitution of tiger nut increased. the carbohydrate values from this study are higher than 4.44-32.79% for high quality cassava flour/tiger nut residue flour chin-chin [28]. energy content of the chin-chin samples ranged from 393.34-401.68kcal. control sample had the highest value while sample substituted with 30% ssf was lowest. the control sample and samples substituted with 40% and 50% ssf were significantly (p<0.05) similar. this indicated that energy content of the chin-chin samples decreased with increasing level of ssf substitution, but beyond 30% level, the energy content was observed to increase significantly (p<0.05). this could be attributed to the protein, fat and carbohydrate contents of the blend constituents which contributed to the energy value of the chin-chin samples. the result from this study implies that the substitution of soursop flour above 40% level could help to boost the calories level of chin-chin. ibidapo, et al. [26] reported energy value of biscuits enriched with carrot flour (448.25-461.02kcal) which is higher that obtained from this study. table-2.proximate composition of chin-chin produced from wheat and soursop flour blends. note: mean values bearing different superscript in the same column differ significantly (p˂0.05). key: sga= wheat flour (100%), sgb=wheat/soursop flour (90:10), sgc=wheat/soursop flour (80:20), sgd=wheat/soursop flour (70:30), sge=wheat/soursop flour (60:40), sgf=wheat/soursop flour (50:50). 3.2. sensory properties of chin-chin produced from wheat and soursop flour blends the sensory scores of the chin-chin samples produced from wheat and soursop flour blends are presented in table 3. appearance ranged from 5.65-7.45 with control sample as most preferred and sample with 40% soursop flour (ssf) substitution as the least. chin-chin substituted with soursop flour at different levels were significantly (p<0.05) similar while control sample was significantly (p<0.05) different. the scores for appearance suggest that the higher percentage of soursop flour, the lower the mean appearance score of the chin-chin. this might be attributed to the addition of soursop flour which resulted to colour darkening of the chin-chin. the low mean scores for appearance of chin-chin substituted with soursop flour could also be due to the known popularity of the panellists with chin-chin prepared from wheat flour. similar findings were also reported by wordu and akusu [30] for wheat and fluted pumpkin flour blend chin-chin. the scores suggest that chin-chin substituted with up to 30% ssf have some level of acceptable appearance as the scores are above 6.00. scores for taste of the chin-chin samples ranged from 5.50-7.50 with control sample as most preferred and sample substituted with 50% ssf as the least. the control sample was significantly (p<0.05) different from all samples but not from chin-chin with 10% ssf substitution. the decrease in mean scores of taste observed as soursop flour substitution increase may also be as a result of the sweet-sour taste of soursop which may have altered the original taste of the chin-chin. similar finding was also reported by zabidi and yunus [13]. the mean sensory scores for crispness and texture of the chin-chin samples ranged from 5.30-7.20 and 5.757.50 with control sample as most preferred and sample substituted with 40% ssf as the least. control samples was significantly (p>0.05) different from all other samples. crispness and texture of the chin-chin samples decreased with increase in percentage of soursop flour. the low preference for crispness in chin-chin substituted with soursop flour as compared to 100% wheat flour chin-chin may be due to moisture uptake by soursop flour. it has been reported that moisture uptake leads to loss of crispness of food products [2]. similar finding was also reported by adebayo-oyetoro, et al. [27] for chin-chin made from wheat and tiger nut flour. the decrease in texture on substitution with soursop flour may be attributed to the high crude fibre content, which makes the texture less tender [2]. this was also reported by kehinde, et al. [16] for chin-chin produced from wheat-tiger nut pomace. the mean scores of crispness and texture of samples substituted with 10-30% soursop flour was above 6 suggesting that these samples may be acceptable. flavour is an important attribute that influences the acceptance of finished food products even before they are tasted. flavour of the chin-chin samples ranged from 5.65-7.25 with control sample as most preferred and sample with 50% ssf substitution as the least. substitution of wheat flour with soursop flour significantly (p<0.05) decreased the flavour of the chin-chin samples. the scores also suggest that chin-chin samples substituted with up to 40% soursop flour have some level of acceptable flavour as the scores were above 6.00. overall acceptability of chin-chin ranged from 5.58-7.38 with 100% wheat flour chin-chin as most preferred. this was followed closely by chin-chin substituted with 10% ssf. sample substituted with 50% ssf was least preferred. the control sample was significantly (p<0.05) different from others while samples substituted with 20, 30, 40 and 50% ssf were significantly (p<0.05) similar. this study is in line with the findings of anozie, et al. [31] and china and ezema [32] who reported that control snack made from 100% wheat flour was most preferred. the overall acceptability of the chin-chin was observed to decrease with increase in the level of soursop flour substitution. this result is in agreement with the findings of ajani, et al. [33] who reported that increased levels of samples moisture (%) ash (%) fat (%) protein (%) crude fibre (%) cho (%) energy (kcal) sca 4.85±0.01d 0.42±0.03b 33.31±1.49c 5.32±0.00b 0.96±0.00b 55.14±0.20a 401.68±1.91a scb 5.70±0.16c 0.84±0.08a 35.31±0.13b 5.36±0.05b 0.95±0.00b 51.76±1.26b 396.84±0.93b scc 6.81±0.12b 0.88±0.01a 36.88±0.30ab 5.68±0.00b 0.99±0.01b 48.77±0.01bc 393.72±1.10c scd 7.02±0.11b 0.88±0.01a 38.35±0.27a 5.75±0.04b 0.98±0.01b 46.98±0.45c 393.34±0.91c sce 7.21±0.00b 0.89±0.01a 38.42±1.13a 7.21±0.00a 0.99±0.01b 45.28±1.37c 399.69±0.96a scf 7.65±0.11a 0.96±0.04a 39.29±1.73a 7.94±0.00a 1.12±0.00a 42.94±1.84d 400.38±0.42a agriculture and food sciences research, 2020, 7(1): 97-104 102 © 2020 by the authors; licensee asian online journal publishing group breadfruit flour in chin-chin resulted in significant decrease in overall acceptability. on the basis of this observation, substitution of soursop flour with wheat flour at the level of 10% could be considered the best from sensory point of view. table-3.mean sensory scores from taste panel of chin-chin from wheat and soursop flour blends. samples appearance taste crispness texture flavour overall acceptance sca 7.45a 7.50a 7.20a 7. 50a 7.25a 7.38a scb 6.70b 6.90a 6.40ab 6.45ab 6.60ab 6.75ab scc 6.15b 5.95b 6.45ab 6.30b 6.40b 6.18b scd 6.25b 5.70b 6.15b 6.00b 6.35b 6.03b sce 5.65b 5.75b 5.30c 5.75b 6.00b 5.88b scf 5.95b 5.50b 5.35c 5.90b 5.65b 5.58b note: mean values bearing different superscript in the same column differ significantly (p<0.05), n=20 key: sca = wheat flour 100%, scb = wheat/soursop flour (90:10%), scc = wheat/soursop flour (80:20%), scd=wheat/soursop flour (70:30%), sce=wheat flour/soursop flour (60:40%), scf = wheat flour/soursop flour (50:50%). 3.3. microbiological status of chin-chin produced from wheat and soursop flour blends 3.3.1. total bacterial counts results of the total bacterial counts (tbcs) of chin-chin samples are shown in table 4. tbcs on week 0 ranged from no growth in control sample to 1.00×102cfu/g in sample substituted with 20% soursop flour. as storage period progressed, an increase in bacterial counts was noted. at the end of three weeks of storage, tbcs were observed to range from 5.20×103-7.00×104cfu/g. samples substituted with soursop flour were observed to have higher tbcs than the control sample. this could be due to the high nutritional value of soursop fruit and its relation with its rapid deterioration [34]. similar trend was also observed by omachi and yusufu [35] who repored increased in the level of microbial contamination due to increased level of proteins and fats. the finding of this study also supports the statement of adams and moss [36] that spoilage organisms grow faster in medium that is highly nutritious. tbcs of chin-chin from this study after storage for 3 weeks were within acceptable limits of 102-104cfu/g established by international commission of microbiological specification of food microorganisms [37] for ready to eat food products. table-4. total bacterial counts (cfu/g) of chin-chin produced from wheat and soursop flour blends during storage. samples storage period (weeks) 0 1 2 3 sca ng 1.00×102 2.00×103 5.20×103 scb 1.00×10 1.00×102 3.00×103 4.50×104 scc 1.00×102 1.40×102 3.00×103 4.00×104 scd 1.00×10 1.10×102 3.80×103 4.00×104 sce ng 1.90×102 3.50×103 6.00×104 scf ng 1.95×102 3.00×103 7.00×104 key: cfu/g= colony forming units per gram; ng= no growth; sca= wheat flour 100%, scb=wheat/soursop flour (90:10%), scc=wheat/soursop flour (80:20%), scd=wheat/soursop flour (70:30%), sce=wheat/soursop flour (60:40%) scf=wheat/soursop flour (50:50%). 3.3.2. total fungal counts results of the total fungal counts (tfcs) of chin-chin samples are presented in table 5. no visible fungal growth was observed on all the chin-chin samples at week 0. however as storage weeks progressed, tfcs were observed to increase to range of 4.00×104-6.00×104cfu/g with control sample having the lowest counts and sample substituted with 30 and 50% soursop flour as highest. the increase in tfcs as substitution of soursop increased could be due to the fact that high moisture content in soursop flour (18.33-24.53%) encourages the microbial action [38]. though there was an increase in the microbial count of the chin-chin, these counts were within the recommended safe limit of microbial guidelines for ready to eat foods such as chin-chin adopted by the international commission of microbiological specification of food which states that the microbial safe limit for ready to eat food should fall between the range of 102-104cfu/ml [35]. fungal growth on foods could lead to deposit of mycotoxins on foods which are a public health concern. table-5.total fungal counts (cfu/g) of chin-chin produced from wheat and soursop flour blends during storage. samples storage period (weeks) 0 1 2 3 sca ng 1.00×102 2.00×103 4.00×104 scb ng 1.00×102 3.00×103 5.00×104 scc ng 1.00×102 4.00×103 5.00×104 scd ng 1.00×102 4.00×103 6.00×104 sce ng 2.00×102 4.50×103 5.50×104 scf ng 2.50×102 4.00×103 6.00×104 key: cfu/g= colony forming units per gram; ng= no growth; sca= wheat flour 100%, scb=wheat/soursop flour (90:10%), scd=wheat/soursop (80:20%), scd=wheat/soursop flour (70:30%), sce=wheat/soursop flour (60:40%) scf=wheat/soursopflour (50:50%). 4. conclusion the present study showed the potentials of utilizing soursop flour for the production of highly nutritious chinchin. the substitution of wheat flour with soursop flour for the production of chin-chin significantly improved the nutritional composition in terms of protein, ash, fat and crude fibre contents while carbohydrate content was observed to decrease. energy content of the chin-chin decreased as substitution of soursop flour increased, but agriculture and food sciences research, 2020, 7(1): 97-104 103 © 2020 by the authors; licensee asian online journal publishing group beyond 30%, level, the energy content was observed to increase significantly. the higher ash, protein, crude fibre and low carbohydrate contents of chin-chin prepared from wheat and soursop flour blends has nutritional advantage over 100% wheat flour chin-chin especially for individuals with health problems requiring protein, fibre and mineral rich foods and low in carbohydrate. sensory results showed that sensory attributes of the control chinchin was more preferred and this was followed closely by chin-chin substituted with 10% ssf. substitution of soursop flour with wheat flour at the level of 10% compared favorably with the control sample suggesting that acceptable chin-chin could be produced at ssf substitution of up to 10%. total bacterial and fungal counts of chinchin increased during storage and upon increased substitution with sour-sop flour. however, all the products presented adequate microbiological conditions and were within recommended safe limit of microbial guidelines. this result therefore indicates that the use of soursop flour for the production of chin-chin would greatly enhance the utilization of this fruit in nigeria, and other developed countries where the fruit has not been optimally utilized, thereby reducing wastage of the fruit and contributing to household food security. references [1] t. t. iombor and a. u. banjo, "evaluation of the potential of soursop (annona muricata) flour in bread production," confluence journal of pure and applied sciences, vol. 2, pp. 19-26, 2018. 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[38] n. p. minh, "production of formulated juice beverage from soursop and grapefruit," international journal of applied engineering research, vol. 12, pp. 15311-15315, 2017. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 7 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 7-15, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.7.15 © 2020 by the authors; licensee asian online journal publishing group weather and crop management impact on crop yield variability rezwanul parvez1 nazea hasan khan chowdhury2 ( corresponding author) 1research and planning associate, community college of denver, denver, usa. 2faculty , department of business & information technology, front range community college, boulder county campus, longmont, usa. abstract this research is primarily designed to examine crop yield variation due to change in weather pattern and crop management activities by exploring a comprehensive list of factors (e.g. environmental, economic etc.). given that existing literature indicates significant effects when farmland value is regressed on weather variables, a natural question is to ask whether major agricultural crop yields responds to change in historical weather pattern. to answer this question, this study relies on a state level panel dataset including agricultural and high-resolution weather data covering the period 1997-2018. using a seemingly unrelated regression (sur) approach, this study estimates how crop spatial distribution patterns have impacted crop yield variation in response to weather in united states greater midwest region. key findings indicate that changes in crop management activities correspond closely to estimate of both crop price and weather effects. additionally, corn yield response to weather change varies with crop spatial distribution pattern, with distinct impacts on the magnitude and even the direction at the state level. further, revenue of corn, soybeans, wheat and their respective lagged yields have positive and significant effect to respective crop yields. finally, a major crop yield does vary across region due to variation of crop prices, precipitation and temperature. these findings have useful implications for agriculture sector on how historical weather trends have affected crop yield and distribution pattern. keywords: crop yield, seemingly unrelated regression, crop spatial distribution, weather pattern, crop management, greater midwest. citation | rezwanul parvez; nazea hasan khan chowdhury (2020). weather and crop management impact on crop yield variability. agriculture and food sciences research, 7(1): 7-15. history: received: 7 november 2019 revised: 10 december 2019 accepted: 13 january 2020 published: 19 february 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ......................................................................................................................................................................................... 8 2. materials and methods ................................................................................................................................................................... 11 3. data description .............................................................................................................................................................................. 11 4. empirical strategy ........................................................................................................................................................................... 11 5. results and discussion ................................................................................................................................................................... 12 6. conclusion ......................................................................................................................................................................................... 14 references .............................................................................................................................................................................................. 15 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.7.15&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.7.15&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1336 https://orcid.org/0000-0001-5167-699x https://orcid.org/0000-0001-8066-7005 agriculture and food sciences research, 2020, 7(1): 7-15 8 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this research is primarily designed to examine crop yield variation due to change in weather pattern and crop management activities by exploring a comprehensive list of factors (e.g. environmental, economic etc.). 1. introduction there has been a growing interest to examine the relation between crop productivity and weather change among advocates of weather change and food security, farmers, resource management professionals, and policy makers. a vast majority of existing literature primarily focused on investigating how crop yield respond to weather conditions, with emphasis on the decadal and interannual weather variability [1-3] weather extremes [4] vapor pressure deficits [5] atmospheric co2 concentration [6] as well as weather variability [7]. agricultural crop yield losses due to weather change are predicted to be as high as 82% by the end of 20th century for some crops [8, 9]. weather change is defined as a temporal and spatial variation in the distribution of temperature and precipitation [10]. according to national weather assessment, the upper midwest (minnesota, wisconsin, michigan, iowa, south dakota, and north dakota) and the northern plains of u.s. have experienced more variation in temperatures and precipitation as compared to southern states [11]. the long-term weather trends picture can effectively signal farmers to adopt suitable cropping pattern. existing literatures have listed multiple adaptation measures (i.e. planting dates shifting, switch crop patterns, develop new crops, shift in crop growing pattern) to mitigate the negative impacts and severity of weather on crop yield and production [12-15]. agricultural producers tend to follow subsequent crop rotations to maximize crop production using lands with optimal water and nutrient storage conditions. thus, it is useful to investigate how changes in crop spatial distribution patterns over time have impacted crop yield response to weather change in u.s. greater midwest? majority of literature have assumed that crop distribution in space remain constant over study period [16-18]. empirical models (both process-based simulation models and statistical models) have been used to assess the potential impacts of weather on crop yields [19]. for example, a comprehensive data-driven analysis is developed to investigate crop pattern between counties in regulating corn yield response to weather change at the state level of u.s [20]. the issue of land use change and agricultural crop acreage response due to economic and biophysical factors is well documented focusing on northern great plains agriculture [21, 22]. thus, it would be useful to investigate how historical crop yields responded to economic and biophysical factors for greater midwest. here, the functional relationships between historical observation of weather patterns and agricultural crop yields (for corn, soybean and wheat) are examined at the state (11) level of greater midwest (i.e. illinois, indiana, kansas, michigan, minnesota, missouri, nebraska, north dakota, ohio, south dakota and wisconsin) of the united states (u.s.). this paper addresses the key question of how agriculture adapts to weather change through crop yield loss patterns. the empirical challenge associated with measuring weather change impacts on crop yields centers around how to organize weather information relevant to farmers’ cropping decision. this research developed a state-level panel of “weather data” to address this. the annual change in weather data for a state reflects knowledge on local weather. the yield variables are regressed on the weather data in a sur panel estimation, controlling for trends and temporal effects to better examine the relation between yields and weather. this study develops a set of weather indices to estimate crop yield response across regions to test the hypothesis of spatially varying weather change impacts. this paper makes three key contributions. first, as an initial rigorous empirical analysis of crop yield response to weather pattern change, it extends the knowledge on biophysical environment change impacts on agriculture and highlights the importance of considering yield variation in projections of future weather change impacts. second, the summary contributes to the understanding of factors driving crop yield response. third, the paper also builds on the economic literature of environmental adaptation by providing evidence of adaptation when the environmental change occurs gradually (e.g., [23, 24]). the yield response model should generate more stable coefficients and yield predictions as compared to models including highly correlated weather variables. the historical pattern of crops (corn, soybean, wheat) price and yield for all 11 states of us greater midwest are presented in figures 111. figure-1. crop price and yield (illinois) from 1997-2018. agriculture and food sciences research, 2020, 7(1): 7-15 9 © 2020 by the authors; licensee asian online journal publishing group figure-2. crop price and yield (indiana) from 1997-2018. figure-3. crop price and yield (kansas) from 1997-2018. figure-4. crop price and yield (michigan) from 1997-2018. figure-5. crop price and yield (minnesota) from 1997-2018. agriculture and food sciences research, 2020, 7(1): 7-15 10 © 2020 by the authors; licensee asian online journal publishing group figure-6. crop price and yield (missouri) from 1997-2018. figure-7. crop price and yield (nebraska) from 1997-2018. figure-8. crop price and yield (north dakota) from 1997-2018. figure-9. crop price and yield (ohio) from 1997-2018. agriculture and food sciences research, 2020, 7(1): 7-15 11 © 2020 by the authors; licensee asian online journal publishing group figure-10. crop price and yield (south dakota) from 1997-2018. figure-11. crop price and yield (wisconsin) from 1997-2018. understanding the connection among major crop yields, crop prices, weather, and other input prices are vital to stakeholders who are concerned with addressing weather change and improving global food security. also, researchers and policy-makers are deeply engaged to better understand the effects of weather change on the agricultural landscape of the greater midwest. the importance of estimating functional relationships between historical observations of weather and yields are well cited in scientific literature. the results discussed here will help forecast future crop yield trends in response to change in weather pattern. further, this research will help policy-makers and producers make better-informed crop production decisions in a changing weather scenario. 2. materials and methods this section empirically examines the effects of weather change (weather factors like monthly average precipitation and temperature, lagged crop prices,) on multiple crops yields (corn, soybeans, wheat) loss. the weather data is constructed to investigate weather trends at the state level. the functional relationship is presented on how crop yields (current year) correlate with current and last year crop prices, lag crop yields, crop revenue, and weather variables like precipitation, temperature, using longitudinal data. here, dependent variables of the stated regression model for each three major crop choices (corn, wheat, and soybean) are measured at yield per bushel. further, a seemingly unrelated regression (sur) approach is employed with spatial autoregressive terms to capture the weather effects on crop yield loss. this is because statistical models including only on spatial variation (i.e. cross-sectional studies) are very prone to omitted variable bias, whereas those including only on temporal variation (i.e. time series studies) are often subject to significant errors in estimates of temperature sensitivity. these errors can happen due to limited temporal variation in temperature compared to other weather variables, as well as strong temporal correlation between temperature and rainfall or radiation. 3. data description the analyses presented in this study compiles agricultural and climatologic data. state-level annual data on crop yields are collected from nass surveys from 1997 to 2018. weather data (e.g. precipitation and temperature) for the month of march, april, may, june, july, and august for last 22 years come from the national climatic data center [25]. information on prices and yields of corn, soybeans, and wheat for all eleven midwest states (e.g. north dakota, south dakota, nebraska, minnesota, iowa, missouri, wisconsin, illinois, kansas, michigan, indiana, and ohio) for the period 1997 to 2018 year are obtained from the census of agriculture (united states department of agricultural, national agricultural statistics service). major crops (e.g. corn, soybeans, and wheat) yield and price data are lagged. prices data are adjusted for inflation using gdp price deflator (2017 as base year). the gdp price deflator data are collected from the federal reserve economic data, economic research division. crops (e.g. corn, soybeans, and wheat) revenue are also calculated using yield and price information. 4. empirical strategy the empirical strategy relies on employing a seemingly unrelated regression (sur) model to examine the marginal effects of weather factor change on agricultural crop yield at the state level. a “full information maximum likelihood” approach is used to examine the effects. the right-hand side of estimated regression equations includes agriculture and food sciences research, 2020, 7(1): 7-15 12 © 2020 by the authors; licensee asian online journal publishing group no endogenous variables and assume correlation of error term among equations. heteroskedasticity is also almost certain since presence of multiple dependent variables. the regression equations can be expressed in the form of stacked regression. 𝑌 = 𝑋𝛽 + 𝑢 (1) where, y is a set of dependent variables of crop (corn, soybeans and wheat) yields. x is a set of independent variables (crop prices, precipitation, and temperature for the month of march, april, may, june, july, and august, crop revenue etc.). further, 𝑢 is error term. 𝛽 is parameter estimates in the proposed model. 5. results and discussion summary statistics are reported in table 1. the estimation results of equation 1 are reported in tables 2-4. the underlying identification assumption is that a state experiencing change in weather pattern would have changed its management activities of agricultural crops differently from a state not experiencing any change, after purging off state-level trends and nation-level shocks. table-1. summary statistics. variable name observation mean standard deviation min max corn_yield 242 143.98 24.45 75.00 210.00 soybeans_yield 242 41.79 8.05 20.00 65.00 wheat_yield 242 54.28 13.82 24.30 89.00 price_corn 242 3.83 1.36 2.18 7.92 price_soybeans 242 9.49 2.65 5.48 15.87 price_wheat 242 5.12 1.54 2.47 9.07 lagged yield_corn 231 142.76 23.80 75.00 201.00 lagged yield_soybeans 231 41.37 7.79 20.00 61.00 lagged yield_wheat 231 54.03 13.82 24.30 89.00 precipitation_march 242 2.07 1.32 0.15 6.81 precipitation_april 242 3.12 1.57 0.50 9.61 precipitation_may 242 3.96 1.51 0.81 9.60 precipitation_june 242 4.28 1.58 1.29 9.44 precipitation_july 242 3.50 1.37 0.75 8.18 precipitation_august 242 3.32 1.21 0.62 6.94 temperature_march 242 36.73 8.13 16.30 58.10 temperature_april 242 48.32 6.08 31.30 60.90 temperature_may 242 59.24 5.05 47.40 72.50 temperature_june 242 68.65 4.32 59.10 78.10 temperature_july 242 73.34 4.19 63.50 84.90 temperature_august 242 71.31 4.16 60.90 83.00 revenue_corn 242 550.62 203.35 245.97 1188.02 revenue_soybeans 242 398.84 138.72 124.43 721.18 revenue_wheat 242 275.30 103.92 109.63 648.94 table-2. results of seemingly unrelated regression (corn equation). dependent variable: corn yield explanatory variables coefficient and standard deviation p-value price_corn -0.29*** (1.33) 0.00 price_soybeans 0.78 (0.80) 0.33 price_wheat 0.16 (0.90) 0.86 lagged yield_corn 0.11*** (0.04) 0.00 lagged yield_soybeans 0.04 (0.12) 0.70 lagged yield_wheat 0.020 (0.07) 0.78 precipitation_march -0.36 (0.45) 0.42 precipitation_april -0.58 (0.39) 0.14 precipitation_may -0.27 (0.33) 0.41 precipitation_june -0.36 (0.34) 0.29 precipitation_july 0.72* (0.38 0.06 precipitation_august 1.07** (0.43) 0.01 temperature_march 0.29*** (0.11) 0.01 temperature_april -0.02 (0.14) 0.92 temperature_may 0.34* (0.18) 0.06 temperature_june 0.13 (0.24) 0.59 temperature_july 0.20 (0.28) 0.46 temperature_august -0.73*** (0.22) 0.00 revenue_corn 0.19*** 0.00 agriculture and food sciences research, 2020, 7(1): 7-15 13 © 2020 by the authors; licensee asian online journal publishing group (0.009) revenue_soybeans 0.002 (0.01) 0.86 revenue_wheat 0.005 (0.01) 0.68 constant 109.85*** (14.40) 0.00 note: the symbols ***, **, and * indicate statistical significance at α=0.01, α=0.05, and α=0.10, respectively. numbers in parentheses are the standard errors of the parameter estimates. table-3. results of seemingly unrelated regression (soybean equation). dependent variable: soybeans yield explanatory variables coefficient and standard deviation p-value price_corn 0.33 (0.36) 0.35 price_soybeans 3.67*** (0.22) 0.00 price_wheat 0.12 (0.24) 0.61 lagged yield_corn -0.010 (0.010) 0.34 lagged yield_soybeans 0.12*** (0.03) 0.00 lagged yield_wheat 0.02 (0.02) 0.26 precipitation_march -0.04 (0.12) 0.76 precipitation_april -0.13 (0.11) 0.21 precipitation_may 0.07 (0.09) 0.43 precipitation_june (0.03) (0.09) 0.74 precipitation_july 0.13 (0.10) 0.20 precipitation_august (0.47) *** (0.11) 0.00 temperature_march 0.04 (0.03) 0.18 temperature_april -0.04 (0.040) 0.30 temperature_may 0.04 (0.05) 0.41 temperature_june 0.09 (0.06) 0.16 temperature_july 0.06 (0.07) 0.45 temperature_august -0.17*** (0.06) 0.00 revenue_corn -0.002 (0.002) 0.44 revenue_soybeans 0.09*** (0.003) 0.00 revenue_wheat -0.004 (0.003) 0.18 constant 32.84*** (3.88) 0.00 note: the symbols *** indicate statistical significance at α=0.01. numbers in parentheses are the standard errors of the parameter estimates. the sur approach used here includes longitudinal panel data for eleven midwest states of three major crop yields over 22 years (19972018). key findings indicated, in all three-crop yield equation, price of soybeans positively related to its respective yields whereas corn and wheat prices are negatively related to its respective yields tables 2-4. also, the relationship between crop yield and prices are statistically significant. in addition, lagged price of corn and wheat also negatively related to their respective yields. last year corn yields positively related to current year corn yields and also statistically significant. furthermore, revenue of corn also positively related to corn yields and also statistically significant. moreover, soybeans revenue and lagged yields also positively related to soybeans yields and statistically significant. further, wheat revenue and lagged yields also positively related to its current year yield and statistically significant. together with regional trends in temperature and precipitation, these estimates can partly explain the crop expansion pattern over the past few decades. for instance, higher yield of corn, soybeans, and wheat have coincided with increase in both temperature and precipitation in the dakotas and the upper midwest. this association has a particularly vital implication given the relatively large marginal effects estimated for this region. these results are also in line with the historical movements of corn location discussed [26, 27]. agriculture and food sciences research, 2020, 7(1): 7-15 14 © 2020 by the authors; licensee asian online journal publishing group table-4. results of seemingly unrelated regression (wheat equation). dependent variable: wheat yield explanatory variables coefficient and standard deviation p-value price_corn -0.26 (0.68) 0.70 price_soybeans 0.96 (0.41) 0.02 price_wheat -8.58*** 0.46 0.00 lagged yield_corn -0.04** (0.020) 0.02 lagged yield_soybeans 0.17*** (0.06) 0.00 lagged yield_wheat 0.18*** (0.03) 0.00 precipitation_march -0.28 (0.23) 0.22 precipitation_april 0.009 (0.20) 0.96 precipitation_may 0.25 (0.17) 0.14 precipitation_june -0.16 (0.17) 0.36 precipitation_july 0.32* (0.19) 0.10 precipitation_august 0.78*** (0.22) 0.00 temperature_march 0.03 (0.06) 0.61 temperature_april -0.04 (0.07) 0.63 temperature_may -0.07 (0.09) 0.44 temperature_june 0.06 (0.12) 0.63 temperature_july -0.003 (0.14) 0.98 temperature_august 0.008 (0.11) 0.94 revenue_corn 0.004 (0.005) 0.37 revenue_soybeans -0.01** (0.006) 0.03 revenue_wheat 0.15*** (0.006) 0.00 constant 39.21*** (7.34) 0.00 note: the symbols ***, **, and * indicate statistical significance at α=0.01, α=0.05, and α=0.10, respectively. numbers in parentheses are the standard errors of the parameter estimates. average precipitation (march, april, may, june) are found to be negatively related to corn yields because preplanting time and during planting time precipitation adversely effects crop yield. in soybean equation, precipitation (for the month of march and april) are negatively related to soybeans yields as we expected. on the other hand, precipitation july and august are positively related to soybeans yields. in wheat equation, precipitation (for the month of march and june) negatively related to wheat yields. precipitation (for the month of july and august) are positively related to wheat yields. temperature also significantly affects crop yield pattern. in corn equation, temperature (for the month of march and may) are positively related and statistically significant to corn yields for current year. the average temperature (for the month of august) are negatively related and statistically significant to corn yield. in soybean equation, average temperature (for the month of april and august) are negatively related to soybeans yield in current year and statistically significant table 3. this is because producers expect positive temperature for optimal growth of soybeans crop and yields before and during planting season. in wheat equation, average temperature (for the month of march and august) are positively related to wheat yield table 4. 6. conclusion the issue of weather change impacts on crop yields variability is well discussed by existing literature. all else equal, the heterogeneity of multiple crop yield responses also implies a change in relative profitability per acre. as weather change persists, this will slowly alter cropping patterns. this study analyzes the importance of taking crop yield effects into consideration when evaluating weather pattern change impacts on agriculture. the scenario of adjustment and adaptation in crop acreage by producers is important and need to be included in policy discussion to avoid imprecise and even unrealistic projections of future crop losses due to weather change. the findings reported here are useful to see how yield responds to different economic and biophysical factors (e.g. crop prices, lagged yields and prices, crop revenue, temperature, precipitation). results also indicate that precipitation before planting season has impact on crop yield. the impact of average temperature on crop yields is evident. further, crop revenue, lagged yields of corn, wheat, and soybeans are positively related to current year yields. moreover, prices of corn and wheat negatively related to current year yield. factors like precipitation (for the month of agriculture and food sciences research, 2020, 7(1): 7-15 15 © 2020 by the authors; 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[27] j. m. beddow and p. g. pardey, "moving matters: the effect of location on crop production," the journal of economic history, vol. 75, pp. 219-249, 2015.available at: https://doi.org/10.1017/s002205071500008x. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.ncdc.noaa.gov/ agriculture and food sciences research issn: 2411-6653 vol. 2, no. 1, 12-18, 2015 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 12 mixolab profile of wheat flour and their correlation with textural properties of hot-press tortilla esther perez-carrillo 1 --ana a. chew-guevara 2 --erick heredia-olea 3 --hector h. velázquez-reyes 4 --sergio o. serna-saldivar 5* 1,2,3,5 centro de biotecnología-femsa, escuela de ingeniería y ciencias, tecnológico de monterrey, av. eugenio garza sada, monterrey, n.l,. mexico 4 universidad autónoma agraria antonio narro, calzada antonio narro, col. buenavista, saltillo, coahuila, méxico abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ......................................................................................................................................................................... 13 2. materials and methods ........................................................................................................................................................ 13 3. results and discussion ......................................................................................................................................................... 14 4. conclusions .......................................................................................................................................................................... 16 5. acknowledgments ................................................................................................................................................................ 16 references ................................................................................................................................................................................ 16 refined wheat flours commercially milled from the same red winter wheat under 10 different commercial conditions were tested for quality with a mixolab and then processed into tortillas using the hot-press forming procedure. tortilla making qualities of the flour samples were evaluated during dough handling, hot-pressing, baking, and the first five days on the shelf at room temperature. the most dominating mixolab variables that were correlated with flour tortilla performance and textural shelf stability were c3 related to starch gelatinization (1.93-2.18), c4 related to amylase activity (1.461.78) and c5 related to starch retrogradation (2.82-3.41). these mixolab parameters influenced tortilla texture after one day storage whereas parameters c3 and c4 influenced tortillas stored for two and five days. hot-press tortillas produced from 03 flour, which had the highest c3, c4 and c5 values, had the worst textural shelf-life (force 12.41 n) and rollability. on the other hand, tortillas produced from 07 flour, that had the lowest c3, c4 and c5 values, exhibited the best textural shelf life (force 6.73 n). mixolab parameters c3, c4 and c5 proved to be useful in predicting the quality of wheat flours intended for hot-press tortilla production. keywords: hot-press tortillas, mixolab, refined wheat flour, wheat flour tortillas. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(1): 12-18 13 1. introduction wheat flour tortillas are unleavened, flat, circular bakery items commonly leavened with baking powder. historically, tortillas have been produced for several centuries practically since the spaniards conquered mexico and introduced wheat to the americas. nowadays, tortillas are consumed practically all over the globe because most fast food chain restaurants offer burritos and tacos in their menus. in the usa, tortillas are considered as the second most popular bakery item after yeast-leavened breads mainly due to their convenience as wraps that suit an on-the-go lifestyle. the key quality attributes that provide the desired functionality in tortillas are extensibility during pressing (diameter) and retention of flexibility over time (shelf stability). most flour tortillas are produced with all-purpose flours with intermediate protein content (10 to 11%). these flours yield rollable tortillas that do not crack and break when reheated and folded [1-3]. however, no wheat cultivars have been identified or bred for this particular use. generally, hard red winter wheats with relatively low protein content are selected and supplemented with various types of additives (i.e. oxidizing and reducing agents, hydrocolloids, emulsifiers or dough conditioners) to achieve the required functionality [4]. the protein of the flour and the rate of starch retrogradation are primary factors affecting tortilla textural shelf-life. it is recognized that tortillas obtained from low protein flour crack and split apart when folded after one day of storage whereas counterparts that contain more protein yield tortillas with improved textural stability [5]. koksel, et al. [6] concluded that the mixolab data could be related with farinograph and alveograph parameters, the zeleny sedimentation test and bread volume. therefore this instrument can be effectively used for flour screening especially in terms of dough strength. hrušková, et al. [7] observed a link among parameters of farinograph, amylograph and mixolab tests, and a principal component analysis confirmed these relationships. a tight associations between farinograph dough development time or stability and mixolab c1 (behavior during mixing), and between mixing tolerance index and the difference c1-c2 (protein quality) were observed. maximum amylograph peak viscosity was connected with c3 (starch gelatinization), c4 (amylase activity) and c5 (starch retrogradation) torque points as well with the differences of c3-c2, c3-c4, and c5-c4. levels of torque points c3, c4, and c5 corresponded to dough consistency changes (i.e., resistance against mixing) and the farinograph dough development time and stability whereas differences c3-c2, c3-c4, and c5-c4 are highly linked to amylase activity and starch gel properties during heating and cooling. despite their popularity, there are no established methods to predict the most suitable flours for tortilla quality and predict their textural shelf life [1]. the mixolab is a rheological instrument recently developed suited to evaluate the quality of wheat flours for different end uses. the instrument introduced in 2004 by chopin technologies measures the consistency of dough over time with a gradual increase in the applied temperature. it is designed to test both protein and starch characteristics of the flour and also provide information about rheological properties of dough [8]. the computerized analysis of the curve yields the mixolab profiler. the mixolab has been successfully used to evaluate flour for bread and cookie making qualities. except for a recent report of posner, et al. [9] there is no information on the use of mixolab to determine the quality characteristics of flours intended for hot-press wheat flour tortillas. therefore this research was undertaken to assess the optimal mixolab parameters for hot-press wheat flour tortillas and their textural shelf-life. 2. materials and methods 2.1. flour samples ten untreated flour samples milled from the same blend of wheats in different commercial mexican mills were donated by the us wheat associates, mexico city, mexico. these flours differed in particle size distribution, color and damaged starch. 2.2. flour characterization and dough tests particle size distribution was determined with the sieving to an end point method [10] on a set of flour sieves (us meshes no. 80, 100, 120 and 140). 100 g sample was placed on the top sieve and rotapped and sieved for 1 min. then, the overs of each sieve were weighed and sieved again. the procedure was repeated until the weight difference between consecutive weights was less than 1 g. flour moisture and falling number were determined using aacc methods 44-15a and 56-81b, respectively [11]. the dough rheological properties dough especially in terms of strength, extensibility and resistance to extension were determined with the alveograph (chopin instruments, villeneuve-la-garenne, france) approved method 54 30a and brabender farinograph following the method 54-21 [11]. starch damage was conducted according to aacc method 76-33.01. the mixolab (chopin, france) was used to analyze the flour samples according to aacc method 54-60.01 [11] and icc 173. the starch gelatinization was followed during the increase of temperature from 35 ° to 90 ° c at a rate of 2°c/min. protein properties related to water absorption, stability, elasticity and weakening were determined. the enzymatic activities and retrogradation as affected by mixing and the temperature increase were also monitored. a mixolab wheat tortilla profile was generated based on these parameters [8]. 2.3. preliminary tortilla trials tortillas were prepared by methods described by bello, et al. [12] and serna saldivar [13] with slight modifications. 200 g flour (14% mb) plus 30 g vegetable shortening (productos lirio, monterrey, nl, mexico), 3 g refined iodized salt (la fina, sales del istmo, coatzacoalcos, ver., mexico), 4 g double acting baking powder (rexal, productos mexicanos, monterrey, nl, mexico), 2 g whole dry milk (nido, nestlé, querétaro, qro, méxico), 0.4 g calcium propionate (tecsa, monterrey, nl, mexico), 0.4 g fumaric acid (primak, monterrey, nl, mexico), 0.4 g carboxymethyl cellulose (piasa, monterrey, nl, méxico), 0.4 g sodium stearoyl-2-lactylate (tecsa, monterrey, nl, mexico), and distilled water were mixed with a predetermined volume of warm water (40 o c) in a 100-200 g dough mixer (national manufacturing co., lincoln, ne). optimum water absorption and mix agriculture and food sciences research, 2015, 2(1): 12-18 14 times were subjectively determined by observing dough handling properties. dough texture was subjectively evaluated using a 1 to 5 rating. a subjective score of 1 meant that the dough was slack or soft and needed less force to extend whereas a score of 5 was assigned to very rough or firm dough that needed high force to extend. water absorption was varied in order to obtain a dough with intermediate properties (2.5 in the subjective score of 1 to 5) suited for hot-press tortillas. experimental doughs were divided into 30 ± 0.25 g pieces, manually rounded and allowed to rest in a proof cabinet (national manufacturing co., lincoln, ne) adjusted to 29 o c and 85% rh for 30 min. each dough ball was flattened using a commercial inclined hot press for 3.13 sec. the temperature of the plates was set at 187°c and the gap between the hot plates adjusted to 1.75 mm. the resulting flattened tortilla discs were baked on a four-pass circular moving griddle set at different temperatures (manufacturas c&d industriales, monterrey, nl, mexico). the raw tortilla was baked on one side for 10.79 sec at 200°c, turned over and baked for another 11.01 s at 260°c, turned over again and baked for 11.04 s at 265°c and finally turned over again and baked at 230°c for 13.60 s [13]. 2.4. pilot plant tortilla trials tortillas were prepared by methods delineated by serna saldivar [13]. 2 kg flour (14% mb) plus 300 g vegetable shortening (productos lirio, monterrey, nl, mexico), 30 g refined iodized salt (la fina, sales del istmo, coatzacoalcos, ver., mexico), 40 g double acting baking powder (rexal, productos mexicanos, monterrey, nl, mexico), 20 g whole dry milk (nido, nestlé, querétaro, qro, méxico), 4 g calcium propionate (tecsa, monterrey, nl, mexico), 4 g fumaric acid (primak, monterrey, nl, mexico), 4 g carboxymethyl cellulose (piasa, monterrey, nl, méxico) and 4 g sodium stearoyl-2-lactylate (tecsa, monterrey, nl, mexico) were mixed with a volume of warm water determined according to preliminary tortilla trials. the dough mixing protocol consisted of first blending dry ingredients with the shortening at slow speed with a hook attachment for 1 min. then, distilled water tempered to 40°c was added and the blend mixed at slow speed for 1 min and then at medium speed until gluten development. resulting doughs were placed in the hopper of an automatic dough cutter and rounder (manufacturas c&d industriales, monterrey, nl, mexico). the speed of the blade was adjusted to yield 30 g pieces and rounded mechanically and immediately placed in the proof cabinet set at 29 o c and 85% rh for 30 min. then, the relaxed dough balls were hot-pressed into tortilla discs and baked as explained above. the baked tortillas were cooled on a wire rack to room temperature (25±2°c) for about 30 min, placed inside sealed polyethylene bags and kept at room temperature for further evaluations [13]. 2.5. evaluation of tortilla properties ten tortillas from each treatment were randomly selected and measured for diameter and thickness. the diameter of tortillas was the average of two diagonal measurements. likewise, two tortillas from each batch were randomly selected and measured for color using the minolta chromameter (model cr-300, minolta camera co., ltd., chuoku, osaka, japan). values for l* (brightness or whiteness), a*(redness and greenness), and b*(yellowness and blueness) were measured. surface color was measured from four different randomly selected spots of each tortilla. tortilla moisture and protein were determined using the aacc methods 44-15a, and 08-03, respectively [11]. texture analyses were conducted after 0, 1, 2 and 5 days of storage at room temperature using a ta.xt2i texture analyzer (stable micro systems, godalming, england) and rollability or dowel technique [13, 14]. a tortilla was rolled around a 1 cm wooden dowel and rated from 1 (no cracks; very flexible) to 5 (break immediately; cannot be rolled). tortillas were considered unacceptable when the rollability score was lower than 3. to assess the reheating functionality after 7 days storage at room temperature, a griddle was heated in an oven to a surface temperature of 232°c. five tortillas from each lot were reheated for 15 sec on one side, turned over and heated for 15 sec on the other side and finally an additional 15 sec on the initial side. immediately after the reheating schedule, the rollability was evaluated using a dowel [13] and a subjective scale of 1 to 5 where 1 was excellent and 5 very poor. 2.6. statistical analysis flour characteristics and tortilla data were analyzed using a randomized experimental design using analysis of variance procedures. tortilla texture and rollability measurements were analyzed with non-parametric tests (friedman). minimum significant differences and tukey tests were applied to determine differences among means (p<0.05). 3. results and discussion 3.1. functional properties of flours the wheat flour milling process involves a series of breaking, middling reduction, and sifting operations. particle size is a relevant concept in flour milling [15] and a critical factor in determining the flour usefulness and functionality. in this study, particle size analysis results were compared for the ten flours produced from the same wheat blend (fig. 1). flours 01 and 04 had a more uniform particle size distribution whereas flours 08 and 10 have higher proportion of fine particles. interestingly, coarser flours had lower water absorptions whereas finer flours had higher water absorptions and starch damage and lower dough stability (table 1). these observations agree with wang and flores [15]. kuakpetoon, et al. [16] observed that in the same type of wheat the particle of fine flours were highly uniform in shape with a spherical form. in contrast, the particles of coarse flours were irregularly shaped and had a rougher surface. fine flours had higher mixolab c2 and c3-c4 values and lower c3, c4, c5, c1-c2, c2-c3 and c4-c5 values compared to counterparts with coarser particles (table 2). flours parameters clearly differed despite that all originated from the same wheat blend (table 1). results indicate that none of the mills processed wheat using the same method and procedures so the refined flours had different functional and rheological properties. falling number (fn) values indicated that flours were adequate for bread making. the 01 and 07 flours showed the highest and lowest fn values, respectively. the difference between agriculture and food sciences research, 2015, 2(1): 12-18 15 these flours were approximately 30%. the 07 flour had the highest farinograph water absorption and ucdc value. the flour 01 showed the highest alveograph w value and farinograph dough stability, this particular flour had the lowest end point percentage at sieve no. 140. the lowest w value was observed in flour 06 which had the highest p/g and p/l values. furthermore, this flour had the highest farinograph water absorption. the lowest p/g and p/l values were observed in the 03 flour which also had the lowest farinograph absorption value. the lowest farinograph stability was associated to flours 03 and 09 which also had the lowest ucdc values. differences in fn indicate that flours differed due to the milling procedures. according to rani, et al. [17] the settings of the milling rolls and combination of different flour streams affect functionality of flours obtained from the same lot of wheat kernels. none of the commercial mills followed methodical grinding rolls adjustment and the operation was based only on the head millers’ experience [9]. the higher alveograph w and p/g values and farinograph stability observed in flour 01 indicated that this flour was capable of forming a stronger gluten network. the starch damage value is the one of the best parameters to evaluate mill grinding adjustments. millers evaluate continuously the starch damage in their flour streams and adjust milling rolls to the optimum required in the final product. typically the starch damage after the break stages should be in the range of 4 to 5% and in the sizing and reduction stages in the range of 5-6% and 8-10%, respectively. waniska, et al. [18] studied 61 wheat flour properties and their effects on tortilla quality and observed that starch damage values for a good tortilla flour should be between 4 and 12 with average of 7.6%. 3.2. mixolab profile the mixolab allows the measurement of dough consistency over time and evaluate in the same assay both the mixing and pasting properties of flour during a gradual increase in temperature [8]. table 2 depicts mixolab parameters of the ten commercial flours obtained from the same wheat blend. the combined effect of the mechanical shear stress and the temperature in the second stage of the mixolab curve produced a decrease in the c2 torque, which is attributed to the weakening of the gluten network. protein weakening is also represented by the difference between points c1 and c2 (c1-c2). codinӑ, et al. [19] observed that the c2 parameter was positively correlated with alveograph w value. the correlation between alveograph w and c2 values was not significant in this study; however, a negative correlation between c1-c2 and w values was observed. the mixolab c3, c4 and c5 parameters are commonly affected by the starch condition and characteristics. water absorption values were similar to the ones obtained from the farinograph; however, in case of higher absorption the mixolab has better precision. the highest dough development was reported for 04 flour and the lowest for 07 and 09 flours. the highest stability and c5-c4 parameter were reported for the 02 flour which also had the highest fn (table 1). however, mixolab stability values did not correlate with farinograph dough stability. the lowest c2 and ucds values were reported for the 04 flour (table 1). the 07 flour had the highest ucd and the lowest c3, c4 c5 and c3-c4 parameters. however, there were not significant correlations among alveograph, farinograph and mixolab parameters. 3.3. plant pilot tortilla trials table 3 summarizes mixing data of dough for tortilla production during tortilla trials. as expected and according to functional tests, the 03 and 04 flours absorbed the lowest water to produce optimum dough. in contrast, flour produced with the 08 flour absorbed higher water than the recommended for hot-press tortillas (tables 1 and 2). mixing properties indicated that the dough rheological properties and gluten content values did not necessarily correlate to practical tortilla dough mixing requirements (tables 1 and 3) probably due to the high shortening addition to the typical formulation. differences in absorption and mixing time coincide with c3, c4, c5, c1-c2 and c3-c2 mixolab parameters (table 2). all types of tortillas produced from 30 g dough balls weighed approximately 26 g after baking and cooling. the 10 flour had the highest protein content and c2 mixolab value related to gluten weakening after achieving dough development time. tortilla diameter is one of the key quality attributes because a tortilla is primarily meant to function as a wrap. diameter, thickness and color parameters differed among the different types of tortillas (table 4). the 08 tortillas had the highest diameter and lowest thickness. the rest of the tortillas did not differ in diameter. in the case of tortilla color, the highest l, a, b values were observed in tortillas produced with the 05, 10 or 03 flours, respectively. the rest of the tortillas had similar color values. waniska, et al. [18] reported similar physical tortilla values whereas ramirez-wong, et al. [20], reported higher diameter (19.93 cm) and lower thickness (0.90 mm) with tortillas weighing 32 g. tortilla color parameters “a” and “b” were similar to data reported by barros, et al. [21] and ramirez-wong, et al. [20]. however, l was lower compared to barros, et al. [21] and higher compared to tortillas obtained by ramirez-wong, et al. [20]. 3.4. tortilla texture the textural shelf-life of tortillas plays a critical role in terms of quality and acceptance. most flour tortillas are expected to last on the shelf at room temperature for at least one week and up to three weeks. staling of flour tortillas is mainly due to the gradual transformation of amorphous starch to a partially crystalline, retrograded state. the reassociation of starch molecules during storage corresponds to loss of freshness and increased structure or firmness of tortilla [22]. the 07 tortillas had the lowest force value throughout five days of storage at room temperature whereas counterparts manufactured from the 03 flour exhibited the toughest texture or highest force values (table 5). these flours absorbed the lowest and highest water during dough mixing and showed important differences in their mixolab profiles. correlation coefficients between tortilla texture (force) and mixolab parameters are presented in table 7. results showed that the c3, c4 and c5 parameters influenced tortilla texture of fresh tortillas (day 1); however, the mixolab parameters c3 and c4 influenced texture of tortillas stored for 2 to 5 days. as expected, fresh tortillas (day 0) had the lowest force and largest extension values (table 5) and the best rollability properties (table 6). most of the loss of texture in all tortillas occurred during the first day of storage. this agrees with a previous research conducted by bejosano, et al. [23] which indicated that most of the changes in flour tortilla texture also occurred during the first day of storage although a progressive hardening occurred during the subsequent 4 days of agriculture and food sciences research, 2015, 2(1): 12-18 16 storage. these authors also concluded that tortilla staling is best estimated by subjective rollability and 2-dimensional extensibility tests. cracking and breaking of tortillas during rolling can be delayed by using flour with higher protein quality or adding vital gluten, hydrocolloids and/or emulsifiers [24]. the force values related to tortilla firmness almost doubled after 24 h of storage. likewise, the tortilla extension values after one day of storage were approximately one half to one third of the values originally observed in fresh tortillas. the higher force and lower extension values are typically observed in bakery products and is mainly attributed to starch retrogradation. a comparison of the 5-day stored tortillas indicated that tortillas made with the 08 flour had the best textural properties. these tortillas had 50% less force compared to counterparts produced from the 03 flour. in terms of extension, there were not differences among tortillas at the same day. it is well known that the optimum protein for tortillas is intermediate because soft wheats usually yield tortillas with limited textural shelf-life and are more prone to lose texture with less reheating capacity. on the other hand, the use of hard wheat flours yields doughs that require more proofing and result in doughy and firmer tortillas [2, 18]. tortilla rupture was also correlated to total flour starch damage. accordingly, one of the major variables that need to be controlled during the milling process is the level of starch damage in flour and the stages where they are generated. a high wet gluten content in relation to starch damage, and optimal water distribution in the flour as affected by particle size apparently had positive effects on final tortilla quality. tortillas from the different mills were tested for rollability 5 days after baking. data shows significant differences in the rollability of the different baked tortillas. the 5-day old tortillas of three of the mills showed rollability values above 4. rollability tortilla values were negatively correlated to total flour starch damage as well as that in the reduction stages, -0.828 and -0.946 respectively. accordingly, an increase and controlled starch damage that affects water absorption and falling number values will guarantee good rollability of the final tortilla. 4. conclusions wheat flours with the highest c3, c4 and c5 parameters produced the best textured tortillas. these mixolab parameters proved to be indicative of the quality of wheat flours intended for hot-press tortilla production. 5. acknowledgments the authors would like to acknowledge us wheat associatesmexico for donating the arrays of flours and chopin technologiesfrance for generating the tortilla flour profiler. references [1] j. n. alviola and j. m. awika, "relationship between objective and subjective wheat flour tortilla quality evaluation methods," cereal chem., vol. 87, pp. 481-485, 2010. 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[24] c. p. friend, r. d. waniska, and l. w. rooney, "effects of hydrocolloids on processing and qualities of wheat tortillas," cereal chemistry, vol. 70, pp. 252-256, 1993. table-1. functional characteristics of refined wheat flours milled from the same wheat blend by ten different types of commercial mills. wheat flour 01 02 03 04 05 06 07 08 09 10 moisture, % 14.40 13.90 13.30 13.10 12.70 12.40 13.10 13.30 13.00 14.20 falling number, (sec.) 483 524 416 415 423 421 403 436 441 440 alveograph w 271 269 216 201 203 174 211 215 185 228 p/g 3.52 3.45 2.48 2.58 2.92 3.88 3.54 2.89 3.63 3.17 p/l 0.77 0.74 0.50 0.54 0.64 0.97 0.80 0.59 0.86 0.67 farinograph absorption 500, % 55.70 56.50 54.80 55.30 56.90 60.30 60.70 60.40 60.20 55.70 absorption 14 % 56.20 56.40 54.00 54.20 55.40 58.50 59.90 59.60 59.00 56.20 stability, min 14.30 13.40 10.50 10.00 8.60 8.50 6.20 8.00 5.60 14.30 starch damage values ucdc (units chopin dubois converted) 15.90 15.00 15.10 12.50 16.00 18.40 19.10 19.20 18.70 15.90 table-2. mixolab parameters of refined wheat flours milled from the same wheat blend by ten different types of commercial mills wheat flour water absorption (% 14) dough development time (min) primary parameters secondary parameters stability (min) c2 (nm) c3 (nm) c4 (nm) c5 (nm) c1c2 (nm) c3-c2 (nm) c3-c4 (nm) c5-c4 (nm) 01 55.0 1.43 9.67 0.49 2.16 1.75 3.36 0.63 1.67 0.41 1.61 02 55.0 1.83 9.82 0.50 2.14 1.73 3.41 0.64 1.64 0.41 1.68 03 53.6 1.48 8.58 0.46 2.18 1.78 3.36 0.65 1.72 0.40 1.58 04 54.5 1.62 9.32 0.45 2.14 1.75 3.30 0.63 1.69 0.39 1.55 05 55.0 1.32 9.42 0.45 2.04 1.71 3.12 0.64 1.59 0.33 1.41 06 56.9 1.53 9.20 0.53 2.06 1.70 3.30 0.57 1.53 0.36 1.60 07 57.6 4.68 9.10 0.53 1.94 1.46 2.82 0.58 1.41 0.48 1.36 08 57.1 5.30 9.83 0.53 1.93 1.52 2.95 0.56 1.40 0.41 1.43 09 57.2 4.68 8.53 0.54 2.02 1.55 2.87 0.56 1.48 0.47 1.32 10 56.0 5.02 9.85 0.56 1.98 1.56 3.10 0.59 1.42 0.42 1.54 a each value is the average of at least three observations. data on 14% mb flour. table-3. mixing water absorption and dough development time of refined wheat flours milled from the same wheat blend by ten different types of commercial mills for tortilla production1. wheat flour water absorption 2,3 , % optimum mix time (min) 01 49.85 12:20 02 47.79 13:00 03 47.39 11:42 04 47.25 12:00 05 48.85 11:00 06 52.27 10:30 07 53.07 10:42 08 52.20 11:00 09 51.71 10:50 10 53.83 11:00 1 each value is the average of at least three observations. data on 14% mb flour. 2 subjectively determined using a 1 to 7 scale where 7 meant that dough was extremely firm and 1 extremely slack. the optimum consistency was rated as 4. 3 water was tempered to 40°c before dough mixing. table-4. protein content and physical characteristics of tortillas produced from 10 different refined flours obtained from different commercial mills1. tortilla moisture protein2 diameter cm thickness mm color parameters l a b 01 25.17dc 11.60ab 12.34ef 2.00c 46.58ab -0.76f 14.49ab 02 27.98ab 11.50 ab 12.92bcde 2.04bc 53.99ab -0.62ef 12.34ab 03 24.10de 9.73b 12.33ef 2.00c 45.76ab 0.29b 15.40a 04 26.23bcd 10.85ab 12.16f 2.04bc 50.39ab -0.31de 12.62ab 05 24.09 de 10.25ab 12.36def 2.02c 59.13a 0.07bcd 14.25ab 06 29.05a 10.59ab 12.97bcd 2.31bc 48.91ab -0.33de 12.28ab 07 27.40abc 11.97ab 13.49b 2.37ab 43.21b 0.25b 11.27b 08 22.10 e 10.60ab 14.64a 1.54d 39.15b 0.20bc 13.52ab 09 25.50bcd 11.99ab 12.63cdef 2.71a 45.96ab -0.18cd 13.24ab 10 26.63abc 12.46a 13.01b 2.08bc 47.66ab 0.83a 13.40ab 1 each value is the average of at least three observations. means with different letter(s) in each column are statistically different (p<0.05). 2 values are expressed on dry matter basis. agriculture and food sciences research, 2015, 2(1): 12-18 18 table-5. textural properties of hot-press tortillas stored for 5 days at room temperature produced form flours produced by ten different types of commercial mills. a each value is the average of at least three observations. data on 14% mb flour. table-6. rollability of hot-press tortillas kept at room temperature for five days or reheated produced from different types of flours1 wheat flour rollability of tortillas at room temperature 2 rollability of reheated tortillas 3 day 0 day 1 day 2 day 5 day 7 01 1.0 a ±0 1.3 a,b ±0.2 2.5 a ±0.3 3.4 a,b ±1 1.7 b,c ±0.3 02 1.0 a ±0 1.5 a,b ±0.2 2.2 a,b ±0.1 2.5 b,c ±0 3.4 a ±0 03 1.0 a ±0 1.4 a,b ±0.2 2.2 a,b ±0.1 2.4 b,c ±0 2.0 b,c ±0 04 1.0 a ±0 1.1 b ±0 n/a n/a 1.5 c ±0.4 05 1.0 a ±0 1.2 a,b ±0.1 2.2 a,b ±0.1 4.4 a ±03 1.8 b,c ±0.2 06 1.0 a ±0 1.6 a ±0.2 1.7 b ±0.3 2.2 c ±0.1 2.2 b,c ±0.4 07 1.0 a ±0 1.4 a,b ±0.1 1.5 b ±0.1 2.1 c ±0.1 1.9 b,c ±0.3 08 n/a n/a n/a n/a n/a 09 1.0 a ±0 1.4 a,b ±0.2 2.1 a,b ±0.6 2.7 b,c ±0.6 2.4 a,b,c ±0.7 10 1.0 a ±0 1.3 a,b ±0.1 2.0 a,b ±0 2.6 b,c ±0.1 2.9 a ±0.7 1 each value is the average of at least three observations. 1 (breaks immediately; cannot be rolled) and 5 (no cracks; very flexible). 2means with different letter(s) in each row or mill of “rollability of tortillas at room temperature” are statistically different (p<0.05). 3means with different letter(s) in the column of “rollability of reheated tortillas” are statistically different (p<0.05). table-7. correlation coefficients between wheat flour mixolab parameters and force (texture parameter) of hot-press tortillas produced from ten different types of flours force at day of storage mixolab parameters c2 c3 c4 c5 0 1 0.878 0.872 0.786 2 0.775 0.656 5 0.120 0.732 (-) not significant. fig-1. particle size distribution of ten flours obtained from at the same wheat blend according to sieving to an end point method. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. 1 © 2023 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 10, no. 1, 1-7, 2023 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v10i1.4839 © 2023 by the authors; licensee asian online journal publishing group optimal design of blade parameters for fracturing tea-picking machine zehui jiang1 yongguang hu2 wenchao wu3 ( corresponding author) 1,2,3key laboratory of modern agricultural equipment and technology, ministry of education jiangsu province, zhenjiang, jiangsu university, china. 1email: 2222016029@stmail.ujs.edu.cn 2email: deerhu@ujs.edu.cn 3email: 2111716005@stmail.ujs.edu.cn abstract the blade is one of the most critical components in the fracturing tea-picking machine, and this study is conducted to optimize the blade's working parameters. in this study, the effects of blade width, blade thickness, and cutting angle on the maximum fracturing force of tea stems were analyzed using the l9 (34) standard orthogonal table, with the maximum fracturing force used as the evaluation index. the results indicate that the main factors affecting the maximum fracturing force (mff) of tea stems are cutting angle (ca), blade width (bw), and blade thickness (bt) in that order. furthermore, microscopic observation of the fracture surface revealed that compared with the thickness of the other two blades, the thickness of 0 mm caused the cross-section uneven and had lots of burrs, correspondingly resulting in the section's oxidation and the deterioration of tea leaf quality. therefore, the optimal combination of design parameters was a cutting angle of 90°, a blade width of 2.0 mm, and a blade thickness of 0.5 mm. the findings of this study can provide reference for blade design to reduce the fracturing force of tea-picking machines, lower the working power consumption, and improve the quality of freshly plucked tea leaves. keywords: blade, famous premium tea, fracturing force, optimal design, orthogonal experiment, tea-picking machines. citation | jiang, z., hu, y., & wu, w. (2023). optimal design of blade parameters for fracturing tea-picking machine. agriculture and food sciences research, 10(1), 1–7. 10.20448/aesr.v10i1.4839 history: received: 13 march 2023 revised: 26 april 2023 accepted: 3 july 2023 published: 18 july 2023 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the china and jiangsu postdoctoral science foundation (grant numbers: 2022m711396 and 2021k614c). institutional review board statement: the ethical committee of the jiangsu university, china has granted approval for this study (ref. no. jsdx20230717001). authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. competing interests: the authors declare that they have no conflict of interest. contents 1. introduction ......................................................................................................................................................................................... 2 2. materials and methods ...................................................................................................................................................................... 3 3. results and discussion ...................................................................................................................................................................... 5 4. conclusion ............................................................................................................................................................................................ 6 references ................................................................................................................................................................................................. 7 mailto:2222016029@stmail.ujs.edu.cn mailto:deerhu@ujs.edu.cn mailto:2111716005@stmail.ujs.edu.cn https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v10i1.4839 https://orcid.org/0009-0000-9017-0406 https://orcid.org/0000-0002-3771-9380 https://orcid.org/0000-0001-6135-8004 agriculture and food sciences research, 2023, 10(1): 1-7 2 © 2023 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study determined the optimal combination of blade parameters for a breaking-type tea harvester using the orthogonal experiment method. currently, there is no research available on the optimization of blade parameters for this type of tea harvester. 1. introduction china is the largest tea producer in the world, with a tea plantation area of 3307.84 thousand hectares in 2021 [1, 2]. the shortage of tea pickers and high labor cost have become common problems for tea production [3]. in view of the non-uniform planting specifications of tea gardens and the complex terrain of tea-growing areas in china, the development of portable tea-picking equipment is of practical significance [4, 5]. to improve the efficiency and quality of tea picking, people have been constantly exploring and innovating [6]. the hand-held tea harvester is a machine that has emerged under this background [7]. compared with traditional manual picking, the hand-held tea harvester can quickly pick tea leaves, greatly improving the efficiency and yield of tea picking [8-10]. the fracturing tea-picking machine working principle is to use a rotating leaf-beating rod and a blade to cut off the tender shoots of tea trees, and then collect them through a collection device [11]. compared with the reciprocating cutting tea picking machine, the hand-held tea harvester has adaptability to be adjusted according to different types of tea trees and crown structures [12] to ensure the quality and efficiency of picking. pan designed a tea stem shear test device using an electronic universal testing machine and a fixed support to obtain the force-deformation curve when the stem is cut by the blade [13]. cao specifically designed a mini-test machine that can adjust the entry angle of the tea harvester blade to measure the maximum shear force when the stem is cut [14]. the orthogonal experiment is to be conducted to analyze the effects of cutting angle, blade width, and blade thickness on the maximum fracturing force of tea stems. their impact level is evaluated through data analysis, and the microscopic structure of the section cut by the blades of different parameters is observed. this study aims to explore the related content of the design experiment of blade parameters for the hand-held tea harvester, providing useful reference and guidance for its research and application. figure 1 shows the fracturing tea-picking machine. figure 2 shows the blade of fracture tea-picking machine. figure 1. fracturing tea-picking machine. figure 2. blade of fracture tea-picking machine. blade agriculture and food sciences research, 2023, 10(1): 1-7 3 © 2023 by the authors; licensee asian online journal publishing group 2. materials and methods 2.1 materials the selected experiment site is maichun tea farm located in danyang, jiangsu province, china (32°02'26" n, 119°67'44" e), which belongs to the hilly tea region of the middle and lower reaches of the yangtze river. the tea variety planted is mao green, and the planting direction is north-south with a row spacing of 1.5 m and a crown width of 1.2 m. the tea garden was moderately pruned on august 10, 2022, and the experiment was conducted on october 12, 2022.five square test areas with equal side lengths of 1.0 m were randomly selected, and 45 stems of similar growth stage and stem diameter were randomly selected in each area for leaf removal and subsequent testing. the stems are placed on the fixed holder by fixture. the maximum fracturing force was measured using a tensiometer (edinburgh, china, accuracy 0.01 n). the microscopic structure of the stem cross-section was observed using a super depth-of-field three-dimensional microscope (keyence, japan, vhx-900f). the position of the tensile tester was adjusted using a lifting platform, and the blade approached the tested stems by a screw push rod. three types of blades with different thicknesses were fixed onto the tensile tester using a clamp, and the stem position was measured using a ruler and fixed using a fixture. the experimental instrument is shown in figure 3. figure 3. experiment materials. note: 1. fixed holder 2. fixture 3. stem 4. blade 5. clamp 6. tensiometer 7. screw push rod 8. lifting platform. 2.2. orthogonal experimental design by assessing whether the design parameters of the blade affecting the maximum cutting force can be controlled quantitatively, the degree of control difficulty, and the control method selected the main influencing factors are: blade width (a), blade thickness (b), and cutting angle (c). to consider the size of the prototype, blade width(bw) range of 2.0-4.0mm, blade thickness(bt) of 0-1.0mm, and cutting angle(ca) of 90-135 °. orthogonal tables are statistical tools used for experimental design and data analysis. common orthogonal tables include l4 (23), l8 (827), l16 (215), l9 (34), etc. take l9 (34) and so on. in these tables, "l" represents the orthogonal table, while the numbers represent specific parameters of the table. for instance, l9 (34) means there are 9 horizontal rows, which corresponds to 9 experiments to be conducted. there are 3 factors, with each factor having 3 levels, and the maximum number of factors allowed to be arranged is 4. the degrees of freedom for each factor are 2. based on the principle that the degrees of freedom of the orthogonal table should be greater than or equal to the sum of the degrees of freedom of each factor and the degrees of freedom of interaction, as well as the principle of choosing the smallest number of orthogonal tables, l9 (34) was selected for the experiment [15]. it can be observed that orthogonal tables have two characteristics: (1) in each column, each level of each factor appears an equal number of times in the total number of experiments. (2) an ordered series in which different levels of any two factor columns appear in pairs, with each pair appearing an equal number of times. therefore, the distribution of each factor level combination in the orthogonal table is balanced dispersed, and neatly comparable among all level combinations. the assignment of the interactions in the orthogonal table header design needs to be done according to the interaction table, which is mentioned in detail in the paper [15]. the level table of experimental factors is shown in table 1.the experiment was started by replacing each factor level code with a specific level setting, and the experimental arrangement and sequence were shown in table 2, and the maximum fracturing force was filled in the result column in table 2 at the end of the experiment. table 1. table of factor levels. level a b c bw/mm bw/mm ca/° 1 2 0 (sharp edge) 90 2 3 0.5 120 3 4 1 135 agriculture and food sciences research, 2023, 10(1): 1-7 4 © 2023 by the authors; licensee asian online journal publishing group table 2. table of orthogonal tests. test number a b c d y 1 1 1 1 1 1.79 2 1 2 2 2 3.32 3 1 3 3 3 4.34 4 2 1 2 3 3.23 5 2 2 3 1 4.31 6 2 3 1 2 2.54 7 3 1 3 2 4.35 8 3 2 1 3 2.83 9 3 3 2 1 4.48 2.3. fracturing experiment stem fracturing experiments were performed on a liftable platform and tensiometer. a clamp is used on the tensiometer to fix the blade for fracturing. the measured internode is fractured on the stem, the specimen is placed on the fixed holder, the blade position and the stem inclination are adjusted, the stem is slowly fractured and the maximum fracturing force is read on the tensiometer. the blade width is the distance between the blade and the fixture fixing the stem, the blade thickness is the thickness of the blade, and the cutting angle is the angle formed between the blade cutting into the stem and the stem in the direction of the stem. each experiment was repeated five times, and the average maximum fracturing force was recorded in the result column of table 2. figures 4-6 show the different cutting angle. figure 4. 90°ca. figure 5. 120° ca. figure 6. 135° ca. 2.4. stem section observation cross-sectional sections of the stems were cut by the conventional sectioning method, and the cross-sectional features were compared under different blade fractures using an ultra-deep field 3d microscope. agriculture and food sciences research, 2023, 10(1): 1-7 5 © 2023 by the authors; licensee asian online journal publishing group 3. results and discussion 3.1. orthogonal test results and analysis for the processing and analysis of experimental results, there are usually two methods. one is the visual analysis method, also called range analysis method; the other is the analysis of variance method. the following two methods are used to compare and analyze the results of orthogonal experiments. 3.1.1. range analysis the analysis steps of range analysis method are as follows: first, the experimental results of level one for each factor of each experiment are calculated, and the mean of k1. then the mean of level two for each factor of each experiment is calculated and the mean of k2, then the mean of level three for each factor of each experiment is calculated, and the mean of k3. then the range r of the mean of the three levels and k1, k2 and k3 is found, and finally, r is sorted from largest to smallest to determine the degree of influence of the factors. the results of range analysis are shown in table 3. the principle of range analysis is the larger range, the more important the influence of the corresponding factor. in addition, the optimal level of each factor can be determined according to the average size of the results corresponding to the level of each factor. if the indicator is expected to be larger and better, the larger the average level is taken; if the indicator is expected to be smaller and better, the smaller the average level is taken. it can be seen that the required indicator in this test is the smaller the better, so the preliminary optimal level derived by the range analysis is a1b1c1. table 3. table of range analysis. test number a b c d y 1 1 1 1 1 1.79 2 1 2 2 2 3.32 3 1 3 3 3 4.34 4 2 1 2 3 3.23 5 2 2 3 1 4.31 6 2 3 1 2 2.54 7 3 1 3 2 4.35 8 3 2 1 3 2.83 9 3 3 2 1 4.48 k1 9.45 9.37 7.16 10.58 / k2 10.08 10.46 11.03 10.21 / k3 11.66 11.36 13 10.4 / k1 3.15 3.12 2.39 3.53 / k2 3.36 3.49 3.68 3.40 / k3 3.89 3.79 4.33 3.47 a1b1c1 range r 0.74 0.66 1.95 0.12 c>a>b the orthogonal tests were performed according to the designed l9 (34) with a total of 9 test combinations, and the obtained test results are shown in table 3. in order to accurately estimate the importance of the influence of the three design parameters of the blade on the maximum fracturing force, especially considering the interaction between the factors, it is necessary to conduct anova on the orthogonal test results. the anova results are shown in table 2. the variances of a, b and c are less than 0.05, so a, b and c all have a significant influence on the test results, and the main order of the factors is: c>a>b. 3.1.2. analysis of variance anova is used to distinguish whether the differences in the investigated factors corresponding to experimental results due to different levels are caused by level changes or by experimental errors, in order to further test which factors have an effect on the results and which don’t and to distinguish which factors are major and which are minor. the analysis steps of anova are as follows: first, the degrees of freedom of each factor, the degrees of freedom of interaction, the degrees of freedom of error, and the total degrees of freedom are listed. then the sum of squares of deviations corresponding to each of these factors is calculated as equation 1. 𝑆𝑆𝑇 = ∑ (𝑥𝑖 − �̅�)2𝑛 𝑖=1 . (1) then the mean sum of squares of deviations corresponding to each factor is found by dividing the sum of squares of deviations by the degrees of freedom (df). equation 2 yields the mean sum of squares of deviations corresponding to a factor. 𝑆𝐴 2 = 𝑆𝑆𝐴 𝑓𝐴 . (2) then the f-value corresponding to each item is found by dividing the average deviation sum of squares by the average deviation sum of squares of errors. equation 3 yields the f-value of a factor. 𝐹𝐴 = 𝑆𝐴 2 𝑆𝑒 2. (3) the p-value is calculated using excel's f.dist.rt function. if the factor's p-value is ≤ α, there is (1-α) certainty that the factor has a significant impact. commonly used values of α are α=0.01, α=0.05, and α=0.10. when pa≤0.01, it indicates an extremely significant impact, noted as "**"; when 0.01≤pa≤0.05, it indicates a significant impact, noted as "*"; and when pa≥0.05, there is no significant impact. table 4 shows the anova results for the orthogonal experiments. factor c had an extremely significant impact, while factors a and b had a significant impact, with the main order of factors being c>a>b. this is consistent with the results of the range analysis. agriculture and food sciences research, 2023, 10(1): 1-7 6 © 2023 by the authors; licensee asian online journal publishing group table 4. analysis of variance (anova) table. source of variance sst df s2 f p significance a 0.864 2 0.432 37.865 0.026 * b 0.662 2 0.331 29.008 0.033 * c 5.885 2 2.942 257.855 0.004 ** d (difference) 0.023 2 0.011 1.000 1.000 / 3.2. analysis of microscopic features of the cross-section the difference in stem fracture between the different groups was in the fracture caused by the blade with a sharp edge and the blade without a sharp edge. the blade with a sharp edge fracture is shown in figure 7, the 0.5 mm thick blade fracture is shown in figure 8, and the 1 mm thick blade fracture is shown in figure 9. under the blade with a sharp edge fracturing picking, the blade squeezed the stem and formed an oval incision. the fracture interface was uneven and showed a large number of burrs. the stem tissue was torn at the fracture point and the fractured interior was also damaged by the squeezing action, resulting in the cavity observed in the wood pith. after fracturing picking, the fractured portion and the damaged internal tissues are extensively oxidized. the oxidation produces substances that darken the tea leaves [9]. fracturing picking with a blade with a sharp edge disrupts the tissue within the stem, and this rupture produces excessive tissue fluid, which further increases the rate of oxidation. in contrast, fracturing with an unopened blade produces a flat, burr-free fracture. the oxidized area in the incisions of hand-picked shoots was significantly smaller than that of the blade fracturing picked ones. after the incisions were oxidized by ambient air, the xylem and pith in the stem tissue took on a brownish-red color, which could affect the color and flavor of the finished tea [16]. figure 7. 0mm blade section. figure 8. 0.5 mm blade section. figure 9.1mm blade section. 4. conclusion (1) through the orthogonal test of three design parameters of the blade, the results show that the main and secondary factors affecting the maximum fracturing force in the order of cutting angle, blade width, and blade thickness. among them, the cutting angle on the maximum fracturing force is very significant, the blade width, and blade thickness on the maximum fracturing force is significant, the preliminary optimal combination of design parameters level for the cutting angle: 90 °, blade width: 2.0 mm, blade thickness: 0 mm. agriculture and food sciences research, 2023, 10(1): 1-7 7 © 2023 by the authors; licensee asian online journal publishing group (2) analysis of the stem sections fractured by different blades revealed that the fracture interface formed by the 0mm blade was uneven relative to the other blades and showed a large number of burrs. the stem tissue was torn at the fracture point and the fractured interior was also damaged by the crushing action, with extensive oxidation of the fractured portion and the damaged tissue inside. the substance produced by oxidation will darken the tea leaves and affect the color and taste of the finished tea leaves. therefore, a blade thickness of 0.5 mm should ultimately be selected. the optimal structural parameter level combination is the optimal structural parameter level combination of cutting angle: 90°, blade width: 2.0 mm, and blade thickness: 0.5 mm. references [1] statistical bureau of the people's republic of china, china statistical yearbook. beijing: china statistics press, 2022. 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[16] k. liu, research on the enzymological characteristics of polyphenol oxidase from tea and the effect of infrared on its activity and conformation. wuxi, china: jiangnan university, 2013. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.14083/j.issn.1001-4942.2015.04.013 https://doi.org/10.13733/j.jcam.issn.2095-5553.2020.11.006 https://pss-system.cponline.cnipa.gov.cn/disclaimer https://pss-system.cponline.cnipa.gov.cn/disclaimer 69 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 69-78, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.69.78 © 2020 by the authors; licensee asian online journal publishing group determination of np fertilizer rates and plant population density on yield and yield related parameters of quality protein maize (zea mays l.) in southwestern ethiopia sisay gurmu jimma agricultural research center, jimma, ethiopia. abstract field studies were conducted in 2015 and 2016 under rain fed conditions in south western ethiopia, jimma zone to determine optimum plant population density and np fertilizer on maize. the experiment had four np fertilizer rates; 69/52, 92/69, 115/86, and 138/104 n/p2o5 kg ha-1 and four plant population density; 44,444(75cm*30cm), 53,333(75cm*25cm), 66,666(75cm*20cm) and 88,888(75cm*15cm) plants per hectare. each treatment was assigned to each plot in split plot design. grain yield and above ground biomass significantly increased as plant populations and np fertilizer rates increased. the number of ears per plant was significantly decreased as plant population density increased and increased with the increased rate of np fertilizer. the greater lodging was recorded in 2015 cropping season when compared with that of 2016 cropping season. the highest grain yield 8752 kg ha-1 and above ground biomass 18.33 ton ha-1 were recorded from 138/104 n/p2o5 fertilizer. regarding plant population density the highest grain yield 9401 kg ha-1 and above ground biomass 19.90 ton ha-1 were recorded from 88,888 plants ha-1 (75cm*15cm). the highest net benefit 30,669etb ha-1 with highest mrr 164% was recorded from 115/86 n/p2o5 kg ha-1. concerning plant population density the highest net benefit 36,705 etb ha-1 with acceptable mrr 585% was obtained from 88,888 plants ha-1 (75*15cm). therefore, plant population density 88,888 plants ha-1 (75*15cm a plant hill-1 or 75 x 30 cm two plants hill-1) with n/p2o5 fertilizer rate of 115/86 kg ha-1 can be advisable for farmers in the study area and adjacent woredas’ with similar agro-ecologies. keywords: grain yield, plant population density, quality protein maize and fertilizer. citation | sisay gurmu (2020). determination of np fertilizer rates and plant population density on yield and yield related parameters of quality protein maize (zea mays l.) in southwestern ethiopia. agriculture and food sciences research, 7(1): 69-78. history: received: 5 february 2020 revised: 10 march 2020 accepted: 13 april 2020 published: 15 may 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group funding: the research was funded by ethiopia institute of agricultural research competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 70 2. materials and methods ................................................................................................................................................................... 70 3. results and discussion ................................................................................................................................................................... 72 4. conclusion and recommendation ................................................................................................................................................ 77 references .............................................................................................................................................................................................. 77 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.69.78&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.69.78&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://asianonlinejournals.com/index.php/aesr/article/view/1634 https://orcid.org/0000-0002-9501-9526 agriculture and food sciences research, 2020, 7(1): 69-78 70 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this research study can find answers to things that are unknown around maize planting, filling gaps in knowledge and changing the way that maize agronomist works in general. 1. introduction globally, maize (zea mays l.) is among the leading cereals in production along with rice and wheat. in africa, ethiopia is the third largest maize producer next to nigeria and egypt [1]. maize ranks second after teff in area coverage and first in total production in ethiopia [2]. in 2016/17 the maize crop area and grain production in ethiopia was 2,135,572 ha and 7, 8471,175 ton, respectively with productivity of 3675 kg ha-1 [2]. normal maize grain has greater nutritional value as it contains 72% carbohydrate, 8.8% protein, 2.15% fiber and 2.33% ash [3]. it is a good source of carbohydrates, fat, protein and some important vitamins (b6, a and e) and minerals (magnesium, potassium and phosphorus), but deficient in essential amino acids viz., lysine and tryptophan that reduces its biological value [4]. the amount of these deficient amino acids has been increased by incorporating opaque-2 gene in quality protein maize (qpm) [5]. it produces 70-100% more of lysine and tryptophan than the most modern varieties of tropical maize [6]. insufficient application of nutrients and poor soil management, along with harsh climatic conditions and other factors, have contributed to the degradation of soils including soil fertility depletion in developing countries, especially in ssa [7]. poor soil fertility is one of the principal factors that limit maize productivity in maize growing areas of ethiopia [8]. degradation of soil physico-chemical properties, soil acidity with high p sorption and soil nutrient depletion due to low chemical fertilizer use by most small-holder farmers who cannot afford the expensive fertilizers leads to declining in maize production in ssa [9]. among plant nutrients nitrogen is a vitally important, a major yield determining nutrient and its availability in sufficient quantity throughout the growing season is essential for optimum maize growth [10]. it is a component of protein, nucleic acids and other compounds essential for plant growth process [11]. whereas phosphorus is the second most important nutrient element (after nitrogen) limiting agricultural production [10]. it is used for growth, utilization of sugar and starch, photosynthesis, metabolic process which leads to higher yield of the crop [12]. maize is commonly planted in rows of varying spaces; less effort has been made to plant at optimum densities to maximize its productivity in different agro-ecologies of ethiopia. summaries of earlier results from different studies on maize plant population densities indicate that better yields were obtained at planting density in range of 6-7.5 plants m-2 [13]. it is being observed that medium and early maize varieties in humid lowlands and low moisture stress area found to be varied in structure and leaf arrangements from known normal maize varieties. these variations in morphology may lead to different planting density to reach at their at maximum yield potentials. plant population density has a significant impact on growth and yield of crops, including maize, a popular c4 cereal crop. therefore, understanding how plants regulate their growth in response to plant population densities has problems, such as determination of optimal sowing density. increased plant populations could lead to increased yields under optimal climatic and management conditions due to greater number of smaller cobs per unit area [14]. hence, for further dissemination and adoption of qpm hybrid there is a need to understand its performance with various agronomic management practices, of which nutrient management and plant stand are vital in influencing the growth and yield of the crop. hence, this study was initiated with the objective to study the effect of plant population density and np fertilizer on the growth and yield of maize. 2. materials and methods 2.1. description of the study area the research activity was performed on farmers’ field in omonadawareda (nada-sadechakebele) of jimma zone, south western ethiopia in main cropping seasons (mid may up to early november) of 2015 and 2016 figure 1. this location is foundon latitude 7º36' n and longitude 37º 15' e and laid at an altitude of 1764 m.a.s.l. ten years average minimum and maximum temperature is 8ºc and 28ºc respectively and reliably receives annual average rainfall of 1200 mm with agro-ecology characterized by humid mid altitude. the farming system of the study site is cereal crops dominated with maize, teff and sorghum also has warm and cold climate, also convenient topography which is very suitable for all agricultural practices. the soil type of the experimental area was eutric-nitisols (reddish brown). figure-1. decadal time scale (2007-2016) of precipitation, eto, 1/2 eto, maximum and minimum temperature at jimma, omonadaworeda. agriculture and food sciences research, 2020, 7(1): 69-78 71 © 2020 by the authors; licensee asian online journal publishing group 2.2. soil physico-chemical properties the soil of the experimental field was characterized for selected physico-chemical properties before the application of the treatments table 1.the average soil ph of the trial site was 5.13, which was strongly acidic [15] and ideal for the production of most field crops. the ph of the soil affects maize growth by suppressing the root development and reducing availability of macronutrients to plants especially phosphorus [16]. the soil total n was 0.19%.these was found to be a medium rate for crop growth and development [17]. the oc content of experimental area was 2.08% and it was found medium rate for crop growth [18]. the bray ii extractable available p was 6.17 mg kg-1 which is below the critical level (8 mg kg-1) for most crop plants as described by tekalign and haque [19]table 1. table-1.selected physico-chemical properties of the soil of the experimental sites before planting. soil characters values rating reference method of analysis ph(1:2.5) 5.13 strongly acidic batjes [15] 1:2.5 ratio water method av p(mg kg-1) 6.17 low tekalign and haque [19] bray ii method [20] tn (%) 0.19 medium berhanu [17] kjeldahl method [21] oc (%) 2.08 medium tekalign [18] walkley and black method [22] c:n ratio 11.05 medium brady and weil [23] om (%) 3.58 high berhanu [17] note: where ph= hydrogen ion concentration, oc=organic carbon, tn=total nitrogen, av. p=available phosphorous, om=organic matter. values are the means of duplicated samples. 2.3. experimental treatments and procedures the experimental field was ploughed and prepared following the conventional tillage practice before planting. the land was leveled using manual power before the field layout was made. the maize was planted during mid may and harvested early november of each year. two maize seeds were planted per hill and then thinned to one plant per hill after good establishment of seedlings so as to maintain a single healthy plant per hill. there were four np fertilizer rates; 69/52, 92/69, 115/86, and 138/104 kg/ha n/p2o5 and four plant population density; 44,444, 53,333, 66,666, and 88,888 plants per hectare. the experimental plot size was 22.95m2 (4.5 m x 5.1 m) for each treatment. the split plot design with three replications was used that plant population densities in main plot and fertilizer rates in subplots. nitrogen and phosphorus fertilizers were calculated as per rates given above and applied per area base. to increase the nitrogen use efficiency, it was split in three equal rates and applied at planting time, knee height and flag leaf initiation stages. all other agronomic practices were applied uniformly to all experimental plots in the study area. 2.4. description of the experimental materials medium maturing maize variety bhqpy545 was used in the present study. it was released by bako agricultural research centre through the national maize research program in 2008. it performs well in agroecology of 1000-2000 m.a.s.l with rainfall of 1000-1200 mm. it can give 8.0-9.5 and 5.5-6.5 t ha-1 grain yields under on-station and on-farm experiments, respectively. it was moderately tolerant to rust, blight and gray leaf spot with maturity date of 138. 2.5. data collected 2.5.1. plant height (cm) it was measured at ground level to terminal stem using measuring stick at the point where the tassel starts branching from six randomly selected plants. 2.5.2. number of ear per plant it was obtained by counting total number of ears in each plot and divided to total number of plant stand harvested. 2.5.3.grain yield (kg ha-1) grain yield per plot was recorded using electronic balance and then adjusted to 12.5% moisture and converted to hectare basis. 2.5.4.above ground biomass (kg ha-1) all above ground biomass was harvested from net plot and weighted, ears were removed and weighted separately, six plants were selected, chopped and oven dried till get uniform weight. 2.5.5. lodging percent it was obtained by counting the total number of stalk and root lodging in each plot and divided to the total number of plant stand at harvesting. 2.5.6. harvest index was calculated as the ratio of grain yield to above ground biomass yield on dry weight basis [24]. hi(%) = economic yield (kg/ha) total biological yield (kg/ha) x100 agriculture and food sciences research, 2020, 7(1): 69-78 72 © 2020 by the authors; licensee asian online journal publishing group 2.6. data analysis analysis of variance (anova) for all collected data was computed using sas version 9.3 statistical software. whenever the anova results showed the significant differences between sources of variation, the means were compared using least significant difference. 2.7. partial budget analysis partial budget analysis was performed to investigate the economic feasibility of the treatments and assess the costs and benefits associated with different treatments of chemical fertilizers and plant population density of the seed rates. the partial budget technique as described by cimmyt [25] was applied. the partial budget analysis was done using the prevailing market prices for inputs at planting and for outputs at the time the crop was harvested. all costs and benefits were calculated on a per hectare basis in ethiopian birr (etb). the inputs and/or concepts used in the partial budget analysis were the mean grain yield of each treatment, the gross field benefit (gfb) ha-1 (i.e., the product of field price and the mean yield for each treatment), the field price of chemical fertilizers and urea kg-1 (the nutrient cost plus the cost of transportation from the point of sale to the farm), cost of labor spent on seed purchase and planting, the total costs that varied (tvc) which included the sum of field costs of fertilizers and their application, and seed purchase and planting. the net benefit (nb) was calculated as the difference between the gfb and the tvc. the marginal rate of return (mrr %) were also calculated. to obtain an estimate of these returns, the mrr (%) was calculated as changes in nb divided by changes in cost. thus, a minimum acceptable mrr of 100% was used; indicating for every one etb expended there is a return of one etb for a given variable input [25] which is suggested to be realistic. this enables the farmer to make recommendations from marginal analysis. the dominance analysis procedure as detailed in cimmyt [25] was used to select potentially profitable treatments from the range that was tested. sensitivity analysis for different interventions was also carried out to test the recommendation made for its ability to withstand price changes. sensitivity analysis simply implied redoing marginal analysis with the alternative prices. through sensitivity analysis, maximum acceptable field price of an input was calculated with the minimum rate of return as described by shah, et al. [26]. 3. results and discussion the effect of different rates of np fertilizer and plant population density on yield and yield related parameters of qpm maize variety and cost benefit analysis were presented and discussed as follows.it has been observed that medium maturing maize variety like bhqpy545 vary in structure and leaf arrangements from other late maturing maize varieties. these variations in morphology may lead to different planting density to reach the maximum yield potentials. table-2. mean square from combined analysis of the effects of np fertilizer rates and plant population density on growth, yield attributes and yield of qpm maize during 2015 and 2016 cropping seasons at omonadaworeda, jimma zone southwestern ethiopia. mean square for source of variation parameter year (1) intra row (3) year*intra row (3) year x np (3) np (3) intra row x np (9) year x intra row x np (9) error a (6) error b (24) plant height(cm) 41242.7** 823.5972* 88.39583ns 52.6875ns 2132.5** 119.44ns 209.340ns 238.079 88.396 ears per plant 1.3085** 0.98015** 0.02850ns 0.02554ns 0.23307** 0.02496ns 0.03228ns 0.0522 0.0285 lodging % 4773.8** 431.436ns 457.346ns 544.517ns 93.628ns 56.884ns 94.989ns 82.85 89.85 grain yield (kg ha-1) 112165917** 48126018** 1241199ns 1692656ns 17101704** 1535100ns 1459915ns 2916763 12411 98 agb (t ha-1) 402.058** 219.1591** 0.5320ns 7.3594ns 54.8910** 6.2721ns 5.2058ns 8.539 0.532 harvest index 0.0011ns 0.000973ns 0.0051ns 0.00011ns 0.001581ns 0.002343n s 0.0017ns 0.0026 0.0051 note: *numbers in parenthesis = degrees of freedom; *= significant (p < 0.05); ** = highly significant (p<0.01) difference; agb= aboveground biomass; ha = hectare. the homogeneity test of the error variances for years indicated that the error variance was homogenous and hence combined analysis of variance was conducted. over year combined analysis effects of np fertilizer rates and plant population density didn’t show significant (p >0.05) interaction effect on plant height, ears per plant, lodging percent, grain yield, aboveground biomass and hi. plant height was significantly (p <0.05) influenced by plant population density and highly significantly (p <0.01) influenced by np fertilizer and year. whereas ears per plant, grain yield and above ground biomass were highly significantly (p <0.01) influenced by np fertilizer rates, plant population density and year. however, lodging percent and harvest index were not significantly (p >0.05) affected by both np fertilizer rates and plant population density, but lodging percent was highly significantly (p <0.01) affected by the year difference table 2. 3.1. effect of np fertilizer and plant population density on growth parameters 3.1.1. plant height at omonada woreda jimma zone, medium maturing maize hybrid (hbqpy545) had exhibited vigorous growth performances and resulted in good response to both np fertilizer and plant population density. the mean significant higher plant height was recorded in 2016 main cropping season. numerically, the longer plant height (247.0cm) was recorded by plant population density of 88,888 plants ha-1 (75*15cm) but its effect was not statistically significant different from plant population density of 53,333 and 66,666 plants ha-1. while the shorter plant height (237.1cm) was recorded by 44,444 plants ha-1 (75*30cm) table 3. these results indicated as plant population increased the plant height also increased and vice versa. the plant height was increased by 4.2% at the highest plant population density as compared to the lowest plant population density.highest plant height in closer intra row spacing might be due to the presence of higher competition for sun light, crowding effect of the plant and agriculture and food sciences research, 2020, 7(1): 69-78 73 © 2020 by the authors; licensee asian online journal publishing group other resources that decrease in the stem diameter and number of green leaves. earlier results explained that the number of plants increased in a given area, the competition among the plants for nutrients uptake and sunlight interception also increased [27]. these finding is in agreement with hassan [28] who revealed that plant height increased with increasing plant density from 47600 to 71400 plants ha-1. regarding the effect of np fertilizer rates the longer plant height (250.5cm) was recorded by 138/104 kg ha-1 n/p2o5 fertilizer. but its effect was statistically at par with 115/86 kg ha-1 n/p2o5 fertilizer,while the shorter plant height (0.90) was recorded from 69/52 kg ha-1 n/p2o5 fertilizer table 3. the results indicated that as np fertilizer increased the plant height was also increased and vice versa. application of 138/104 kg ha-1 n/p2o5 fertilizer increased plant height by 6.7% over 69/52 kg ha-1 n/p2o5 fertilizer. the increase in plant height with increasing rate of np fertilizer could be due to their major effect in crop growth and development. nitrogen is considered as one of the major limiting nutrients in plant growth and adequate supply of it promotes the formation of chlorophyll which in turn resulted in higher photosynthetic activity, vigorous vegetative growth and taller plants. whereas phosphorous is required for shoot and root development where metabolism is high and cell division is rapid [29]. these results were in line with the findings of adekayode and ogunkoya [30]who explained that there was very high significant difference in maize plant height in plots treated with higher fertilizer compared with nil application. also kumar, et al. [31] reported that growth and yield of maize plants in terms of plant height varied significantly due to various fertility levels. he reported the highest plant height was recorded from application of 100% npk (120n:26.2p:33.2k) with 10 ton fym ha-1 over control. 3.2. effect of np fertilizer and plant population density on yield and yield related parameters 3.2.1. number of ears per plant number of ear per plant was determined by prolific ability of the bhqpy545 maize variety as described by adefris, et al. [32] and the growth behavior of the crop which is dependent upon management practices and edaphic and climatic factors. the number of ears per plant was higher (1.43) during 2016 main cropping season as compared with 2015 cropping season. the maximum number of ear per plant (1.50) was recorded from 44,444 plants ha-1 (75*30cm) which was statistically at par with 53,333 plants ha-1 (75*25cm) while the minimum number of ears per plant (1.17) was recorded from 88,888 plants ha-1 (75*15cm) figure 2. these results indicated as plant population density increased number of ears per plant linearly decreased and vice versa. the number of ears per plant was increased by 28.2% at the lowest plant population density (44,444 plants ha-1) as compared to the highest plant population density (88,888 plants ha-1). this might be due to in the closest spacing there is high inter stem competition to nutrients and sun light so, it caused lack of nutrient for each plant and consequently compelling the plants to undergo less reproductive growth. these finding are in agreement with hashemi-dezfouli and herbert [33] who reported a significantly higher number of ear per plant at lower plant density as compared to higher plant density. concerning the response of number of ears per plant to np fertilizer rates the maximum number of ears per plant (1.41) was recorded from 138/104 kg ha-1 n/p2o5 fertilizer. but its effect was statistically at par with 92/69 and 115/86 kg ha-1 n/p2o5 fertilizer, while the minimum number of ears per plant (1.26) was recorded from 69/52 kg ha-1 n/p2o5 fertilizer figure 2. these result indicated as np fertilizer rate increased to 138/104 kg ha-1 n/p2o5 fertilizer, the number of ears per plant was increased and vice versa. application of 138/104 kg ha-1 n/p2o5 fertilizer increased number of ears per plant by 11.9% over application of 69/52 kg ha-1 n/p2o5 fertilizer. the improvement of the soil conditions or enrichment with nutrients due to soil-added materials might be responsible for better cob production under plots treated with higher np fertilizer. nitrogen can trigger vegetative growth and development and it is an integral part of chlorophyll, which is the primary absorber of light energy needed for photosynthesis. these results were in line with the result of singh and nepalia [34] who reported the application of higher dose of chemical fertilizer improved the number of cobs plant-1 in qpm hybrid significantly over the lower nutrient application. agriculture and food sciences research, 2020, 7(1): 69-78 74 © 2020 by the authors; licensee asian online journal publishing group figure-2.effect of np fertilizer and plant population density on number of ear per plant at jimma, omonadaworeda in 2015 and 2016 cropping seasons. 3.2.2. lodging percent during 2015 main cropping season the lodging percent of qpm maize variety was higher than during 2016 cropping season. it was not significantly (p>0.05) affected by both np fertilizer rates and plant population density table 2. 3.2.3. grain yield hybrid qpm maize bhqpy545 had exhibited vigorous growth performances and resulted in good response to np fertilizers and plant population density in season 2015 and significant increase in grain yield was recorded table 3. over season mean indicated, the higher plant population density (88,888 plant ha-1) gave higher grain yield (9401.2 kg ha-1) while the lower grain yield (7046.0 kg ha-1) was recorded from 44,444 plants ha-1 (75*30cm)table 3. the result indicated as the plant population density increased the grain yield of qpm maize variety linearly increased and vice versa. the grain yield of the crop was increased by 33.4% at the highest plant population density (88,888 plants ha-1) as compared to lowest plant population density (44,444 plants ha-1) and this is due to low plant population harvested ha-1 in wider spacing intra row spacing. thus, balanced growth and development of plants need optimum plant density because optimum density enables plants efficient utilization of available nutrients, soil water and better light interception coupled with other growth influencing factors. therefore, results of this contradicted the previous recommendation 75x30cm inter and intra row spacing resulted in the production of grain yield. these results were supported by emam [35] who verified that kernels ear-1 and kernels row ear-1 were the most important yield adjustment components in response to plant population density in maize. these finding was in agreement with farnham [36] who reported that maize grain yield increased as plant density increased from 59,000 to 89,000 plant ha-1. regarding the effect of np fertilizer rates 138/110 kg ha-1 n/p2o5 fertilizergave significantly higher grain yield of 8751.6 kg ha-1 which was statically at par with 115/86 kg ha-1 n/p2o5 fertilizerrate while, the lowest grain yield 7442.2 kg ha-1 was recorded from 69/52 kg ha-1 n/p2o5 fertilizerratetable 3. the grain yield advantage of 15.2% was obtained from 115/86 kg ha-1 n/p2o5 when compared to 69/52 kg ha-1 n/p2o5 fertilizerrate. the responses of maize grain yield to increased np fertilizer application rates were simulated via a linear regression model, at four densities over two years figure 3. the result indicated as np fertilizer rate increased the grain yield was increased and vice versa. reduction of grain yield might be due to nutritional imbalance and deficiency of certain important plant growth elements at various important growth stages and also due to reduced leaf area development resulting in lesser radiation interception and, consequently, low efficiency in the conversion of solar radiation. the higher doses of chemical fertilizers increased grain yield as nutrients are the main driving force to produce high yield of maize [37].it is clear from the result that grain yield increased in response to increasing rate of np fertilizer application possibly due to higher number of ears per plant.these increase the nutrients in the soil and modification of soil environments that resulted in better vegetative growth which in turn enabled the crop to produce greater photo-assimilate. the n has synergistic effects on growth and yield attributes resulting in greater translocation of photosynthates from source to sink, beneficial effect on physiological process, plant metabolism, growth and it is the major ingredient of proteins, enzymes, amino acids, amides and nucleic acids [38] and there by leading to higher grain yield. the p supply is particularly important for stimulating early root formation and growth, functions in plant macromolecular structures as a component of nucleic acids and phospholipids, with crucial roles in energy metabolism, participation in signal transduction path ways via phosphorylation and controlling key enzyme reactions [39]. agriculture and food sciences research, 2020, 7(1): 69-78 75 © 2020 by the authors; licensee asian online journal publishing group figure-3. maize grain yields as a function of np application rate for four plant population densities over the year study. planting densities were 44,444 (75x30cm), 53,333 (75x25cm), 66,667 (75x20cm) and 88,888 (75x15cm) in panels a–d respectively. 3.3. aboveground biomass yield the significant higher above ground biomass was recorded during the 2015 main cropping season than 2016 cropping season. the maximum above ground biomass 19.90 ton ha-1 was recorded from plant population density of 88,888 plants ha-1 (75x15cm) while, the minimum above ground biomass yield was obtained from plant population density of 44,444 plants ha-1 (75x30cm) table 3. the result indicates that as plant population density increased up 88,888 plants ha-1 the above ground biomass linearly increased and vice versa. it was increased by 33.3% by planting the highest plant population rate than 44,444 plants ha-1. the result showed that biological yield was increased by increasing plant density due to high grain yield, number of ears per plant and plant height in the treatment of high plant population density of 88,888 plants ha-1 (75x15cm) and low biomass yield was obtained in the treatment of lower plant population density 44,444 plants ha-1 (75x30cm). the aboveground biomass yield was decreased in wider spacing due to minimum plant height and decreased ability of plants for capturing resources which was reflected as evident in their decreased biomass production. these results were in agreement with bullock, et al. [40] who reported that narrow row spacing made more efficient use of available light and shaded the surface soil more completely during the early part of the growing season while the soil is still moist and therefore, narrow row spacing are more effective in producing biomass. 3.4. harvest index the harvest index was not significantly (p>0.05) affected by cropping season, plant population density and np fertilizer rate. but, the gained data was in the acceptable range of 0.4 0.6 for maize [41]. 3.5. economic analysis the official prices of chemical fertilizer (nps=13.5 birr kg-1) and urea (10 birr kg-1) were calculated based on stock sale prices of omonadafarmers cooperative in may, 2015 and 2016. the sale of grain maize at omonada open market average price was 4.6 etb kg-1 in january, 2016 and 2017.the cost of labor spent on chemical fertilizer application and transport, seed planting and purchase were also used for partial budget analysis. the cost of application and transport of fertilizer was taken to be 15 birr 100 kg-1. grain yield was adjusted by 10% for management difference to reflect the difference between the experimental yield and the yield that farmers could expect from the same treatment [25, 42]. the dominance analysis procedure as detailed in cimmyt [25] was used to select potentially profitable treatments. dominance analysis led to the selection of treatments ranked in increasing order of total variable costs table 5. for each pair of ranked treatments, the percent marginal rate of return (mrr) was calculated. the mrr (%) between any pair of un-dominated treatments were the return per unit of investment in chemical fertilizer and plant population density. it was calculated by dividing the change in net benefit to the change in variable costs. 100% mrr means for every 1 birr invested in different cost of fertilizer and maize seed, farmers can expect to recover 1 birr and obtain an additional1 birr [25]. agriculture and food sciences research, 2020, 7(1): 69-78 76 © 2020 by the authors; licensee asian online journal publishing group table-3. main effects of intra row spacing, np fertilizer rates and year on growth and yield component of bhqpy-545 at jimma zone, omonadaworeda over 2015 and 2016 cropping season. treatment plant height (cm) lodging % grain yield (t ha-1) agb (t ha-1) hi year 2015 228b 28.4a 8950a 18.75a 0.48 2016 257a 18.4b 7421b 15.85b 0.47 lsd (0.05) 4.398 4.159 486.830 0.833 0.015 f-test ** ** ** ** ns plant population density (plantha-1) 88,888 (75x15cm) 247a 21.0 9401a 19.90a 0.47 66,667 (75x20cm) 243a 20.6 8485b 18.00b 0.47 53,333 (75x25cm) 243a 26.6 7809b 16.37c 0.48 44,444 (75x30cm) 237b 25.3 7046c 14.93d 0.47 lsd (0.05) 6.11 13.86 727.7 0.474 0.046 cv (%) 6.36 38.9 20.87 16.89 10.81 f-test * ns ** ** ns n/p2o5 fertilizer rates(kg ha-1) 69/52 235c 22.0 7442c 15.91c 0.47 92/69 241b 22.4 7974bc 17.05bc 0.47 115/86 245ab 24.2 8574ab 17.91ab 0.48 138/104 251a 25.0 8752a 18.33a 0.48 lsd (0.05) 6.22 5.88 688.5 1.178 0.021 cv (%) 3.87 40.5 13.61 4.22 15.19 f-test ** ns ** ** ns note: lsd= least significant difference; cv=coefficient of variation; ns=non significant; hi= harvest index; agb=above ground biomass; values followed by the same letter within a column are not significantly different at p< 0.05. the highest net benefit 36,705.0 etb ha-1 was obtained from plant population density of 88,888 plants ha-1 (75*15cm) with acceptable mrr 584.5%. whereas, the lowest net benefit 28,062 etb ha-1 was obtained from the lower plant population density 44,444 plants ha-1table 4. due to planting of 88,888 plants ha-1 (75*15cm), there was a net benefit increase by 30.8% (8,643 etb ha-1) when compared with that of lower plant population density 44,444 plants ha-1 (75*30cm). table-4. partial budget analyses of np fertilizer rates and plant population density on grain yield of medium maturing maize variety qpybh545 at jimma zone during 2015-2016. n/p2o5 (kg ha-1) gy (kg ha-1) adj.gy (kg ha-1) gfb (etb ha-1) tvc (etb ha-1) nb (etb ha-1) 69/52 7442.2 6698.0 30810.71 2933.6 27877.11 92/69 7973.6 7176.2 33010.7 3885.3 29125.4 115/86 8574.1 7716.7 35496.77 4827.5 30669.27 138/104 8751.6 7876.4 36231.62 5807.3 30424.32 plant population density 88888(75*15cm) 9401.2 8461.1 38920.97 2216 36705.0 66666(75*20cm) 8485.2 7636.7 35128.73 1662 33466.7 53333(75*25cm) 7809.2 7028.3 32330.09 1317.6 31012.5 44444 (75*30cm) 7046 6341.4 29170.44 1108 28062.4 note: *gy= grain yield; gfb = gross field benefit; tcv = total cost that varied; nb = net benefit; etb = ethiopian birr; price of chemical fertilizer = 13.5birr kg-1; price of urea = 10 birr kg-1; wage rate = 30 birr man-day-1; retail price of grain = 4.6 birr kg-1. table-5. partial budget with dominance analysis and mrr (%) of np fertilizer and plant population density of medium maturing maize variety qpybh545 at jimma zone in 2015-2016. n/p2o5 (kg ha-1) gy (kg ha-1) adj.gy (kg ha-1) gfb (etb ha-1) tvc (etb ha-1) nb (etb ha-1) domi nance mrr (%) 69/52 7442.2 6698.0 30810.71 2933.6 27877.11 ---- 92/69 7973.6 7176.2 33010.7 3885.3 29125.4 ud 131.2 115/86 8574.1 7716.7 35496.77 4827.5 30669.27 ud 163.9 138/104 8751.6 7876.4 36231.62 5807.3 30424.32 d -- population density 44444 (75*30cm) 7046.0 6341.4 29170.44 1108.0 28062.4 ---- 53333(75*25cm) 7809.2 7028.3 32330.09 1317.6 31012.5 ud 1407.5 66666(75*20cm) 8485.2 7636.7 35128.73 1662.0 33466.7 ud 712.6 88888(75*15cm ) 9401.2 8461.1 38920.97 2216.0 36705.0 ud 584.5 note: *gy= grain yield; gfb = gross field benefit; tcv = total cost that varied; nb = net benefit; etb = ethiopian birr; price of chemical fertilizer = 13.5birr kg-1; price of urea = 10 birr kg-1; wage rate = 30 birr man-day-1; retail price of grain = 4.6 birr kg-1. regarding the effect of np fertilizer rates the highest net benefit 30,669etb ha-1 was obtained from 115/86 kg ha-1 n/p2o5 fertilizer with acceptable mrr 163.9%. whereas, the lowest net benefit (27,877 etb ha-1) was obtained from the lower np fertilizer rate 69/52 kg ha-1 n/p2o5table 5 and figure4. due to application of 115/86 kg ha-1 n/p2o5 fertilizer, there was net benefit increase by 10% (2792 etb) when compared with 69/52 kg ha-1 n/p2o5 fertilizer. agriculture and food sciences research, 2020, 7(1): 69-78 77 © 2020 by the authors; licensee asian online journal publishing group figure-4. total variable cost, net benefit and mrr % of np fertilizer rate and plant population density rate of qpybh545 maize variety at jimma zone in 2016 and 2017 cropping season. market prices are ever changing and as such a recalculation of the partial budget using a set of likely future prices (i.e., sensitivity analysis) was essential to identify treatments which may likely remain stable and sustain satisfactory returns for farmers despite price fluctuations. the sensitivity analysis study indicates an increase in the field price of the total variable costs, and a fall in the price of maize grain, which represented a price variation of 15%. the price changes are realistic under market conditions prevailing at jimma area which were above the minimum acceptable mrr of 100% for application of 115/86 np2o5 fertilizer. concerning plant population density 88,888 plants ha-1 (75*15cm), 66,666 plants ha-1 (75*20cm) and 53,333 plants ha-1 (75*25cm) are realistic which were above the minimum acceptable mrr of 100% table 6. table-6. sensitivity analysis of maize production based on a 15% rise in total cost and maizeprice of gross field benefit fall. n/p2o5 fertilizer tvc (etb ha-1) nb (etb ha-1) raised cost raised benefit mrr (%) 69/52 3373.64 23695.54 --------- 92/69 4468.095 24756.59 1094.46 1061.05 96.95 115/86 5551.625 26068.88 1083.53 1312.29 121.11 population density 44444 (75*30cm) 1274.2 23853.1 --------- 53333(75*25cm) 1515.24 26360.6 241.04 2507.54 1040.30 66666(75*20cm) 1911.3 28446.7 396.06 2086.10 526.71 88888(75*15cm ) 2548.4 31199.2 637.10 2752.50 432.04 note: *tcv = total cost that varied; nb = net benefit; etb = ethiopian birr; mrr= marginal rate of return; price of chemical fertilizer = 13.5 birr kg-1; price of urea = 10 birr kg-1, wage rate = 30 birr man-day-1; retail price of grain = 6 birr kg-1. 4. conclusion and recommendation lately innovated maize varieties such as quality protein maize (qpm) versions vary in stature and leaf arrangements from known normal maize varieties. these variations in morphology may lead to different planting density to reach at their at maximum yield potentials. in view of this, plant population density study on qpm hybrid (bhqpy 545) was proposed with two objectives to maximize yield potential and to help small holder farmers to feed their families with high protein diet. accordingly, rigorous research efforts were made on research station fields of jimmajimma research center (melko and mettu) and on farmers field in vicinity of the center for three cropping seasons. summary of results from across seasons and sites indicated that at highest plant population density, grain and biomass yields were significantly increased. similarly, the higher fertilizer doses produced significantly higher biomass and grain yields. to increase certainty and further use of optimum plant density evaluation criteria was set by farmers and subjected to choose or reject any density not technically appropriate. thus, for the future production of maize hybrid, bhqpy545 in omonadawareda and adjacent waredas with similar agro-ecologies, a plant population density of 88,888 plant ha-1 (75 x 15cm a plant hill-1 or 75 x 30 cm two plants hill-1) in complement with np fertilizer rate of 115/86 kg ha-1 can be recommended. references [1] faostat, "statistical database of the food and agriculture organization of the united nations. retrieved from faostat3.fao.org/browse/q/qc/e," 2016. 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[42] a. getachew and b. taye, "on-farm integrated soil fertility management in wheat on nitisols of central ethiopian highlands," ethiopian journal of natural resources, vol. 7, pp. 141-155, 2005. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 30 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 1, 30-35, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i1.5486 © 2024 by the authors; licensee asian online journal publishing group agro-morphological analysis of yield and yield attributing traits of wheat under heat stress condition pragyan bhattarai1 prashant gyanwali2 netra prasad pokharel3 parbin bashyal4 rasmita mainali5 renuka khanal6 ( corresponding author) 1,2,3,4,5,6institute of agriculture and animal science, paklihawa campus, rupandehi, tribhuvan university, nepal. 1email: pragyanbhattarai29@gmail.com 2email: prashantgyawali7@gmail.com 3email: netrapokharel73@gmail.com 4email: parbinbashyal56@gmail.com 5email: 2057mainalirasmita@gmail.com 6email: renukhanal57@gmail.com abstract wheat is the most important cereal crop worldwide and ranks third in nepal. improvements in wheat yield can be done effectively by selection for yield attributing traits. in this experiment, twenty wheat genotypes were evaluated in the terai region of nepal at paklihawa, rupandehi in alpha lattice design under heat stress conditions. the characters were evaluated to find their correlation and direct and indirect effects on yield. positive significant correlation of grain yield with no. of spikes m-2 (0.405) and harvest index (0.647) were found whereas spike weight (0.322) showed a significant negative correlation with grain yield. similarly, path analysis showed that the harvest index (0.5511) and no. of spikelets per spike (0.3365) had a high direct effect, whereas thousand kernel weight, spike m-2, and plant height showed a lower positive direct effect on grain yield. ten spikes weight, spike length, and no. of grains per spike showed low negative direct effects. the conclusions drawn from this analysis can be useful for breeding programs under heat stress by providing information on which characteristics significantly affect the yield. however, multi-locations and multi-year trials need to be done for further verifications on the selection of such traits for improving yield. keywords: correlation, heat-stress, path-coefficient analysis, plant breeding, selection, wheat parameters. citation | bhattarai, p., gyanwali, p., pokharel, n. p., bashyal, p., mainali, r., & khanal, r. (2024). agro-morphological analysis of yield and yield attributing traits of wheat under heat stress condition. agriculture and food sciences research, 11(1), 30–35. 10.20448/aesr.v11i1.5486 history: received: 27 december 2023 revised: 30 january 2024 accepted: 15 february 2024 published: 21 march 2023 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by institute of agriculture and animal science, pakhlihawa campus, tribhuvan university (grant number: 21-1104). institutional review board statement: the ethical committee of the institute of agriculture and animal science, pakhlihawa campus, tribhuvan university, nepal has granted approval for this study on 21 august 2023 (ref. no. 15 0823). transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 31 2. materials and methods ................................................................................................................................................................... 31 3. result and discussion ..................................................................................................................................................................... 32 4. conclusion ......................................................................................................................................................................................... 34 references .............................................................................................................................................................................................. 34 mailto:pragyanbhattarai29@gmail.com mailto:prashantgyawali7@gmail.com mailto:netrapokharel73@gmail.com mailto:parbinbashyal56@gmail.com mailto:2057mainalirasmita@gmail.com mailto:renukhanal57@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i1.5486 https://orcid.org/0009-0001-9687-4479 https://orcid.org/0000-0001-6428-491x https://orcid.org/0009-0001-9237-3992 https://orcid.org/0009-0002-2929-0868 https://orcid.org/0009-0006-7150-5577 https://orcid.org/0009-0001-6929-4284 agriculture and food sciences research, 2024, 11(1): 30-35 31 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this paper builds on the existing knowledge of wheat breeding and helps guide wheat breeding by providing information regarding the major parameters that have the maximum effect on wheat yield. it helps highlight the parameters to look out for and prioritize during selection. 1. introduction 1.1. background wheat is one of the most important crops in the world, providing a staple diet for 2 billion people [1]. it accounts for 28% of world cereal production and 41.5% of all cereal trade globally [2]. it was cultivated on 215.9 million hectares worldwide in 2019, yielding 765.8 million tonnes with productivity of 3.54 mt/ha [3]. wheat is the third most cultivated cereal after rice and maize in nepal. the area, productivity, and production of wheat in nepal are 711ha, 2.99 mt/ha, and 2,127,276 mt respectively [4]. the wheat productivity in nepal is far behind compared to a nation like new zealand with a high productivity status [5]. wheat accounts for about 55% of all carbohydrates and 20% of all food calories consumed worldwide [1]. the demand for wheat is expected to rise by 60% in 2050 [6]. increment in wheat production by 2% annually is needed to fulfill this demand [7]. the developing nations should increase their wheat production by 77% to meet future demands [8]. this emphasizes various crop improvement practices for increment of both production and productivity as a necessity of the 21st century [9]. this can be accomplished by producing high-yielding, climatesmart, and stress-tolerant varieties of wheat by intense selection under field circumstances [10]. the average global temperature has increased by 1.04°c between 1880 to 2019 [11]. the temperature is estimated to increase further by 1.5°c in the next twenty years [12]; so, global wheat production is predicted to fall by 6% for every 1°c rise in temperature [13]. the impact of heat stress is determined by the length and severity of the stress, along with genetic factors [14]. terminal heat stress in wheat is observed when the mean temperature exceeds 31°c at the grain-filling stage [15]. increment of 1°c average temperature during march and april causes a seven-day reduction in the length of the wheat crop and a yield decrease of around 400 kg/ha [16]. the terai region of nepal has a significant temperature increase starting in mid-march, which coincides with anthesis and is subject to scorching winds from the west [17]. it was reported that the maximum temperature rose by 1 °c in the terai region of nepal over the previous 25 years which will be detrimental to wheat cultivation in the future [18]. yield is a complex quantitative trait that is considerably affected by, and depends on the environment. the selection of genotypes based on various components of yield is effective in comparison to the selection regarding yield alone [19]. the study aims to understand the relationship between various components of 20 wheat genotypes and the yield in heat stress conditions. the associations will help determine the most important characters affecting the yield and set a standard selection guide of traits to look out for selection. 1.2. statement of problem the major physiological functions like chlorophyll degradation and photosynthesis are affected by heat stress condition [20]. the grain filling period , pollen abortion, poor seed set are also observed under this condition [21]. it is estimated that a 1°c increase in temperature would result in a 6% decrease in global wheat yield [5]. increased temperature in the future will hugely impact the yield of wheat worldwide [22]. research activities exploring the relationship between yield and yield attributing characters among various genotypes have been conducted by numerous researchers, but only a few have been conducted under a heat-stress environment, especially in the western region of nepal. this will help us in creating an efficient selection of genotypes under heat stress condition. 1.3. rationale of study heat stress resulted in a total of 36.03 billion in wheat yield loss in the year 2020 in the context of nepal [9]. the development of wheat genotypes with high yield potential and the capacity to sustain output in all conditions, including heat stress, is one of the key objectives of wheat breeders. but, the practice of unilateral selection for agronomic features and inadequate understanding of the interactions between multiple characters usually leads to unsatisfactory results in wheat breeding. hence, this study is performed to determine which characteristics have a significant impact on wheat output. plant breeders would then be able to prioritize these traits for further yield enhancement, as the study of yield-contributing components concerning their genetic mechanism is very important for yield improvement. 2. materials and methods twenty wheat genotypes including 15 nepal-lines (nl), 3 bhairahawa-lines (bl) and 2 check varieties i.e. bhrikuti and gautam, were collected from national wheat research program, bhairahawa. the research was conducted in paklihawa campus, rupandehi, during 2021-2022. yield and yield attributing traits like plant height (ph), spike length (sl), spike weight (sw), spikes per m2 area (spms), number of grains per spike (ngps), number of spikelets per spike (nsps), thousand kernels weight (tkw) and harvest index (hi) were noted from ten randomly selected plants per plot. agro-meteorological data were recorded at the experimental site from 26th december 2021 to 17th april 2022, and average was found as in figure 1. line sowing of seeds was done on 26th december, so as to reach the reproductive stage of wheat in the hot weather of april inducing heat stress condition. agriculture and food sciences research, 2024, 11(1): 30-35 32 © 2024 by the authors; licensee asian online journal publishing group figure 1. agro-meteorological data of temperature and relative humidity recorded at the site of experiment through crop period. the experiment was carried out in alpha-lattice design replicated twice with five blocks figure 2. one meter space was maintained between two replications. the plot size was maintained at 4m*2.5m each. figure 2. (alpha lattice design) layout of the field. data entry was done by using microsoft excel spreadsheet software. spss version 25 and microsoft excel were used to perform co-relation and path coefficient analysis. 3. result and discussion 3.1. correlation the association of grain yield and yield attributing traits are presented in table 1. plant height showed a positive correlation with nsps, sw, spms and a negative correlation with ngps, tkw, gy, and hi. dwarf wheat varieties increase lodging resistance which leads to higher nitrogen use efficiency. wheat plants tend to use stem reserve when they are under heat stress thus developing a positive correlation with yield [23]. it was found that selecting dwarf plants with thick stems may boost yield when coping with late heat stress [23]. plant height was found to have a positive correlation with all studied traits except maturity, grain yield, and harvest index for both genetic and phenotypic levels [24]. number of grains per spike showed a highly positive significant correlation with nsps (0.68), and sw (0.58) and a non-significant positive correlation with sl, and spms. it showed a significant negative correlation with tkw (-0.33) and a non-significant negative correlation with ph, gy, and hi. it was found that ngps showed a positive and highly significant correlation with sw and nsps [25]. agriculture and food sciences research, 2024, 11(1): 30-35 33 © 2024 by the authors; licensee asian online journal publishing group table 1. correlation of different wheat parameters. parameters ph ngps nsps sl sw tkw gy spms hi ph 1 -0.044 0.071 0.299 0.125 -0.232 -0.026 0.026 -0.114 ngps -0.044 1 0.685** 0.290 0.587** -0.330* -0.177 0.045 -0.216 nsps 0.071 0.685** 1 0.442** 0.460** -0.212 0.037 0.177 -0.175 sl 0.299 0.290 0.442** 1 0.510** 0.104 -0.172 -0.222 -0.091 sw 0.125 0.587** 0.460** 0.510** 1 0.119 -0.322* -0.293 -0.252 tkw -0.232 -0.330* -0.212 0.104 0.119 1 0.229 -0.160 0.263 gy -0.026 -0.177 0.037 -0.172 -0.322* 0.229 1 0.405** 0.647** spms 0.026 0.045 0.177 -0.222 -0.293 -0.160 0.405** 1 0.123 hi -0.114 -0.216 -0.175 -0.091 -0.252 0.263 0.647** 0.123 1 note: ph= plant height, ngps= number of grains per spike, nsps= number of spikelets per spike, sl= spike length, sw= ten spikes weight, tkw= thousand kernels weight, gy= grain yield, spms= number of spikes per m2. ** correlation is significant at the 0.01 level (2-tailed). * correlation is significant at the 0.05 level (2-tailed). number of spikelets per spike showed a significantly high positive correlation with sl (0.44), sw (0.46), and ngps (0.68) and a non-significant positive correlation with ph, gy, and spms. it showed a negative correlation with tkw, and hi. a similar relationship of nsps with sw and ngps was found by thapa, et al. [25]. spike length showed a significantly high positive correlation with nsps (0.44) and sw (0.51). it showed a nonsignificant positive correlation with ph, ngps, and tkw. it exhibited a negative correlation with gy, spms, and hi. it was shown that sl had a positive and highly significant correlation with sw but in contrast, it had a negative and highly significant correlation with nsps [25]. spike weight showed a highly significant positive correlation with ngps (0.58), nsps (0.46), and sl (0.51) and a non-significant positive correlation with tkw, and ph. it showed a significant negative correlation with gy (-0.32) and non-significant negative correlation with hi. heat stress accelerates the spike development process resulting in fewer grains and reduced harvest index [26]. thousand kernel weight showed a significantly negative correlation with ngps (-0.33) and a non-significant negative correlation with ph, nsps, and spms. it showed a positive correlation with sl, sw, gy, and hi. there is a decrement in the life cycle of the wheat crop under heat stress [26]. this results in a reduction of the grain filling period [27]. this lowers the thousand kernel weight and grain size, degrading the quality of the seed and its yield [28]. grain yield showed a significantly negative correlation with sw (-0.32) and a non-significant negative correlation with ph, ngps, and sl. it showed a highly significant positive correlation with spms (0.40) and hi (0.64) and a non-significant positive correlation with tkw. in another research, it was found that grain yield showed a significantly positive correlation with harvest index and spike per meter square which is similar to our finding [29]. number of spikes per m2 showed a negative correlation with sl, sw, and tkw. it showed a highly significant positive correlation with gy (0.40) and a non-significant positive correlation with ph, ngps, nsps, and hi. harvest index showed a negative correlation with ph, ngps, nsps, sl, and sw. it showed a highly significant positive correlation with gy (0.64) and a non-significant positive correlation with tkw, and spms. it was found that the harvest index showed a significant negative correlation with nsps but in contrast, it showed a significant negative correlation with grain yield [30]. 3.2. path coefficient analysis the path coefficient analysis helps us determine the direct and indirect effects of the yield parameters and quantifies their role over one another. the positive and negative values of parameters determine the direction of yield, whether it increases or decreases when there are changes in the parameters. table 2. path coefficient analysis of different wheat parameters. parameters ph ngps nsps sl sw tkw spms hi correlation with yield via ph 0.146 -0.006 0.01 0.044 0.018 -0.034 0.004 -0.017 -0.026 via ngps 0.001 -0.026 -0.018 -0.008 -0.016 0.009 -0.001 0.006 -0.177 via nsps 0.024 0.23 0.336 0.149 0.155 -0.071 0.06 -0.059 0.037 via sl -0.052 -0.05 -0.077 -0.174 -0.089 -0.018 0.039 0.016 -0.172 via sw -0.028 -0.129 -0.101 -0.112 -0.22 -0.026 0.065 0.055 -0.322 via tkw -0.06 -0.085 -0.055 0.027 0.031 0.259 -0.041 0.068 0.229 via spms 0.005 0.009 0.038 -0.047 -0.063 -0.034 0.213 0.026 0.405 via hi -0.062 -0.119 -0.096 -0.05 -0.139 0.145 0.068 0.551 0.647 note: ph= plant height, ngps= number of grains per spike, nsps= number of spikelets per spike, sl= spike length, sw= ten spikes weight, tkw= thousand kernels weight, gy= grain yield, spms= number of spikes per m2. direct effect of yield attributing character (bold in table 2) showed that hi had the highest positive direct effect (0.55114) on the grain yield. following the hi, exhibiting the second highest positive direct effect is nsps (0.33647). likewise, tkw (0.25882), spms (0.21340), and ph (0.14572) had similar direct positive effects. confirmatory similar positive results were also obtained by meena, et al. [31]. the other parameters showed a negative direct effect on yield, where ngps showed the lowest direct effect (-0.0266), followed by sl (-0.17366) and sw (-0.22019). contradicting results regarding ph and sl were obtained by mohanty, et al. [23]. likewise, positive direct effects by ngps and ph on grain yield were obtained by oliveira, et al. [32]. the direct effects have a greater possibility of determining the outcome while selecting the yield as they do not depend upon the other parameters. the traits or parameters whose direct effect value is close to the correlation coefficient value, usually have little to no indirect effect on the grain yield. this suggests that selection for such traits is beneficial and much more effective in observing the improved grain yield [33]. the traits having positive correlation with the yield but agriculture and food sciences research, 2024, 11(1): 30-35 34 © 2024 by the authors; licensee asian online journal publishing group no positive direct effect is not considered while selecting for the improved yield since other traits are responsible for the yield indirectly [34]. the indirect effects weren’t as pronounced as the direct effects, with the majority having a nearly negligible magnitude. the highest positive indirect effect on yield was shown by ngps via nsps (0.23048), followed by sw via nsps (0.15478), sl via nsps (0.14872), and tkw via hi (0.14495). the highest negative indirect effect on yield was shown by sw via hi (-0.13889), followed by ngps via sw (-0.12925), ngps via hi (-0.11905), sl via sw (-0.1123), and nsps via sw (-0.10129). the other indirect effects on yield had low values. partial similar results were obtained by kayastha, et al. [35]. 4. conclusion the grain yield has a positive correlation with spms and harvest index, which also showed high direct effects through the path coefficient analysis. therefore, selection should be done in favor of these yield parameters to increase the overall yield of the wheat crop in the given environment condition. grain yield has a negative correlation with spike weight, plant height, number of grains per spike & spike length. the path coefficient analysis showed the highest negative direct effects on grain yield by spike weight. therefore, it is suggested that we avoid selecting genotypes with higher value of sw during selection. the indirect effects on grain yield weren’t profound in magnitude as the direct effects, however, the highest positive and negative indirect effects were shown by the number of grains per spike via number of spikelets per spike, and spike weight via harvest index. references [1] a. k. maurya, r. yadav, a. k. singh, a. deep, and v. yadav, "studies on correlation and path coefficients analysis in bread wheat (triticum aestivuml.)," journal of pharmacognosy and phytochemistry, vol. 9, no. 4, pp. 524-527, 2020. 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licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 2, 155-165, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.62.155.165 © 2019 by the authors; licensee asian online journal publishing group measuring the climate variability impact on cash crops farming in india: an empirical investigation ajay kumar singh1 bhim jyoti2 ( corresponding author) 1assistant professor (economics), department of humanities and social sciences, dit university, dehradun, uttarakhand-248009, india 2assistant professor (seed science and technology), v.c.s.g., uuhf, college of forestry, ranichauri, tehri garhwal. uttarakhand, india abstract this study investigates the climate variability impact on potato, cotton, groundnut and sesame crops in indian states. thereupon, it estimates the technical efficiency (te) of climatic and nonclimatic factors in crops farming in indian states during 1971-2014. production, yield and area sown of aforementioned crops are considered as dependent variables and regressed with socioeconomic and climatic variables using state-wise panel data employing c-d pfm. sates-wise te of cash crops is estimated using sfpfm under non-parametric condition. estimates indicate that climate variability show negative impact on production, yield and area sown of aforesaid cash crops. thus, it is essential to adopt effective policy to mitigate the negative consequences of climate variability in cash crop farming in india. cropped area, production and yield of cash crops are negatively impacted due to climatic variability in india. so there needs to adopt crop specific policies to mitigate the negative impact of climate variability in cash crop farming. finally, it estimates the technical efficiency (te) of cash crop using sfpfm across indian states. estimated values of te for production, yield and area sown of associated crops imply that there is significant variation in te due to climate variability and huge diversity in socio-economic characteristic across indian states. thus, most of states have potential opportunity to increase production and yield of cash crops through enlightening technical efficiency of inputs in cultivation. keywords: commercial crops, yield, production, cropped area, technical efficiency, sfpfa, c-d pfm, climate change. citation | ajay kumar singh; bhim jyoti (2019). measuring the climate variability impact on cash crops farming in india: an empirical investigation. agriculture and food sciences research, 6(2): 155-165. history: received: 25 march 2019 revised: 2 may 2019 accepted: 6 june 2019 published: 18 july 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. background ..................................................................................................................................................................................... 156 2. climate sensitivity and cash crops farming........................................................................................................................... 156 3. research methodology ................................................................................................................................................................. 157 4. analytical viewpoint of stochastic frontier production function approach (sfpfa) ................................................. 158 5. empirical analysis ......................................................................................................................................................................... 158 6. statistical inferences on regression results ........................................................................................................................... 159 7. conclusion and policy implications ........................................................................................................................................... 163 references ............................................................................................................................................................................................ 164 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.155.165&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.155.165&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/857 https://orcid.org/0000-0003-0429-0925 https://orcid.org/0000-0002-6960-5097 agriculture and food sciences research, 2019, 6(2): 155-165 156 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the study contributes to the existing literature by estimating the technical efficiency of production, yield and area sown of major cash crops across indian states. 1. background agriculture is the backbone for indian economy and it is prime source to sustain food security and raw material for various industries like textile, oilseed, sugar and other industries. indian agriculture is highly vulnerable due to climate change. in this context, numerous of studies have assessed the impact of climate change on yield and production of food-grain and cash crops in different regions/states in india [1-11]. in brief: singh, et al. [12]; kumar and sharma [1]; ayyogari, et al. [13]; kumar, et al. [6]; kumar, et al. [14]; yadav, et al. [15]; singh, et al. [9] have investigated the impact of climate change on potato yield, hundal and kaur [16]; ashalatha, et al. [17]; kumar and sharma [18]; kumar and sharma [1]; birthal, et al. [19]; kumar, et al. [14]; yadav, et al. [15]; singh, et al. [9]; ramachandran, et al. [11] have assessed the climate change impact on groundnut yield, nath, et al. [20]; kumar and sharma [1]; kumar, et al. [14]; singh, et al. [9] have observed the climate change impact on sesame productivity, and masters, et al. [21]; sankaranarayanan, et al. [22]; moorthy, et al. [23]; deshmukh and lunge [24]; ashalatha, et al. [17]; kumar and sharma [1]; hebbar, et al. [2]; raju, et al. [25]; thakare, et al. [26]; kumar, et al. [14]; singh, et al. [9] have explored the climate change impact on cotton crop productivity in india. descriptive and empirical results of aforesaid studies were based on time series and district-wise/state-wise panel data, which were included crop productivity as a dependent variable. few studies have incorporated per hectare output (in monetary) of a cash crop as a dependent variable and it was regressed with climatic and nonclimatic factors using district-wise/state-wise panel data in india [7, 9, 10, 14, 19, 27]. most researchers have used agricultural production per hectare land (in monetary) of selected food-grain and cash crops as dependent variables, which was regressed with climatic and non-climatic factors using state-wise panel data in india [3, 4, 6, 10, 14, 27, 28]. few studies have segregated the agricultural productivity (in monetary) of rabi and kharif crops to assess the impact of climate change on it using state-wise and district-wise panel data in india [1, 4-6, 10, 27]. earlier studies have provided empirical evidence that climate change have a negative and significant impact on productivity of cash crops in india. agricultural productivity is also negatively impacted due to climate change in india [1, 5, 6, 27, 28]. however, limited studies assessed the impact of climate variability on all aspects (i.e., cropped area, production and yield) of cash crops farming in india. existing researchers also could not estimate the technical efficiency (te) of climatic and non-climatic factors in cash crop farming in india. it would be useful to formulate crop specific policy to increase the te of inputs in cash crop farming. most studies clearly indicate that agricultural productivity or net revenue of most cash crops decrease due to climate change in different regions of india. most studies analyzed the climate change impact on agricultural productivity or net revenue of cash crops for a particular regions of india. therefore, the present study address the few research questions which are given as: • how climate variability does affect area sown, production and yield of cash crops in different periods of crops in india? • is technical efficiency of cash crop farming varied across indian states? • is possible to mitigate the adverse effective of climate variability in cash crop farming in india? relevance to aforementioned research questions, the present study is achieved following objectives: • to investigate the impact of climate variability on various cash crops (i.e., potato, cotton, groundnut and sesame) in india. • to estimate the te of climatic and non-climatic factors in cash crops farming across indian states. • to provide the conclusive policy implications to mitigate the adverse effects of climate change in cash crop faming. 2. climate sensitivity and cash crops farming it is observed that productivity of cash and food-grain crops are negatively impacting due to variability in minimum temperature, maximum temperature and change in rainfall pattern in india ayyogari, et al. [13]; yadav, et al. [15]. hijmans [29] detected that potato yield decreases by 18% to 32% as temperature increases by 1 to 1.40c respectively in potato producing economies. furthermore, potato yield is expected to be increased in algeria, china, morocco and south africa [29]. potato crop is passing through various physiological processes during sowing, growing and harvesting time [13]. hence, variability in climatic factors have a negative impact on potato growth and yield [12-15, 29]. hundal and kaur [16] estimated that groundnut yield is likely to be declined by 23.2% and 36.2% as temperature increases by 1.2 and 30c respectively in punjab (india). nath, et al. [20] investigated the impact of climatic variations on sesame productivity during sowing time in kalyani (india). it shows that climate variability during various days in sowing time have a negative impact on sesame yield and the effect of climate variability on sesame yield varies across days. masters, et al. [21] reported that growing season of cotton crop increases in moderate temperature, thus cotton yield would be improved. the study show that climate change would have a negative impact on cotton yield in near future. sankaranarayanan, et al. [22] also reviewed that climatic factors have a negative impact on production and yield of cotton crop in india. deshmukh and lunge [24] explored that association of maximum and minimum temperatures and rainfall with cotton yield in vidarbha (maharashtra) india. it found that climatic factors have a significant impact on cotton yield. moorthy, et al. [23] projected the effect of historical climate change on yield of cotton, sugarcane, rice, wheat and sorghum crops. it shows that temperature and precipitation did show insignificant impact on yields of cotton, while climate factors have a non-linear relationship with it. ashalatha, et al. [17] analyzed the drought effect on crop yield and farmer’s understanding towards climate change in dharwad karnataka (india). it observed that groundnut and cotton yields decreases by 34.09 and 59.96 kg/ha respectively in rainfed areas due to low rainfall. agriculture and food sciences research, 2019, 6(2): 155-165 157 © 2019 by the authors; licensee asian online journal publishing group kumar and sharma [1] observed that yield of cotton and sesame crops were negatively associated with maximum temperature, while yields of potato and linseed crops are expected to be declined as increase in minimum temperature in india. hebbar, et al. [2] assessed the climate change impact on cotton yield in different climate change scenarios (i.e., a2, b2 and a1b). it reported that cotton yield has a probability to be declined by 477kg/ha and 268kg/ha under a2 and b2 scenario respectively, while cotton yield was statistically in-significant under a1b scenario. singh, et al. [12] attain the farmer’s understanding on climate change and assess the climatic impact on cotton yield and production in rajasthan, andhra pradesh and maharashtra. this study concluded that climate change have a negative impact on cotton yield, water quality and fabric quality. singh, et al. [30] examined the impact of climate change on groundnut yield in anantapur and mahboobnagar districts of andhra pradesh and junagadh districts of gujarat in india. results of the study based on cropgrogroundnut model indicate that groundnut yield decreases as increase in temperature. however, the effects of temperature, rainfall and co2 emissions on groundnut yield would vary in aforementioned districts. thakare, et al. [26] assessed the variability in cotton yield due to change in rainfall pattern, and maximum and minimum temperature in surat district of gujarat in india. it found that rainfall variability has a negative impact on crop physiology and cotton yield. raju, et al. [25] measured the impact of temperature and rainfall on cotton yield in andhra pradesh (india). it point out that cotton yield decreases by 13kg/ha and 0.7kg/ha as increase in 10c maximum temperature and 10 mm rainfall respectively. birthal, et al. [19] explored the variability in climatic factors (i.e., temperature and rainfall) during 1969-2005 and climate impact on yield of food-grain crops in india. it observed that groundnut yield decreases by 9% as 10c increase in maximum temperature, while minimum temperature and rainfall showed a positively impact on groundnut yield. however, rainfall could not compensate the reduction in groundnut yield due to increase in maximum temperature. kumar, et al. [14] examined the climate change impact on mean yield and yield variability of cotton, groundnut, potato, sesame and linseed crops using stochastic production function approach in india. it observed that mean yield of cotton and sesame crops are likely to be declined by 2.03% and 2.80% respectively as 10c increase in maximum temperature. productivity of potato and linseed may be declined by 0.47% and 0.84% respectively due to 10c increase in minimum temperature. singh, et al. [9] evaluated the impact of climatic and non-climatic factors on cropped area, yield and production of cash crops across indian states. it observed that cropped area under groundnut and sesame crops decrease by 13.87% and 23.50% respectively, while groundnut and cotton production declines by 9.83% and 41.09% respectively; and productivity of potato, groundnut, sesame and cotton crops declines by 3.71%, 10.31%, 4.51% and 7.20% respectively due to 1% variability in climatic factors. ramachandran, et al. [11] reported that productivity of potato and groundnut crops are expected to be declined by 3-7% as increase in 10c temperature in india. 3. research methodology 3.1. introduction of study area separate state-wise panel data (during 1971-2014) was compiled for potato, cotton and groundnut crops from fifteen states of the country refer table 1. climatic and non-climatic factors are considered in this analysis. table-1. detail of crops which are included in each state-wise panel. name of states name of crops undertaken from respective states andhra pradesh potato, cotton, groundnut, sesame assam potato, cotton, sesame bihar potato, cotton, groundnut, sesame gujarat potato, cotton, groundnut, sesame haryana potato, cotton, groundnut, sesame karnataka potato, cotton, groundnut, sesame kerala cotton, groundnut, sesame madhya pradesh potato, cotton, groundnut, sesame maharashtra potato, cotton, groundnut, sesame odisha potato, cotton, groundnut, sesame punjab potato, cotton, groundnut, sesame rajasthan potato, cotton, groundnut, sesame tamil nadu potato, cotton, groundnut, sesame uttar pradesh potato, cotton, groundnut, sesame west bengal potato, cotton, groundnut, sesame source: author's compilation based on undertaken data of different states for this study. 3.2. description on data sources information on production, yield, area sown, irrigated area and consumption of fertilizer for each crop are derived from centre for monitoring indian economy (cmie). forest area is also taken from cmie. value of production of each crop is estimated at farm harvest prices with constant level as 1993-94 base year. farm harvest price of cash crop is taken from the different publications of directorate of economics and statistics department of agriculture, cooperation and farmers welfare, ministry of agriculture (goi). agricultural labour and cultivators are taken from the different publication of census (goi), which is available in decadal periods (i.e. 1971, 1981, 1991, 2001 and 2011). rural literate population is taken from the planning commission (goi), which is available aforesaid periods. minimum temperature, maximum temperature and precipitation related data are derived from the indian meteorological department (imd) (goi) database, geographic information system and official website of indian institute of tropical meteorology, new delhi. climatic data is available on daily intervals with latitude and longitude information of monitoring stations. so, city-wise geographical locations or stations are identified to derive the data on climatic factors for all cities in a state. thereafter, groups of different geographical location in a state is integrated to arrive at state-wise data [8, 9]. thence, these data are converted in monthly averages citywise, after that data is transformed in state-wise monthly maximum temperature, minimum temperature and agriculture and food sciences research, 2019, 6(2): 155-165 158 © 2019 by the authors; licensee asian online journal publishing group precipitation [9]. aforesaid process is done through the c++ statistical software, while spss statistical software is used to extract and bring data to excel format. minimum temperature, maximum temperature and precipitation in three growth periods i.e., sowing time, growing time and harvesting time of each crop is integrated in regression models [8-10]. interpolation and extrapolation techniques are used to fill the missing values to complete the time series during 1971-2014 in each state-wise panel data [1, 3, 4, 6-10, 14]. 4. analytical viewpoint of stochastic frontier production function approach (sfpfa) existing researchers have used sfpfa to explore the effect of climatic and non-climatic factors on crop productivity and to estimate the technical efficiency (te) in agricultural production analysis in different economies [31-37]. sfpfa model can be applied in parametric and non-parametric condition [34, 36, 37]. it comprises parameters of production and inefficiency function simultaneously in a model [38]. so, in this study sfpfa model is applied to investigate the te of cash crops farming. production, yield and area sown of cash crops are considered as dependent variables and recursively regressed with climatic and non-climatic factors [5]. cobb-douglas production function model is used to estimate the regression coefficients of climatic and non-climatic variables in empirical models [4, 14]. it is observed that crop productivity and production are significantly associated with variability in climatic and non-climatic factors [1-11, 17, 21, 23, 24]. thus, the sfpfa model assumes that variability in climate factors have a significant association with production efficiency of crops [39]. climate change have a negative impact on technical efficiency of production, yield and cropped area in crop farming [39]. te of a production unit can be measured through sfpfa [31-37]. in the present study sfpfa approach is used, which comprises two terms: non-negative and systematic distribution. it may be presented in following functional forms [34, 36, 37]: yi = f (xst, β) exp(est) (1) here, yi is the production or productivity of each crop, xit is the vector of independent variables; β is the vector of regression coefficients of associated explanatory variables; and est is the error term in equation 1. yi = f (xst, β) exp(vst-ust) (2) here, yst is the output of a specific crop in state (s) in year (t); x is vector of climatic and non-climatic variables; β is the vector of regression coefficients of explanatory variables [36, 37]; est (=vst – ust) is random error term that includes ust and vst in equation 2. it is identically and normally distributed [n(0, σ2)] term [36, 37]. ust may be considered truncated-normal distribution with zero mean (μ) with constant variance (σ2) while ust and vst are also normally distributed for different states [38]. sfpfa model may be used in terms of time-invariant and timevarying models. this study is used time-invariant model to assess the regression coefficients of variables and to estimate te. assessment of technical efficiency (te): te of a state can be estimated using following formula [38]: test = exp(-ust) (3) here, test is the technical efficiency of a state, and ust is estimated through equation 3. if the value of te of a states is equal to 1 then it can be concluded that the state has no technical inefficiency in crop production [38]. 5. empirical analysis 5.1. creation of empirical model for crop farming log-linear (cobb-douglas) (c-d) production function model is considered to measure the impact of climate and non-climatic on production of undertaken crops. for this, total production of crops are used as dependent variables and regressed with climatic and non-climatic factors under the sfpfa. following functional form of empirical model is applied [9, 40]: ln(tp)st= β0 +β1 ln(as)st+β2 ln(srlpca)st +β3 ln(sfaca)st +β4 ln(ia)st +β5 ln(fcca)st +β6 ln(ralcca)st +β7 ln(vprc)st +β8 ln(vamaxtst)st +β9 ln(vamaxtgt)st +β10 ln(vamaxtht)st +β11 ln(vamintst)st +β12 ln(vamintgt)st +β13 ln(vamintht)st +β14 ln(vapcpst)st +β15 ln(vapcpgt)st +β16 ln(vapcpht)st +β17 ln(cvdapcp)st + β18 ln(cvdamint)st + β19 ln(cvdamaxt)st + (vstust) (4) here, tp-production (in 000 tonne) of respective crop in state (s) in time (t); ln-natural logarithms of associated variables; as-area sown (in 000 ha); srlpcashare of rural literature population with cropped area (in number); sfacashare of forest area with cropped area (in 000 ha); iairrigated area (in 000 ha); fcca-fertilizer consumption on cropped area (in 000 tonnes); ralccashare of rural agricultural labour and cultivator with cropped area (in number); vprc-value of production of respective crop at constant prices of 1993-94 (in inr); vamaxtst, vamaxtgt and vamaxtht (in %)-variability in maximum tempereture during sowing time, growing time and harvesting time respectively; vamintst, vamintgt and vamintht (in %)-variability in minimum temperature during sowing time, growing time and harvesting time respectively; vapcpst, vapcpgt and vapcpht (in %)-variability in precipitation during sowing time growing time, growing time and harvesting time respectively; cvdapcp, cvdamint and cvdamaxt (in %)-coefficient variation in daily precipitation, minimum tempereture and maximum temperature respectively during crop period. β0 is the constant coefficient; β1,… β19 are the regression coefficients of associated explanatory variables; vst and ust are the symmetric error term and non-negative random variable respectively in the equation 4. agriculture and food sciences research, 2019, 6(2): 155-165 159 © 2019 by the authors; licensee asian online journal publishing group also, land productivity (yield) or production per hectare land of cash crops was used as depended variables and regressed with climatic and non-climatic factors. for this, following functional form of empirical model was used [9, 40]: ln(lp)st= α0 +α1 ln(as)st+α2 ln(srlpca)st +α3 ln(sfaca)st +α4 ln(ia)st +α5 ln(fcca)st +α6 ln(ralcca)st +α7 ln(vprc)st +α8 ln(vamaxtst)st +α9 ln(vamaxtgt)st +α10 ln(vamaxtht)st +α11 ln(vamintst)st +α12 ln(vamintgt)st +α13 ln(vamintht)st +α14 ln(vapcpst)st +α15 ln(vapcpgt)st +α16 ln(vapcpht)st +α17 ln(cvdapcp)st + α18 ln(cvdamint)st + α19 ln(cvdamaxt)st + (vstust) (5) here, lp-land productivity (yield) of respective crop in state (s) in time (t); ln-natural logarithms of associated variables. the detail description of all explanatory variables are presented in earlier equation 5. α0 is the constant coefficient; α1,… α19 are the regression coefficients of corresponding explanatory variables. the present study assess the impact of climate variability on cropped area of cash crops, therefore area sown of respective crop is considered as dependent variables and regressed with climatic and non-climatic factors. for this, the proposed model accept that area sown of a crop get impacted due to variability in climatic factors in sowing time only [9, 40]. it also considered that area sown of a crop has a functional relationship with srlpca, sfaca, ia, ralcca, vprc, vamaxtst, vamintst, vapcpst, cvdapcpst, cvdamintst and cvdamaxtst. after converting it into c-d production functional model, said relationship can be written in following form [9, 40]: ln(as)st= γ0 +γ1 ln(srlpca)st +γ2 ln(sfaca)st +γ3 ln(ia)st + γ4 ln(ralcca)st +γ5 ln(vprc)st + γ6 ln(vamaxtst)st + γ7 ln(vamintst)st + γ8 ln(vapcpst)st + γ9 ln(cvdapcpst)st + γ10 ln(cvdamintst)st + γ11 ln(cvdamaxtst)st +(vstust) (6) here, as-area sown of respective crop in state (s) in time (t); cvdapcpst, cvdamintst and cvdamaxtst (in %)coefficient variation in daily precipitation, minimum tempereture and maximum temperature respectively during sowing time. γ0 is the constant coefficient; γ1,… γ11 are the regression coefficients of corresponding independent variables in equation 6. signification on other variables has presented in equation 5. regression coefficients and technical efficiencies of yield, production and area sown functions of all crops are estimated through stata statistical software. 5.2. consistency and viability of dependent and explanatory variables dependent variables: the present study was used production (tp) (in 000 tonne), yield (lp) (in kg/ha) and area sown (as) (in 000 ha) as a dependent variables in three different types of empirical exercises [1, 3, 8, 9, 41]. non-climatic and control variables: area sown (as) (in 000 ha), irrigated area (ia) (in 000 ha), fertilizer consumption on cropped area (fcca) (in tonne), share of rural literature population with cropped area (srlpca) (in number), share of rural agricultural labour and cultivator with cropped area (ralcca) (in number) and value of production of respective crop (vpcc) (in rs) are considered as explanatory variables [1, 3, 4, 9, 10, 41]. ecological factor: forest area is an important factor to maintain sustainability in environmental factors, therefore it might be ecosystem adaption based techniques to mitigate the negative consequences on climate change in crop farming. hence, share of forest area with cropped area (sfaca) (in 000 ha) was used to capture the impact of ecological factor on cash crop farming [1, 41]. variability in climatic factors: maximum tempereture, minimum tempereture and precipitation are the crucial weather factors which are significantly associated with growth, development and productivity of crops. so these factors are considered as a climatic factors [1, 5, 8, 10, 19, 25, 26, 41]. to estimate the variability in climatic factors, the original value of a particular climatic factor is subtracted from the mean value of respective climatic factor for time period 1971-2014. then, square root of this climatic factor is used to capture the climatic variability impact on production, productivity (yield) and cropped area of cash crops [9, 41]. hence, climate variability in each factor is estimated as: 𝑉𝑎𝑟𝑖𝑎𝑏𝑖𝑙𝑖𝑡𝑦 = √[(𝑋𝑠𝑡 − 𝑋)] 2 (7) here, xit is original value of climatic factor in state (s) and year t; and x is mean value of particular climatic factor for time period 1971-2014 in equation 7 [9, 38]. coefficient variation is used to assess the impact of daily variation in climatic factor on production, yield and cropped area cash crops [9, 10, 14]. 5.3. limitation of the study solar radiation, sunshine, wind speed, hail storm, humidity, fog, co2 concentration, soil moisture and other weather patterns, which directly or indirectly affect the crop production, yield and cropped area of crops. though, these variables are not included in this study. furthermore, it also assumes that all varieties of a crop is similarly get affected due to climate change, while farmers are cultivating various varieties of a crop. also, it accepts that a particular state lies in the same agro-ecological zone, while india is divided in 15 broad agro-climatic zones with 127 sub agro-climatic zones. many states (e.g. uttar pradesh, madhya pradesh, bihar, punjab, haryana, orissa, rajasthan etc.) are divided in various agro-ecological zones. however, this study could not capture the zone-wise effects on crop productivity, production and cropped area of crops. 6. statistical inferences on regression results 6.1. association of crops production and climatic and non-climatic factors regression coefficients of climatic and non-climatic factors in production function of cash crops are estimated through time-invariant regression model. impact of climatic and non-climatic factors on production of potato, cotton, groundnut and sesame crops are given in equations 8, 9, 10 and 11 respectively. regression coefficients of area sown with production of all crops are found positive and statistically significant. thus, production of these crops have a high potential to be increased as area sown increases under [9]. literate population have a lower agriculture and food sciences research, 2019, 6(2): 155-165 160 © 2019 by the authors; licensee asian online journal publishing group interest in cultivation as compared to unskilled labours. so, rural literate population have a negative association with crop production. forest area would be useful to mitigate the adverse effect of climate change in potato and groundnut farming [1, 38]. conversely, forest area have a negative implication on cotton and sesame crop farming. irrigated area have a higher yielding capacity than non-irrigated area [1, 3, 4, 6, 7, 9, 14]. thus, irrigated area have positive association with production of cotton and groundnut crops, it imply that production of these crops would be increased as irrigated area increases. extensive fertilizer application in cultivation, contribute high ghgs emission in atmosphere, thus it has a negative impact on environmental factors [42].1 thus, fertilizer application may be significant driver to increase variability in climatic factors. subsequently, fertilizer application have a negative impact on crop production [1, 3, 4, 6, 7, 9, 14, 27]. estimates specify that fertilizer consumption on cropped area have a negative impact on production of potato, cotton, groundnut and sesame crops. application of agricultural labour and cultivator show positive impact on production of all cash crops. estimates suggest that production would improve as utilization of agricultural labour and cultivator in cash crop farming. as indian farmers are small and marginal who have farm size with less than 1 acre in which mechanization may be costly. thus, application of human skill would be important to increase crop production. appropriate price of crop is useful to increase crop production as farmer gives more preference to those crops that give them greater financial benefits [27]. it also increases the decision power of farmers to choose an appropriate crop for cultivation. value of production is a crucial factor to increase the farmer’s consciousness to grow a specific cash crop. it improves farmer’s economic capacity to use another inputs like organic fertilizer, pesticides, better quality of seed, advance technology and better irrigation in cultivation. thus, value of production have a positive impact on production of all crops. few climatic factors have a statistically significant impact on production of cash crops. maximum temperature during sowing time is appeared negative impact on production of potato and cotton crops. estimates can be interpreted that germination of these crops decrease as variability in maximum temperature during sowing time, therefore production of these crops would decline. variability in maximum temperature during growing time have a negative impact on production of potato and cotton crops [9, 15, 20]. crop production of potato and cotton crops in harvesting time is also negatively impacted due to increase in maximum temperature during harvesting time. minimum temperature during growing period have a positive and negative impact on production of cash crops. estimates specify that minimum temperature during sowing time have a negative impact on potato and groundnut crops. it recommended that output of these crop grow in soil, therefore these crops require moderate minimum temperature during sowing, growing and harvesting time. cotton and sesame crops get benefit due to increase in variability in minimum temperature during sowing time. minimum temperature during growing time is seen negative impact on productivity of undertaken cash crops. precipitation during sowing and harvesting time have a negative impact on production of cash crops. estimates are consistent with earlier studies which argued that change in rainfall pattern have a negative impact on crop productivity [1, 6, 10, 14, 23, 27]. coefficient variation in daily precipitation also show negative impact on production of cash crops. coefficient variation in daily maximum and minimum temperature have a positive and negative impact on production of these crops. potatoln(tp)st= -3.8643* +0.7621* ln(as)st -0.5505* ln(srlpca)st +0.0357 ln(sfaca)st -0.0485 ln(ia)st -0.1503* ln(fcca)st +0.3748* ln(ralcca)st +0.5547* ln(vprc)st -0.0049 ln(vamaxtst)st -0.00156 ln(vamaxtgt)st -0.0059 ln(vamaxtht)st -0.0045 ln(vamintst)st 0.0015 ln(vamintgt)st -0.0094 ln(vamintht)st -0.0141 ln(vapcpst)st -0.0022 ln(vapcpgt)st -0.0029 ln(vapcpht)st -0.0491 ln(cvdapcp)st + 0.0688 ln(cvdamint)st + 0.0046 ln(cvdamaxt)st +(vstust) [(n=578) (wald chi2=6858.73*) (log likelihood = -14.8610) (8) cottonln(tp)st= -7.4639*+0.4845* ln(as)st -0.6311* ln(srlpca)st -0.0944** ln(sfaca)st + 0.0547*** ln(ia)st -0.1231* ln(fcca)st +0.7567* ln(ralcca)st +0.5670*ln(vprc)st -0.0072 ln(vamaxtst)st -0.0189*** ln(vamaxtgt)st -0.0036 ln(vamaxtht)st +0.0025 ln(vamintst)st -0.0050 ln(vamintgt)st +0.0010 ln(vamintht)st -0.0004 ln(vapcpst)st +0.0145 ln(vapcpgt)st -0.0039 ln(vapcpht)st +0.2088** ln(cvdapcp)st -0.1824** ln(cvdamint)st +0.0374 ln(cvdamaxt)st +(vstust) [(n=583) (wald chi2=6871.17*) (log likelihood = -66.9939)] (9) groundnutln(tp)st= -2.1741* +1.0863*ln(as)st -0.6311* ln(srlpca)st +0.0770* ln(sfaca)st +0.0676** ln(ia)st -0.2156* ln(fcca)st +0.1982 ln(ralcca)st +0.5730* ln(vprc)st +0.0128 ln(vamaxtst)st -0.0096 ln(vamaxtgt)st +0.0166*** ln(vamaxtht)st -0.0069 ln(vamintst)st -0.0066 ln(vamintgt)st -0.0006 ln(vamintht)st -0.0046 ln(vapcpst)st -0.0019 ln(vapcpgt)st -0.0065 ln(vapcpht)st 0.1580** ln(cvdapcp)st +0.0211 ln(cvdamint)st -0.0384 ln(cvdamaxt)st +(vstust) [(n= 581) (wald chi2= 13115.87*) (log likelihood = -62.3242)] (10) sesameln(tp)st= -3.1582* +1.1245* ln(as)st -0.5765* ln(srlpca)st +-0.0332 ln(sfaca)st -0.0333 ln(ia)st -0.2467* ln(fcca)st +0.2625* ln(ralcca)st +0.5340* ln(vprc)st +0.0083 ln(vamaxtst)st -0.0019 ln(vamaxtgt)st -0.0028 ln(vamaxtht)st +0.0066 ln(vamintst)st -0.0145* ln(vamintgt)st -0.0111 ln(vamintht)st -0.0039 ln(vapcpst)st -0.0059 ln(vapcpgt)st -0.0049 ln(vapcpht)st 0.1682* ln(cvdapcp)st +0.0584 ln(cvdamint)st 0.0864 ln(cvdamaxt)st +(vstust) [(n= 626) (wald chi2= 6699.65*) (log likelihood = -71.4120)] (11) 6.2. association of yield of crops with climatic and non-climatic factors regression results which estimate the impact of climatic and non-climatic factors on yield of cash crops are given in equation 12, 13, 14 and 15. area sown is an important factor to decide output per hectare land and yield is negatively impacted due to increase in cropped area of cash crops [7, 43]. it has observed that yield have a nonlinear relationship with cropped area of cash crops [9, 43]. so, yield of potato, cotton and sesame crops have a negative associated with area sown. yield of cash crops have a negative relationship with rural literate population. it may be due to that literate population have an insignificant contribution in farming. regression coefficient of forest area with cropped area of potato and groundnut crops are positive, which imply that forest area might be useful to increase yield of potato and groundnut crops [1, 38]. 1 http://www.epa.gov/climatechange/science/causes.html. http://www.epa.gov/climatechange/science/causes.html agriculture and food sciences research, 2019, 6(2): 155-165 161 © 2019 by the authors; licensee asian online journal publishing group irrigated area would be useful to increase yield of cotton and groundnut crops, while yield of potato and sesame crops are negatively associated with irrigated area [1, 6, 9, 10]. regression coefficient of fertilizer consumption with yield of cash crops are found negative, it imply that yield of these crops would be declined as fertilizer application increases. agriculture labour and cultivators and monetary value of crops have a positive impact on yield of cash crops. better return on production of cash crop is helpful to increase the economic capacity of farmers, therefore they would be in a better position to apply advance methods in cultivation. consequently, yield of cash crop would be increased as return on per hectare land increases. few climatic factors have statistically significant impact on yield of cash crops. maximum temperature, minimum temperature and precipitation have a negative impact on potato yield. maximum temperature during all crop growth period have a negative impact on cotton yield, while, minimum temperature during sowing and growing time are positively associated with cotton yield. in contrary, cotton yield have a negative association with precipitation during sowing and harvesting time. estimates also indicate that daily variation in climatic factors also show negative impact on cotton yield. as groundnut crop needs moderate minimum temperature during sowing, growing and harvesting time. thus, any variability in minimum temperature would have a negative impact on growth and yield of groundnut crop. groundnut crop requires less water as compared to other crops, thus increase in precipitation have a negative impact on groundnut yield. sesame yield is negatively influenced due to variability in climatic factors during sowing, growing and harvesting time. furthermore, estimates show that variability in climatic factors during sowing, growing and harvesting time have a negative impact on yield of potato, cotton, groundnut and sesame crops. potatoln(lp)st= 3.0434* -0.2379** ln(as)st -0.5505* ln(srlpca)st +0.0357 ln(sfaca)st -0.0485 ln(ia)st -0.15032* ln(fcca)st +0.3748* ln(ralcca)st +0.5547* ln(vprc)st -0.0049 ln(vamaxtst)st -0.0016 ln(vamaxtgt)st -0.0059 ln(vamaxtht)st -0.0045 ln(vamintst)st -0.0015 ln(vamintgt)st -0.0094 ln(vamintht)st -0.0141 ln(vapcpst)st -0.0022 ln(vapcpgt)st -0.0029 ln(vapcpht)st 0.0491 ln(cvdapcp)st +0.0688 ln(cvdamint)st + 0.0046 ln(cvdamaxt)st +(vstust) [(n= 578) (wald chi2= 1304.91*) (log likelihood = -14.8654)] (12) cottonln(lp)st= -2.3281* -0.5155*** ln(as)st -0.6311* ln(srlpca)st -0.0944** ln(sfaca)st +0.0548*** ln(ia)st -0.1231* ln(fcca)st +0.7567* ln(ralcca)st +0.5670* ln(vprc)st -0.0072 ln(vamaxtst)st -0.0189*** ln(vamaxtgt)st -0.0036 ln(vamaxtht)st +0.0025 ln(vamintst)st -0.0050 ln(vamintgt)st +0.0010 ln(vamintht)st -0.0004 ln(vapcpst)st +0.0145 ln(vapcpgt)st -0.0039 ln(vapcpht)st +0.2088** ln(cvdapcp)st -0.1824** ln(cvdamint)st + 0.0373 ln(cvdamaxt)st +(vstust) [(n= 583) (wald chi2= 1435.51*) (log likelihood = -66.9969) (13) groundnutln(lp)st= 4.7359* +0.087 ln(as)st -0.6313* ln(srlpca)st +0.0771* ln(sfaca)st +0.0667** ln(ia)st -0.2151* ln(fcca)st +0.1978* ln(ralcca)st +0.5730* ln(vprc)st +0.0128 ln(vamaxtst)st -0.0096 ln(vamaxtgt)st +0.0167*** ln(vamaxtht)st -0.0069 ln(vamintst)st -0.0067 ln(vamintgt)st -0.0006 ln(vamintht)st -0.0046 ln(vapcpst)st -0.0019 ln(vapcpgt)st -0.0065 ln(vapcpht)st 0.1581** ln(cvdapcp)st +0.0212 ln(cvdamint)st -0.0384 ln(cvdamaxt)st +(vstust) [(n= 581) (wald chi2= 2154.19*) (log likelihood = -62.3299) (14) sesameln(lp)st= 3.7526* +0.1248*** ln(as)st -0.5762* ln(srlpca)st -0.0332 ln(sfaca)st -0.0331 ln(ia)st -0.2467* ln(fcca)st +0.2619* ln(ralcca)st +0.5340* ln(vprc)st +0.0083 ln(vamaxtst)st -0.0019 ln(vamaxtgt)st -0.0028 ln(vamaxtht)st +0.0066 ln(vamintst)st -0.0145*** ln(vamintgt)st -0.0111 ln(vamintht)st -0.0039 ln(vapcpst)st -0.0059 ln(vapcpgt)st -0.0049 ln(vapcpht)st -0.1681* ln(cvdapcp)st +0.0585 ln(cvdamint)st +0.0864 ln(cvdamaxt)st +(vstust) [(n= 626) (wald chi2= 1408.82*) (log likelihood = -71.4405) (15) 6.3. association of cropped area of crops with climatic and non-climatic factors regression coefficients of area sown of cash crops with climatic and non-climatic factors are given in equation 16, 17, 18 and 19. estimates indicate that area sown are positively associated with rural literate population, forest area, irrigated area, and agricultural labour and cultivators. irrigated area is a crucial determinant to increase areas sown, therefore it have positive impact on area sown. hence, it is proposed that irrigated area would be useful to increase cropped area of cash crops [9, 38]. agricultural labour and cultivators are useful to prepare land during sowing time of crops, thus agricultural labours and cultivators are positively associated with cropped area. value of production of all crops have a negative impact on cropped area of respective crops. monetary value of crop production would be useful to increase the farmer’s economic capacity to buy new technology, high yielding varieties of seed, and other inputs, subsequently it may be useful to increase crop productivity. however, high cost of cultivation of cash crops may be another reason to reduce the farmer’s consciousness to enlarge area under cash crops. regression coefficients of climatic variables shows that variability in maximum temperature and minimum tempereture during sowing time have a positive impact on area sown of potato, groundnut and sesame crops. estimates infer that area sown positively reflects due to increase in maximum and minimum temperature during sowing time. variability in precipitation have negative and statistically significant impact on cropped area of potato, groundnut and sesame crops. also, variability in daily precipitation during sowing time is also negatively associated with area sown of cash crops. cropped area of cotton crop is positively associated with precipitation. for positive association of precipitation in sowing time for this crop can be interpreted that the large cropped area of this crop comes under rainfed areas. while, variation in daily maximum and minimum temperature during sowing time show positive impact on cropped area of these crops. potatoln(as)st= -3.8397*+0.1453* ln(srlpca)st +0.1041* ln(sfaca)st +0.0633* ln(ia)st +0.6113* ln(ralcca) -0.0380* ln(vprc)st +0.0033 ln(vamaxtst)st +0.0151* ln(vamintst)st -0.0097** ln(vapcpst)st -0.0177 ln(cvdapcpst)st +0.0784** ln(cvdamintst)st +0.1496* ln(cvdamaxtst)st +(vst ust) [(n= 607) (wald chi2= 18488.19*) (log likelihood = -489.2042)] (16) agriculture and food sciences research, 2019, 6(2): 155-165 162 © 2019 by the authors; licensee asian online journal publishing group cottonln(as)st= -1.4850*+0.1179* ln(srlpca)st +0.2678* ln(sfaca)st +0.0914* ln(ia)st +0.4983* ln(ralcca) -0.0380* ln(vprc)st 0.0056 ln(vamaxtst)st -0.0022 ln(vamintst)st +0.0023 ln(vapcpst)st -0.1012** ln(cvdapcpst)st +0.0165 ln(cvdamintst)st +0.1133** ln(cvdamaxtst)st +(vst ust) [(n= 603) (wald chi2= 19483.01*) (log likelihood = -377.3423)] (17) groundnutln(as)st= -2.2501*+0.0568*** ln(srlpca)st +0.1915* ln(sfaca)st +0.1152* ln(ia)st +0.6293* ln(ralcca) -0.0402* ln(vprc)st +0.0022 ln(vamaxtst)st +0.0014 ln(vamintst)st -0.0138* ln(vapcpst)st -0.1208* ln(cvdapcpst)st +0.0601** ln(cvdamintst)st +0.0480 ln(cvdamaxtst)st +(vst ust) [(n= 604) (wald chi2= 21497.47*) (log likelihood = 349.16462)] (18) sesameln(as)st= -2.4808*+0.0326*** ln(srlpca)st +0.1551* ln(sfaca)st +0.0837* ln(ia)st +0.6680* ln(ralcca) -0.0256* ln(vprc)st +0.0003 ln(vamaxtst)st +0.0006 ln(vamintst)st +-0.0099** ln(vapcpst)st -0.1351* ln(cvdapcpst)st +0.0468*** ln(cvdamintst)st +0.0053 ln(cvdamaxtst)st +(vstust) [(n= 649) (wald chi2= 15074.98*) (log likelihood = -420.6113)] (19) 6.4. explanation on estimated technical efficiency of cash crops state-wise mean value of estimated technical efficiency (te) during 1971-2014 for production, yield and area sown for potato, cotton, groundnut and sesame crops are presented in figure 1, 2, 3 and 4. west bengal have a highest te (95%) of production and yield of potato crop. gujarat, uttar pradesh and tamil nadu have 86%, 75% and 71% te of production in potato crop refer figure 1. so, these states are able to produce better return in potato farming. it is observed that rajasthan and haryana have high efficiency to maintain the area sown under potato crop. estimates conclude that all states have a high variation in technical efficiency to produce potato production and to maintain planting area under potato crop. kerala is observed technically efficient state to produce optimum cotton production with te of 90% refer figure 2. other states have less than 50% te of cotton production. thus, these states are seen technically inefficient to produce cotton production and maintain higher yielding capacity. haryana and punjab are seemed most technically efficient states to utilize cropped area under cotton crop, while other states are appeared technically inefficient to increase or sustain planting area under this crop. hence, it is suggested that other states need to apply technological advancement to increase technical efficiency of production and cropped area in cotton crop. the value of te is found 95% for groundnut production and yield in kerala, thus the state is technically efficient to produce optimum groundnut production refer figure 3. haryana has 87% te of groundnut, imply that haryana is second technically efficient state to produce groundnut output. other states have value of te less than 62%, thus these states are technically inefficient in groundnut farming. andhra pradesh, gujarat, karnataka, madhya pradesh, maharashtra and rajasthan have less than 25% value of te, therefore these states are in poor position and unable to produce desire production of groundnut. haryana have a 97% value of te for cropped area, thus the state is in better position to utilize cropped area in groundnut farming, while punjab is second technically efficient state to maintain cropped area under groundnut crop. other states have a less than 90% value of te, thus these states needed to increase technical efficiency in production and cropped area of groundnut crop. highest value of te (94%) for sesame production and yield in kerala shows that this state is technically efficient to provide better return in sesame crop refer figure 4. other states have low value of te (less than 90%), therefore these states are technically inefficient. there is needed to improve technical efficiency of sesame crop in these states. haryana, madhya pradesh, rajasthan, uttar pradesh, maharashtra states have a less than 30% value of te, thus these states are unable to produce desire output. punjab and haryana have 92% and 89% value of te respectively for cropped area for sesame crop. so, these states have an ability to maintain cropped area under sesame crop. remaining states have a low value of te for area sown of sesame crops, thus these are technically ineffective to sustain cropped area under this crop. figure-1. state-wise technical efficiency of potato farming during 1971-2014. source: estimated by authors. agriculture and food sciences research, 2019, 6(2): 155-165 163 © 2019 by the authors; licensee asian online journal publishing group figure-2. state-wise technical efficiency of cotton farming during 1971-2014. source: estimated by authors. figure-3. state-wise technical efficiency of groundnut farming during 1971-2014. source: estimated by authors. figure-4. state-wise technical efficiency of sesame farming during 1971-2014. source: estimated by authors. 7. conclusion and policy implications this study is assessed the impact of climate variability on production, yield and cropped area of crucial commercial crops in india. for this, sfpfa model is employed using state-wise panel data during 1971-2014. regression coefficients of climatic and non-climatic factors in proposed models are estimated through cobbdouglas production function (log-linear regression model). accordingly, state-wise technical efficiency of production, yield and cropped area for each crop was estimated using time in-variant decay model under nonparametric condition. empirical results show that variability in maximum temperature, minimum temperature and precipitation during sowing, growing and harvesting time have a negative and statistically significant impact on cropped area, production and yield of potato, cotton, groundnut and sesame crops. here, it is concluded that cash agriculture and food sciences research, 2019, 6(2): 155-165 164 © 2019 by the authors; licensee asian online journal publishing group crops farming is in stress due to variability in climatic factors in india. as potato, cotton, groundnut and sesame are crucial commercial crops which provide the raw material to agro-industries like textile, oilseed, and others. therefore, agro-industries based on these crops would be in alarming position due to climate variability in india. thus, it is advised that agro-industries are essential to take an effective action to mitigate the negative impact of climate change on cash crop farming. otherwise, agro-industries would be in serious position in near future. it would also cause to decrease employment opportunities and production in industries, and supply of goods in domestic market. estimated state-wise technical efficiency of each crop infer that there is existence high variation in production activities of cash crops across indian states. most states are found technically inefficient to produce optimum production, which are also unable to use cropped area under cash crops efficiently. kerala is not largely agrarian state of the county, despite that the state have highest technical efficiency in cotton, groundnut, and sesame crop production. west bengal and punjab are technically efficient states to produce optimum potato production. haryana and punjab are observed technically efficient states to better use of cropped area under potato, cotton, groundnut and sesame crops. thus, it recommended that all states are required to improve their technical efficiency to get better returns in cash crop farming. non-climatic factors such as irrigated area, forest area and appropriate prices of crops would be helpful to mitigate the negative impact of climate variability in cash crop farming in india [41]. also, change in plating time and crop varieties may be better adaption technique to reduce climate change impact on cash crop farming [2, 6, 9, 19, 29]. moreover, modern breeding techniques of crops also would be favorable to mitigate the adverse effect of climate change in cultivation [5]. to sustain forest area, reforestation and water conservation techniques would be essential to mitigate the climate change impact in indian cash crop farming in near future [10, 19]. also, adoption of micro-irrigation techniques through sprinkler and drip irrigation may be useful to sustain water for irrigation in near future [3, 8, 10, 19, 44]. application of renewable energy, green technology and suggested application of fertilizer may be valuable to reduce ghgs emission from agricultural activities [9, 10]. extensive fertilizer application in cultivation would be caused to reduce soil nutrients contents. thus, crop production and productivity has a tendency to be declined as extensive application of fertilizer in cultivation. as extensive utilization of fertilizer in cultivation contributes more ghgs emission in atmosphere, resulting it would turn as significant cause to increase more variability in climatic factors. so, recommended fertilizer application in cultivation would be helpful to adapt the climate change impact in cash crop farming. also, application of technological change in cultivation have a better adaptability to resolve the climate change impact in farming [8, 9, 14, 17, 19, 23, 25]. moreover, implementation of drip-irrigation would be useful to maintain for soil quality and actual nutrition contents of soil [22, 44, 45]. intercropping, mixed and multi-tier cropping pattern must be preferred by farmers to reduce climate change impact in cultivation [10, 19, 22]. to promote more organic farming may be advantageous to mitigate the adverse effect of climate change in agriculture [17, 45]. irrigation facilities are useful to reduce the maximum temperature impact in cultivation [3, 5, 8, 10, 19, 44]. thus, water conservation technologies must be adopted by farmers to meet the irrigation requirement in agriculture, which would be useful to mitigate the negative consequences of climate change in cultivation in india [25]. there is essential to increase adoption of improved quality of seed, and to reduce contamination of soil and water surface to improve cash crops productivity [12]. crop insurance policies would be useful to recover the cost of production for farmers, if crop production decreased or damaged due to climate variability [1, 3, 8, 17, 26]. for this, agro-industries desires to provide appropriate financial support to farmers to increase their consciousness to grow commercial and industrial crops like cotton, sugarcane, groundnut, potato, sesame and other crops. government of india is also needs to provide financial support to agricultural scientists and researchers to do more research in agricultural related activities. thereby, scientific research community would be in better position to develop drought resistant and high temperature tolerant crops to mitigate the negative effect of climate change in indian cash crop farming [17, 19]. references [1] a. kumar and p. sharma, "impact of climate 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[45] k. r. kranthi, "cotton in the climate trap," cotton statistics & news, vol. 6, pp. 6-9, 2014. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://hpccc.gov.in/pdf/agriculture/climate%20change%20and%20crop%20yields%20in%20india.pdf, 182 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 2, 182-197, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.62.182.197 © 2019 by the authors; licensee asian online journal publishing group quality of peach (prunus persica l.) genotypes packed in ldpe plastic packaging under different storage conditions tajebe mosie1 kebede woldetsadik2 mulualem azene3 ( corresponding author) 1holeta agricultural research center; temperate and indigenous fruits research program, holeta, ethiopia. 2haramaya university, college of agriculture and environmental science, school of plant sciences, dire dawa, ethiopia. 3ambo university, college of agriculture and veterinary science, department of horticulture, ambo, ethiopia. abstract peach marketing in ethiopia is complicated by postharvest losses like other horticulture produce. although peach varieties have been introduced and studied for their productivity, their postharvest quality has not been evaluated at holeta. therefore, the experiment was done to assess influence of polyethylene plastic packaging and storage conditions on postharvest quality and storage life stability of peach genotypes. it was conducted at holeta agricultural research center from january to february, 2018. peach genotypes ‘bonnigold, early grande, 90-19h and topic beauty’ packed in perforated and non-perforated low density polyethylene plastic (ldpe) and stored under two storage conditions i.e. ambient (6.4°c to 20.2 °c; 40%–71 % rh) and cold storage (-1°c to1 °c; 85–99 % rh). genotypes had different response to packaging materials and storage conditions. perforated bag packaging with cold storage was found more effective as compared to non-perforated and the control fruits in maintaining firmness. non-perforated bag packaging’s combined with cold storage extended shelf life of peach fruits 10 – 20 days compared to other treatment combinations. the non-perforated plastic packaged fruits stored in the cold storage also maintained more tss, aa, ta, tss/ta, ph, marketability and specific gravity content and lowered weight loss and decay. therefore, non-perforated plastic packaging with cold storage condition can be considered in maintaining its quality and extending the shelf life of peaches mainly for commercial producers, traders as well as research stations. keywords: packaging films, peach, quality, storage condition, genotypes, perforated plastic, shelf life. citation | tajebe mosie; kebede woldetsadik; mulualem azene (2019). quality of peach (prunus persica l.) genotypes packed in ldpe plastic packaging under different storage conditions. agriculture and food sciences research, 6(2): 182-197. history: received: 5 april 2019 revised: 15 may 2019 accepted: 25 june 2019 published: 20 august 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: the authors express their heartfelt gratitude for holeta agricultural research center staff members particularly horticulture case team for their valuable contribution for the accomplishment of this research. funding: this study is funded by ethiopian institute of agricultural research. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 183 2. materials and methods ................................................................................................................................................................. 183 3. statistical data analysis............................................................................................................................................................... 185 4. results and discussion ................................................................................................................................................................. 185 5. conclusions ..................................................................................................................................................................................... 192 references ............................................................................................................................................................................................ 195 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.182.197&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.182.197&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/929 https://orcid.org/0000-0001-7985-7928 agriculture and food sciences research, 2019, 6(2): 182-197 183 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to literature by evaluating the quality and shelf life stability of peach (prunus persica) genotypes packed in ldpe plastic packaging. 1. introduction peach (prunus persica (l.) batsch) is mainly grown in temperate regions and is the most important among the stone fruits [1]. it is a versatile fruit with good source of antioxidants byrne [2] and cantin, et al. [3]. peaches consumption has increased due to high sources of sugars, organic acids, minerals and vitamins. ethiopian highlands are endowed with a mosaic of soils and climate which are suitable for the production of many temperate fruits and nut crops. the country is characterized by having diverse topography and agroecological zones, of which over 50% of the total area is highland with elevation between 2000 – 4500 masl with adequate water resources and low temperature during winter that can favor many temperate fruit crops to grow [4]. ethiopia’s share of peach and nectarine production in the world is negligible (172 tons and area coverage of 31 hectare). however, equations show increasing in area as well as production compared to previous years [5]. peach is a climacteric fruit that have short shelf life under room temperature due to their high respiratory rate and fast ripening process. the ripening of peaches is complex that cannot be measured by a single factor. moreover, its marketing in ethiopia is complicated by high postharvest losses like other fresh produces. a postharvest loss of about 30% is reported for fresh produce in ethiopia [6]. this postharvest loss is due to poor storage facilities coupled with improper packaging and transportation [7]. peaches are highly perishable that do not able to prolonged storage. generally, peach varieties have a very short storage potential due to fast ripening process; which results in a limited time for commercialization. high temperature combined with low relative humidity during harvesting and marketing are the major factors which reduce the post-harvest life of peaches. under ambient storage conditions the life of peach does not exceed 3-5 days [8]. refrigerator storage is chosen for extending the commercial life of peach fruits [9]. sound, well matured peach fruits can be stored for 14 42 days under -0.6 0 oc temperature range [10]. on the other hand, if stored too long at or near 0 oc, they are subjected to chilling injury. the start of such symptoms determines the postharvest storage capacity since chilling injury can reduce consumer acceptance [11]. storage at low temperatures, mainly with the range of 2.2-5 oc, is prone to physiological disorder known as internal breakdown [12] which is a symptom associated with chilling injury [12]. many activities have been studied to reduce the incidence of physiological disorder in peach cultivars [12] including cold room with controlled atmosphere (ca) storage. ca storage has shown the reduction of mealiness in peach and nectarine cultivars, mainly with high co2 and reduced o2 levels [13]. packages during harvesting and postharvest handling of fresh produce are used to reduce damage and postharvest loss [14]. packing of fruits in polymeric films creates modified atmospheric conditions (map) inside the package which results in reduced rate of respiration, transpiration, delaying ripening and other metabolic processes of fruits [15]. this packaging technique is accepted now as the technology of future. changes that result in loss in quality occur during postharvest storage of fresh produce as a result of internal and external factors. therefore, use of environmentally friendly technologies such as cold storage, waxing, calcium dipping, packaging and use of plant extracts can be used to delay the adverse effects [16]. postharvest factors responsible for development of browning are the picking date, the duration of the cooling period, the co2 and o2 partial pressure, the storage temperature and condition [17]. although peach varieties have been introduced and evaluated for their productivity, their postharvest quality was not evaluated in our country. besides, efforts have not been done to evaluate the influence of postharvest handling practices to maintain peach fruit quality and shelf life at holeta agricultural research center; local producers store the fruits in pure water till transported to market. thus the objective of this study was to evaluate the quality of peach genotypes packed in plastic packaging under different storage conditions at holeta, central ethiopia. 2. materials and methods 2.1. experimental sites this study was conducted at holeta agricultural research center (harc) which is situated at an altitude of 2400 masl 09`n latitude and 38`29e longitude. the area is characterized by mean annual rainfall of 1041.4 mm and relative humidity of 58.7%. the main rainy season is june to september, which accounts for 70% of the rainfall while the remaining 30% is from february to april. the average annual maximum and minimum temperature is 21.7 °c and 6.7 °c, respectively [18]. 2.2. treatments and experimental design the treatments consisted of 4x3x2 factorial combinations of peach genotypes (tropic beauty, 90-19h, bonnigold and early grande), ldpe plastic packaging (perforated plastic, non-perforated plastic and nonpackaged) and storage conditions (cold and ambient). completely randomized design (crd) with factorial arrangement with three replications was used. the experiment was conducted between january and february 2018 at holeta agricultural research center horticulture laboratory. 2.3. experimental procedure peach fruits were obtained from orchard of holeta agricultural research center. fruits were harvested from 13 years old trees that received all the necessary management practices (pruning, training, fertilization and irrigation) uniformly. fruits were harvested when surface color changed from green to yellow and shape with fullness of shoulder and suture [19]. agriculture and food sciences research, 2019, 6(2): 182-197 184 © 2019 by the authors; licensee asian online journal publishing group 2.4. data collection three fruits of sample per treatment were taken to laboratory for chemical analysis at five days interval. ten fruits were kept for non-destructive evaluation per treatment. 2.4.1. physiological loss in weight (plw) physiological loss in weight was determined using the methods described by mohammed, et al. [20]. weight loss was calculated using the formula: 𝑤𝑒𝑖𝑔ℎ𝑡 𝑙𝑜𝑠𝑠 (%) = 𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑤𝑒𝑖𝑔ℎ𝑡−𝑓𝑖𝑛𝑎𝑙 𝑤𝑒𝑖𝑔ℎ𝑡 𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑤𝑒𝑖𝑔ℎ𝑡 𝑥100 (1) 2.4.2. total soluble solid it was determined in the laboratory by extracting the juice of three peach fruits flesh using juice extractor. the reading of tss was taken from hand held refractmeter with a reading of 0-32 obrix. 2.4.3. phand titratable acidity (ta) the ph was measured with ph meter and ta was obtained by using the method of garner, et al. [21]. the acidity was calculated with the following formula: %𝑎𝑐𝑖𝑑𝑖𝑡𝑦 = [𝑚𝑙𝑠 𝑜𝑓 𝑁𝑎𝑂𝐻]𝑥[0.1𝑁 𝑁𝑎𝑂𝐻][𝑚𝑖𝑙𝑙𝑖𝑒𝑞𝑢𝑖𝑣𝑎𝑙𝑒𝑛𝑡 𝑓𝑎𝑐𝑡𝑜𝑟][100] 𝑔𝑟𝑎𝑚𝑠 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 (2) where, a stone fruit predominant acid is malic acid and its milli equivalent factor is 0.067. 2.4.4. tss/acid ratio (ripening index): it was determined by dividing total soluble solid taken from refractometer reading (obrix) by percent acidity obtained in titratable acidity. 2.4.5. ascorbic acid (aa) it was measured by using a modified method of bessey and king [22]. concentration of vitamin c in peach flesh juice was determined by the following equation: conc. of vitamin c in juice ( mg ml ) = 𝐷𝐶𝑃𝐼𝑃 (𝑚𝑙)𝑢𝑠𝑒𝑑 𝑡𝑜 𝑡𝑖𝑡𝑟𝑎𝑡𝑒 𝑝𝑒𝑎𝑐ℎ 𝑗𝑢𝑖𝑐𝑒 𝐷𝐶𝑃𝐼𝑃 (𝑚𝑙)𝑢𝑠𝑒𝑑 𝑡𝑜 𝑡𝑖𝑡𝑟𝑎𝑡𝑒 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑 𝑥1𝑚𝑔/𝑚𝑙 (3) ascorbic acid content was expressed on a fresh weight basis as mg 100 g-1 of juice. 2.4.6. firmness firmness of peach fruit was assessed using the subjective method by pressing the fruit with the thumb and forefinger [23]. the firmness of fruit was measured based on the resistance of finger pressure and scored by partially trained panelists, using a 1 to 5 scale which represents five firmness classes. the firmness classes was represented as 1 = extra soft, 2 = soft, 3 = firm, 4 = hard, and 5 = extra hard. 2.4.7. fruit marketability was assessed subjectively using the procedure described by mohammed, et al. [20]. the descriptive quality attributes were determined by observing the level of decay, color, surface defects and shriveling. a 1-9 ratings, with 1=unusable, 3=unmarketable, 5=fair, 7=good and 9=excellent, was used to evaluate the fruit quality. fruits receiving a rating of five and above were considered as marketable, while those rated less than five were unmarketable. the number of marketable fruits was used as a measure to calculate the percentage of marketable fruits during storage. percentage marketability = 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑎𝑟𝑘𝑒𝑡𝑎𝑏𝑙𝑒 𝑓𝑟𝑢𝑖𝑡𝑠 𝑡𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑓𝑟𝑢𝑖𝑡𝑠 𝑥100 (4) 2.4.8. percentage decay percentage decay is the number of decayed fruits due to micro-organisms infection which was recorded and calculated as a percentage of the total number of fruits using the following equation. fruit was considered as rotten when more than 5% of surface area was decayed, blackened or rotten [24]. percentage decay = 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑑𝑒𝑐𝑎𝑦𝑒𝑑 𝑓𝑟𝑢𝑖𝑡 𝑡𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑓𝑟𝑢𝑖𝑡 𝑥100 (5) 2.4.9. specific gravity (sg) was determined by converting total soluble solid reading expressed with obrix using the following simple equation [25]: sg = 1 + (0.004 x obrix) (6) where, obrix= tss reading, sg= specific gravity and 0.004 gravity point (constant). 2.4.10. flesh color browning fruits were visually evaluated for flesh color browning and scored by sensory panelists, using a 1-6 scale which presents six flesh browning stages of peach. the six browning stages was represented as 1=none, 2=very slight browning in the pit cavity, 3=slight browning in the pit cavity and surrounding tissue, 4=moderate browning on less than 50% of the flesh, 5=severe browning on 50-75% of the flesh, and 6=extreme browning covering most of the flesh [26]. agriculture and food sciences research, 2019, 6(2): 182-197 185 © 2019 by the authors; licensee asian online journal publishing group 3. statistical data analysis analysis of variance (anova) for the parameters was done using sas statistical software and comparison of treatment means was made by least significance difference test at 5% probability level. 4. results and discussion 4.1. the storage conditions temperature (t0) and relative humidity (rh) of both ambient and cold storage conditions were recorded during the storage period. under ambient condition temperature were varied between 6.4 °c and 20.2 °c with the average of 13.3 °c while the cold storage temperature was maintained between -1 and 1 °c. the relative humidity of the storage environment ranged between 40% and 71% under the ambient conditions, with average 55.5% while it ranged between 85% and 99% in cold store with the average being 92%. 4.2. physiological loss in weight there was highly significant (p≤0.001) difference in physiological loss in weight (plw) due to the combined effect of packaging and storage condition on peach fruit genotypes throughout the storage period table 1. the weight loss values varied between 0.01 to 6.3% on day 5 and from 0.30% to total loss on day 10. generally, weight loss of peach fruits was increased during the storage period both under cold and ambient conditions. on day 5, weight loss of non-packaged fruits of early grande variety stored at ambient condition was 6.30 % which was significantly (p≤0.001) higher than weight loss of peach fruits subjected to all other treatments on the same date. fruits packaged with perforated plastic and stored at ambient conditions had a relatively higher weight loss next to non-packaged ones. the highest weight loss was recorded from the control fruits of early grande variety stored at cold storage, whereas the lowest was for fruits of 90-19h without packaging and stored in the cold storage on day 10. from day 10 onwards, fruits of all cultivars kept ambient condition without packaging were unmarketable whereas all cultivars kept in cold store were in good condition up to the last storage period (30 days) with pwl ranging from 0.3% (tropic beauty packaged in perforated plastic) to 3.37% in cultivar 90-19h packaged in perforated plastic. non-perforated plastic in cold store kept fruit pwl less than 1% while in the perforated plastic combined with cold store it reached 2.14% (tropic beauty) to 3.37% (90-19h) on day 30 of storage. in general, peach fruits packed with non-perforated plastic bags showed lower weight loss both under ambient and cold storage compared to perforated plastic and control in respective storage conditions. this could be due to less availability of o2 inside non-perforated packed fruits for respiration. therefore, reduction in the rate of respiration which ultimately caused decrease and increase in o2 and co2 levels, respectively, as storage time advances. the balanced levels of co2 and o2 inside the package after initial storage could cause marked changes in the activities of specific enzymes [27]. after day 5, nearly all non-packed fruits stored under ambient conditions were unmarketable while those fruits in cold storage kept well for over 10 days. generally, weight loss of fruits was higher under ambient condition than in the cold storage which is varied with genotypes. lower weight loss in non-perforated polyethylene packed fruits might be due to less availability of oxygen for respiration. the present results are in agreement with the findings of calvo, et al. [28] and tijskens and vollebregt [29]. weight loss differences among the treatments also appear to be due to differences in temperature and relative humidity among the storage conditions. hence reduced rate of respiration and transpiration at lower temperature and higher relative humidity (rh) could be the reason for such reduced rate of weight loss of fruits in the cold storage [30]. about 10.0% physiological loss in weight is considered as an index of termination of shelf life of commodities[31]. high storage temperature leads to accelerated water loss and subsequently to shriveling and softening of the fruit [32]. high temperature contributed to high water loss of peach in ambient condition which is associated with faster metabolism; increased cell wall degradation and higher membrane permeability leading to easy evaporation [33]. furthermore, lower weight loss of fruits in the package could be due to slow rate of ripening and prevention of excessive moisture loss. similar results were also presented by tilahunsiyum and kebede [34] and nath, et al. [35]. relatively lower water vapor transmission rate of non-perforated plastic may also contribute for the development of relative higher humidity inside the package mathooko [36]. according to ben-yehoshua [37] the main function of packaging is to reduce respiration rate and water loss by transpiration which could affect the fruits metabolism. this might have led to the extension of shelf life of peach fruits in non-perforated plastic as compared to control and perforated packed fruits. the present findings are in agreement with the previous findings of ribeiro, et al. [38] and drake, et al. [39]. 4.3. total soluble solids the values in total soluble solids (tss) of peach fruits during the storage periods are displayed in table 2. there was significant (p≤0.05) interaction effect of storage condition, genotype and packaging on the tss values of fruits which varied between 8.3–13.33 °brix in the cold storage and from 8.33–12.4 °brix under ambient conditions. there is high consumer acceptance of peaches with high soluble solids; for mid-season peach cultivars a minimum of 11.0% tss is considered optimum [40]. tss value showed increment with storage duration in respective treatments. acceptable tss values were attained in bonnigold and early grande on day five while the remaining cultivars required 10 to 15 days to reach similar levels. in general, non-packaged fruits had relatively higher tss while lower values were recorded in non-perforated plastic packed fruits. in cold storage, bonnigold variety with perforated plastic packs had the highest tss content (13.33) while least value was recorded in 90-19h (11.07) at the end of storage. increase in tss during storage might be associated with starch hydrolysis of fruits [41]. agriculture and food sciences research, 2019, 6(2): 182-197 186 © 2019 by the authors; licensee asian online journal publishing group table-1. influence of plastic packaging and storage condition on plw of peach genotypes during the storage period. treatments storage period(days) genotypes plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 0.00 0.44ij 1.01i 1.17h 1.83d 2.07d 2.14d ambient 0.00 0.53i 1.23h 1.78f nonperforated cold 0.00 0.01m 0.30m 0.31l 0.50i 0.59h 0.62h ambient 0.00 0.17klm 0.41l 0.47k 0.72g nonpackaged cold 0.00 2.38e 10.54b ambient 0.00 4.08c 90-19h perforated cold 0.00 0.29jk 2.46e 2.60c 3.29a 3.35a 3.37a ambient 0.00 0.77h 3.47d 3.95a nonperforated cold 0.00 0.02lm 0.60k 0.68j 0.74g 0.78f 0.80f ambient 0.00 0.12klm 0.81j 1.25h 1.44e nonpackaged cold 0.00 3.53d 10.35c ambient 0.00 5.66b bonnigold perforated cold 0.00 0.81h 1.19h 2.53c 2.90b 2.94b 2.96b ambient 0.00 0.12klm 1.83f 2.27d nonperforated cold 0.00 0.01m 0.49l 0.55k 0.90f 0.94e 0.96e ambient 0.00 0.13klm 0.85j 1.63g 1.82d nonpackaged cold 0.00 1.29g 10.42c ambient 0.00 5.60b early grande perforated cold 0.00 0.23jkl 1.58g 2.00e 2.11c 2.18c 2.20c ambient 0.00 0.60hi 2.48e 2.98b nonperforated cold 0.00 0.02lm 0.47l 0.56k 0.59h 0.67g 0.69g ambient 0.00 0.19klm 0.84j 0.91i 0.95f nonpackaged cold 0.00 1.63f 10.95a ambient 0.00 6.30a se± 0.08 0.03 0.03 0.02 0.02 0.02 cv (%) 9.10 1.88 3.54 2.57 1.97 1.53 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. on day 5 the control fruits of bonnigold stored at ambient conditions had the highest tss value (11.33 °brix) while tropic beauty had the lowest tss value (9.27). likewise, on day 10, control fruits of bonnigold stored in the cold storage attained the highest tss content (13.33 °brix) while tropic beauty under similar condition had the lowest tss content (9.53).this could be due to accelerated ripening because of higher temperature at ambient conditions and free access of the non-packed fruits to o2which increase respiration rates, resulting in faster conversion of starch to soluble sugars [30]. the variations of values among the cultivars show differences in their genetic makeup and response to storage conditions. perforated ldpe bag packaged fruits followed by non-perforated ldpe packed ones maintain their tss value better than the control under cold storage condition whereas non-perforated ldpe packed fruits followed by perforated ldpe packed ones maintained their tss value better than control under ambient condition. tss of fruits kept in perforated and non-perforated packages and stored in the cold storage increased slowly and reached their maximum on 30th day of storage. slow increment in tss of fruits in non-perforated plastic package during storage could be due to production of higher levels of co2, which may lead to less physiological processes and slow ripening of fruits. those fruits packaged using perforated plastic and stored under ambient conditions attain their maximum tss on day 15. on the other hand, non-perforated packaged fruits stored at ambient conditions reached their maximum tss on day 20. slight rises in the tss levels were observed under both cold and ambient storage conditions regardless of the packaging types. however, the tss of peach fruits was maintained at lower level in the cold storage than under ambient condition. high temperature enhances climacteric respiration, which leads to shorter shelf life of fruits [42]. slow changes in tss of fruits stored in the cold storage is in agreement with the findings of aliye, et al. [43]for mango and hirut, et al. [44] for tomato. this may be partly attributed to lower temperature and higher relative humidity maintained in the cold storage that could have resulted in slow conversion of starch in to water soluble sugars [42]. in general, packing of peach fruits in plastic bags combined with cold storage showed better maintenance of the tss towards the end of storage time. in agreement with this result, increases occurred in tss values with the prolonging storage period with map storage of peach and nectarine cultivars [45]. the changes reputed in tss determined during the storage of fruits in a study on map of different peach and nectarine cultivars carried out by agar, et al. [46] and fernandez and artes [47] support the results obtained from this study. the possible atmospheric modification, that is, reduced o2 and increased co2 created in the package, combined with lower temperature in the cold storage might have delayed ripening of the fruits as a result of reduced respiration rate [36]. hence, packed fruits do not rapidly deplete their soluble solids as those of the control fruits as observed in this study. 4.4. ph the ph values of peach fruit genotypes subjected to different packaging and storage treatments for 30 days displayed in table 3. its values varied from 3.72–3.97 when stored in the cold storage and from 3.74–3.96 when stored under ambient conditions. under both storage conditions, packed fruits had higher ph values compared to their respective control treatments. on day 5, cultivar 90-19h packed in non-perforated plastic and stored at ambient had the highest ph value (3.91). agriculture and food sciences research, 2019, 6(2): 182-197 187 © 2019 by the authors; licensee asian online journal publishing group table-2. influence of plastic packing and storage condition on tss of peach genotypes during 30 days of storage. treatments storage period(days) variety plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 8.46a 9.07f-j 9.40hi 10.00fg 10.87e 11.00d 11.13d ambient 8.23a 8.67ij 9.07i 10.27fg nonperforated cold 8.26a 8.47ij 8.73i 9.73g 10.13f 10.60e 10.73e ambient 8.40a 8.33j 9.47hi 9.80g 10.40ef nonpackaged cold 8.60a 9.20f-j 9.53hi ambient 8.80a 9.27f-i 90-19h perforated cold 8.46a 8.30j 9.27i 10.40fg 10.80e 10.93de 11.07de ambient 8.33a 8.73hij 10.10gh 10.40fg nonperforated cold 8.46a 8.80hij 9.27i 10.07fg 10.80e 10.87de 11.00de ambient 8.66a 8.93g-j 9.27i 10.53f 10.87e nonpackaged cold 8.46a 9.97def 10.53fg ambient 8.66a 9.33f-i bonnigold perforated cold 8.66a 9.80d-g 11.17c-f 12.87a 12.93a 13.13a 13.33a ambient 8.40a 10.53cd 11.20c-f 11.53cde nonperforated cold 8.46a 10.33de 10.80d-g 12.33ab 12.47ab 12.67b 12.87b ambient 8.73a 9.37fg 11.53dc 12.00bcd 12.20bc nonpackaged cold 8.50a 11.30bc 13.33a ambient 8.40a 11.33bc early grande perforated cold 8.33a 10.67cd 11.40cde 12.13bc 12.40ab 12.53b 12.67b ambient 8.86a 11.70ab 11.97bc 12.40ab nonperforated cold 8.36a 10.33de 10.67efg 11.27e 11.47d 11.67c 11.93c ambient 8.40a 9.60e-h 10.80d-g 11.33de 11.67cd nonpackaged cold 8.53a 12.27a 12.73ab ambient 8.40a 10.60cd se± 0.27 0.33 0.28 0.25 0.20 0.13 0.12 cv (%) 5.62 5.75 4.63 3.90 3.09 1.92 1.82 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. table-3. influence of plastic packing and storage condition on ph of peach genotypes during 30 days of storage. treatments storage period(days) variety plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 3.72a 3.81de 3.84f-i 3.85d 3.89bc 3.91bc 3.95b ambient 3.71a 3.81de 3.82hi 3.84de nonperforated cold 3.72a 3.85b 3.89c 3.90b 3.90b 3.92ab 3.95b ambient 3.71a 3.91a 3.92a 3.94a 3.95a nonpackaged cold 3.71a 3.84b 3.86def ambient 3.71a 3.82cde 90-19h perforated cold 3.71a 3.73ij 3.77j 3.85d 3.87cd 3.92ab 3.95b ambient 3.71a 3.85b 3.83ghi 3.93a nonperforated cold 3.72a 3.86b 3.89bc 3.90b 3.91b 3.94a 3.97a ambient 3.71a 3.91a 3.91ab 3.94a 3.96a nonpackaged cold 3.72a 3.84bc 3.85efg ambient 3.71a 3.79fg bonnigold perforated cold 3.71a 3.81de 3.84f-i 3.86cd 3.89bc 3.90bc 3.93c ambient 3.72a 3.74hi 3.82hi 3.83e nonperforated cold 3.71a 3.72j 3.77j 3.84de 3.86d 3.89c 3.93c ambient 3.71a 3.81de 3.85e-h 3.88bc 3.91b nonpackaged cold 3.71a 3.75hi 3.78j ambient 3.71a 3.81de early grande perforated cold 3.70a 3.81de 3.83ghi 3.88bc 3.89bc 3.91bc 3.93c ambient 3.71a 3.76h 3.82hi 3.89b nonperforated cold 3.71a 3.83bcd 3.88cd 3.89bc 3.90bc 3.91bc 3.93c ambient 3.71a 3.76gh 3.93a 3.95a 3.96a nonpackaged cold 3.71a 3.77gh 3.87cde ambient 3.70a 3.81de se± 0.01 0.01 0.01 0.01 0.01 0.01 0.01 cv (%) 0.42 0.42 0.37 0.41 0.37 0.33 0.24 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. on day 10 of storage time, early grande packed in non-perforated plastic and kept at ambient had the highest ph (3.93). on day 30, genotype 90-19h packaged in non-perforated plastic and stored in cold storage had the highest ph value (3.97) while the least value (3.93) was recorded with bonnigold and early grande that were packed in both perforated and non-perforated plastic. the higher ph of fruits under ambient storage conditions at initial stage could be associated with the breakdown of acids and sugar in respiration process at faster rate under ambient than in the cold condition [48]. hence, lowering the storage temperature can reduce respiration rate and delay senescence of peach fruits. relatively lower ph values of packed fruits could be explained by reduced respiration rate in the package. reduced o2 and increased co2 which could be created as a result of respiration could delay the rate of respiration in the package and keep ph values low [36]. the rate of use of acids as respiratory substrates may increase at higher temperature and result in depletion of acid content of fruits stored at ambient condition [48]. ph values of peach agriculture and food sciences research, 2019, 6(2): 182-197 188 © 2019 by the authors; licensee asian online journal publishing group fruits, generally, increased with ripening of the fruits and this could be attributed to the fact that fruits with ripening process diminish its predominant malic acid [49]. the increase in ph value of peach with advance in storage time is in agreement with the findings of agozzino, et al. [50]. 4.5. titratable acidity the interaction between packing materials, genotypes and storage conditions had highly significant (p≤0.001) effect on the titratable acidity of peach fruits table 4. titratable acidity of the sample fruits, irrespective of the treatments, decreased linearly throughout the storage period, as opposed to ph value which showed increasing trend. the ta value varied from 0.82% in 90-19h genotype packed in plastic and stored in cold storage on day 30 to 1.21% in early grande cultivar packed in perforated plastic and stored under ambient condition on day 5. this trend is in agreement with the findings of pongener, et al. [51] that the titratable acidity of peach fruits packed under polythene films showed a linear declining trend with the advancement of storage period which could be due to its use by the cells as a respiration substrate [49]. on day 5 and 10 of storage, the ta of early grande variety under perforated plastic package and stored at ambient condition was significantly higher than ta values in all other treatments. table-4. influence of plastic packaging and storage condition on ta of peach genotypes during 30 days of storage. treatments storage period(days) variety plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 0.72a 1.09cde 1.06cd 1.02cd 0.98ab 0.93ab 0.88bc ambient 0.73a 1.11cd 1.05cd 0.99cd non-perforated cold 0.72a 1.06def 1.02cd 0.99cd 0.96b 0.94ab 0.89bc ambient 0.73a 1.04ef 1.01d 0.96def 0.89c non-packaged cold 0.73a 1.10cde 1.07cd ambient 0.73a 0.82j 90-19h perforated cold 0.73a 1.03fg 1.00d 0.93efg 0.87c 0.84c 0.82d ambient 0.72a 0.96h 0.89e 0.89g non-perforated cold 0.73a 0.95h 0.87e 0.88g 0.86c 0.84c 0.82d ambient 0.72a 0.96h 0.90e 0.89g 0.86c non-packaged cold 0.72a 1.09cde 1.05cd ambient 0.72a 0.95h bonnigold perforated cold 0.72a 1.17ab 1.16ab 1.12a 1.02a 0.96a 0.93a ambient 0.72a 1.07c-f 1.05cd 1.05bc non-perforated cold 0.73a 1.06def 1.04cd 1.01cd 0.99ab 0.95a 0.92ab ambient 0.73a 1.12bc 1.09bc 1.05bc 1.01a non-packaged cold 0.72a 1.04ef 1.02cd ambient 0.72a 1.03fg early grande perforated cold 0.72a 1.07c-f 1.04cd 1.04bc 0.99ab 0.92ab 0.88bc ambient 0.73a 1.21a 1.21a 1.10ab non-perforated cold 0.72a 0.96h 0.91e 0.99cd 0.96b 0.90b 0.84cd ambient 0.72a 0.97gh 0.91e 0.92fg 0.90c non-packaged cold 0.73a 1.12bcd 1.09bc ambient 0.73a 0.88i se± 0.01 0.02 0.03 0.02 0.01 0.02 0.02 cv (%) 1.59 3.50 4.55 3.99 2.57 3.36 3.02 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. perforated plastic packaged fruits of bonnigold variety in the cold storage had higher ta values starting from day 15 to 30, indicating varietal difference and effect of storage condition on this parameter. likewise, non perforated plastic packaging tended to have ta values relatively lower than perforated packaged fruits and the control under their respective storage conditions and varieties. previous report also shows that ldpe non-perforated packed fruits could maintain a higher level of acidity [35]. it might be due to reduced respiration rate in the later stage of storage as affected by film permeability to atmospheric gas. the titratable acidity of fruits was an indicator of potential storage quality and declined gradually over the storage period [52]. the lower ta value of polyethylene bag packed fruits in the cold storage could be explained by the reduced rate respiration which results in slow use of organic acids as a substrate of respiration. the higher loss of ta in control fruits could be due to depletion of organic acids as a result of relatively faster respiration and ripening rate of fruits at ambient storage zerbini [53]. kader [54] the atmospheric modification created when fruits are packed with bags may delay respiration and consumption of respiration substrates such as organic acids and sugars are reduced. consequently, as the fruit respires, the o2 level decreases and the co2 level increases in the bags [54]. under these atmospheric conditions, the respiration rate of the fruit decrease which is helpful since high acidity in fruit has been suggested to contribute in part to the flavor retention of ripened fruit [55]. generally, ta content of peach fruit had been maintained higher in the cold storage compared with ambient condition. 4.6. tss/ta (ripening index) the interaction effect between packing materials and storage conditions had significant (p≤0.05) effect on the ripening index of peach genotypes table 5. the tss/ta values of peach fruits showed an increase with storage time, which also varied with genotype and storage condition. tss/ta ratio has important roles to play as parameters that can be used to tell maturity of peach fruit, especially for small-scale growers [56]. the ripening index is important for fruit flavor with lower values makes fruits more favorable [57]. on day 5, early grande fruits without package and stored at ambient had the highest tss/ta ratio (12.09). compared to rest of the cultivars, tropic beauty recorded relatively lower tss/ta values except in non-packaged agriculture and food sciences research, 2019, 6(2): 182-197 189 © 2019 by the authors; licensee asian online journal publishing group ones kept at ambient storage. on the 10th day of storage, non-packaged fruits of bonnigold kept in cold storage resulted in the highest tss/ta value (13.03). table-5. influence of plastic packaging and storage condition on tss/ta of peach genotypes during 30 days of storage. treatments storage period(days) variety plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 10.9b 8.30gh 8.84hi 9.84f 11.13ef 11.79b 12.65c ambient 11.4ab 7.82h 8.66i 10.33def nonperforated cold 12.0ab 8.01h 8.62i 9.83f 10.53f 11.30b 12.12c ambient 11.6ab 7.99h 9.41ghi 10.22ef 11.73de nonpackaged cold 11.9ab 8.37gh 8.95hi ambient 12.0ab 11.37ab 90-19h perforated cold 11.5ab 8.10h 9.27ghi 11.25bcd 12.47abc 12.98a 13.51b ambient 11.4ab 9.11fg 11.32bc 11.69abc nonperforated cold 11.5ab 9.31fg 10.70cde 11.44bc 12.52abc 12.94a 13.47b ambient 11.9ab 9.32fg 10.34def 11.88abc 12.59abc nonpackaged cold 11.6ab 9.14fg 10.09defg ambient 11.9ab 9.80ef bonnigold perforated cold 11.8ab 8.38gh 9.61fgh 11.49abc 12.72ab 13.54a 14.29a ambient 12.2a 9.84def 10.73cde 11.05cde nonperforated cold 11.7ab 9.72f 10.42def 12.17ab 12.56abc 13.29a 13.99ab ambient 11.9ab 8.34gh 10.60cde 11.45bc 12.08bcd nonpackaged cold 11.6ab 10.83bcd 13.03a ambient 11.6ab 11.01bc early grande perforated cold 11.5ab 10.01c-f 10.93bcd 11.71abc 12.48abc 13.58a 14.34a ambient 12.3a 9.64f 9.93efg 11.31bc nonperforated cold 11.6ab 10.77b-e 11.73b 11.34bc 11.90cd 13.01a 14.16ab ambient 11.5ab 9.87def 11.82b 12.40a 12.99a nonpackaged cold 11.7ab 10.98bc 11.64b ambient 11.5ab 12.09a se± 0.39 0.36 0.39 0.32 0.24 0.24 0.24 cv (%) 5.85 6.57 5.85 4.95 3.39 3.21 3.03 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. towards the end of storage, bonnigold and early grande cultivars either in perforated or non-perforated plastics had higher tss/ta values (13.99 ― 14.34) while the lowest was recorded in tropic beauty under similar storage condition. generally, there were increasing trend of tss/ta towards the end of storage time due to increased tss and decreased ta for respective treatment under both storage conditions which was similar with the findings of mkhathini, et al. [56]. similarly, the decrease in o2 and elevated co2 due to packaging delay the ripening [58] and resulted in lower tss/acid ratio [59]. the negative correlation between firmness and tss/ta reflect an increase in tss and decrease in ta with decreasing firmness upon ripening [59]. 4.7. ascorbic acid (vitamin c) the interaction of packaging materials, genotypes and storage condition showed highly significant effect (p≤0.001) on the ascorbic acid (aa) content of peach fruit table 6. vit. c content of peach fruits varied from 11.446.44 mg100−1 g which is in agreement with the range of 10.2–6.2 mg 100−1g aa reported by mahajan, et al. [60]. ascorbic acid content of the fruit generally showed linear decline during storage. similar findings were also reported by soliva-fortuny and martín-belloso [52] in pear. on day 5 and 10 bonnigold cultivar packaged in perforated plastic and kept at ambient had higher ascorbic acid content, which was at par with values of fruits of early grande cultivar kept in perforated plastic and stored in cold storage; after 15 days of storage, bonnigold fruits with perforated plastic packages stored in cold store recorded higher ascorbic acid values, with the values from early grande being at par on day 15 and 20 and bonnigold fruits in non-perforated fruits stored in cold store. all treatments showed decreasing trend towards the end of the storage time both at cold and ambient conditions. activities of oxidizing enzymes might reduce in packed fruits and stored in cold result in higher vitamin c retention up to last day of storage. variation in vitamin c retention in different treatments might be due to genetic differences and levels of oxidation as affected by temperature and plastic permeability to environmental oxygen. lower loss of ascorbic acid content might be due to low o2 permeability, which reduces respiration rate. during storage, higher temperature and oxygen favor activities of oxidizing enzymes like ascorbic acid oxidase, peroxidase, catalase and polyphenol oxidase to act on substrates of respiration, including acids, and hasten loss of ascorbic acid of the fruits [61]. the vitamin c content decreased rapidly which was higher under ambient than in the cold storage. as the storage time increased, the trend was changed. peaches stored in the cold storage showed more vitamin c content than those stored under ambient conditions. packed fruits maintain and showed higher vitamin c content than the control. this might be due to reduced rate of respiration at lower temperature and inside the package that retards respiration and depletion of acids. furthermore, the reduction in internal o2 and ethylene concentration might explain the maintenance of higher value of aa in packed fruits although there is delay in respiration and ripening [54]. agriculture and food sciences research, 2019, 6(2): 182-197 190 © 2019 by the authors; licensee asian online journal publishing group table -6. influence of plastic packing and storage condition on vitamin c content of peach genotypes during 30 days of storage. treatments storage period (days) variety plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 9.44a 9.11ghi 9.11cde 8.78ef 8.44b 7.89de 7.56de ambient 8.99a 8.22ijk 8.00fg 7.33h nonperforated cold 9.55a 9.22fgh 9.00de 8.78ef 8.44b 7.89de 7.44de ambient 9.11a 9.55d-h 9.11cde 8.56f 7.44cd nonpackaged cold 9.11a 8.67hij 7.67g ambient 9.11a 8.67hij 90-19h perforated cold 9.11a 7.66k 7.67g 7.44gh 7.22d 6.67f 6.44f ambient 8.89a 8.22ijk 7.89g 7.44gh nonperforated cold 8.99a 8.78hij 8.78ef 8.44f 7.89c 7.44e 7.33e ambient 9.00a 9.89c-g 9.33cde 9.33bcd 8.44b nonpackaged cold 8.77a 8.00jk 7.44g ambient 8.89a 7.44k bonnigold perforated cold 9.11a 9.7c-g 9.67bcd 9.89a 9.56a 9.33a 8.78a ambient 9.22a 11.44a 10.78a 9.44abc nonperforated cold 9.11a 10.22b-e 9.89bc 9.67ab 9.33a 9.00ab 8.44ab ambient 9.22a 10.11b-f 9.44b-e 8.89def 7.78c nonpackaged cold 8.99a 10.34bcd 9.22cde ambient 9.22a 10.44bcd early grande perforated cold 9.11a 10.56abc 10.22ab 9.89a 9.44a 8.56bc 8.11bc ambient 9.11a 10.89ab 9.89bc 8.89def nonperforated cold 9.11a 9.33e-h 9.22cde 9.11cde 8.78b 8.22cd 7.89cd ambient 9.22a 8.78hij 8.11fg 7.89g 7.11d nonpackaged cold 9.11a 9.56d-h 9.11cde ambient 9.22a 8.22ijk se± 0.34 0.33 0.28 0.19 0.17 0.17 0.16 cv (%) 6.51 6.20 5.37 3.73 3.60 3.73 3.52 note: n:b: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. 4.8. fruit firmness the interaction of packaging, storage and genotype had high significant (p≤ 0.001) effect on firmness of peach fruits during most of the storage time. firmness of peach fruits varied between 1.17 in early grande in perforated plastic stored under ambient condition on day 20 to 4.67 kg cm−2 in cultivar 90-19h non-packaged and stored at ambient table 7. in general, firmness decreased with storage time; but, variation among genotypes was observed under the different storage conditions. this might be due to activity of enzymes and degradation of pectic substances present in the fruits and loss of water. galvis, et al. [62] and drake, et al. [39] reported similar findings in pear fruits. table-7. influence of plastic packaging and storage condition on firmness of peach genotypes during 30 days of storage. treatments storage period(days) variety plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 4.25a 3.67c 3.67fg 3.33cd 3.17b 2.67a 1.92bc ambient 4.50a 4.42ab 3.50g 3.12d non-perforated cold 4.50a 4.42ab 4.25abc 4.00a 3.58a 2.50ab 1.67cd ambient 4.50a 4.67a 4.25abc 4.17a 3.33ab non-packaged cold 4.33a 3.33cde 2.42j ambient 4.41a 3.17ef 90-19h perforated cold 4.41a 4.25b 4.33ab 3.67b 3.17b 2.25bc 1.58cde ambient 4.50a 3.42cde 3.17h 2.92e non-perforated cold 4.50a 4.67a 4.08bcd 3.67b 2.75cd 2.00c 1.42de ambient 4.41a 4.67a 3.92def 3.33cd 2.75cd non-packaged cold 4.41a 3.58cd 3.17h ambient 4.41a 3.58cd bonnigold perforated cold 4.50a 4.58ab 4.33ab 3.50bc 3.17b 2.50ab 2.33a ambient 4.50a 3.25de 3.08h 2.83e non-perforated cold 4.50a 4.50ab 4.00cde 3.50bc 3.08bc 2.58a 2.17ab ambient 4.58a 4.50ab 3.75efg 3.42c 2.67d non-packaged cold 4.41a 2.67gh 1.33k ambient 4.41a 3.08ef early grande perforated cold 4.50a 4.67a 4.42a 3.67b 2.67d 2.08c 1.50de ambient 4.50a 2.83fg 2.50ij 1.25g non-perforated cold 4.41a 4.42ab 4.33ab 3.42c 2.67d 2.17c 1.25e ambient 4.41a 4.50ab 2.75i 1.92f 1.17e non-packaged cold 4.41a 2.58gh 1.42k ambient 4.58a 2.33h se± 0.16 0.15 0.09 0.08 0.12 0.11 0.11 cv (%) 6.20 6.58 4.70 4.32 7.32 7.85 11.43 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. agriculture and food sciences research, 2019, 6(2): 182-197 191 © 2019 by the authors; licensee asian online journal publishing group the control recorded least firmness than the packed fruits in all varieties on days 5 and 10. until day 15 storage time, significant difference was observed in the different packaging materials; tropic beauty and 90-19h packaged with non-perforated bag were the most firm. early grande fruits packaged with perforated as well as non-perforated plastic lost firmness sharply after 20 days while tropic beauty, bonnigold and 90-19h kept in perforated and non-perforated bags remained firmer relatively longer. under ambient storage environment varieties packed with non-perforated plastic were relatively firmer than perforated packed and the control. in general, softening of fruits was experienced as the storage time advanced which might be due to texture modification due to degradation of polysaccharides that take place during ripening [42]. texture changes in fruits are consequences of modifications by component polysaccharides that, in turn, give rise to disassembly of primary cell wall and middle lamella structures due to enzyme activity on carbohydrate polymers [63]. therefore, the differences in decrease of firmness of peach fruits between treatments might be due to differences in rate of respiration that affect depolimerization of pectin’s during ripening [64]. perforated package kept firmness than non-perforated package and control in cold storage up to the end of storage period. however, the least fruit firmness in packages with non-perforated indicates high co2 or low o2 injury that may result in fruit softening [65].the non-perforated package also retain firmness fruits than the control whereas at the ambient storage condition non-perforated packages maintain firmness better than perforated package and control at all days. such influence of package may contribute to their retardation effects of ripening thorough reduction of oxygen concentration around the fruits as well as through reduction of water loss and keeping tissues more turgid [63]. better firmness of fruits in cold storage could be due to the presence of higher relative humidity and lower temperature which in turn retard its rate of respiration and transpiration. likewise, lower softening rate in packed fruits could be due to the packages in lowering its rate of respiration, ripening process and moisture loss [65].the decrease in the fruit firmness with increment of storage could be due to hydrolysis of starch [66]. 4.9. marketability (%) interaction of packages, storage conditions and genotype had highly significant (p≤0.001) effect on marketability of peach fruits table 8. the marketability (%) of peach fruits remained 100% in all genotypes kept in non-perforated bags in the cold storage while fruit stored in perforated under cold storage were 100% marketable for up to day 25. non packaged fruits kept under ambient condition had only 86-90% marketability on day 5, which were discarded thereafter. however, perforated bags extended 100% fruit marketability up to day 10 under ambient storage while non-perforated bags extended by additional five days. under ambient condition and with no packaging, genotype 90-19h and early grande had more marketable fruits (90%) than the other two which had 86.67% marketable fruits. bonnigold fruits stored at ambient in nonperforated bags had 7-10 percent more marketability compared to the rest on day 20 of storage time. the shelf life termination of peaches stored at ambient condition was due to shriveling, over ripening, discoloration (decay) and mould growth. shriveling and mould development were dominant in the control fruits and polyethylene packed fruits, respectively, with mould being more in perforated packages. faster transpiration rate at higher temperature could result in shriveling of unpackaged fruits. mainly, high respiration rate at high temperature can lead to senescence because of the stored food reserve exhaust [67]. table-8. influence of plastic package and storage condition on marketable percentage of peach genotypes during the storage. treatments storage period (days) genotypes plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a 86.67c ambient 100.00a 100.00a 100.00a 86.67b non-perforated cold 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a ambient 100.00a 100.00a 100.00a 100.00a 76.67c 13.33c non-packaged cold 100.00a 100.00a 86.67bc ambient 100.00a 86.67c 90-19h perforated cold 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a 86.67c ambient 100.00a 100.00a 100.00a 80.00c non-perforated cold 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a ambient 100.00a 100.00a 100.00a 100.00a 76.67c 10.00c non-packaged cold 100.00a 100.00a 83.33c ambient 100.00a 90.00b bonnigold perforated cold 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a 93.33b ambient 100.00a 100.00a 100.00a 90.00b non-perforated cold 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a ambient 100.00a 100.00a 100.00a 100.00a 83.33b 26.67b non-packaged cold 100.00a 100.00a 90.00b ambient 100.00a 86.67c early grande perforated cold 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a 90.00bc ambient 100.00a 100.00a 100.00a 86.67b non-perforated cold 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a 100.00a ambient 100.00a 100.00a 100.00a 100.00a 73.33c 13.33c non-packaged cold 100.00a 100.00a 83.33c ambient 100.00a 90.00b se± 0.96 1.83 1.18 1.92 1.67 2.04 cv (%) 1.70 3.25 2.12 3.60 4.0 3.74 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. agriculture and food sciences research, 2019, 6(2): 182-197 192 © 2019 by the authors; licensee asian online journal publishing group peach fruits in cold storage remain fresh and firm for reasonable period of time. this could contribute to lower rate of respiration and transpiration due to relatively lower temperature and higher rh. since high respiration decreases storage life [33] lower temperature storage is known to extending the storage life of perishable produce [68]. generally, marketability of peaches stored in cold storage was better than those stored under ambient condition. similar reports were stated in mango [34] and aliye, et al. [43] and tomato [44]. packed fruits had more marketability percentage than the control under both storage conditions while nonperforated packages had more percent marketable fruits than perforated packed ones. these effects could be due to modified atmosphere created inside the package as well as reduction in water loss [69]. lower rates respiration and ethylene production, reduced action of ethylene, reduced ripening and senescence, slow growth of decay causing pathogens due to modification of gas atmosphere inside the package and low temperature that is not suit for microorganisms might be the reason to extend storage life of the fruits [70]. 4.10. percent decay the result for decay index is shown in table 9. in non-packed fruits stored at ambient condition, the decay started after 5 days and reached maximum (100%) on 20th day of storage period. perforated plastic packed fruits stored at ambient delayed total decay loss to 25 days. no decay was recorded in fruits packed with non-perforated plastic till 30 days of cold storage; this may be due to less chance of microorganism entry and lower rate of respiration that results in less moisture inside the packaging. fruits in non-perforated bags had less decay than those in perforated ones. this lower decay might be due to limited permeability of gases (co2 and o2) and water vapour, which can interplay with physiological processes [29, 52]. the index also varied with the varieties where bonnigold variety showed 7-15% more marketable fruits than the remaining ones in non-perforated bags stored at ambient after 25 days of storage. packages can protect from contamination, damage and most importantly, against excess moisture loss; hence, maintained cell wall stability and middle lamella of the fruits [71]. similar findings of plum fruits at low temperature were also reported [72]. 4.11. specific gravity the interaction effect between packaging materials, peach fruit genotypes and storage conditions had significant (p≤0.05) effect on the specific gravity table 10. in this study, the sg value varied from 1.033 to 1.053. the highest specific gravity was recorded for non-perforated plastic packed and control (unpacked) treatments of bonnigold variety under cold storage condition in perforated plastic bag on day 30 of storage time, whereas the lowest was for fruits of 90-19h with perforated plastic packed and stored in the cold storage and tropic beauty with non-perforated and stored in ambient on day 5. however, fruits of tropic beauty under all treatments and 90-19h in perforated as well as non-perforated bags had statistically similar low value on 5th day of storage. in general, there was an increasing trend with storage time for each treatment, which is directly related with increase of total soluble solid of the fruits. there was highly significant (p ˂ 0.001) difference between varieties throughout the storage period. at the end of the storage period bonnigold (1.053), followed by early grande showed highest specific gravity of fruits packed and stored in cold storage. with increasing of obrix reading there was positive relation of specific gravity; similar result was observed by ting and blair [73]. 4.12. flesh browning cultivars showed very different flesh browning values table 11 after day 20 of storage. genotype 90-19h and early grande with non-perforated package had the highest values on day 25 and 30. non-perforated plastic packed fruits were highly affected with flesh browning irrespective of variety except bonnigold which did not show browning until the last storage time. this high browning of the fruits might be due to fruits high total phenolic content and polyphenol oxidase activities, since both of them are said to be the main factors responsible for enzymatic browning [74]. it has been shown that the rate of browning of fruit products depends on the nature, concentration and interaction of the phenolic compounds that are co-present in tissues [75]. bonnigold didn’t show any flesh color browning throughout the storage period of cold storage both on packed as well as nonpacked. a certain difference between varieties for these parameters was observed by some authors on peach [75] apple [76] and pear [77]. for all cultivars of perforated and non-perforated packed fruits, except bonnigold variety, browning rose after day 20 of cold storage. 5. conclusions non-perforated packed fruits had reduced weight loss compared to the control under both storage conditions. towards the end of storage, non-perforated plastic packaged fruits in cold storage maintained more tss, aa, ta, tss/ta, ph and specific gravity content. non-perforated bag with cold storage were generally more effective compared to perforated package and the control in maintaining the quality of the fruit. except tropic beauty and early grande, non-perforated packaged fruits had higher tss than perforated packed fruits at day 15 of ambient environment storage. as the storage time progressed ph increased irrespective of treatment applied. tss/ta increased throughout the storage period irrespective of treatments due to increased tss and decreased ta values. ascorbic acid content decreased as the storage time advanced irrespective of treatments in which cold storage maintains better than ambient condition. overall, packaging combined with cold storage maintained the freshness, firmness and maintained the shelf life stability of peach fruits. agriculture and food sciences research, 2019, 6(2): 182-197 193 © 2019 by the authors; licensee asian online journal publishing group table-9. influence of plastic package and storage condition on percent decay of peach genotypes during 30 days of storage treatments storage period (days) genotypes plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 0.00 0.00 0.00 0.00 0.00 0.00 13.33(21.14b) ambient 0.00 0.00 0.00 13.33(21.14ef) 76.67(61.22b) 100.00(90.00a) 100.00(90.00a) non-perforated cold 0.00 0.00 0.00 0.00 0.00 0.00 0.00 ambient 0.00 0.00 0.00 0.00 23.33(28.77ef) 86.67(68.85b) 100.00(90.00a) non-packaged cold 0.00 0.00 13.33(21.14b) 70.00(56.99c) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) ambient 0.00 13.33(21.14a) 80.00(63.93a) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) 90-19h perforated cold 0.00 0.00 0.00 0.00 0.00 0.00 13.33(21.14b) ambient 0.00 0.00 0.00 20.00(26.56e) 80.00(63.93b) 100.00(90.00a) 100.00(90.00a) non-perforated cold 0.00 0.00 0.00 0.00 0.00 0.00 0.00 ambient 0.00 0.00 0.00 0.00 23.33ef 90.00(71.56b) 100.00(90.00a) non-packaged cold 0.00 0.00 16.67(23.36b) 80.00(63.93b) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) ambient 0.00 10.00(18.43a) 70.00(56.99a) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) bonnigold perforated cold 0.00 0.00 0.00 0.00 0.00 0.00 6.67(12.29c) ambient 0.00 0.00 0.00 10.00(18.43f) 56.67(48.85d) 100.00(90.00a) 100.00(90.00a) non-perforated cold 0.00 0.00 0.00 0.00 0.00 0.00 0.00 ambient 0.00 0.00 0.00 0.00 16.67(23.85f) 73.33(59.00c) 100.00(90.00a) non-packaged cold 0.00 0.00 10.00(18.43b) 53.33(46.92d) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) ambient 0.00 13.33(12.29a) 70.00(53.07a) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) early grande perforated cold 0.00 0.00 0.00 0.00 0.00 0.00 10.00(18.43b) ambient 0.00 0.00 0.00 13.33(21.14ef) 66.67(54.78c) 100.00(90.00a) 100.00(90.00a) non-perforated cold 0.00 0.00 0.00 0.00 0.00 0.00 0.00 ambient 0.00 0.00 0.00 0.00 26.67(30.99e) 86.67(68.85b) 100.00(90.00a) non-packaged cold 0.00 0.00 16.67(23.85b) 73.33(59.00bc) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) ambient 0.00 10.00(18.43a) 73.33(59.00a) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) 100.00(90.00a) se± 3.36 3.62 2.14 1.82 1.11 1.62 cv (%) 33.09 15.70 6.59 4.75 2.27 3.71 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant difference at p=0.05. agriculture and food sciences research, 2019, 6(2): 182-197 194 © 2019 by the authors; licensee asian online journal publishing group table-10. influence of plastic packaging and storage condition on specific gravity of peach genotypes during 30 days of storage. treatments storage period(days) variety plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 1.034a 1.036f-i 1.037gh 1.040gh 1.043de 1.044d 1.045d ambient 1.033a 1.035ghi 1.036h 1.041gh nonperforated cold 1.033a 1.034hi 1.035h 1.039h 1.041e 1.042e 1.043e ambient 1.034a 1.033i 1.038gh 1.039h 1.042e non-packaged cold 1.034a 1.037e-i 1.038gh ambient 1.035a 1.037e-i 90-19h perforated cold 1.034a 1.033i 1.037gh 1.041gh 1.043de 1.043de 1.044de ambient 1.033a 1.035ghi 1.040fg 1.041gh nonperforated cold 1.034a 1.035ghi 1.037gh 1.040gh 1.043de 1.043de 1.044de ambient 1.035a 1.036f-i 1.037gh 1.042fg 1.043de non-packaged cold 1.034a 1.040cde 1.042ef ambient 1.035a 1.037e-i bonnigold perforated cold 1.035a 1.039c-f 1.045b-e 1.051a 1.051a 1.052a 1.053a ambient 1.034a 1.042bc 1.045b-e 1.046cde nonperforated cold 1.034a 1.041cd 1.043c-f 1.049ab 1.049abc 1.051b 1.051b ambient 1.035a 1.038d-g 1.046bc 1.048bcd 1.047bc non-packaged cold 1.034a 1.045ab 1.053a ambient 1.034a 1.045ab early grande perforated cold 1.033a 1.043bc 1.046bcd 1.048bcd 1.050ab 1.050b 1.051b ambient 1.036a 1.047a 1.048b 1.050ab nonperforated cold 1.034a 1.041cd 1.043def 1.045ef 1.046cd 1.046c 1.048c ambient 1.034a 1.038d-g 1.043c-f 1.045ef 1.047bc non-packaged cold 1.034a 1.049a 1.051a ambient 1.034a 1.042bc se± 0.034 0.001 0.001 0.001 0.001 0.001 0.0004 cv (%) 0.18 0.22 0.18 0.16 0.17 0.08 0.08 note: se=standard error, cv=coefficient of variation, different letters in a column indicate significant differences at p=0.05. table-11. influence of plastic packaging and storage condition on flesh color browning of peach genotypes during 30 days of storage. treatments storage period (days) genotypes plastic packaging storage 0 5 10 15 20 25 30 tropic beauty perforated cold 1.00 1.00 1.00 1.00 1.00 1.17bcd 1.58d ambient 1.00 1.00 1.00 1.00 non-perforated cold 1.00 1.00 1.00 1.00 1.00 1.25bc 2.08b ambient 1.00 1.00 1.00 1.00 1.00 non-packaged cold 1.00 1.00 1.00 ambient 1.00 1.00 90-19h perforated cold 1.00 1.00 1.00 1.00 1.00 1.08cd 1.33e ambient 1.00 1.00 1.00 1.00 non-perforated cold 1.00 1.00 1.00 1.00 1.00 1.33ab 1.83c ambient 1.00 1.00 1.00 1.00 1.00 non-packaged cold 1.00 1.00 1.00 ambient 1.00 1.00 bonnigold perforated cold 1.00 1.00 1.00 1.00 1.00 1.00d 1.00f ambient 1.00 1.00 1.00 1.00 non-perforated cold 1.00 1.00 1.00 1.00 1.00 1.00d 1.00f ambient 1.00 1.00 1.00 1.00 1.00 non-packaged cold 1.00 1.00 1.00 ambient 1.00 1.00 early grande perforated cold 1.00 1.00 1.00 1.00 1.00 1.33ab 1.75cd ambient 1.00 1.00 1.00 1.00 non-perforated cold 1.00 1.00 1.00 1.00 1.00 1.50a 2.58a ambient 1.00 1.00 1.00 1.00 1.00 non-packaged cold 1.00 1.00 1.00 ambient 1.00 1.00 se± 0.08 0.07 cv (%) 11.17 6.93 note: se=standard error, cv=coefficient of variation, means with different letters in a column indicate significant differences at p=0.05. the percentage marketability of peach fruits remained 100% in all genotypes kept in non-perforated bags in the cold storage while fruit stored in perforated under cold storage were 100% marketable for up to day 25. all genotypes with non-perforated packaging under cold storage did not show any sign of decay while perforated packaging exposed fruits to decay. in non-packaged fruits stored under ambient condition severity of decay started from day five onwards reaching 100% starting from day 15. flesh color browning was not experienced only in bonnigold in perforated and non-perforated packaging under cold store. based on the result of the present study, peach genotypes had different response to packaging materials and storage environment. over all, use of non-perforated plastic packaging and cold store extend shelf life by maintaining the quality of peach during post-harvest storage compared to perforated packaging and control. agriculture and food sciences research, 2019, 6(2): 182-197 195 © 2019 by the authors; 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licensee asian online journal publishing group feasibility and economic analysis of bread production in gashua, yobe state, nigeria adekoyeni oludare olumuyiwa1 abdulhamid ellawule2 ( corresponding author) 1department of home science and management, federal university gashua, yobe, nigeria. 2department of accounting, federal university gashua, yobe, nigeria. abstract the feasibility and economic analysis of bread production for a modern cottage bread production business in gashua, yobe state was analysed. this assessment involved analysis of the project to determine the viability, cost, and benefits associated with a bakery project before financial resources are allocated. the capital investment for the bakery establishment was estimated at ₦10,316,303.00 with maximum capacity to utilise 10 bags of flour (50 kg) per day. the production capacity is to increase at 50, 60, 75, 85 and 90 mi efficiency for five years respectively. the cost of production ranged between ₦30,776,550 to 45,059,946.85 while the profit after tax ranged from ₦12,783,071 to 18,878,298.79 for five years. the non current asset schedule annual depreciation was estimated at #655,000. the cash flow and breakeven point were at ₦41,395,161 and 27,705 respectively. bakery business in gashua is worthwhile for entrepreneur as profit making venture. keywords: feasibility study, economic analysis, bakery, depreciation, cash flow, breakeven point. citation | adekoyeni oludare olumuyiwa; abdulhamid ellawule (2020). feasibility and economic analysis of bread production in gashua, yobe state, nigeria. agriculture and food sciences research, 7(2): 125-130. history: received: 4 may 2020 revised: 15 june 2020 accepted: 17 july 2020 published: 30 july 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 126 2. description of methodology and data collection .................................................................................................................. 126 3. the project ..................................................................................................................................................................................... 127 4. manufacturing process of bread ................................................................................................................................................. 127 5. financial analysis .......................................................................................................................................................................... 128 6. conclusion and recommendation .............................................................................................................................................. 130 references ............................................................................................................................................................................................ 130 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.72.125.130&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.72.125.130&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1953 agriculture and food sciences research, 2020, 7(2): 125-130 126 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to the existing literatures by providing information on costs and return analysis associated with bread production including the resource use efficiency. this also makes data available for entrepreneur who is interested in bread production and related business in decicion making and planning. 1. introduction bread is a staple fermented confectionery produced by baking dough recipes largely based on or containing significant quantities of wheat or other cereal flour which are blended with other ingredients [1]; [2]. the breadmaking process has undergone dramatic development over the ages, from traditional baking which involves bread production using wheat flour, yeast (saccharomyces cerevisiae), salt, and water followed by a series of processes such as mixing, kneading, proofing, shaping and baking to commercial industrial production on a large-scale basis that involves the use of enzymes and additives to improve qualities of bread [3]. several varieties of bread are available in nigerian market wheat-meal bread, multigrain and kibbled bread, rye bread and fruit bread. the wheat-meal bread comprises whole wheat and white bread usually referred to as sandwich or sugar bread. the white bread is made with wheat flour from which the bran and germ layers have been removed [4]. the bread business in nigeria is dominated by the white bread while the whole bread is sparingly demanded by people with health challenges. it constitutes one of the most important sources of nutrients such as carbohydrate, protein, fibre, vitamins and minerals in the diets of many people worldwide. the consumption of bread in nigeria is on a steady increase because it is convenient and ready-to-eat food. the desire for more nutritious baked goods is creating a new form of products in the specialized baked goods market. bread is the most staple bakery food eaten by both the low and high social classes in nigeria. it is relatively cheap, affordable and easy to find in the streets. it is widely consumed across the country by almost all citizens, ethnics, and religious groups. according to ohimain [5]; malomo, et al. [6]; odedeji and adeleke [7] the consumption of bread has increased considerably in nigeria due to population increase and urbanization, and the changing preference for convenience foods. nigerian population of 198 million and which is expected to be the third most populous country in the world by 2050 [8]. the country has an estimated national population growth rate of 5.7 per cent per annum and an average economic growth rate of 3.5 per cent per annum in the past five years. in 2016, the value growth of bread in nigeria was put at 12 per cent while other baked products were 11 per cent [9]. it is evident that investment in bread business is increasing. however, the investors require certain eminent information that would assist in the business projection for profitability. economic feasibility assessment of a proposed business involves a cost/ benefits analysis of the project, helping organizations determine the viability, cost, and benefits associated with the project before financial resources are allocated [10]. it also serves as an independent project assessment and enhances project credibility—helping decision-makers determine the positive economic benefits to the organization that the proposed project will provide. it takes into account both quantitative and qualitative factors for analysis of the value for money for a particular project or investment opportunity. therefore, it is imperative to carry out such research before embarking on any business as entrepreneurs. aside from economic feasibility, financial planning is also very important to handle the different operations of the organization within the budget limits. it covers assessment of total capital requirements, sales and prices, break-even outputs, amount of sales required to attain profit in the business. it helps entrepreneurs to get an idea about how much money is required for handling a business project successfully. however, economic feasibility studies and financial planning for a particular business may vary based on certain factors such as geographical location, culture, religion and so on. this paper provided an economic feasibility study of bread production business in gashua, yobe state, nigeria with emphasis on the costs and return associated with bread production including the resource use efficiency. this will make data available for an entrepreneur who is interested in bread production in gashua and its environ. 2. description of methodology and data collection primary data was used for the study. two out of three existing bakeries in gashua were selected for preliminary information on bread business in gashua. the information on the cost of production and other analysis was based on laboratory scale production at the home science and management laboratory, federal university gashua. the bread considered was the family size bread of between 0.85kg±0.05 dough before the baking process. the bread contains 25 slices in a loaf. the price of fixed assets and consumables were based on a market survey at alaba international market, lagos and cost at arrival in gashua. 2.1. potential for bread and bakery business in gashua gashua is a town in bade local government. yobe, nigeria located at 12.87⁰n latitude and 11.04⁰e longitude. it is situated at elevation 339 meters above sea level with an area of 772 km². the estimated nigeria population is 198 million and yobe state with 2,321,339 people out which 125, 817 people live in gashua. yobe state is regarded as one of the states with low consumption of bread with zamfara state at the bottom [11]. however, the steady increase of the nigeria population would automatically affect the population of gashua as a whole which might have either increased arithmetically or geometrically. the bread bakery landscape in nigeria continues to be dominated by unpackaged/artisanal producers with a slight edge over the packaged/industrial baked bread. artisanal producers continued to lead baked goods in 2013, 2014, and 2015 accounting for 53%, 52.2 %, and 50.91% respectively of total current value sales. special bread consumption in nigeria is about 5 % and the figure was expected to rise to about 10.5 % and 11.5 % in 2014 and 2015 respectively. the change in the configuration of people in gashua who are elite would increase demand for special/good bread. according to omeh [12] 10 million of loaves of bread are consumed in lagos every day, 3 million in jigawa which are related to the population of the two states; approximately 16 million and 5 million respectively. it could be estimated that 2 million loaves are consumed daily in yobe state with a population of approximately 3,173,200 million and 130,000 loaves consumed in bade local government. agriculture and food sciences research, 2020, 7(2): 125-130 127 © 2020 by the authors; licensee asian online journal publishing group gashua has a good advantage of notable nearby cities such as nguru, hadejia, potiskum, and azare. damaturu which is the state capital is about 188 km from gashua town. gashua has the market for commercial activities for farm produce and cattle. this population is part of the market strength of the proposal. there are notable federal government agencies in gashua such as prison, military battalion just some few distance and federal university institution of learning and a state college of education with a large population from different parts of the country. these categories of workers provide a ready market for the bakery products especially bread. factors such as population size, population projections, demographic and socioeconomic characteristics of the population, income and existing competition would have a positive change on bread consumption in gashua. 3. the project the bakery project is to be located in gashua, bade local government, yobe state. it is a type of a cottage modern bread-shop with the capacity of producing 5 bags of 50 kg per day at the start and maximum production of 10 bags. plant production increases as marketing mileage is achieved. other confectioneries such as special niche bread (whole wheat bread, banana composite flour bread, cakes, pies, biscuits in the same processing line) shall also be produced. the bakery is a profit-making venture to produce premium bread. it is anticipated that the project will start with 5 bags of flour which is 50% of the installed capacity in the first year of operation, increasing to 60%, 75%, 85% and 90% in second, third, fourth, and fifth-year respectively. 3.1. profit potential in bread making information about the financial cost of production and returns from a modern bakery revealed that it costs about ₦160.00 to produce medium-size standard sliced family bread in nigeria. the bakers sell at ₦230.00 to suppliers at a gain of ₦70.00 per bread. the suppliers sell the bread to retailers at ₦260.00 and make a profit of ₦30.00 per loaf while the retailers sell to the final consumer at the rate of ₦300.00 per loaf. the market analyses have revealed that ₦60.00 ₦70.00 is the average gain that could be made on a family size bread (a bread of 25 slices) by the producer per bread in the market distribution. a typical standard bakery with good supply sales sell at least 20,000 loaves of sliced bread weekly on average and makes a profit of ₦1,400,000.00 per week (₦70.00 x 20,000). this translates that a full-blown bakery produces approximately 34 bags of flour per day excluding 1 day of the week. however, to attain maximum production there is a need for structured marketing strategies, needs enhancements in the areas of packaging and value addition to bakery products [13]. the most artisanal bakery sells directly to the retailers at less cost. the same family size bread sold that usually sold at ₦230.00 by modern bakery can go for ₦200.00 to retailers and the retailers sell it at the rate of ₦250.00 but a lower cost of production such as ₦140.00 per unit. the variation is based on the technology and quality of the bread. the estimated cost of running the bakery per week below is based on 20,000 loaves per week as shown below. therefore, the financial and profits and returns on bakery is impressive and worthy of investing in it. 4. manufacturing process of bread the manufacturing process fits into the production process of any type of bread with adjustment on the processing operation. other baked products passed through similar processing unit operations. straight dough method was used for the bread production as described by ijah, et al. [2] with some adjustment based on the laboratory scale production. the bread-making process is mainly based on 3 steps: dough formation, fermentation and baking. the first step in preparation for mixing is assembling and weighing the ingredients. some ingredients require special preparation. the yeast, whether compressed or dry must be suspended in water according to the manufacturer's instructions. this is usually written on the packaging material. the baking formula was 60.80% wheat flour, 21.90 % water, 9.70 % sugar, 3.70 % skim milk powder, 1.22 % salt, 4.87 % fat, 3.7 % preservative, 0.5 % improver, and 1.22 % yeast. all ingredients are mixed in a kenwood mixer (model a 907d) for 10 min. the purpose of mixing the dough is to distribute the yeast cells throughout the dough, distribute food for the yeast, and to form and develop the gluten. gluten is formed when the two proteins of the flour, gliadin and glutenin come in contact with water. the time required to develop the gluten depends on the strength of the flour and the speed of the machine. the dough was fermented in bowls, covered with wet clean muslin cloth for 55 min at a warm temperature. to ensure alcoholic fermentation which is most desirable, a temperature range between 25.5 -27.5 ⁰c was maintained during the processing. the dough was punched, scaled to 0.9 g dough pieces, proofed in a proofing cabinet at 30°c for 90 min and 85% relative humidity, and baked at 250°c for 30 min. the intense baking heat dries out the part exposed to the air and causes a crust to form. the golden-brown colour of the crust is the result of chemical changes in the starch, sugar and milk known as a browning reaction (maillard reaction) also known as caramelisation. the baked bread samples are then depanned, cooled at ambient temperature and packaged in a polyethene bag. 4.1. functions of the ingredients • flourprovides most of the bulk of the baked item. the wheat flour is high in gluten (protein) as this substance gives bread its fine texture and supports the ingredients during rising. flour from another source can also be used as a substitute such as cassava flour, dry mushroom flour etc. • yeast: this is a microorganism that feeds on starch and sugar releasing co2, alcohol and water. the co2 bubble gives the dough light and airy texture. • fat gives a softer texture and helps prevent the co2 bubbles from escaping from the mixture too soon. • sugar: provides a direct food source for the yeast improving its action. • vitamin c: shortens the time for the dough to mature. agriculture and food sciences research, 2020, 7(2): 125-130 128 © 2020 by the authors; licensee asian online journal publishing group • egg: act as a binder and beaten egg white, like fat, helps to retain gas bubbles. • baking powder: this comprises of baking soda with acid added. it neutralizes the basic condition by producing co2 according to the following equation: nahco3 + h+ → na+ + h2 o + co2 the cost of production of bread using 5 bags of flour is provided in table 1. the cost of materials was based on market price of the consumables at gashua, yobe state. table-1. cost of production of 5 bags of flour per day. materials weight (kg) price (₦) price (₦) for 5 bag/labour flour 5 (goldenpenny) 10,500.00 52,500.00 sugar 4.0 (golden penny) (₦14,00.00 for 50 kg) 1,120.00 5,600.00 yeast 0.20 (royal) (₦1,500.00 for 500g) 600.00 3,000.00 shortening 2.00 (crisco) (₦1,000.00 for 1kg) 2,000.00 10,000.00 salt 0.50 (₦200.00 for 1kg) 100.00 500.00 preservative 0.20 (probake cp) (₦23,000.00 for 25kg. 184.00 920.00 improver 0.20 (₦5,500.00 for 1kg) 1,100.00 5,500.00 water 25 kg 100.00 500.00 packaging material 1 pack of 100 units 50.00 250.00 total 21,618.00 78,770.00 labour i supervisor (₦30,000) and 2 attendants (₦40,000) 70,000.00 2,333.00 cost of fuel generator and tricycle 1,500.00 1,500.00 cost of gas source of heat for baking 1,000.00 repair and maintenance 300.00 damages/depreciation damages/depreciation and other miscellaneous 600.00 total cost of production 83,903.00 5. financial analysis 5.1. investment capital investment cost of the bakery covers the acquisition of all capital items as reflected in the project cost below. table 2 provides estimated project cost for the take off of the bakery. the cost of equipment are based on the landing cost in gashua, yobe state. table-2. estimated project cost. specification ₦ land and development a plot of land 60 x 120 ft 500,000.00 borehole water 500,000.00 building direct labour 3,000,000.00 electric oven with gas this is an electric oven that can also use gas as source heat with a capacity of baking 50 kg (100 standard loaves of 900g) 1,800,000.00 mixer spiral mixer with the capacity to mix 50kg at a time 800,000.00 dough divider 250,000.00 dough moulder 800,000.00 bread slicer top table slicer 250,000.00 measuring scale 100,000.00 working table stainless table 6 x 3m2 50,000.00 bread shelf/rack stainless steel 20,000.00 gen. set (11 kva) lister generator may also be used 500,000.00 auxiliary office equipment/furniture 100,000.00 one tricycle tricycle with extended housing 700,000.00 laboratory equipment and reagents 100,000.00 product registration with nafdac 150,000.00 sub –total 9,620,000.00 add contingency (2% of total cost) 192,400.00 add manpower maintenance salary/wages for 6 months 420,000.00 add initial working capital 83,903.00 total project cost total 10,316,303.00 5.1.1. contingency the contingency provision of ₦192,400.00 was made for items, not yet acquired. this is about 2% of the investment cost. 5.1.2. financial plan the total project cost at ₦10,316,303.00 5.2. financial analysis for five years of operation for the proposed bakery in gashua the mixer and oven which are the major equipment can mix/prepare 50 kg of flour (1 bag) and baked the same quantity respectively in 1 (one) hour. the cost of bread is related to quality and quantity. mathematically, a bag (50 kg) of flour will produce 99 loaves of approximately 800 g of medium size bread. such a loaf is sold at a market price of ₦300.00. half of such agriculture and food sciences research, 2020, 7(2): 125-130 129 © 2020 by the authors; licensee asian online journal publishing group loaf goes for ₦200.00. the bakery at inception shall focus on the production of standard size of 800g sold at ₦300.00 (market price) and 400g loaf sold at a market price of ₦200.00. the estimated cost of producing standard bread sold at ₦300.00 is ₦170.00 and distributed to retailers at the rate of ₦250.00 per loaf who is expected to make a profit of ₦50.00 per loaf. the small loaf is half of the standard bread at ₦180.00 per loaf with ₦100.00 cost of production. the retailers receive the loaves at ₦140.00 per loaf. the bakery is projected to start with 5 bags of 50 kg flour. an average of ₦40.00 gain is made from a loaf of ₦180.00 and ₦80.00 from the loaf of ₦300.00. there is no significant variation in profits. since two small loaves are almost equal to a big loaf, therefore, production of a standard (big) loaf is used for the financial analysis. 5.3. non-current assets schedule the lifespan and annual depreciation of assets of the bakery is projected in table 3. table-3. non-current assets schedule. item cost lifespan annual depreciation ₦ years ₦ 1 building 1.1 factory building 3,000,000.00 25 120,000.00 2 plant and machinery 2.1 mixer 800,000 10 80,000.00 2.2 rotary oven 1,800,000 10 180,000.00 2.3 dough divider 250,000 10 25,000.00 2.4 dough moulder 250,000 10 25,000.00 2.5 bread slicer 250,000 10 25,000.00 2.6 gen. set 500,000 5 50,000.00 2.7 measuring scale 100,000 10 10,000.00 3 utilities 3.1 lab. chemicals 100,000.00 5 20,000.00 3.2 furniture and fittings 100,000.00 5 20,000.00 3.3 tricycle 700,000.00 7 100,000.00 4 total 7,850,000.00 655,000.00 table-4. cash flow analysis. bread project investment appraisal year description cashflow df @15% pv ₦ ₦ 0 equipment (10,232,400) 1.0000 (10,232,400.00) 0 working capital (83,903) 1.0000 (83,903.00) 1 cash profit 8,709,560 0.8696 7,573,530.43 2 cash profit 10,612,232 0.7561 8,024,372.02 3 cash profit 13,466,240 0.6575 8,854,271.39 4 cash profit 15,368,912 0.5718 8,787,225.32 5 cash profit 16,320,248 0.4972 8,114,047.62 5 recovery of working capital 83,903 0.4972 41,714.62 31,078,858.40 table-5. income projection cost-volume. statement of income projection cost-volume analysis for bread production for 5 years year 1 year 2 year 3 yea 4 year 5 production capacity 50% 60% 75% 85% 90% units of loaves 180,675 216,810 271,013 307,148 325,215 sales price/unit (₦) 230 230 230 230 230 ₦ ₦ ₦ ₦ ₦ total revenue 41,555,250 49,866,300 62,332,875 70,643,925 74,799,450 cogp: material expense 29,663,550 35,596,260 44,495,325 50,428,035 53,394,390 labour cost 840,000 840,000 840,000 840,000 840,000 30,503,550 36,436,260 45,335,325 51,268,035 54,234,390 gross profit 11,051,700 13,430,040 16,997,550 19,375,890 20,565,060 expenses: repairs and maintenance 109,500 109,500 109,500 109,500 109,500 miscellaneous 219,000 219,000 219,000 219,000 219,000 depreciation 655,000 655,000 655,000 655,000 655,000 983,500 983,500 983,500 983,500 983,500 pbt 10,068,200 12,446,540 16,014,050 18,392,390 19,581,560 tax @ 20% 2,013,640 2,489,308 3,202,810 3,678,478 3,916,312 net profit 8,054,560 9,957,232 12,811,240 14,713,912 15,665,248 cashflow 8,709,560 10,612,232 13,466,240 15,368,912 16,320,248 variable cost (vc) 29,663,550 35,596,260 44,495,325 50,428,035 53,394,390 fixed cost 1,823,500 1,823,500 1,823,500 1,823,500 1,823,500 vc/unit 164 164 164 164 164 break-even point 27,705 27,705 27,705 27,705 27,705 agriculture and food sciences research, 2020, 7(2): 125-130 130 © 2020 by the authors; licensee asian online journal publishing group a project of this nature involves a huge sum of money and as such the management must ensure that it is consistent with the company’s long-term plan and also ensure that that the project provides adequate investment returns. because of the huge sum of money involved, it is advised that the project be evaluated before it is undertaken to determine its viability. the evaluation of the project shows cash flow of ₦41,395,161 and it is ₦31,078,858 above the initial capital investment which is a viable project to undertake table 4. also, from the statement of income and break-even analysis below table 5, it shows that the project has a good margin of safety. the project generates an estimated ₦8,054,560 profit in its first year and ₦15,665,248 in the 5th year which will continue unless the project cannot produce up to 27,705 loaves in a year which is the break-even units. 6. conclusion and recommendation the feasibility and the economic-financial analysis revealed that bakery business could be successful in gashua, yobe state. the profit is considerable and is worth investing on. increase in production will also translate into a high-profit base on demand. the bakery will also meet the goal by setting the standard in bakery business especially in producing safe bread and providing job opportunities. any investment put into the project will be a worthy investment that will yield much return. it is, therefore, hitherto recommended for funding. references [1] o. o. adekoyeni, a. f. adegoke, and a. e. ayano, "nutritional, functional, sensory and microbial qualities of wheat-tomato seed flour bread," carpathian journal of food science and technology, vol. 10, pp. 47-56, 2018. [2] u. j. j. ijah, h. s. auta, m. o. aduloju, and s. a. aransiola, "microbiological, nutritional, and sensory quality of bread produced from wheat and potato flour blends," international journal of food science, pp. 1-6, 2014. available at: https://doi.org/10.1155/2014/671701. [3] a. t. yahaya, t. r. shittu, m. a. k. ogunjobi, c. o. jayeola, and a. o. williams, "gross margin analysis of cocoa bread production," international journal of horticulture, agriculture and food science, vol. 1, pp. 18-21, 2017. [4] c. guthrie, "the different types of bread from around the world. retrieved from https://www.oola.com/day-to-daylife/2493632/what-to-watch-on-tv/," 2018. [5] e. i. ohimain, "the prospects and challenges of composite flour for bread production in nigeria," global journal of human-social science, vol. 14, pp. 1-11, 2014. [6] s. malomo, a. eleyinmi, and j. fashakin, "chemical composition, rheological properties and bread making potentials of composite flours from breadfruit, breadnut and wheat," african journal of food science, vol. 5, pp. 400-410, 2011. [7] j. odedeji and r. adeleke, "pasting characteristics of wheat and sweet potato flour blends," pakistan journal of nutrition, vol. 9, pp. 555-557, 2010. available at: https://doi.org/10.3923/pjn.2010.555.557. [8] g. i. agbara, a. bade, s. h. ali, and a. m. fannah, "evaluation of production, consumption and nutritive value of gurasa, an indigenous flat bread of north-western nigeria," international journal of scientific research and management, vol. 6, pp. 1-7, 2018. available at: https://doi.org/10.18535/ijsrm/v6i5.ft01. [9] statista, "nigerian baked goods and bread value growth 2016 statista research department. retrieved from. statista research department. https://www.statista.com/statistics/856356/nigeria-baked-goods-and-bread-value-growth," 2016. [10] a. saeed, "economic feasibility study: preparation and analysis, sultan qaboos-academic publication and outreach department," sultnate of oma, pp. 1-7, 2019. [11] nigeria bureau of statistics, "consumption pattern of bread in nigeria. nigeria data portal retrieved from https://nigeria.opendataforafrica.org/yzqsxpd/consumption-pattern?tsid=1000090 " 2014. [12] d. omeh, "food processing-manufacturing: how to start a bread bakery business in nigeria. retrieved from https://www.wealthresult.com/food-processing/how-start-bread-bakery-business," 2019. [13] s. kiumarsi, k. jayaraman, m. i. salmi, and v. asra, "marketing strategies to improve the sales of bakery products of smallmedium enterprise (smes) in malaysia," international food research journal, vol. 21, pp. 2101-2107, 2014. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.oola.com/day-to-day-life/2493632/what-to-watch-on-tv/, http://www.oola.com/day-to-day-life/2493632/what-to-watch-on-tv/, http://www.statista.com/statistics/856356/nigeria-baked-goods-and-bread-value-growth, http://www.wealthresult.com/food-processing/how-start-bread-bakery-business, 12 © 2025 by the author; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 12-18, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6554 © 2025 by the author; licensee asian online journal publishing group sustainable biomass sourcing through agroforestry systems based on poplar and eucalyptus as fast-growing trees niels thevs independent researcher, gluckstrasse 2, 53115 bonn, germany. email: niels.thevs@giz.de abstract global biomass demand is expected to increase during the next decades, which bears risks of accelerating food insecurity and biodiversity loss. therefore, guidelines and standards have been developed to ensure sustainable biomass sourcing as feedstock for material or energetic purposes. this review addresses the question of how fast-growing trees in agroforestry systems increase biomass production and serve as sustainably sourced biomass feedstock. the land equivalency ratio (ler), a measure of productivity gains by agroforestry, was positively correlated with the number of trees per hectare (r = 0.561) and with the spacing between tree rows (r = 0.26). the former corresponds to a high wood yield of the given agroforestry systems, while the latter corresponds to high crop yields within the agroforestry systems. the ler of tree windbreak systems (spacing between tree rows >100 m) was 1.1-2.1. tree windbreak systems adhered to principles of sustainable biomass sourcing, while other agroforestry systems often provided lower food crop yields compared to the corresponding crop monoculture. still, such agroforestry systems help to diversify incomes, have the potential to protect croplands against erosion, and improve the microclimate. depending on local conditions, biomass from those agroforestry systems can be considered sustainable, too. keywords: alley cropping, bioeconomy, biomass demand, land equivalency ratio, material use, tree windbreak systems, woody biomass. citation | thevs, n. (2025). sustainable biomass sourcing through agroforestry systems based on poplar and eucalyptus as fast-growing trees. agriculture and food sciences research, 12(1), 12–18. 10.20448/aesr.v12i1.6554 history: received: 19 february 2025 revised: 24 march 2025 accepted: 28 march 2025 published: 4 april 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. contents 1. introduction ...................................................................................................................................................................................... 13 2. methods ............................................................................................................................................................................................. 13 3. results ................................................................................................................................................................................................ 14 4. discussion .......................................................................................................................................................................................... 15 5. conclusion ......................................................................................................................................................................................... 17 references .............................................................................................................................................................................................. 17 mailto:niels.thevs@giz.de https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6554 https://orcid.org/0000-0003-1923-4280 agriculture and food sciences research, 2025, 12(1): 12-18 13 © 2025 by the author; licensee asian online journal publishing group contribution of this paper to the literature: this study, for the first time, pools together data on wood and crop yields of agroforestry systems with fast-growing trees. these data are discussed to assess agroforestry systems as biomass feedstock in light of the emerging issue of sustainable biomass sourcing for the bio-economy. 1. introduction global biomass demand is expected to increase during the next decades, as shown by, for instance, [1], who assumed that the global biomass demand will increase from 12.14 gt in 2011 to between 17.13 gt and 20.36 gt by 2050, in order to satisfy the needs for increased food production as well as for the material and energetic use of biomass. biomass is primarily used to feed livestock (58% of global biomass use), bioenergy (amounting to 16%), food (14%), material use (10%), and biofuels (biodiesel and bioethanol for transport account for 1%) [1]. in the course of reducing the use of fossil carbon sources for material uses, it is claimed that the overall consumption of materials needs to be reduced, recycling needs to be increased, and bio-based products need to be used for the amount of production of virgin materials (such as chemicals, but also construction materials) that cannot be covered by the reduction of consumption and recycling [2]. this is underlined by unep [3], which states that a sustainable and circular bioeconomy must be based on prioritizing the use of biomass to maximize well-being, while the conversion of biodiversityand carbon-rich natural systems must be avoided. yet, the production and sourcing of an increasing amount of biomass for human demands bear high risks of accelerating food insecurity, biodiversity loss, and climate change. therefore, guidelines, safeguards, and standards have been developed to ensure sustainable biomass sourcing as feedstock for material or energetic purposes across different feedstocks [4, 5], as well as for specific feedstocks, such as sugar cane [6], soy [7], or palm oil [8]. to address the concerns regarding food insecurity, biodiversity loss, and climate change, the bfa [4], for instance, stipulated the following principles that sustainably sourced biomass must fulfill: i) it is derived from renewable biomass whose production is sustainably managed, ii) it does not adversely impact food security and affordability and maintains or improves social and economic conditions along with ecosystem services in producing communities, iii) it does not directly or indirectly result in the destruction of critical ecosystems or loss of high conservation value habitats, and iv) it contributes to landscape resilience and is resilient to the impacts of climate change. other standards, such as rsb [5], adhere to very similar principles. agroforestry (af) systems, which according to fao [9] deliberately integrate woody perennials (trees, shrubs, palms, bamboos, etc.) with agricultural crops and/or animals, are acknowledged to increase overall biomass productivity compared to respective monocultures of trees and crops, while providing a number of benefits to improve food insecurity, contribute to halting biodiversity loss, and help mitigate and adapt to climate change. furthermore, af has the potential to improve soil fertility, reduce erosion, improve water quality, sequester carbon, increase aesthetics [10], and enhance the microclimate for crops [11]. in particular, fast-growing trees can contribute substantially to increasing the productivity of af systems. hence, in the first step, this paper assesses the productivity gains of af systems from fast-growing trees, followed by a discussion of how far the criteria for sustainable biomass sourcing are met. thereby, this paper focuses on af systems with the two fast-growing tree species, poplar and eucalyptus, which are listed among the most widely used fast-growing tree species in af globally [12]. in this paper, the jurisdictional and societal principles are not studied, such as sustainably sourced biomass must adhere to labor laws and must economically benefit local and rural communities. the land equivalency ratio (ler) after [13] is a widely used metric to express productivity gains of agroforestry (af) systems versus corresponding monocultures of crops and trees. the ler is calculated as follows. 𝐿𝐸𝑅 = 𝑌𝑖𝑒𝑙𝑑 𝑡𝑟𝑒𝑒 𝑎𝑔𝑟𝑜𝑓𝑜𝑟𝑒𝑠𝑡𝑟𝑦 𝑌𝑖𝑒𝑙𝑑 𝑡𝑟𝑒𝑒 𝑟𝑒𝑓𝑒𝑟𝑒𝑛𝑐𝑒 + 𝑌𝑖𝑒𝑙𝑑 𝑐𝑟𝑜𝑝 𝑖𝑛 𝑎𝑔𝑟𝑜𝑓𝑜𝑟𝑒𝑠𝑡𝑟𝑦 𝑌𝑖𝑒𝑙𝑑 𝑐𝑟𝑜𝑝 𝑟𝑒𝑓𝑒𝑟𝑒𝑛𝑐𝑒 yield tree reference is a plantation of the given tree, while yield crop reference refers to a field plot of the given crop with no kind of intercropping. thus, ler only considers biomass yield changes (wood yields from the trees and crop yields), regardless of their economic value or further benefits beyond sheer biomass. crop-related ler (lerc) after [14] focuses on crop yield changes by af systems and therefore highlights the benefits associated with crops rather than trees. hence, lerc captures the impact of af systems on food supply. 𝐿𝐸𝑅𝑐 = 𝑌𝑖𝑒𝑙𝑑 𝑐𝑟𝑜𝑝 𝑖𝑛 𝑎𝑔𝑟𝑜𝑓𝑜𝑟𝑒𝑠𝑡𝑟𝑦 − 𝑌𝑖𝑒𝑙𝑑 𝑐𝑟𝑜𝑝 𝑟𝑒𝑓𝑒𝑟𝑒𝑛𝑐𝑒 𝑌𝑖𝑒𝑙𝑑 𝑐𝑟𝑜𝑝 𝑟𝑒𝑓𝑒𝑟𝑒𝑛𝑐𝑒 2. methods the productivity gains by agroforestry (af) of fast-growing trees were assessed through a literature review using the search terms "land equivalency ratio" and "poplar" and "poplar agroforestry" as well as "land equivalency ratio" and "eucalyptus" and "eucalyptus agroforestry," which were used in google scholar and web of science. papers were screened for absolute crop yield data for crops within af in comparison to the same crop grown as a monoculture to be able to calculate lerc. those papers that, in addition to crop yield data, contained tree yield data for the af trees and trees from plantations were used to calculate ler values. crop yields and tree yields were converted into t ha-1 yr-1. thereby, tree yields referred to above ground woody biomass, mostly stem wood biomass, which was divided by the given tree age. part of the studies on poplar-based af systems reported tree or stem volumes, which were converted into biomass with a wood density of 0.35 t m-3 after [15]. next to the crop and tree yield data, the following data were extracted from the selected papers, as far as they were available. • site data: geographical location (by name), climate classification (köppen-geiger), mean air temperature, and annual precipitation. • af system data: type of af system, tree species (clone, if available), spacing between tree rows and within rows, number of trees per hectare, tree height, and crop. agriculture and food sciences research, 2025, 12(1): 12-18 14 © 2025 by the author; licensee asian online journal publishing group ler and lerc values were grouped by tree (poplar and eucalyptus), af system, and crop in order to calculate the respective averages and standard deviations. the differences between averages were tested for significance at α < 0.05 through a t-test (the averages by tree) and by an analysis of variance, respectively. the studies of the af experiments reviewed did not contain tree windbreaks or field boundaries of wide spacing (100 m and more), so this gap was closed with crop yield data from [16] and our own tree yield data. those own tree yield data stem from a dataset of poplar-based (p. nigra, local clone mirza terek) tree windbreaks in kyrgyzstan, jalalabad region, (spacing of 200 m) and plantations from the same poplar clone in that region. the selected tree windbreaks and plantations consisted of 10-year-old trees. 3. results a total of 23 field experiments were screened for this study, which yielded a total of 100 ler datapoints, of which 58 and 42 datapoints were from poplar and eucalyptus af systems, respectively. the vast majority (87%) of those datapoints stemmed from experiments that were laid out as alley cropping systems or contiguous plantations with the given crop as understory vegetation, while only 13% were oriented around tree rows as field boundaries. the tree spacing within the contiguous plantations was 3m x 3m, while most of the alley cropping had spacings between tree rows of less than 8 m. the majority of all datapoints (70%) stemmed from af systems with trees aged 5 years and younger. nine additional datapoints allowed for the calculation of lerc only, because no tree data were reported. these nine additional datapoints stem from experiments laid out as alley cropping, with eight datapoints from poplar af systems and one from a eucalyptus af system. the ler and lerc averaged across all data points were 1.51 and -0.29, respectively. the lers divided by af tree species were 1.57 for poplar and 1.42 for eucalyptus af systems, whereby the two did not show significant differences (table 1). the lers divided by af system did not differ significantly either (table 2). additionally, ler by major crops did not show significant differences (table 3). out of the 100 data points, only six show an ler < 1; these six data points all stem from eucalyptus af systems, while the minimum ler among the poplar af systems was 1.14 (table 1). table 1. land equivalency ratio (ler) of af systems by poplar and eucalyptus. tree ler average ± standard deviation ler maximum ler minimum number of data points poplar 1.57 ± 0.37 2.78 1.14 58 eucalyptus 1.42 ± 0.4 2.19 0.68 42 all data points 1.51 ± 0.39 2.78 0.68 100 table 2. land equivalency ratio (ler) of the different af systems: alley cropping, contiguous plantations with the given crop as understory vegetation, and tree rows as field boundary. tree ler average ± standard deviation ler maximum ler minimum number of data points alley cropping 1.48 ± 0.36 0.68 2.78 81 contiguous plantations 1.56 ± 0.19 1.27 1.83 6 field boundary 1.62 ± 0.57 0.85 2.23 13 table 3. land equivalency ratio (ler) of af systems by crops, poplar and eucalyptus pooled together under each crop. crop ler average ± standard deviation ler maximum ler minimum number of data points wheat 1.51 ± 0.47 2.78 0.85 35 switchgrass (panicum virgatum l.) 1.46 ± 0.18 1.71 1.26 6 sorghum 1.55 ± 0.24 2.01 1.18 24 fodder grass 1.76 ± 0.19 1.94 1.45 5 cowpea 1.5 ± 0.16 1.64 1.23 5 barley 1.58 ± 0.66 2.77 1.22 5 the lerc values of poplar and eucalyptus agroforestry (af) systems did not differ significantly (table 4). divided by af system, the lerc of field boundary af systems was significantly higher than the lerc values of alley cropping and plantation with crops as understory vegetation (table 5). table 4. crop related land equivalency ratio (lerc) of af systems by poplar and eucalyptus. tree lerc average ± standard deviation lerc maximum lerc minimum number of data points poplar -0.23 ± 0.23 0.35 -0.74 66 eucalyptus -0.37 ± 0.24 0 -0.82 43 all data points -0.29 ± 0.24 0.35 -0.82 109 table 5. crop related land equivalency ratio (lerc) of the different af systems: alley cropping, contiguous plantations with the given crop as understory vegetation, and tree rows as field boundary. tree lerc average ± standard deviation lerc maximum lerc minimum number of data points alley cropping -0.3 ± 0.24a 0.35 -0.82 89 contiguous plantations -0.48 ± 0.21a -0.17 -0.73 7 field boundary -0.08 ± 0.13b 0.14 -0.25 13 note: the letters a and b indicate significant differences of the averages of lerc (α = 0.05). agriculture and food sciences research, 2025, 12(1): 12-18 15 © 2025 by the author; licensee asian online journal publishing group the large majority of the ler datapoints (94%) and all nine additional lerc datapoints showed lower crop yields inside the af system compared to the corresponding monoculture crop yields. this crop yield reduction increased with tree age and thus tree height (see data points by kimura et al. [17]). among the experiments listed here, crop yields decreased more pronouncedly the narrower the tree spacing had been laid out (see data points by sirohi et al. [18]). the datapoints with crop yields higher inside af than the corresponding monoculture refer to af systems with spacing between tree rows of 25 m, 40 m, and 48 m, respectively, which is wide spacing compared with the majority of the datapoints included in this study. ler was significantly positively correlated with the number of trees per hectare (r = 0.561) and with the spacing between tree rows (r = 0.26). the former corresponds to a high wood yield of the given agroforestry (af) systems due to a high tree density, while the latter corresponds to high crop yields within the af systems. lerc was not significantly correlated with the number of trees per hectare, but it was positively and significantly correlated with the spacing between tree rows (r = 0.5), which underlines that a wider spacing between tree rows impacts crop yields to a lesser extent. lerc was significantly and negatively correlated with the tree yield in the given af systems (r = -0.24), which indicates a trade-off between crop yields and tree biomass yield for af systems with a narrow spacing between tree rows. in contrast to the experiments listed here, crop yields on average increased in tree windbreak systems, i.e., agroforestry (af) systems with distances between tree rows of 100 m or several hundred meters, which is reflected in positive lerc values across crops (table 7) [16]. tree yield data of poplars in tree windbreaks versus plantations were drawn from our own data in kyrgyzstan (table 6). the ler of those systems also indicated an overall increase in biomass yields inside af systems compared to separated monocultures (table 7). table 6. tree age, dbh, height, volume, biomass, and tree yield of tree windbreaks with 200 m spacing (single row) and corresponding plantations in the jalalabad region, kyrgyzstan. tree windbreaks and plantations consisted of p. nigra, local clone mirza terek. tree parameter (average) tree wind break plantation tree age [yr] 10 10 dbh [cm] 21 17.8 height [m] 22.3 19.5 tree volume [m³] 0.32 0.2 tree biomass [kg] 127 79 trees per ha 100 2500 biomass [t ha-1] 12.7 198.4 yield [t ha-1 yr-1] 1.3 19.8 table 7. crop yield increases, ler, and lerc of tree windbreak systems. crop crop yield increase [%] of crop within tree wind break system, after [16] ler lerc spring wheat 8 1.1 0.08 winter wheat 23 1.3 0.23 barley 25 1.3 0.25 oats 6 1.1 0.06 rye 19 1.3 0.19 millet 44 1.5 0.44 corn 12 1.2 0.12 alfalfa 99 2.1 0.99 4. discussion 4.1. ler and lerc values experimental studies of agroforestry (af) systems with tree species other than poplar or eucalyptus revealed ler values larger than one; for example, [19] reported an ler of 1.36 for the af system of alder and wheat. further studies also underlined that crop yields shrank under agroforestry (af) systems with a high tree density [20], which is due to the shading of the crops. pardon et al. [21] and zhao et al. [22] found that crop yields, compared to corresponding monocultures, shrank the closer the crop grew to af trees and the taller those trees were. this is compounded by findings from [23] that the photosynthetically active radiation significantly drops close to af trees. despite the lower crop yields in narrowly spaced af systems, those systems, in most cases, show an increased land equivalent ratio (ler). this can be explained by the high wood yield associated with the high tree density, which overcompensates for the loss of crop yields. in other words, the wood biomass yielded from such af systems implies a trade-off in terms of crop yield, which in most cases are food or fodder crops. this trade-off is underlined by the mostly negative values of lerc across af systems and crops, which indicate lower crop yields inside af systems compared to the respective monocultures. in the context of tree windbreaks, crop yields were found to be reduced in the neighborhood of windbreaks to a distance of 1 x tree height [24], 2 x tree height [25], and 3 x tree height [26], respectively, also due to the shade that impacted the given crops. in the case of tree windbreaks with tree row spacing of 100 m and more, crop yields do increase on average, while the annual growth of the single agroforestry (af) trees is also larger than that of trees in corresponding plantations. therefore, tree windbreak systems offer land equivalent ratio (ler) values of larger than one, albeit lower than the reviewed alley cropping systems, but without facing the trade-off of sacrificing food or fodder crop yields, which is underlined by positive lerc values across all crops (table 7). 4.2. discussion of agroforestry and sustainable biomass sourcing this second part of the discussion examines the changes in crop and tree yields of agroforestry (af) systems compared to the respective monocultures in light of the principles for sustainable biomass sourcing [4] that are listed in the introduction. wood biomass sourced from af systems is a renewable biomass because trees can be regrown after the preceding harvest of af trees. if the af systems are managed according to so-called good agricultural practices, agriculture and food sciences research, 2025, 12(1): 12-18 16 © 2025 by the author; licensee asian online journal publishing group which do not degrade soils and water resources, then the production of the wood biomass can be considered sustainably managed [5] to adhere to the first principle of bfa [4] for sustainable biomass sourcing. according to the results of the agroforestry (af) experiments listed in this study, food or fodder production is compromised when a given crop monoculture is converted into an af system with narrow spacing between trees, high tree density, and hence high yields of woody biomass. this is also reflected in the negative land equivalent ratio (lerc) values of those af systems, as listed in the results section. on the other hand, wood biomass offers an additional income to farmers, which may improve food affordability and the social and economic situation of local and rural communities, and therefore helps diversify farmers’ income [12]. if the overall social and economic situation of the local population is improved, then this second principle would be adhered to. the tree windbreak systems (table 7), however, do adhere to the second principle of bfa [4] for sustainable biomass sourcing, because food and fodder production is not compromised, but often even increased compared to a respective monoculture, while obtaining an additional benefit from the tree biomass. if plantations of fast-growing trees are planned in areas that have not been producing food or fodder, then those plantations can be enriched with an understory crop to form af systems that also adhere to the second principle, because additional food or fodder production is added to a given area and its population. af systems must not be established within the area of critical ecosystems in order to avoid directly causing the loss of critical ecosystems or habitats of high conservation value. af systems with negative lerc values, which compromise food and fodder production, may lead to land conversion to compensate for the loss of food or fodder production, which may indirectly result in the destruction of critical ecosystems or losses of high conservation value habitats. therefore, af systems with negative lerc values are prone to violate the third principle by bfa [4]. af systems offer a number of benefits to landscape resilience and are resilient to the impacts of climate change themselves. the potential of af systems to improve soil fertility, reduce erosion, improve water quality, and enhance the microclimate for crops [10, 11] strengthens landscape resilience and is delivered by af systems with both high and low ler or lerc values alike. af systems with a high tree density, which tend to deliver reduced crop yields compared to respective monocultures, offer a high potential to protect soils from erosion. gupta et al. [27] showed that water erosion of soils was significantly reduced under a poplar af system with tree spacing of 6.5 m x 4.3 m in punjab, india. additionally, in new zealand, poplars and willows have been planted on pastures to form an af system to control water erosion on slopes for the past 60 years [28, 29]. young eucalyptus plantations (tree age between 3 and 7 years) in brazil reduced soil erosion by more than 50% compared with bare soils on slopes of 20° steepness [30]. as a result, the older and larger the eucalyptus tree grew, the lower the soil losses became. furthermore, such systems can improve soil properties, such as infiltration for water, thereby contributing to mediating the water cycle and improving water quality [10]. af systems with high tree density and reduced crop yields compared to the corresponding crop monocultures tend to protect the soil better against erosion. if, in a given region, erosion control on cropland is an overarching goal, then introducing af and careful harvesting of its wood biomass would seem justifiable, as long as the affordability of food for the local population is not compromised. programs to control soil erosion, such as china’s so-called green for grain program, incentivized tree planting on mainly marginal cropland prone to erosion (see [31]). on the other hand, poplars and eucalyptus trees are substantial water consumers, so newly established af systems with high tree density may over-exploit local water resources and cause water stress in a given landscape, as was shown, for instance, for afforestation programs with poplars in northern china [32, 33]. af systems change the microclimate under which the accompanying crops grow through shade (reduced radiation), lower temperatures, increased air humidity, and reduced wind speed compared with respective monocultures. in particular, tree windbreaks are designed to reduce wind speed and thereby decrease crop water consumption, mechanical damage to crops, and wind erosion of cropland soils (alemu [34]). thevs et al. [35] showed that tree windbreaks of poplars at a spacing of 100 m or more reduced the water consumption of the whole af system compared to the corresponding crop grown as a monoculture. these effects on the microclimate make the af systems themselves more resilient to climate change, specifically increased temperatures, enhanced water stress, and extreme weather events, than corresponding crop monocultures, as detailed by quandt et al. [36]. thus, shading lowers air temperatures and crop water consumption, minimizes soil erosion and soil loss during heavy rainfall and/or flood events, and protects crops from strong winds. reduced wind speeds also correspond with lowered crop water consumption and a lower risk of crops suffering from water stress (weninger et al. [37]). the compliance with the principles for sustainable biomass sourcing by bfa [4] af systems is summarized in table 8. table 8. compliance of af systems with negative and positive lerc with the principles for sustainable biomass sourcing by bfa [4]. af system refers to af system. principle af system with high tree density and negative lerc af system with low tree density (tree stand breaks, field boundaries) and positive lerc. biomass is derived from renewable sources, and its production is sustainably managed. adheres to principle. adheres to principle. biomass sourcing does not adversely impact food security and affordability, and it maintains or improves social and economic conditions along with ecosystem services in producing communities. compromises food/fodder production but may provide additional income and, hence, improve social and economic conditions. ecosystem services are often improved (see explanation in the text to principle 4). adheres to principle biomass sourcing does not directly or indirectly result in the destruction of critical ecosystems or the loss of high conservation value habitats. adheres to the principle, as long as the af system is not established inside the area of critical ecosystems. it may result in indirect destruction of critical ecosystems or loss of high conservation value habitats. adheres to the principle, as long as the af system is not established inside the area of critical ecosystems. biomass sourcing contributes to landscape resilience and is resilient to the impacts of climate change. largely adheres to principles. may cause water stress in drylands. adheres to principle. agriculture and food sciences research, 2025, 12(1): 12-18 17 © 2025 by the author; licensee asian online journal publishing group 5. conclusion literature data on poplar and eucalyptus-based agroforestry (af) systems were reviewed to calculate land equivalent ratio (ler) and corrected land equivalent ratio (lerc) values across a range of regions, climates, crops, and af systems, which included alley cropping, plantations with a crop as understory vegetation, field boundaries, and tree windbreaks. af systems with narrow spacing and high tree density, i.e., alley cropping, plantations with a crop as understory vegetation, and field boundaries, as reviewed in this study, provide high wood biomass yields but often lower crop yields than the corresponding crops and trees grown as monocultures. in total, those af systems deliver more biomass than the corresponding crops and trees would deliver when grown as separate monocultures, which results in high ler values, as listed above in this study. yet, the lerc values of most of those af systems are negative, which is a result of the reduced crop yields within those af systems compared to the corresponding crop monocultures. in contrast, tree windbreak systems, in which the spacing between tree lines is much wider than the af systems above, often 100 m and more, show ler > 1 and lerc > 0. this indicates that the tree windbreak systems produce more total biomass than the corresponding crops and trees would deliver when grown as monocultures and that crop yields do not decrease within those af systems. given that tree windbreak systems also contribute to soil protection and often improve the microclimate, biomass sourced from those agroforestry (af) systems has a high potential to fulfill the criteria of sustainably sourced biomass. additionally, the af systems with negative lerc values have the potential to fulfill the criteria of sustainably sourced biomass if the overall income and socio-economic situation of the local population is improved by, for example, additional income from the woody biomass. if af systems with high tree density contribute substantially to landscape resilience, such as soil protection or regulating the water cycle, they also have the potential to fulfill the criteria of sustainably sourced biomass. if landscape resilience is an overarching goal in a given region, then af systems with high tree density should be considered to fulfill the criteria of sustainably sourced biomass. references [1] s. piotrowski, m. carus, and r. essel, "global bioeconomy in the conflict between biomass supply and demand," industrial biotechnology, vol. 11, no. 6, pp. 308-315, 2015. https://doi.org/10.1089/ind.2015.29021.stp [2] o. arodudu, b. holmatov, and a. voinov, "ecological impacts and limits of biomass use: a critical review," clean technologies and environmental policy, vol. 22, no. 8, pp. 1591-1611, 2020. https://doi.org/10.1007/s10098-020-01911-1 [3] unep, "global resources outlook 2024: bend the trend – pathways to a liveable planet as resource use spikes, international resource panel: nairobi," 2024. retrieved: https://www.resourcepanel.org/reports/global-resources-outlook-2024. 2024. 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https://doi.org/10.1590/s1413-70542013000200007 https://doi.org/10.1007/s40333-020-0091-9 https://doi.org/10.1111/j.1365-2664.2008.01605.x https://doi.org/10.1016/j.jaridenv.2020.104257 https://doi.org/10.3390/land8110167 https://doi.org/10.1016/j.cosust.2022.101244 https://dx.doi.org/10.1088/1748-9326/ac1d0d 10 © 2021 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 8, no. 1, 10-14, 2021 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2021.81.10.14 © 2021 by the authors; licensee asian online journal publishing group the effect of planting distance and bokashi from several types of organic fertilizers on the growth and yield of elephant ginger variety (zingiber officinale var. roscoe) desak ketut tri martini1 desak putu raka paramita2 ( corresponding author) 1faculty of agriculture, universitas udayana, indonesia. 2department of tropical biology, universitas gadjah mada, indonesia. abstract given the large market opportunity for ginger to meet consumption, it is necessary to seek various methods of increasing production, both in terms of quality and quantity. to achieve these result, ginger production in indonesia must be expanded, intensified, and diversified in an appropriate and economically sustainable manner. the goal of this study was to see how spacing and bokashi from various organic fertilizers affected the growth and yield of elephant ginger. this experiment used a factorial design with a randomized block design (rbd) of two factors. the first factor is the spacing (j), which has three levels: 40 cm x 20 cm spacing (j1), 40 cm x 30 cm spacing (j2), and 40 cm x 40 cm spacing (j3). the second factor is organic fertilizer bokashi (b), which is made up of four experiments: no bokashi (bt), pig manure bokashi (bb), cow dung bokashi (bs), and green manure bokashi (bh). the experimental results were statistically analyzed using analysis of variance. results showed that the interaction effect of distances and the type of bokashi with organic fertilizer has no significant difference (p 0.05) in the growth and yield of elephant ginger. the average net assimilation rate 90-105 dap, as well as the average plant growth rate 7590 dap were compared. the highest yield of fresh rhizomes per hectare was obtained in the at a 40 cm x 20 cm spacing. keywords: planting distance, organic fertilizers, growth, variance analysis, zingiber officinale var. roscoe, bokashi citation | desak ketut tri martini; desak putu raka paramita (2021). the effect of planting distance and bokashi from several types of organic fertilizers on the growth and yield of elephant ginger variety (zingiber officinale var. roscoe). agriculture and food sciences research, 8(1): 10-14. history: received: 10 march 2021 revised: 13 april 2021 accepted: 18 may 2021 published: 7 june 2021 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: both authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 11 2. material and methods ..................................................................................................................................................................... 11 3. result .................................................................................................................................................................................................. 11 4. discussion .......................................................................................................................................................................................... 12 5. conclusion and suggestion ............................................................................................................................................................ 14 references .............................................................................................................................................................................................. 14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2021.81.10.14&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2021.81.10.14&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/2968 https://orcid.org/0000-0003-0105-1005 https://orcid.org/0000-0002-9663-1215 agriculture and food sciences research, 2021, 8(1): 10-14 11 © 2021 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study provided a piece of information about the effect and significance of planting distance and the usage of bokashi on the growth and yield of elephant ginger variety (zingiber officinale var. roscoe). 1. introduction the ginger plant (zingiber officinale var. roscoe) is a pseudo-stemmed plant that grows upright and belongs to the zingiberaceae family. the economic value of this plant is found in its rhizome. whether fresh or processed, ginger rhizome can be used for various purposes at a low cost [1]. given the immense market opportunity for ginger to meet consumer demand, it is necessary to seek various production methods, both in quality and quantity. ginger production in indonesia must be expanded, intensified, and diversified in an appropriate and economically sustainable manner [2]. setting the spacing needs to be considered because it is one way to increase the yield of broad unity rhizomes. the spacing is closely related to the plant's need for nutrients and solar radiation. in selecting the spacing, it must be considered that plant competition may occur, especially in nutrients, water, and radiation, which in turn can affect plant growth. in addition to spacing, fertilization is required to increase ginger yield, which aims to increase soil fertility, specifically in organic fertilizer bokashi, where the addition of organic matter into the soil is required for the life of microorganisms in the soil. bokashi results from em4 technology fermenting organic matter (green manure, manure fertilizer, compost, or plant residues), which can then be used as organic fertilizer to fertilize the soil and increase plant growth and production [3]. plant growth is helped by proper spacing and the application of organic fertilizer bokashi. although nutrient availability is limited, the spacing between individual plants determines the level of competition. meanwhile, the use of organic fertilizer bokashi and directly adding nutrients to the soil increases nutrient availability. plants can thus use the nutrients in the soil to grow. this study aimed to see how spacing and bokashi from various organic fertilizers affected the growth and yield of elephant ginger. the following hypotheses can be drawn from the objective: setting the spacing at 40 cm x 20 cm will increase the growth and yield of elephant ginger, giving bokashi cow dung can increase the growth and yield of elephant ginger, and the interaction of 40 cm x 20 cm spacing, bokashi, and cow dung can increase the growth and yield of elephant ginger. 2. material and methods 2.1. study area and material this is a paddy field experiment conducted in pitera village, penebel district, tabanan regency at an altitude of approximately 500 meters above sea level. 9-month-old elephant ginger rhizome, green manure bokashi, pig manure bokashi, cow dung bokashi, and soil as a planting medium were used. thiodan 36 bc and dithane m-45 are used to control pests and diseases. this is a factorial experiment with two factors and a basic randomized block design (rbd). 2.2. experimental set up the study area is a paddy field experiment conducted in pitera village, penebel district, tabanan regency, at approximately 500 meters above sea level. 9-month-old elephant ginger rhizome, green manure bokashi, pig manure bokashi, cow dung bokashi, and soil as a planting medium were used. thiodan 36 bc and dithane m-45 are used to control pests and diseases. data analysis is a factorial experiment with two factors and a basic randomized block design (rbd). the variables to be observed were as follows: the number of tillers per clump, the fresh weight of rhizomes per clump, the weight of oven-dry stems per clump, the weight of oven-dry leaves per clump, the weight of oven-dry roots per clump, the weight of oven-dry roots per clump, the weight of oven-dry roots per clump, the weight of oven-dry roots per clump, the weight of oven-dry roots per clump, the weight of oven clumps, harvest index, wet rhizome yield per hectare, oven-dry rhizome yield per hectare, leaf area index, average swallowed assimilation rate. average plant growth rates are all variables to consider. at the ages of 75, 90, 105, and 120 days, observations were made regularly. 2.3. data analysis the experimental results were statistically analyzed using analysis of variance. if there is a significant difference (f count> f table 5% or 1%), the lsd test is used to determine a different treatment [4]. regression analysis was used to determine the relationship between the various spacing treatments [4]. correlation analysis was performed to determine the closeness of the relationship between the treatment given and the observed variables or the close relationship between the variables. 3. result according to the findings of the statistical analysis, the interaction between plant spacing and organic fertilizer bokashi type (jxb), spacing (j), and bokashi type (b) had a significant effect on the observation variable of the elephant ginger plant. agriculture and food sciences research, 2021, 8(1): 10-14 12 © 2021 by the authors; licensee asian online journal publishing group table-1. the significance of the interaction effect between planting distance with organic fertilizer bokashi type (jxb) on the observation of elephant ginger’s variable no observation variable treatment jxb j b 1 number of tillers per clump (fruit) ns ** ** 2 leaf area index (ild) a. age 75 dap b. age 90 dap c. age 105 dap d. age 120 dap ns ns ns ns ** ** ** ** ** ** ** ** 3 average net assimilation rate (lab) a. age 75-90 dap b. age 90-105 dap c. age 105-120 dap ns * ns ** ** ** ** ** ** 4 average plant growth rate (lpt) a. age 75-90 dap b. age 90-105 dap c. age 105-120 dap ** ns ns ** ** ** ** ** ** 5 fresh rhizome weight per clump (gram) ** ** ** 6 fresh rhizome yield per hectare (ton) ns ** ** 7 weight of oven dry rhizome per clump (gram) ns ** ** 8 yield of oven dry rhizome per hectare (ton) ns ** ** 9 oven dry rod weight per clump (gram) ns ** ** 10 weight of oven dry leaves per clump (gram) ns ** ** 11 weight of oven dry roots per clump (gram) ns ** ** 12 oven dry stable weight per clump (gram) ns ** ** 13 harvest index (%) ns ** ** information: note: ns = no significant effect (p> 0.05). * = significant (p≤0.05). ** = very significant effect (p≤0.01). dap= days after planting. according to table 1, the interaction between plant spacing and organic fertilizer bokashi (jxb has no significant effect (p> 0.05) on most of the observed growth and yield variables of ginger, with the exception of the weight of fresh rhizomes per clump. the average net assimilation rate between 90 and 105 dap, as well as the average growth rate between 75 and 90 dap. the spacing treatment (j) and the organic fertilizer bokashi treatment (b) had a significant (p0.05) to very significant (p0.01) effect on all observed variables. 4. discussion except for fresh rhizome weight per clump, the average net assimilation rate (lab) of age 90-105 dap, and the average plant growth rate (lpt) of age 75-90 dap, the interaction effect of spacing and organic fertilizer bokashi type (jxb) showed no significant difference (p> 0.05) on most of the observed variables. by giving bokashi cow dung (bs) with a spacing of 40 cm x 40 cm (bsj3), the average weight of fresh rhizomes per clump, the average net assimilation rate (lab) at 90-105 dap, and the average plant growth rate (lpt) aged 75-90 dap were obtained in the spacing (j) and organic fertilizer bokashi treatment (b), namely 247.56, 0.69, and 0.21 grams. the interaction occurs because the level of competition against light, nutrients, water, and space is lower at the 40 cm x 40 cm spacing and is also supported by cow dung bokashi, which has a higher nutrient content compared to bokashi pig manure and green manure bokashi. as a result, the combined influence appears to affect the growth of the elephant ginger plant positively. the spacing effect on oven-dry rhizome yield per hectare was highly significant (p0.01). the highest average yield of fresh rhizomes per hectare was at a spacing of 40 cm x 20 cm (j1), namely 20.49 tons or a 36.60 percent and 57.40 percent increase, respectively, over the treatment spacings of 40 cm x 30 cm (j2) and 40 cm x 40 cm (j3). the greater the spacing, the lower the weight of fresh young harvest rhizomes per hectare. the spacing effect on oven-dry rhizome yield per hectare was highly significant (p0.01). the highest average yield of fresh rhizomes per hectare was at a spacing of 40 cm x 20 cm (j1), namely 20.49 tons or a 36.60 percent and 57.40 percent increase, respectively, over the treatment spacings of 40 cm x 30 cm (j2) and 40 cm x 40 cm (j3). the greater the spacing, the lower the weight of fresh young harvest rhizomes per hectare. meanwhile, the spacing treatment with rhizome yield per hectare showed a quadratic relationship with the equation y = 0.0175 x2 1.4267 x + 42.009 and a coefficient of determination (r2) of 0.911. with a high population, there will be maximum utilization of solar radiation at the start of growth to affect the increase in the rate of photosynthesis, and the formation of more dry matter. furthermore, high yields at denser spacing or higher plant populations per hectare influence the high leaf area index. the increase in population is followed by an increase in shade between plant leaves, as evidenced by the increasing average leaf area index (ild) with narrower spacing. the average leaf area index (ild) values at 75, 90, and 105 days after planting were higher at a spacing of 40 cm x 20 cm (j1), i.e., 0.45, 0.50, and 0.61, compared to a spacing of 40 cm x 30 cm (j2), that is 0.40, 0.48, and 0.60, and a spacing of 40 cm x 40 cm (j3), i.e., 0.39, 0.50, and 0.59. the results showed that the plant population at a spacing of 40 cm x 20 cm (j1) is higher than other spacings, because in a dense population, it allows for the best use of light at the beginning of growth. with a high population at the start of growth, there will be maximum utilization of solar radiation to affect the increase in the rate of photosynthesis, and the formation of more dry matter. the high leaf area index is influenced by the high yield at closer spacing. the relationship between fresh rhizome yields per hectare and average leaf area indexes aged 75, 90, 105, and 120 dap (r = 0.645 **, 0.443 **, 0.433 **, and 0.533 **) demonstrates this. the level of relative competition was the lowest at 40 cm x 40 cm (j3) due to a high number of fresh weight and oven-dry weight of rhizomes per clump at a wider space j3. this resulted in better development of plant agriculture and food sciences research, 2021, 8(1): 10-14 13 © 2021 by the authors; licensee asian online journal publishing group organs. the higher average oven-dry weight of roots, leaves, and stems per clump in plants planted with a spacing of 40 cm x 40 cm (j3) demonstrates this. plant growth, particularly leaf growth, will result in a large leaf area per plant. the presence of numerous leaves, supported by healthy roots and the availability of adequate growth factors, will result in increased photosynthetic activity and photosynthesis. it can be seen from the net assimilation rate and growth rate of plants aged 75-90, 90-105, and 105-120 dap at a spacing of 40 cm x 40 cm (j3), which is relatively higher than the spacings of 40 cm x 30 cm (j2) and 40 cm x 20 cm (j1). furthermore, good growth will support high yield per plant, as evidenced by the very significant positive correlation between the average net assimilation rate and the average growth rate of plants aged 90-105 and 105-120 dap oven-dry weight of rhizomes per plant. clumps, namely r = 0.330 *, 0.339 * *, and 0.491 * *, 0.483 * *. increased average leaf area index (ild) can improve net assimilation rate (lab) and plant growth rate (lpt) in plants aged 75-90, 90-105, and 105-120 dap, with the highest average obtained at a spacing of 40 cm x 40 cm (j3). the high lpt was supported by the high average oven-dry leaf weight per clump, oven-dry stem weight per clump, and oven-dry root weight per clump at a spacing of 40 cm x 40 cm (j3), which were 4.56, 3.16, and 3.77 grams, respectively. a significant correlation between oven-dry leaf weight per clump and average plant growth rate (lpt) aged 90-105, 105-120 dap (r = 0.654 * * and 0.585 * *) was found, the weight of oven-dry stems per clump with a plant growth rate (lpt) of 90-105, 105-120 dap (r = 0.670 * * and 0.646 * *) and oven-dry root weight per clump with a plant growth rate (lpt) of 90-105, 105-120 dap (r = 0.592 * * and 0.602 * *). according to monteith [5] plant growth is equivalent to photosynthesis rate. meanwhile, the rate of photosynthesis is determined by the amount of light that plants are able to intercept [6]. plants will produce more dry matter as a result of the relationship between plant growth rate and leaf area index. the effect of organic fertilizer type bokashi on fresh rhizome yield per hectare was significant (p0.01), as was the effect on oven-dried rhizome yield per hectare (p0.05). cow dung bokashi (bs) produced the highest average yield of fresh rhizomes per hectare, namely 19.40 tons, representing an increase of 22.48 percent, 24.74 percent, and 39.70 percent, respectively, when compared to pig manure bokashi (bb), bokashi green manure (bh), and no bokashi (bt). the high weight of fresh rhizomes per clump, which was also obtained in the treatment of cow dung bokashi (bs), supported the increase in yield of fresh rhizomes per hectare, i.e., 218.79 grams. when compared to pork dung bokashi (bb), green manure bokashi (bh), and no bokashi (bt), the number increased by 19.56 percent, 24.10 percent, and 34.63 percent, respectively. it was demonstrated by a highly significant positive correlation (r = 0.657 * *) between fresh rhizome yield per hectare and fresh rhizome weight per clump. the increased number of tillers per clump aided the increase in fresh rhizome weight per clump. when cow dung bokashi (bs) was used, the number of tillers per clump increased by 5.25 percent, 27.71 percent, and 40.00 percent, respectively, when compared to pork manure bokashi (bb), green manure bokashi (bh), and no bokashi (bt).) whose respective scores were 4.44, 4.11, and 3.75. it was demonstrated by a highly significant positive correlation (r = 0.838 * *) between the number of tillers per clump and the weight of fresh rhizomes per clump. cow dung bokashi (bs) produced the highest yield of oven-dried rhizome per hectare, namely 0.60 tons, representing an increase of 16.30 percent, 21.67 percent, and 32.54 percent, respectively. in comparison, 0.52 tons of pork manure bokashi (bb), 0.50 tons of green manure bokashi (bh), and 0.46 tons of no bokashi (bt) were produced. the increased oven-dry rhizome yield and fresh rhizome yield per hectare were supported by a 5.29gram increase in oven-dry rhizome weight per clump in cow dung bokashi (bs). the number has increased by 7.24 percent, 21.35 percent, and 32.91 percent, respectively, when compared to pork manure bokashi (bb), bokashi green manure (bh), and without bokashi (bt). in the bokashi treatment of cow dung (bs), the increase in oven-dry rhizome weight per clump was supported by an increase in fresh rhizome weight per clump of 218.79 (r = 0.465 * *). increases in the average leaf area index (ild) of 75, 90, 105, and 120 dap in the treatment of organic fertilizer bokashi (b) influenced the weight of leaves, roots, and oven-dry stems per clump in the treatment of cow dung bokashi (bs), as well as the height of fresh rhizome weight and oven-dry rhizome weight per clump in the treatment of cow dung bokashi (bs). as evidenced by a highly significant positive correlation between oven-dry leaf weight per clump and average leaf area index (ild) at 75, 90, 105, and 120 dap (r = 0.517 **, 0.635 **, 0.724 **, 0.713 * *), there was a significant correlation between oven-dry stem weight per clump and the mean leaf area index (ild) at 75, 90, 105 and 120 days after planting (r = 0.582 **, 0.668 **, 0.744 **, 0.735 * *), there was a significant correlation between fresh weight of rhizomes per clump and the mean leaf area index (ild) aged 75, 90, 105 and 120 dap (r = 0.470 **, 0.553 **, 0.504 **, 0.476 ** ), a significant correlation between oven dry rhizome weight per clump with mean leaf area index (ild) at 75, 90, 105 and 120 dap (r = 0.681 **, 0.433 **, 0.335 **, 0.470 ** ). increased light interception from a higher leaf area index (ild) would boost net assimilation. cow dung bokashi (bs) had the highest average net assimilation rate (lab) aged 75-90, 90-105, 105-120 dap of 0.31, 0.69, 0.46 mg/cm2/hr1 or increased respectively to 29.07 percent, 40.82 percent, and 12.20 percent compared to bokashi pig manure (bb) 34.78 percent, 43.75 percent, 15 percent compared to green manure bokashi (bh) and 40 (bt). in addition to the increase in the average leaf area index, the increase in oven-dry leaf weight per clump in the treatment of cow dung (bs), namely 5.82 grams or in percentage i.e., 48.85 percent, 50.0 percent, and 80.0 percent compared to the provision of green manure bokashi (bh), was also due to the increase in oven-dry leaf weight per clump in the treatment of cow dung (bs), namely 5.82 grams or an increase of 48.85 percent (bt). the use of bokashi, an organic fertilizer, can help plants grow faster. the increase in plant growth (lpt) rate reflects this. when cow dung bokashi (bs) was provided, the average plant growth rate (lpt) at the age of 75-90 dap increased by 40 percent, 75 percent, and 110 percent, and the plant growth rate (lpt) at the age of 90-105 dap increased by 40 percent, 75 percent, and 110 percent, respectively. compared to the treatment of bokashi pig manure (bb), bokashi green manure (bh), and no bokashi, plant growth rate (lpt) aged 105-120 dap increased by 41.38 percent, 51.85 percent, and 95.24 percent, respectively (bt). plant growth rates increased by cow dung bokashi (bs) can increase net assimilation rate and leaf area index. the amount of dry matter produced by plants increases as the plant growth rate (lpt) increases. increased leaf weight, roots, stems, and oven-dry stems per clump in the organic fertilizer bokashi treatment also contributed to the increase in average plant growth rate (lpt). evidenced by a significant correlation between oven-dry leaf agriculture and food sciences research, 2021, 8(1): 10-14 14 © 2021 by the authors; licensee asian online journal publishing group weight per clump with the average plant growth rate (lpt) aged 90-105 dap and 105-120 dap (r = 0.654 **, and 0.585 **), a very positive correlation. the oven-dry root weight per clump and the average plant growth rate (lpt) of 90-105 dap and 105-120 dap (r = 0.592 ** and 0.602 **), the positive correlation was very significant between oven-dry weight per clump. clumps with an average plant growth rate (lpt) of 90-105 dap and 105-120 dap (r = 0.670 **, and 0.646 **), and a significant correlation between oven-dry weight per hill plant growth (lpt) 90-105 dap and 105-120 dap (r = 0.652 **, and 0.620 **). the availability of nutrients required by plants to carry out physiological and biochemical processes in the plant body encourages good vegetative growth. the use of manure can provide macro and micronutrients to plants while also developing the lives of microorganisms in the soil [7]. furthermore, the presence of these microorganisms is said to accelerate the weathering process in the soil, increasing soil fertility. it is aided by the high organic matter content of manure. furthermore, bokashi cow manure improves soil physical properties by increasing groundwater buffering capacity, soil water content, soil permeability, and aeration while decreasing the influence of surface runoff and erosion. cow dung bokashi can increase yield due to cow dung's high organic matter content in the soil. cow dung fertilizer contains more nutrients than pig and green manure, particularly nitrogen content, where the nitrogen content in the experimental soil is shallow, requiring plants to rely heavily on cow dung bokashi. nitrogen is required to produce amino acids, the formation of proteins, the formation of new cells, cell division, cell enlargement, tissue growth, stem elongation, leaf growth, and vegetative growth in general—nitrogen aids in forming leaf chlorophyll molecules, which serve as a platform for the photosynthesis process. the activity level of microorganisms in each treatment varies due to differences in chemical composition or nutrient content in the organic matter [8]. moreover, variations in the activity of these microorganisms will affect the rate of carbon dioxide release and the organic compounds produced. giving bokashi can boost the population of beneficial microorganisms in the soil to the point where it outnumbers harmful microorganisms. with this dominant population, these microorganisms will play a more significant role in organic matter decomposition and increase the availability of nutrients that plants can use to grow. 5. conclusion and suggestion based on the results and discussion, it is possible to conclude that the interaction between plant distance and organic fertilizer type has no significant effect on all observation variables of elephant ginger, with the exception of fresh rhizome weight per clump. the average net assimilation rate (lab) is 90-105 dap, with a 75-90 dap plant growth rate. cow dung bokashi yields the highest fresh rhizome yield per hectare. elephant ginger has a high growth rate of about 20.5 percent. the yield of oven-dry rhizomes increased by 36.60 percent, 57.40 percent, and 33.20 percent, 60.33 percent when compared to spacings of 40 cm x 30 cm and 40 x 40 cm. the discussion and conclusions that have been described can be given and or with a spacing of 40 cm x 20 cm to obtain high yields of elephant ginger and cow dung bokashi. as a consideration, more research should be conducted, paying special attention to the dose of bokashi cow dung used and the treatment of narrower spacing on the same land conditions. references [1] f. b. paimin and murhananto, cultivation, processing, ginger trade. jakarta: penerbit penebar swadaya, 1991. [2] h. b. santoso, elephant ginger. yogyakarta: kanisius, 1994. [3] g. j. umpel, experience of effective microogranism technology application. jakarta: national seminar on agriculture, 1997. [4] i. m. n. tenaya, "determining the optimal population using the fan design method on cabbage in lowlands," scientific report, faculty of agriculture, udayana university, unpublished, 1986. [5] j. l. monteith, "climate and the efficiency of crop production in britain," philosophical transactions of the royal society of london. b, biological sciences, vol. 281, pp. 277-294, 1977.available at: https://doi.org/10.1098/rstb.1977.0140. [6] b. l. siregar, "effect of population level and nitrogen dosage on growth and production of ginger (zingiber officinale rosc.)," rhino harvested young, scientific report, faculty of agriculture, bogor agricultural university (ipb)1991. [7] m. m. sutedjo and a. g. kartosapoetra, fertilizer and fertilization methods. jakarta: pt bina aksara, 1988. [8] w. agata, "the characteristics of dry matter and yield production in sweet potato under field conditions," in sweet potatoproceedings of the first international symposium, r.e. villareal and t.d. griggs, eds. taiwan: avrdc, 1981, pp. 119–127. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 46 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 46-50, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.46.50 © 2020 by the authors; licensee asian online journal publishing group faba bean variety development for yield, quality, and disease resistance for water logged vertisol areas-registration of a faba bean variety named ‘ashebeka’ deressa tesfaye1 gizachew yilma2 gebeyew achenif3 temesgen abo4 tadese sefera5 tamene temesgen6 (corresponding author) 1,2,3,4,5ethiopian institute of agricultural research, kulumsa agricultural research center, asella, ethiopia. 6international livestock research institutes, addis abeba, ehtiopia. abstract a faba bean (vicia faba l.) variety named ashebeka with a pedigree designation of eh01075-4 has been released in 2015 by kulumsa agricultural research center in ethiopia. the variety is best adapted to waterlogging vertisol environments of ethiopia with altitudes ranging from 1900 to 2800 meters above sea level. the variety was developed through hybridization between n86108-5 and bpl1297-1. the candidate variety has been tested at four locations (arsi robe, sagure, ambo and inewari) representing major waterlogging vertisol environments of the country during 2012 and 2013 main cropping seasons making eight test environments. the variety is mainly characterized by its higher grain yield and heavier seed size (885g 1000 seeds-1) than the standard checks and all the remaining genotypes in the trail. the variety ashebeka showed 34% seed size advantage over hachalu, the popular best standard check recently released for water logging vertisol production areas. based on selected a parametric method that includes yield rank and stability, ashebeka showed better performance stability across the test environments and over years than the standard checks, hachalu and walki. the variety showed better overall agronomic performance, and moderately resistance to major faba bean disease including root rot, rust and chocolate spot than the standard checks in the trial, and could be cultivated across mid to high altitude agro-ecologies with waterlogging vertisol conditions of the country. keywords: disease resistance, national yield trail, water-logging, variety development, vertisol, vicia faba l. citation | deressa tesfaye; gizachew yilma; gebeyew achenif; temesgen abo; tadese sefera; tamene temesgen (2020). faba bean variety development for yield, quality, and disease resistance for water logged vertisol areas—registration of a faba bean variety named ‘ashebeka’. agriculture and food sciences research, 7(1): 46-50. history: received: 14 january 2020 revised: 19 february 2020 accepted: 24 march 2020 published: 16 april 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: we would like to thank staff members of the breeding and genetics research units of kulumsa, holeta, ambo and debre-birhan agricultural research centers for their unreserved efforts in field trail management and data collection during the experimental period. we are thankful to ethiopian institute of agricultural research and alliance for a green revolution in africa (agra) for funding the research throughout the varietal development process. we would also like to thank the international center for agricultural research in the dry areas (icarda) for providing us with one of the parental lines. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 47 2. materials and methods ................................................................................................................................................................... 47 3. result and discussions ................................................................................................................................................................... 48 4. variety maintenance ....................................................................................................................................................................... 50 5. conclusion ......................................................................................................................................................................................... 50 references .............................................................................................................................................................................................. 50 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.46.50&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.46.50&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://asianonlinejournals.com/index.php/aesr/article/view/1518 https://orcid.org/0000-0003-1713-9693 https://orcid.org/0000-0001-8655-6015 https://orcid.org/0000-0002-3860-9837 https://orcid.org/0000-0002-0449-033x https://orcid.org/0000-0001-9060-5756 agriculture and food sciences research, 2020, 7(1): 46-50 47 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the study was conducted at four location (arsi robe, sagure, enewari and ambo) agricultural research center to assess the genetic potential among the faba bean lines/ genotypes using rcbd designs. 1. introduction faba bean (vicia faba l.) is the most important pulse crop in terms of both area coverage and volume of annual production in ethiopia. presently, it occupies about 427,697 hectares of land with an annual national production of 878,011 tons, with a productivity of 2.05 tons ha-1 [1]. ethiopia is the first producer of faba bean in africa and the second in the world next to the people’s republic of china [2]. the crop is mainly cultivated in mid and highaltitude areas, with an elevation ranging from 1800-3000 meters above sea level [3, 4]. faba bean were also known by n fixation crop which is considered as high among the grain legumes [57]. it is cultivated for the purpose of both human food and animal feed, especially for humans as nutritional and medicinal values [8, 9]. the crop contributes to the smallholder farmers as a source of protein in both rural and urban areas [8] and is an important source of income for farmers. it is so far that, the commencement of faba bean breeding in ethiopia was in the 1950’s with the establishment of arsi rural development unit (ardu) followed by the then alemaya college of agriculture, now haramaya university [10]. production of faba bean in the country couldn’t attain the maximum yield potential of the crop because of biotic and abiotic stresses that collectively cause great yield losses. the major constraints to increased faba bean production are lack of improved varieties, chocolate spot (botrytis fabae), root-rot, faba bean gall and rust diseases, insect, pests, broad leaf and grass weeds, water-logging, and cold and drought weather conditions [11-14]. therefore, faba bean research program in ethiopia has been focused on increasing production and productivities of the crop through developing and promoting improved cultivars with high and stable yield, and resistant/tolerant to biotic and abiotic stresses [7, 15]. the potential of faba bean production in ethiopia is its suitability as rotation crop in the cereal based system of not only potential production areas, but also in water-logged vertisol production environments of the country that has been mainly with a problems of root rot disease on faba bean. so considering the water-logged vertisol production environments alone, faba bean is among the most important pulse crop occupying 14.6% of the cultivated vertisols in the country. however, water logging due to a nature of soil and poor drainage are predominant factor constraining an increase in production on this soil. breeding of productive genotype under water logging was the only option to improving the productivity of faba bean under water logging condition. in order to convene the demand of the farmer’ lived in comparable agro ecologies, considerable efforts have recently been paid by faba bean breeding program to develop a variety resistant/tolerant to water-logging conditions leading to a release of a number of varieties. the purpose of this paper is therefore; to describe the merits of recently released faba bean (vicia faba l.) variety named ‘ashebeka’, developed for water-logging vertisol production environments of the country. 2. materials and methods 2.1. breeding procedures crossing was done between n86108-5 x bpl1297-1 introduced from icarda and resulted pedigree designation of eh01075-4 at holeta agricultural research center during 2002 cropping season. screen houses were routinely used in the early generations, i.e., f1, f2, f3 and f4, of a breeding cycle to prevent insects from causing cross-pollination. during these phases, pedigree selection was made for traits with high heritability such as seed size, grain yielding ability, plant habit, time of flowering and resistance to major diseases such as black rootrot, chocolate spot and rust. twenty-two elite individual lines selected from the f5 generation were promoted and evaluated at multilocations for yielding ability, seed size, major disease reactions and stability in a preliminary variety trial (pvt) conducted during the 2010/11 cropping season. from this trial, 12 promising genotypes were promoted and evaluated in a national yield trial (nyt) along with two recently released standard checks walki and hachalu at four representative waterlogging vertisol production areas of the country during 2012 and 2013 main cropping seasons. the locations where the trials conducted include sagure, ambo, arsi robe and enewari. each year at each location was considered as a separate environment, making a total of eight test environments for this study. biophysical description of the four test locations are given in table 1. finally, eh 00101-2 and eh01075-4 were selected as the most promising candidate varieties and evaluated along with two best standard checks, hachalu and walki, on 10 m x 10 m plots by the national variety release technical committee at 4 water-logging vertisol locations, each one on-research station and two on-farm fields during the 2014 cropping season. eventually, eh01075-4 was recommended for commercial production and named ashebeka. 2.2. description of experimental site table-1. biophysical description of the ten experimental locations used for national yield trial. test locations geographical position altitude (m.a.s.l) average rainfall (mm) temperature (oc) soil texture soil ph latitude longitude min max sagure 07012’n 39020’e 2300 620 5.8 23.6 clay-loam 6.2 a/robe 08005’n 39010’e 2200 820 10.5 22.8 dark-clay loam 6.0 enewari 07005’n 39030’e 2780 1010 7.9 16.6 clay-loam 5.0 ambo 08058’n 38014’e 2400 975.5 6.05 22.41 dark-clay 4.9 2.3. experimental layout the experimental layout for multilocation national yield trial (nyt) was arranged in a randomized complete block design with four replications. each experimental plot has 4 rows with 4 m long and arranged with inter and intra block spacing of 1.5 m and 0.6 m, respectively. the seeds were sown at the rate of 50 seeds per row with 8 cm agriculture and food sciences research, 2020, 7(1): 46-50 48 © 2020 by the authors; licensee asian online journal publishing group plant to plant and 40 cm row to row spacing. fertilizer was applied to each plot at the rate of 18 kg n and 46 kg p2o5 ha-1 in the form of diamonium phosphate at planting. other agronomic practices were treated as nonexperimental variables and applied uniformly to the entire experimental area. for data analysis, grain yield from net plot size of 3.2 m2 was converted into kg ha-1 at 10% standard grain moisture contents. 2.4. statistical analysis data for grain yield and other important agronomic traits were subjected to analysis of variance (anova) using the proc glm procedure of sas version 9.0 [16] to determine the existence of significant differences between faba bean genotypes. proc mean procedure of sas was used to generate means of traits by genotypes and environments. comparison of the main effect was performed using lsd at 5% probability level. selected stability parameters that encompass both yield and stability were selected and computed to estimate the genotypic yield stability across test environments. for example, a new approach, known as genotypic selection index (gsi), was used by taking the ammi stability value (asv) and mean grain yield into consideration. another nonparametric stability method, a stratified ranking technique of fox, et al. [17] where a genotype usually found in the top third for mean performance compared to all genotypes tested across environments is considered. all the stability parameters were computed using c a comprehensive sas stability program developed by hussein, et al. [18]. 3. result and discussions result of analysis of variance shows highly significant differences (p≤0.01) among genotypes for gain yield, seed size, pods plant-1 to significant (p≤0.05) for plant height table 2. likewise, test environments exerted highly significant (p≤0.01) effects on the important agronomic traits including grain yield, 1000 seed weight, number pods plant-1 and plant height. the interaction effects of environments and genotypes were highly significant (p≤0.01) for grain yield and seed size and significant at (p≤0.05) probability level for number of pods plant-1 and plant height table 2. significant differences were observed between genotypes across environments for chocolate spot (p≤0.01), root rot (p≤0.01), and rust (p≤0.05). moreover, highly significant differences (p≤0.01) observed among the test environments for all fusarium root-rot, chocolate spot and rust diseases table 2 indicating that the test environments were differently influenced the reaction of the test genotypes towards the diseases. table-2. mean square significance of grain yield, 1000 seed weight, number of pods plant-1, plant height, and root-rot, chocolate spot and rust diseases of 12 faba bean genotypes tested across four locations during 2012 and 2013 cropping seasons. source of variation mean squares grain yield (kg ha-1) seed weight (g) pod per plant plant height (cm) root rot chocolate spot rust environment 86956763** 103324** 2130** 29552 ** 25.9** 11.6** 58.6** block(environment) 810897** 5149ns 134** 486** 1.6** 2.4** 0.8ns genotype 919193** 278879** 84** 258* 1.6** 3.3** 1.2* genotype x environment 774559** 6210** 24* 140* 0.7ns 0.7* 0.5ns mean 3455 855 13.3 123 2.5 4.5 2.9 cv (%) 16.9 7.1 31.2 8.0 32.2 15.2 28.2 r2 0.89 0.85 0.82 0.90 0.62 0.65 0.73 3.1. varietal characteristics the newly released faba bean variety ashebeka is characterized by an indeterminate growth habit having white with black spots flower color. the seed coat and cotyledon colors are light green and ceramic respectively. its average seed weight is 885g 1000 seeds-1, which is heavier than the seeds of all other improved faba bean varieties released so far for water logging vertisol production areas of the country table 3. the average number of days required by the variety to reach its 50% flowering and 95% physiological maturity were 58 and 151, respectively table 3. the average plant height and number of pods plant-1 of ashebeka are 125 cm and 13, respectively table 3. the appropriate planting date for this variety would range from mid-june to early july. for a better harvest the variety must receive 46 kg ha-1 p2o5 and 18 kg ha-1 n at sowing. table-3. mean performance of selected agronomic traits of 12 faba bean genotypes tested across eight environments during 2012 and 2013 cropping seasons. genotypes dtf dtm pht ppl tsw chs r rr eh 01075-4 58 151 125 13 885 4.48 2.71 2.13 eh 00098-2 58 151 126 13 856 4.96 3.04 2.46 eh 00131-3 57 150 120 13 882 4.92 3.08 2.75 eh 00101-2 57 145 123 14 905 4.63 2.92 2.58 eh 01036-2 58 152 121 12 806 4.63 2.83 2.50 ekcsr 010 57 150 123 13 901 4.75 2.75 2.88 ekls 0101 58 151 124 13 894 4.83 3.29 2.92 eklss 020 57 144 127 13 857 4.38 2.79 2.17 eh 06007-4 60 145 124 10 1075 3.83 2.46 2.33 eh 06028-1 59 149 116 14 861 3.83 2.63 2.71 hachalu 58 151 124 16 662 4.58 3.04 2.42 walki 58 149 123 17 677 4.63 2.83 2.46 mean 58 149 123 13 855 4.54 2.86 2.52 lsd (<0.05) 1.32 3.0 4.81 2.04 34.66 0.39 0.46 0.46 cv (%) 4.3 3.8 7.9 31.1 7.1 15.2 28.2 32.2 note: keys: dtf= days to flowering, dtm= days to maturity, pht= plant height, ppl= number of pod per plant, tsw= thousand seed weight (g), gydh= grain yield (kg ha-1), chs = chocolate spot (1-9 scale), r=rust (1-9 scale), and rr= root rot (1-9 scale) agriculture and food sciences research, 2020, 7(1): 46-50 49 © 2020 by the authors; licensee asian online journal publishing group 3.2. yield and quality performance the average grain yield performance across environments of ashebeka is ranged from the lowest of 1245 kg ha-1 at arsi-robe in 2012 to the highest of 6438 kg ha-1 at enewari in 2013 cropping season, with mean overall environment of 3756kg ha-1 which is significantly higher than the best standard check, hachalu (3470kg ha-1), in the trial table 4. the newly released variety has yield advantage of 286 kg ha-1 (8%) and 393 kg ha-1 (12%) over the two standard checks, hachalu and walki, respectively table 4. ashebeka has produced 223g (33%) and 208g (31%) seed size advantage over the best standard checks, hachalu and walki, respectively table 4. table-4. mean grain yield (kg ha-1) of the released faba bean variety ashebeka tested over 8 environments during (2012-2013) cropping season as compared to the two standard checks. genotypes sagure 2012 arsi-robe 2012 inewari 2012 ambo 2012 sagure 2013 arsi-robe 2013 inewari 2013 ambo 2013 mean ashebeka 4652 1245 3610 3274 4597 2965 6438 3264 3756 hachalu 3890 1345 2983 3505 4154 2816 5440 3627 3470 walki 4249 1281 2514 3406 4073 2980 5074 3327 3363 mean 4392 1313 2932 3164 4298 2697 5723 3007 3455 lsd (< 0.05) 791 437 864 623 1226 780 869 890 287 cv (%) 12.2 23.1 20.5 13.7 19.8 20.1 10.6 20.6 16.9 3.3. performance stability and adaptation domain the variety ashebeka was released for the mid-to-high altitude agro-ecologies of the country with water logged vertisol condition and receiving 700-to-1100 mm average annual rainfall. it is well adapted to water logging areas with an altitude range of 1900 to 2800 meters above sea level such as ambo, arsi-robe, inewari, sagure and similar agro-ecologies. based on parameters that comprise both yield rank stability, ashebeka showed relatively better performance stability across a range of water logging environments. additive main effect and multiplicative interaction (ammi) is gaining popularity and is currently the main alternative multivariate approach to determine genotypic performance stability in many breeding programs. another approach called the ammi stability value (asv), which is based on the first and second interaction principal component axis (ipca) scores of the ammi model for each genotype, has also been developed recently. ammi stability value (asv), genotypic stability index (gsi), and a stratified ranking (ft3) of fox, et al. [17] has been generated to evaluate performance stability of the genotypes across test environments. genotypes with the lowest asv and gsi values are considered as having stable performance across the test environments. accordingly, upon its tremendous seed size advantage, the newly released variety ashebeka revealed lowest values of asv and gsi table 5 and identified as the most stable variety among the tested genotypes. similarly, fox, et al. [17] proposed that genotypes which are ranked in the top third of the genotypes in largest proportion of the test environments as having most stable performance than the remaining. based on the stratified ranking technique of the top third (ft3) parameter, ashebeka was ranked as top yielding variety in the top third of the test genotypes in 50% of the test environments table 5, hence considered as the most stable variety. table-5. stability status of the genotypes as revealed by asv, gsi and ft3 stability parameters. no entry yield asv ry rasv gsi top vipc1 vipc2 1 eh 01075-4 3756 9.5 1 2 3 50.0 3.624 -8.113 2 eh 00098-2 3403 21.1 8 7 15 25.0 14.670 -6.606 3 eh 00131-3 3480 35.9 5 10 15 50.0 26.179 1.531 4 eh 00101-2 3671 8.2 2 1 3 62.5 5.455 -3.371 5 eh 01036-2 3363 36.1 10 11 21 37.5 -21.466 -20.876 6 ekcsr 010 3492 13.5 4 4 8 12.5 9.374 -4.301 7 ekls 0101 3191 22.5 12 9 21 12.5 -10.289 -17.555 8 eklss 020 3596 21.7 3 8 11 50.0 -14.627 8.225 9 eh 06007-4 3480 41.6 6 12 18 25.0 -22.826 27.487 10 eh 06028-1 3197 10.0 11 3 14 0.0 -6.270 -5.109 11 hachalu 3470 20.0 7 6 13 37.5 10.177 14.307 12 walki 3363 16.6 9 5 14 37.5 6.000 14.379 note: key: asv = ammi stability value, ry = yield rank, rasv = ammi stability value rank, gsi = genotypic stability index, ft3 = % of the environments where genotypes fall in in the top third high yielding genotypes, vipc1 = varietal interaction principal component one, vipc2 = varietal interaction principal component two. 3.4. reaction to major diseases black root-rot caused by fusarium solani complex is the most destructive faba bean disease causing up to 70% yield losses under sever condition in water logging production areas. agronomic practices such as use of raised seedbed, crop rotation, planting date adjustment, fungicide applications and use of certified seeds may provide partial protection against the foliar and root diseases of faba bean. however, the most effective and eco-friendly disease control method is through developing resistant or tolerant cultivars. developing resistant or tolerant varieties to major diseases such as chocolate spot (botrytis fabae), black root-rot (fusarium solani), and rust (uromyces viciae-fabae) are among the major objectives of the national faba bean breeding program. chocolate spot, fusarium root-rot, and rust diseases have been scored using on (1-9) scale and were analyzed to determine the reaction of the released variety ashebeka to these diseases as compared to the recently released standard checks. consequently, the released variety ashebeka showed an average reaction of 4.48, 2.13 and 2.71 compared to 4.58, 2.42 and 3.04 for that of the best standard check, hachalu, in the trial for chocolate spot, fusarium root-rot and rust, respectively table 3, and is characterized as moderately resistant to these major diseases. agriculture and food sciences research, 2020, 7(1): 46-50 50 © 2020 by the authors; licensee asian online journal publishing group 4. variety maintenance the breeder and foundation seed will be maintained by kulumsa and holeta agricultural research centers. 5. conclusion as fusarium root-rot of faba bean is reported highly relevant pathogen causing significant yield losses in water logging vertisol production areas that covers 13% of the country potential highland arable land, increasing grain yield along with seed size is a recent prime objective in faba bean breeding programs of the country. so far, large scale field phenotyping of the host reactions to fusarium root-rot disease using sick plots has been done and resulted in the development of highly resistant breeding materials among which only few small-to-medium seeded varieties has been released and recommended for commercial use. the current variety, ashebeka, has combined high yield with heavier seeds has produced 33% and 8% seed size and yield advantage, respectively, over the best standard check, hachalu, in the trial and believed to meet the seed quality required for export market. therefore, wide cultivation of ashebeka variety will enhance productivity and marketability of the crop and believed to play significant role in improving farmers’ livelihood in water logging agro-ecologies. references [1] central statistical agency (csa), "report on area and production of major crops (private peasent holdings, meher season)," statistical bulletin, vol. 532, pp. 10-14, 2017. [2] i. a. u. fao, wfp and who (2017) the state of food security and nutrition in the world 2017: building resilience for peace and food security. rome: food and agriculture organization of the united nations (fao), 2018. [3] j. mussa and k. gemechu, vicia faba l. in: brink, m. and belay, g. (eds.). plant resources of tropical africa 1: cereals and pulses. wageningen, netherlands/backhuys: prota foundation, 2006. [4] t. e. kubure, c. v. raghavaiah, and i. hamza, "production potential of faba bean (vicia faba l.) genotypes in relation to plant densities and phosphorus nutrition on vertisols of central highlands of west showa zone, ethiopia, east africa," advances in crop science and technology, vol. 4, p. 214, 2016. [5] e. j. landry, c. j. coyne, r. j. mcgee, and j. hu, "adaptation of autumn-sown faba bean germplasm to southeastern washington," agronomy journal, vol. 108, pp. 301-308, 2016. available at: https://doi.org/10.2134/agronj2015.0028. [6] g. keneni, a. fikre, and m. eshete, "reflections on highland pulses improvement research in ethiopia," ethiopian journal of agricultural sciences, vol. 1, pp. 17-50, 2016. [7] m. tadele, "breeding achievements of faba bean (vicia faba l.) and its impact in the livelihood of ethiopian farmers," international journal of agriculture and bioscicence, vol. 8, pp. 263-269, 2019. [8] a. m. a. osman, a. b. hassan, g. a. osman, n. mohammed, m. a. rushdi, e. e. diab, and e. e. babiker, "effects of gamma irradiation and/or cooking on nutritional quality of faba bean (vicia faba l.) cultivars seeds," journal of food science and technology, vol. 51, pp. 1554-1560, 2014. available at: https://doi.org/10.1007/s13197-012-0662-7. [9] f. etemadi, m. hashemi, b. xing, and h. mashayekhi, "accumulation trend of l -dopa in different parts of faba beans varieties," in in: asa, cssa, & sssa international annual meetings. california, 2019, p. 281. [10] j. m. cohen, integrated rural development: the ethiopian experience and the debate book published with support from the swedish international development authority (sida). uppsala: nordic africa institute(scandinavian institute of african studies), 1987. [11] d. yohannes, faba bean (vicia fabae) in ethiopia. addis ababa, ethiopia: institute of biodiversity, conservation and research, 2000. [12] b. bekele, g. muhammad, t. galano, and t. belayneh, "faba bean and field pea disease research in ethiopia. in: ali k, keneni g, ahmed s, malhotra r, beniwal s, makkouk k, halila mh (eds) food and forage legumes of ethiopia: progress and prospects," in proceedings of a workshop on food and forage legumes. september 2003, addis ababa, ethiopia. icarda, aleppo, syria, 2006, p. 351. [13] g. yitayih and y. azmeraw, "adaptation of faba bean varieties for yield, for yield components and against faba bean gall (olpidium viciae kusano) disease in south gondar, ethiopia," the crop journal, vol. 5, pp. 560-566, 2017. available at: https://doi.org/10.1016/j.cj.2017.05.007. [14] e. hailu, g. getaneh, t. sefera, n. tadesse, b. bitew, a. boydom, d. kassa, and t. temesgen, "faba bean gall; a new threat for faba bean (vicia faba) production in ethiopia," advances in crop science and technology, vol. 3, p. 117, 2014. [15] k. gemechu, j. mussa, and w. tezera, "faba bean (vicia faba l.) genetics and breeding research in ethiopia: a review. in: kemal, a., gemechu, k., seid, a., malhotra, r., beniwal, s., makkouk, k. and halila, m.h. (eds.). food and forage legumes of ethiopian: progress and prospects," in proceedings of a workshop on food and forage legumes.22-26 sept. 2003, addis ababa, ethiopia. icarda, aleppo, syria. isbn 92-9127-185-4, 2006, pp. 42-52. [16] s. sas, guide for personal computers, version 9.0 edition. cary, nc: sas institute usa, 2002. [17] p. fox, b. skovmand, b. thompson, h.-j. braun, and r. cormier, "yield and adaptation of hexaploid spring triticale," euphytica, vol. 47, pp. 57-64, 1990. available at: https://doi.org/10.1007/bf00040364. [18] m. a. hussein, a. bjornstad, and a. aastveit, "sasg× estab: a sas program for computing genotype× environment stability statistics," agronomy journal, vol. 92, pp. 454-459, 2000. available at: https://doi.org/10.2134/agronj2000.923454x. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 203 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 2, 203-210, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.62.203.210 © 2019 by the authors; licensee asian online journal publishing group lettuce (lactuca sativa) growth performance in saltwater, soil and aquaponic system yap, q.c1 teo, s.s.2 ( corresponding author) 1,2department of biotechnology, faculty of applied sciences, ucsi university, kuala lumpur, malaysia. abstract soil erosion is a major problem in most agricultural field where it threatened the whole ecosystem. conventional planting included heavy applies of pesticides and fertilizer had given negative impact to the environment. aquaponics of combining aquaculture and hydroponics in one integrated system is a useful revolutionary food production system. salinity can inhibit the plant growth but saltwater aquaponics is studied as an extension from aquaponics recently. lettuce is selected as the study plant species. a study had carried out by comparing the lettuce seeds germination and plants growth in soils and aquaponics, and lettuces growth with and without salt treatment in freshwater and saltwater aquaponics. the result showed no significance difference on seeds germination and plant growth between sandy soil and coconut coir soil (p > 0.05). lettuces in aquaponics had the highest total water content, mean plant height increment, and most leaves. germination of lettuce seeds that treated with different salt concentration was inhibited and the one treated with 0.2m salt solution had no germination. growth of lettuces from saltwater were inhibited whether it was preliminary treated with salt or not whereas those from freshwater grew very well. in conclusion, aquaponics was more effective in growing lettuces than soils. freshwater is more suitable to use for growing lettuces than saltwater. saltwater inhibited lettuce growth whether with or without salt treatment. waste product from aquaculture had successfully utilised as a nutrient source for growing crops. keywords: lactuca sativa, aquaponics, saltwater, soil. citation | yap, q.c.; teo, s.s. (2019). lettuce (lactuca sativa) growth performance in saltwater, soil and aquaponic system. agriculture and food sciences research, 6(2): 203-210. history: received: 10 april 2019 revised: 20 may 2019 accepted: 2 july 2019 published: 22 august 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: both authors contributed to the conception and design of the study. funding: this research was supported by centre of excellent for research value innovation and entrepreneurship (cervie) (proj-in-fas-026) and faculty of applied sciences of ucsi university. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 204 2. materials and methods .................................................................................................................................................................. 204 3. results and discussion ................................................................................................................................................................. 205 4. conclusion ....................................................................................................................................................................................... 210 references ............................................................................................................................................................................................ 210 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.203.210&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.203.210&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/939 https://orcid.org/0000-0002-3053-5052 agriculture and food sciences research, 2019, 6(2): 203-210 204 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes in the existing literature by comparing the lettuce seeds germination and plants growth in soils and aquaponics, and lettuces growth with and without salt treatment in freshwater and saltwater aquaponics. 1. introduction lettuce, also known as lactuca sativa, is a type of leafy vegetable from asteraceae family, which produces crisp leaves loosely arranged on the stalk. it is easily cultivated and it grown as a hardy annual. besides, it favors low temperature surroundings to prevent from flowering quickly. moreover, the germination of lettuce seed usually takes around 2 to 8 days. leaf lettuce will normally produce a crop within 50 to 70 days, while most heading varieties might require 60-80 days to mature [1]. lettuce was considered to possess some medicinal qualities during medieval times in europe [2]. lettuce is riches in various vitamins and minerals, it also has several benefits such as anti-inflammatory properties, low cholesterol level, antioxidant properties, improve insomnia, antioxidant, antimicrobial, anti-cancer properties and control anxiety. traditional planting is the ancient food production system as it has been practiced for thousands of years. it is not only the oldest planting method in the world but also the main source of improving phase of planting technology such as conventional, modern and organic planting, that involved crops plantation using soils which required very large land space. aquaponics is an integration of hydroponics and aquaculture, where crop plants and aquatic species can be grown together in a soilless water-based system [3, 4]. it is a soilless agricultural system that are highly productive, suitable for urban areas, and can address the shortage of land in relation to growing demand for food production. aquaponics can be set-up in vertical integration systems in indoor urban settings, which address the limitations of soil quality and space availability [5]. it also helps cleanses the water through so called biofilter treatments. the nutrients that released from fish excreta and microbial breakdown of organic wastes had utilized by plants in aquaponic systems [6]. salinity had also played an important role when it comes to crops plantation. different level of salt concentration can affect the seed germination and growth of plants, especially to those plants that are weak in tolerating salts level [7]. recently, the demand for seafood products on the global market is rising especially in asian countries due to the increase in affluence and appreciation of the health benefits of seafood. saltwater aquaponic system had been studied as an extension of aquaponics to grow seafood and high salt tolerance crops [8]. this study aimed to compare lettuce seeds germination and plants growth in soils and aquaponics, and lettuces growth with and without salt treatment in freshwater and saltwater aquaponics. 2. materials and methods 2.1. lactuca sativa (l. sativa) preparation and germination the lactuca sativa (l. sativa) seeds were soaked with 5% naocl solution for 5 minutes to sterilize surface of the seeds. the seeds were washed with distilled water, drained and dried with tissue paper. leca (light expanded clay aggregate), soil mixture (sandy soil, quartz, limestone, sandstone and shale) and coconut coir soil were used as medium for plant grow. l. sativa seeds were germinated in different media (soils, sandy soil and coconut coir soil) under different temperature (15°c, 20°c, 25°c and 30°c) to screen for the l. sativa with the best growth performance and further compared with those in aquaponics. the plants were water twice a day. the l. sativa seedlings were transferred to a larger pot for growth observation after 10 days of germination. 2.2. aquaponic system 2.2.1. lettuce seeds with salt treatment in saltwater salinity experiment was to pre-treated lettuce seeds with different salt concentrations. lettuce seeds were soaked with 0.025m, 0.05m, 0.1m and 0.2m nacl solution in petri dishes respectively. after the germination count for around 10 days, the lettuce seedlings were transferred to saltwater aquaponics system for growth observation. 2.2.2. lettuce seeds without salt treatment in freshwater and saltwater lettuce seeds were firstly germinated for 10 days. the lettuce seedlings were transferred to freshwater and saltwater aquaponic system for growth observation. the growth result of lettuces was used to compare with lettuces growth in sandy soil, coconut coir soil, and saltwater aquaponic system. 2.3. transfer lettuce from soil to aquaponics after one month, the lettuces in soils that grow better were transferred from soils system to aquaponic system and allowed to continue grow for another 2 weeks. 2.4. data collection the seed germination count was collected, final germination percentage, germination rate reduction of germination percentage, and seed viability index [9, 10] were calculated as followed: 𝐹𝑖𝑛𝑎𝑙 𝑔𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑖𝑜𝑛 𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 = 𝑆 𝑇×100 𝐺𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑖𝑜𝑛 𝑟𝑎𝑡𝑒 = 𝑁1 𝐷1 + 𝑁2 𝐷2+⋯ + 𝑁𝑖 𝐷𝑖 where s is the number of germinated seeds, t is the total number of seeds and ni number of germinated seeds, per day (di). reduction of germination percentage = (1 − the number of germinated seeds conditions salinity / the number of germinated seeds conditions control) × 100. agriculture and food sciences research, 2019, 6(2): 203-210 205 © 2019 by the authors; licensee asian online journal publishing group 𝑆𝑒𝑒𝑑 𝑣𝑖𝑎𝑏𝑖𝑙𝑖𝑡𝑦 𝑖𝑛𝑑𝑒𝑥 = 𝐹𝑖𝑛𝑎𝑙 𝑔𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑖𝑜𝑛 𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 × 𝑎𝑣𝑒𝑟𝑎𝑔𝑒 𝑠𝑒𝑒𝑑𝑙𝑖𝑛𝑔 𝑙𝑒𝑛𝑔𝑡ℎ(𝑚𝑚) 100 plant height, fresh weight, dry weight, leaf number, leaf color and weekly water quality analysis were measured and recorded. 2.5. statistical analysis all the collected data were analysed with the means were compared using ibm spss statistics 22. 3. results and discussion 3.1. lettuce seeds germination in sandy soil and coconut coir soil seeds germination in 15°c, 20°c and 25°c was not significantly different from 30°c. figure 1 showed that germination rate of lettuce seeds in sandy soil was slightly higher than that in coconut coir soil. from figure 2, final germination percentage had shown an adverse result whereby there is a higher germination percentage of lettuce seeds in coconut coir soil than sandy soil, with germination percentage of 90%-100%, that is nearly all lettuce seeds had germinated. consequently, sandy soil allowed a more rapid seed germination whereas coconut coir soil was able to provide a higher chance of success for more seeds to be germinated. lettuce seeds grew the slowest at 15°c where the leaves opened very slowly while only shoot were elongated normally. on the other hand, the germination of lettuce seed in both soils was completely inhibited at 30°c. at 15°c, the germination and growth of seeds will be slowed down. temperature of 30°c was very high for lettuces which leads to rapid dry out in soil although watering was applied. as a result, the sprouting process of seed was greatly inhibited due to overheating. the high temperature not only harmed the plant but also altered the taste of lettuce [11]. from the results, it showed that lettuces favoured growth under 25°c. moreover, high soil strength will impeded seedlings emergence and root growth, it also makes tilling more difficult and more energy demanding [12]. the loose structure in sandy soil provided large pore spaces which allow roots easy to move through the soil and shoots can emergence easily [13]. hence, the seeds were able to germinate faster provided with favourable temperature. however, coconut coir soils contains lignin and cellulose that rich in potassium and various micronutrients such as fe, mn, zn, and cu, where it supplied more nutrients to seeds than sandy soil and allowed more seeds to germinate. the overall growth performance of seeds from sandy soil and coconut coir soil under 25°c was the best hence were chosen for comparison with those from freshwater aquaponics. figure-1. germination rate of lettuce seeds with different temperatures (°c) in sandy soil and coconut coir soil in 10 days. means that are not followed by different letters were not significantly different at p < 0.05 level. figure-2. final germination percentage (%) of lettuce seeds with different temperatures (°c) in sandy soil and coconut coir soil in 10 days. means that are not followed by different letters were not significantly different at p < 0.05 level. agriculture and food sciences research, 2019, 6(2): 203-210 206 © 2019 by the authors; licensee asian online journal publishing group 3.2. lettuce growth rate in sandy soil, coconut coir soil and aquaponic system overall, lettuce growth in sandy soil and coconut coir soil was significantly than that in aquaponics as shown in figure 5. however, there is a drastic increase in mean height increment of lettuce in aquaponics than soils as shown in figure 3. aquaponics grew crops a lot faster than those in soil. in addition, there is significance difference between total water content of lettuces that grew in coconut coir soil, sandy soil and aquaponic system. lettuces from aquaponics had highest total water content, whereas the total water content in sandy soil was lower than that in coconut coir soil figure 4. based on table 1, lettuces from aquaponics was largest in size and had the most average number of leaves. lettuces in soils grew a lot slower than lettuces from aquaponics as the plant was very small, the leaves were still tiny even grew for a month. growth of lettuces in sandy soil was the poorest among all, the leaves started to turn yellow while lettuces in other systems remained as fresh green colour. lettuces from coconut coir soil was slightly better had chosen and transferred to aquaponics for 2 weeks, the mean plant height of lettuce had increased, with 41.0mm difference and total water content of 174.9mg had indicated improvement. table 2 showed the average number of leaves also increased, faster than in soil. physical structure of soil is the important properties where it greatly influenced all physical processes that take place such as thermal conductivity, water holding capacity, permeability, and aeration [12]. soil structure can affect water supply to plants and influences nutrients availability. coconut coir fibre possessed better water and nutrient retention ability therefore it produced crops with higher water content. sandy soil has very weak water retention ability, low organic matter content, and poor soil fertility. large particle properties made sandy soil does not hold much water hence applied water will be drained away [14]. in addition, it had low nutrient holding capacity, lead to rapid leaching of native or added nutrient [15]. therefore, sandy soil produced less healthy lettuces. although coconut coir soil riches in many nutrients, however it has low cation exchange capacity (21-30 meq./l) where it does not retain cations or buffer against ph well [16]. even coco coir has the capacity to hold nutrients, but it can also “lock” them up at the same time which leaded to nutrient deficiencies. aquaponics supported the growth of lettuces more efficiently hence the plants were able to grow very fast in short time interval. the water content indicated can potentially be achieved by the plant. hence, aquaponics is built in a closed, recirculating system [17] itself utilized the water and nutrient source directly from fish tank which provided sufficient nutrients to lettuces and no drying issue will occur. figure-3. mean height increment of lettuces in sandy soil, coconut coir soil and aquaponic system after a month. means that are not followed by different letters were not significantly different at p < 0.05 level. figure-4. total water content of lettuces in sandy soil, coconut coir soil and aquaponic system after a month. means that are not followed by different letters were not significantly different at p < 0.05 level. agriculture and food sciences research, 2019, 6(2): 203-210 207 © 2019 by the authors; licensee asian online journal publishing group figure-5. growth of lettuces in sandy soil (right), coconut coir soil (middle) and aquaponic system (left) after a month. figure-6. growth of lettuces before and after transferred from coconut coir to aquaponic system for 2 weeks. table-1. average leaf number and leaf colour of lettuces in aquaponics system,coconut coir soil, and sandy soil after a month. parameters planting system aquaponics coconut coir soil sandy soil average leaf number 7 4 3 leaf colour fresh green fresh green yellow table-2. mean plant height, total water content, average leaf number and leaf colour in lettuces before and after transferred from coconut coir soil to aquaponic system for 2 weeks. parameters planting system before transfer after transfer mean plant height (mm) 58.0 99.0 total water content (mg) 22.9 174.9 average leaf number 4 7 leaf colour fresh green fresh green 3.3. salt stress on lettuce seeds germination germination of lettuce seeds treated with different salt concentrations were inhibited, both the germination rate and final germination percentage were lower than that of the control. seed germination of lettuce seeds that treated in 0.025m, 0.05m and 0.1m salt solutions was not significantly different figure 8 and germination percentage were all reduced to 85% figure 9. although the seeds were affected by salt stress but the level of affection was considered not too severe. there is no germination in lettuce seeds treated with 0.2m salt. from the result, it was indicated that lettuce seeds able to tolerate salt stress below 0.2m salt concentration. reduction of germination percentage in lettuce seeds from different salt concentrations level also showed greatest reduction in 0.2m figure 10. furthermore, salt stress also resulted in loss of seed viability index. as expected, seeds treated with 0.2m salt solution loss seed viability index the most figure 11. growth of lettuce seeds with salt treatment was mostly slightly shorter in length than control. indeed, high salt level will inhibit the germination of new seeds. seeds were still able to grow when salinity is not too high but slower than the seeds with tap water only, due to the salinity that acted as drought on plants which prevents roots from performing their usual osmotic activity. moreover, water and nutrients will travel from an area of low concentration into an area of high concentration. water and nutrients unable to migrate into plant roots, preventing water uptake by germinating seeds [7]. as salinity levels increased, stress on germinating seedlings will also increase. agriculture and food sciences research, 2019, 6(2): 203-210 208 © 2019 by the authors; licensee asian online journal publishing group figure-7. germination rate of lettuce seeds in different nacl concentrations (m) in 10 days. means that are not followed by different letters were not significantly different at p < 0.05 level. figure-8. final germination percentage (%) of lettuce seeds in different nacl concentrations (m) in 10 days. means that are not followed by different letters were not significantly different at p < 0.05 level. figure-9. reduction of germination percentage (%) of lettuce seeds in different nacl concentrations (m) in 10 days. means that are not followed by different letters were not significantly different at p < 0.05 level. figure-10. seed viability index (mm) of lettuce seeds in different nacl concentrations (m) in 10 days. means that are not followed by different letters were not significantly different at p < 0.05 level. 3.4. lettuce growth rate in freshwater and saltwater aquaponics system from figure 11, there was no significance difference of lettuces growth in saltwater whether lettuces were treated with salt or no salt treatment as there were no changes in mean height increment after a month. total water content in lettuces with and without salt treatment was 3.7mg and 3.3mg respectively, which was lower than control as shown in figure 12. lettuce from freshwater showed normal growth while there was no growth in lettuce from saltwater. lettuces from saltwater were thin and tiny, no change of height and no sign of growing new leaf, however no significance difference in leaf colour with control table 3. texture of lettuce without salt treatment was soft and mushy while the lettuce with salt treatment was slightly rigid. based on weekly water agriculture and food sciences research, 2019, 6(2): 203-210 209 © 2019 by the authors; licensee asian online journal publishing group quality analysis in table 4 and figure 13, since saltwater aquaponics is of seawater, na+ concentration is very high (8267ppm) that greatly reduced lettuce growth. high na+ concentration indicated increase starvation of essential nutrients from plants, it may also increase the chance for root diseases to occur. high k+ concentration in saltwater had induced deficiencies of nutrients such as nitrogen, calcium and magnesium [18]. saltwater also has high ca2+ concentration that had caused lock up of various nutrients such as potassium, magnesium, manganese and iron. excess ca2+ ion in water tank came in contact with sulphur, it will precipitated and remained at bottom of the tank. it also decreases the uptake of cation nutrients such as phosphorus, boron, copper and zinc [19]. on the other hand, the conductivity in saltwater was too high and had prevent efficient water absorption in the root zone by plant. however, low levels of phosphate in saltwater had limited plant and algae growth [20]. the soft and mushy texture in plants indicated loss of turgidity as they were no longer can maintain themselves in shape, ultimately culminating in cell death [21]. besides, whether the plants were preliminary treated with salt or no salt solution had no effect on growth since both of them were inhibited. salinity tolerance level in lettuce was not capable of growing in saltwater aquaponics, it is still preferred to grow in freshwater aquaponic system. figure-11. mean height increment of lettuces in freshwater aquaponic system (control), and saltwater aquaponic system (lettuces with salt treatment and without salt treatment) after a month. means that are not followed by different letters were not significantly different at p < 0.05 level. figure-12. total water content of lettuces in freshwater aquaponic system (control), and saltwater aquaponic system (lettuces with salt treatment and without salt treatment) after a month. means that are not followed by different letters were not significantly different at p < 0.05 level. figure-13. weekly water quality test for fresh water aquaponic system andsaltwater aquaponic system. table-3. average leaf number and leaf colour of lettuces in freshwater aquaponic system and saltwater aquaponic system (lettuces with salt treatment and without salt treatment) after a month. parameters aquaponics system control with salt treatment without salt treatment average leaf number 7 2 2 leaf colour fresh green fresh green fresh green agriculture and food sciences research, 2019, 6(2): 203-210 210 © 2019 by the authors; licensee asian online journal publishing group table-4. weekly water quality test for freshwater aquaponic system and saltwater aquaponic system. parameters aquaponics system freshwater saltwater temperature (°c) 22.1a 22.3a ph 7.53a 7.65a do (mg/l) 6.12a 6.86a na+ (ppm) 340a 8267b k+ (ppm) 24a 233b ca2+ (ppm) 123a 333b no3(nitrate) (ppm) 272a 273a conductivity (µs/cm) 3.2a 33.5b salt content (ppt) 1.5a 19.8b po4 3(mg/l) 2.0a 0.5b note: means that are not followed by different letters were not significantly different at p < 0.05 level. 4. conclusion in a nutshell, people started to use soil-less based planting method to grow lettuces as they would want to save more space and increase crop yield. from the study, sandy soil and coconut coir soil had exerted insignificant effect (p > 0.05) on lettuce growth. it involved the mixing of different composition of soils in order to improve the crop growth, where it was complex and time wasting. contrary to the previous method, a simple setting of aquaponic system was easily to be monitored and controlled. the result shown was also obviously more effective where it grew healthier lettuces in shorter time interval than the soil does. lettuce was moderate salt tolerance plant where it can only withstand water salinity up to around 0.2m. therefore, freshwater aquaponics was more suitable in growing them rather than saltwater aquaponics due to extreme high salt level that lead to osmotic inhibitory in plants. saltwater aquaponics will inhibited the growth of lettuces whether with or without preliminary salt treatment. more research is required in the future to develop this aquaponic system with valuable cash crops that could thrive in sea water. aquaponic system had successfully aquacultured the waste materials. waste product from fishes had to be eliminated from the system as it contains high amount of ammonia which will cause toxic effect that will lead to illness to the fishes. however, the nitrifying bacteria had turned this toxic material into nitrate that act as a food source for plants to grow. future research may investigate the lettuce yield with different system design of aquaponics, specific patterns of nutrient availability and uptake, cost-benefit analysis of lettuce productivity, and different aspects of lettuces and other crops under aquaponics or other soilless systems. references [1] s. y. zee and l. h. hui, hong kong food plants. united states: urban council, 1990. [2] c. p. khare, indian herbal remedies: rational western therapy, ayurvedic, and other traditional usage, botany. germany: springer science & business media, 2004. [3] d. e. seawright, r. r. stickney, and r. b. walker, "nutrient dynamics in integrated aquaculture–hydroponics systems," aquaculture, vol. 160, pp. 215-237, 1998. available at: https://doi.org/10.1016/s0044-8486(97)00168-3. [4] j. e. rakocy, t. m. losordo, and m. p. masser, "recirculating aquaculture tank production systems: integrating fish and plant culture, southern region," aquacul center publication, vol. 454, pp. 1-16, 2006. [5] f. orsini, r. kahane, r. nono-womdim, and g. gianquinto, "urban agriculture in the developing world: a review," agronomy for sustainable development, vol. 33, pp. 695-720, 2013. [6] h. r. roosta and m. hamidpour, "effects of foliar application of some macro-and micro-nutrients on tomato plants in aquaponic and hydroponic systems," scientia horticulturae, vol. 129, pp. 396-402, 2011. [7] m. m. azooz and p. ahmad, plant-environment interaction: responses and approaches to mitigate stress. united states: john wiley & sons, 2016. [8] v. nozzi, g. parisi, d. d. crescenzo, m. giordana, and o. carnevali, "evaluation of dicentrarchus labrax ,eats and the vegetable quality of beta vulgaris var. cicla farmed in freshwater and saltwater aquaponic systems," water, vol. 8, pp. 1-14, 2016. [9] m. azizi, m. chehrazi, and s. m. zahedi, "effects of salinity stress on germination and early growth of sweet william (dianthus barbatus)," asian journal of agricultural sciences, vol. 3, pp. 453-458, 2011. [10] m. j. seghatoleslami, "effect of salinity on germination of satureja hortensis l., cichorium intybus l. and cynara scolymus l," iranian journal of agricultural research, vol. 8, pp. 818-823, 2010. [11] l. shepherd, the complete guide to growing vegetables, flowers, fruits, and herbs from containers: everything you need to know explained simply. united states: atlantic publishing company, 2011. [12] r. lal, encyclopedia of soil science. united states: crc press, 2006. [13] victorian department of agriculture, the journal of agriculture vol. 73:. united states: victorian department of agriculture 1975. [14] k. t. osman, soils: principles, properties and management. germany: springer science & business media, 2012. [15] r. s. jackson, wine science: principles and applications, 4th ed. netherlands: elsevier, 2014. [16] k. a. handreck, "properties of coirdust, and its use in theformulation of soillesspotting media," communications soil & plant anal, vol. 24, pp. 349-363, 1993. [17] r. v. tyson, d. d. treadwell, and e. h. simonne, "opportunities and challenges to sustainability in aquaponic systems," hort technology, vol. 21, pp. 6-13, 2011. available at: https://doi.org/10.21273/horttech.21.1.6. [18] r. d. merrill, a. a. shamim, a. b. labrique, h. ali, k. schulze, m. rashid, p. christian, and k. p. west jr, "validation of two portable instruments to measure iron concentration in groundwater in rural bangladesh," journal of health, population, and nutrition, vol. 27, pp. 414-418, 2009. available at: https://doi.org/10.3329/jhpn.v27i3.3384. [19] s. arora, a. k. singh, and y. p. singh, bioremediation of salt affected soils: an indian perspective. germany: springer, 2017. [20] d. willis, l. may, y. gutema, and n. whittlesey, measuring economic benefits of water pollution abatement in an irriated river basin. united states: diane publishing, 1992. [21] b. l. querijero and a. l. mercurio, "water quality in aquaculture and non-aquaculture sites in taal lake, batangas, philippines," journal of experimental biology and agricultural sciences, vol. 4, pp. 109-115, 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. a study of innovative design and development of hand-made diet product series agriculture and food sciences research issn: 2411-6653 vol. 2, no. 2, 39-42, 2015 http://www.asianonlinejournals.com/index.php/aesr 39 a study of innovative design and development of handmade diet product series rui-lin lin 1 1 department of commercial design chienkuo technology university no. 1, chiehshou north road, changhua, taiwan abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ............................................................................................................................................................................... 40 2. literature review ...................................................................................................................................................................... 40 3. research methods ..................................................................................................................................................................... 40 4. design results ............................................................................................................................................................................ 40 5. conclusions ................................................................................................................................................................................ 41 references ...................................................................................................................................................................................... 42 this study had planted flowers, fruits, and vegetables on a farmland in toubiankeng on county highway 136, taiping dist., taichung city, without any chemical, and designed hand-made diet products including jam, honey, minced pork with radishes, coffee, salt pork, osmanthus fragran, jasmine, rose, chrysanthemum, and raspberry with natural food ingredients without any preservative or artificial flavoring. this study stressed the balance between nature and health. the materials of the chinese medicine were blended carefully and made with the traditional bakery technology using local soil and through continuous cultivation with the desire for the purity of the nature and the fragrance of flowers. the mellow and rich mouthfeel of the tea was integrated into people’s lives, containing the attentive care for human body health and delivering the message of valuing this land. with the idea of giving back to the land what’s taken from it, this study designed a series of creative products, so that people can, through eating and drinking, regain health of their body. keywords: innovative design, design and development, hand-made diet, product series, creative product, creative design. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(2): 39-42 40 1. introduction in respect that people often wish to find a place to relax a little bit and spend time with family members during their leisure time as they are usually under great pressure at work, this study accept the case from a client in toubiankeng on county highway 136, taiping dist., taichung city. this case was about using the quiet and beautiful environment in toubiankeng and rich local resources to design products with flowers, fruits, and vegetables planted on this land without any agricultural chemical. in sunny days, this place was comfortable and pleasant as if spraying ink in the sky. and the client called his own house a “cottage”. thus, this place was named “ink cottage”. the area was almost 1000 pings (about 3306 square meters). the space could be used by artists, calligraphers, and writers to exhibit or sell their works. it could also be used by musicians to play a solo or hold a concert. with various kinds of scented tea for tasting and healthy foods for sale, the atmosphere of cultures and arts and the easiness of recreation of this space can be conveyed. among various flowers and plants grown in this place, this study selected osmanthus fragran, jasmine, rose, chrysanthemum, and raspberry, the ones going well with drinks, plucked them, cleaned them, dried them, decomposed them, and preserved them. this series of handmade scented beverage can be mixed or brewed according to drinkers’ preferences. one of the features is that these flowers and plants were grown without any agricultural chemical, so that the soil texture was not damaged and there was no chemical at all on the crops grown in this place. this series was the most natural and healthy choice, which also showed the love for this land and delivered the idea of giving back to the land what’s taken from it. secondly, this study also combined rich local products to make jam, honey, minced pork with radishes, coffee, and salt pork using traditional methods. the principles adhered to include producing only handmade products and making innovation. lastly, this study selected the materials used to create the containers for these handmade foods and drinks and suitable external designs. also, the posters and packages for these products were designed. these products can be given to others as a gift or used for oneself. 2. literature review this study used other non-poisonous ways to expelling inspects and snakes instead of applying agricultural chemicals to planted crops. thus, a lot of attention was paid to innovation management of new technology. here technology means science and technology. there are various definitions and research scales of different levels for this term. in a broad, it is generally called technology. in a narrow sense, the focus is on applications based on innovative and r&d technologies [5]. technology management is the largest part of innovation. and technology innovation is the most common part of technology management. it can be classified in detail into product innovation, process innovation, and administrative innovation. based on the degree of innovation, technology innovation can be categorized into product improvement, new product development, and breakthrough, in the order from the lowest degree to the highest. technology innovation management gives products a chance to be designed and developed [2]. r&d is the key to success in product innovation. if sales and various professional services related to a product can be offered with consumers’ demands being taken into consideration, then the popularity of the product can be increased correspondingly, as well as the enterprise’s profits [4]. novelty of product innovation is an important factor in technology development and cooperation process. it should not be ignored [1]. a lot of companies featuring innovation management or technology innovation have proposed various innovation policies based on the viewpoint of capability and conducted all kinds of promotional campaigns showing various innovative features of their products through internet systems [7]. 3. research methods through field study and field interviews, this study got to know the plan of the owner of the place. issues regarding handmade food materials, visual packaging, and other things were discussed and the conclusions were recorded as important references for the innovative designs and development of a product series of foods and drinks. 3.1. the field study method field study is a method applied to obtain and record information regarding the background, current status, or interaction with environment of individuals, groups, organizations, or communities. for new and unknown research territories, it helps to obtain rich results and interpret large research statistical data to make up the insufficiency of the data. it is very useful for research in the field of social science. however, due to the small sample size, the results are often not representative enough [6]. 3.2. the field interview method an interview is a type of conversation. the structure and the purpose are decided by the interviewer one-sidedly. an interview is a professional interaction, beyond spontaneous viewpoint exchanges in daily conversations [3]. the interview method helps to collect in-depth data related to a research topic. with topic planning in advance, this method can be applied to retrieve necessary information, which can be recorded, summarized, analyzed, and compared to obtain results related to the topic. the purpose is to give interviewees free space to answer interview questions. without any limitation, they can fully express their feelings about, attitudes toward, and value judgments regarding the questions [6]. after several visits and recording, this study was able to further understand the client’s wishes regarding the usage of the internal and external space, floor arrangement, and overall visual style. this information was a great help to the innovative design and development of the product series of handmade foods and drinks. 4. design results although the retention periods of handmade foodstuffs without any preservative or artificial flavoring made of healthy and natural food materials without any agricultural chemical are usually limited, they are still very attractive agriculture and food sciences research, 2015, 2(2): 39-42 41 to consumers. if a dealer wants to promote its non-poisonous products and make them popular in the market, the only way to convince consumers is to insist on the principle of honesty. 4.1. poster and package design nowadays, consumerism is highly valued. to promote the handmade foods, proper poster designs regarding the non-poisonous food materials for the purpose of advertisement is very important. the poster designs and package designs for the handmade foods are presented below (figure 1): figure-1. poster designs and package designs 4.2. the handmade flower product series this study selected the flowers and plants which are suitable to make drinks and followed the procedure to make drinks. these drinks can be mixed or brewed according to drinkers’ preferences. this study offered the service of customized product innovative design and development. the design results are presented below (figure 2). figure-2. the handmade flower product series 5. conclusions when working on the case of the handmade product series of foods and drinks, the design team of this study followed the principles which the client insisted on, including using natural food materials, keeping the original agriculture and food sciences research, 2015, 2(2): 39-42 42 flavors, not adding any additive or preservative, making healthy and safe foods by hand, and environmental protection. the features of the products are summarized below: (1) one principle was no agricultural chemicals on the land where the flowers, fruits, and vegetables were planted. other scientific methods were applied to expel inspects and snakes. this way, it was assured that there was no agricultural chemical left on the crops, and the foods and drinks are non-poisonous and can help with human body circulation and metabolism. (2) the original flavors of the handmade foods were kept. the products featured their natural and healthy taste and small packages and cold storage, which resolved the issue of short retention periods. (3) the containers were made of glasses, so that one can see clearly what is inside. these containers are nonpoisonous, clean, and high-barrier. thus, the processing was rather easy, and the designs could be flexible. there were plenty of raw materials available, and they are eco-friendly for they can be recycled. (4) the product gift boxes were packed with paper materials. the ink printing was replaced by embossing seals, to reduce the usage of ink. these packing materials are eco-friendly and easy to recycle. references [1] f. t. zhang, management of science and technology. taipei: hkbookcity publishing co., ltd, 2008. [2] b. l. zhang and z. x. wu, technology management. taipei: wu-nan book, 2010. [3] z. ma, m. yu, c. gao, j. zhou, and z. yang, "institutional constraints of product innovation in china: evidence from international joint ventures," journal of business research, vol. 68, pp. 949-956, 2015. [4] f. o. marta, m. r. beatriz, j. i. l. sánchez, and j. f. menéndez, "product innovation: when should suppliers begin to collaborate?," journal of business research, vol. 68, pp. 1404-1406, 2015. [5] a. l. raquel, c. l. josé, g. f. nieves, m. r. javier, and p. v. miguel, "fostering product innovation: differences between new ventures and established firms," technovation, vol. 41-42, pp. 25-37, 2015. [6] z. c. ye and l. c. ye, research methods and research paper writing. taipei: shinning culture publishing co., ltd, 2011. [7] y. c. chen, the field interview method. taipei: weber publication international ltd, 2010. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. 113 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 2, 113-124, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.72.113.124 © 2020 by the authors; licensee asian online journal publishing group combining ability analysis of maize inbred lines in ethiopia abenezer abebe tefera1 legesse wolde beyene2 wosene gebreselassie abtew3 ( corresponding author) 1,2department of crop science, ethiopian institute of agricultural research, holetta agricultural research center, holeta, ethiopia. 3department of horticulture and plant science, jimma university, college of agriculture and veterinary medicine, jimma, ethiopia. abstract the study was initiated to estimate combining ability of maize inbred lines and crosses using line by tester analysis. fifty entries consists 48 f1 single crosses developed from 24 inbred lines and 2 testers using line x tester design and two commercial check hybrids used in the study. the experiment was conducted using alpha lattice design with two replications. analysis of variance revealed existence of significant genetic variation among genotypes for all studied traits except for plant aspect (pa). location x entry interaction for most of the traits was not significant which suggests hybrid performance was consistent across tested locations. line x tester analysis of variance showed that mean squares due to gca of lines were significant (p< 0.01 or p< 0.05) for all studied traits. mean squares of tester gca and sca were significant for most of studied traits. this indicates that both additive and non-additive gene effects had contributed for the variation of the crosses. however, higher proportional contribution of additive gene action for all studied traits was obtained. several lines and crosses were identified as good general and specific combiners for yield and yield related traits. lines l23, l11, l15 and crosses l2xt1, l3xt1, l8xt1, l11xt1, l23xt1 and l13xt2 were found to be good general and specific combiners, respectively. in conclusion, the stated inbred lines with desirable gca effects and cross combinations with desirable sca effects for grain yield and yield related traits could be used as useful genetic material. keywords: combining ability, general combining ability, specific combining ability, inbred lines, line by tester, maize. citation | abenezer abebe tefera; legesse wolde beyene; wosene gebreselassie abtew (2020). combining ability analysis of maize inbred lines in ethiopia. agriculture and food sciences research, 7(2): 113-124. history: received: 30 april 2020 revised: 10 june 2020 accepted: 13 july 2020 published: 28 july 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 114 2. materials and method ................................................................................................................................................................... 114 3. results and discussion ................................................................................................................................................................. 116 4. conclusion ....................................................................................................................................................................................... 121 references ............................................................................................................................................................................................ 121 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.72.113.124&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.72.113.124&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1946 agriculture and food sciences research, 2020, 7(2): 113-124 114 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the study was initiated to estimate combining ability of maize inbred lines and crosses using line by tester analysis. 1. introduction maize (zea mays l.) is a diploid (2n = 20) crop belongs to the family of grasses poaceae and tribes maydeae and naturally it is cross pollinated crop. the crop is grown over a wide range of environmental conditions. maize has great world-wide importance as human food, industrial raw material and animal feed. because of its high yield potential and wider adaptation, maize is one of the strategic crops for the achievement of food security. prasanna, et al. [1] noted that the crop is a vital source of calorie, protein, and some important vitamins and minerals to billions of people world-wide, particularly in africa, south america and asia. approximately 88% of maize produced in ethiopia is consumed as food, both as green and dry grain. the per capita consumption is 50 kg per annum [2]. maize is cultivated globally as one of most important cereal crops and ranks third next to wheat and rice. the total world production of maize is 1,068.79 mt, with the united states producing 384.78 mt, china 219.55 mt, brazil 97.00 mt, european union 60.71 mt, south africa 16.40 mt and ethiopia harvesting 6.35 mt of the total production of maize foreign agricultural service (fas) [3]. central statistical agency (csa) [4] reported that in ethiopia by 2016/17 main cropping season out of the total grain crop area (12,574,107.33 hectare), 81.27% was under cereals of which maize share as large area as 16.98%, after tef (24%). regarding total annual production, cereals contributed 87.42% (253,847,239.63 quintals) in which maize ranked first 27.02% (78,471,746.57 quintals) [4]. over 5.8 million hectares of potential suitable land was identified for the highland maize hybrids in the country [5]. the crop is increasingly grown to the highlands of ethiopia where it has been a minor crop in the past. zeng, et al. [6] pointed out that increased productivity and production of maize has significant positive impact on poverty reduction in sub-saharan african countries including ethiopia. considering its importance, wide adaptation, total production and productivity, maize is one of the high priority crops to feed the increasing population of the country. however, national average yield in ethiopia is still as low as 3.675t ha-1[4] compared to that of the developed world 10.96t ha-1[3] and this warrants the increasing maize productivity as high national priority issue. for such yield gap, a number of production constraints are responsible. the shortages of high yielding varieties or potential parent materials and the effect of biotic and abiotic stresses are the major constraints limiting maize production and productivity [2]. this implies the need for developing high yielding maize varieties from suitable parents and crosses which will perform well under stress and non-stress conditions. identification of suitable inbred lines and superior cross combinations require knowledge of combining ability. information from combining ability (gca and sca) analysis can show the type of gene action involved in controlling quantitative characters thereby assisting breeders in selecting suitable parent materials and crosses [7]. parental lines selection can be performed by particular mating designs such as line x tester, north carolina (nc) designs i, ii and iii, and diallel. line x tester analysis as suggested by kempthorne [8]is useful in deciding the relative ability of female and male lines to produce desirable hybrid combinations [9]. lines x tester programs have been applied to provide a systematic approach for the detection of suitable parents and crosses for investigated characters [10]. combining ability has been studied in ethiopia for different sets of new maize inbred lines [9, 11]. however, it is always mandatory for any breeding program to generate such information for any new batch of inbred lines developed or received outside of the program. in line with this, highland maize breeding program at ambo agricultural research center (aarc) in collaboration with cimmyt recently developed inbred lines and crosses whose genetic information has not been studied. hence, the study was conducted to evaluate the combing ability of inbred lines and their testcrosses. 2. materials and method description of study site: the experiment was conducted at ambo and holeta agricultural research centers of the ethiopian institute of agricultural research (eiar) during the main cropping season of 2017. holeta agricultural research center (harc) is located at 09o04’12”n latitude and 38o29’45”e longitudes and an elevation of 2400 m.a.s.l. the center receives an average rainfall of 1102 mm per annum. the maximum and minimum temperatures of this site are 6oc and 22oc, respectively. the center has nitosols and vertisols soil types with ph of 6.0 [12]. ambo agricultural research center (aarc) is located at 8o57’ n l and 37º 51ꞌ e and the elevation is 2225 m.a.s.l. the site receives an average rainfall of 1115mm. the maximum and minimum temperatures of this site are 11.7oc and 25.4oc, respectively. the soil type of ambo is clay (heavy vertisols) with a ph of 7.8 [13]. 2.1. experimental materials the experiment consisted of 50 maize entries which include 48 testcrosses and two hybrid checks (amh853 kolba and amh851-jibat). the testcrosses (48) were generated from crossing of 24 inbred lines (female parents) with two testers (male parents) in line x tester mating design during 2015/2016 cropping season at ambo agricultural research center. the inbred lines were developed at ambo agricultural research center from cymmyt materials using ear-to-row selection and subsequent selfing until they attain homozygosity. the testers used for the formation of the testcrosses were fs59 (tester 1) and fs67 (tester 2) table 1. the first tester was from heterotic group “b”, while the second was from heterotic group “a”. the checks used in the study were amh851 (jibat) and amh853 (kolba) which are three-way cross hybrid varieties released by ambo agricultural research center, highland maize breeding program in 2011 and 2015, respectively. they take about 178 days for grain mature at ambo and similar environments. besides, hybrid checks are high yielding, tolerant/resistance to major maize disease in the country and well adapted to the altitude ranging from 1800-2600m in the highland sub-humid agro-ecological conditions of the country [14]. agriculture and food sciences research, 2020, 7(2): 113-124 115 © 2020 by the authors; licensee asian online journal publishing group table-1.list and pedigree of parents and hybrid checks used for the study. sn line code pedigree origin 1 l1 (cml442*/ofp4)-b-4-2-2-b-b-b-# amb16n42-29/amb16n42-144 2 l2 (cml495*/ofp14)-7-1-5-1-1-b-b-# amb16n42-107/amb16n42-144 3 l3 (cml442*/ofp4)-b-17-1-1-b-b-b-# amb16n42-32/amb16n42-144 4 l4 (cml495*/ofp6)-b-27-1-1-b-# amb16n42-142/amb16n42-144 5 l5 (cml539*/ofp14)-2-1-1-2-2-b-b-# amb16n42-16/amb16n42-144 6 l6 (cml442*/ofp4)-b-17-5-1-b-b-b-# amb16n42-36/amb16n42-144 7 l7 (cml395*/ofp105)-1-1-1-1-1-b-b-# amb16n42-38/amb16n42-144 8 l8 (cml395*/ofp105)-1-2-3-1-1-b-b-# amb16n42-39/amb16n42-144 9 l9 cml539*/ofp1)-b-11-2-2-b-b-b-# amb16n42-20/amb16n42-144 10 l10 (cml444*/ofp23)-6-3-1-1-1-b-b-# amb16n42-44/amb16n42-144 11 l11 (lpsc7-f96-1-2-1-1-b-b-b*/ofp9)-3-2-1-1-1b-b-# amb16n42-2/amb16n42-144 12 l12 (cml444*/ofp14)-3-2-4-1-2-b-b-# amb16n42-47/amb16n42-144 13 l13 (cml444*/ofp4)-b-4-1-1-b-b-b-# amb16n42-50/amb16n42-144 14 l14 (cml444*/ofp4)-b-6-1-1-b-b-b-# amb16n42-51/amb16n42-144 15 l15 (cml537*/ofp106)-6-1-3-1-2-b-b-# amb16n42-53/amb16n42-144 16 l16 (cml537*/ofp106)-7-1-2-1-2-b-b-# amb16n42-56/amb16n42-144 17 l17 (cml491*/ofp4)-b-10-1-2-b-b-b-# amb16n42-88/amb16n42-144 18 l18 cml546-# amb16n42-61/amb16n42-144 19 l19 ([syn-usab2/syn-elib2]-12-1-1-1-b*4-bb-b*/ofp105)-4-2-1-1-2-b-b-# amb16n42-62/amb16n42-144 20 l20 ([cml312/[tuxpseq]c1f2/p49-sr]f245-3-2-1-bb//inta-f2-192-2-1-1-1-bbbb]1-5-1-1-1-bbb-b-b-b*/ofp106)-1-2-2-2-1-bb-# amb16n42-75/amb16n42-144 21 l21 ([cml444/cml395//dtpwc8f31-1-1-2-2bb]-4-2-2-1-2-bb-b-b-b*/ofp105)-1-4-3-3-2b-b-# amb16n42-65/amb16n42-144 22 l22 ([cml444/cml395//dtpwc8f31-1-1-2-2bb]-4-2-2-1-2-bb-b-b-b*/ofp105)-2-1-1-2-1b-b-# amb16n42-66/amb16n42-144 23 l23 (lpsc7-f71-1-2-1-2-b-b-b*/ofp2)-b-1-3-2b-b-b-# amb16n42-8/amb16n42-144 24 l24 [cml444/cml395//dtpwc8f31-1-1-2-2bb]-4-2-2-2-1-b*7-b-# amb16n42-69/amb16n42-144 tester 25 t1 fs59 heterotic group 26 t2 fs67 heterotic group checks 27 jibat 3-way hybrids 28 kolba 3-way hybrids 2.2. experimental design and procedure the experimental materials along with two hybrid checks were grown during the 2016/2017 main cropping season using alpha lattice design [15] with two replications, 10 incomplete blocks and 5 plots per the incomplete blocks at both locations. each entry was planted in a single row plot of 5.25m length with a spacing of 75cm between rows and 25cm between plants. seeds were planted with two seeds per hill and later thinned to one plant at four leaf stage. 2.3. data collection and analysis data were collected days to 50% anthesis (ad), days to 50% silking (sd), anthesis-silking interval (asi), ear aspect (ea), plant aspect (pa), grain yield (gy), number of ear per plant (epp) and thousand kernel weight (tkw) on plot basis. on plant basis data were collected on plant height (ph), ear height (eh), ear length (el), ear diameter (ed), number of kernel rows per ear (krpe) and number of kernels per row (kpr). plant aspect and ear aspect were measured in 1 to 5 scales. the data obtained for different traits from field measurements were organized and analyzed using sas statistical package [16]. analysis of variance across location was conducted with proc glm procedure by considering location, replication and blocks as random and entry/genotype as fixed factors with statement of rondom and test option. the significance of mean squares for entries, crosses, and location in combined analysis were tested against the mean squares for their corresponding interaction with location as error term, while their interaction with location were tested against their corresponding pooled error. based on general analysis of variance, traits that showed significant differences among the entries were further analyzed according to line x tester analysis [8] using analysis of genetic designs with r (agd-r) version 3.0 procedures for individual and combined data [17]. 2.4. estimation of combining ability effects genotypic means of individual locations were used for the determination of gca and sca. the gca effects of lines (l) and testers (t), the sca effect of lxt, and their interactions with the environment were determined following the method stated by kempthorne [8] assuming the following model. yijke= µ + le + r (l) ke +vij+ (lv) ije +ԑijke where, yijke = observed value from each experimental unit; µ = grand mean; le = location effect; rke = replication effect within each location vij = f1 hybrid effect = gi+gj+sij, where, gi = general combining ability of ith lines; gj = general combining ability of jth tester; sij = specific combining ability of ijthf1 hybrids; (lv) ije = interaction effect of ijth f1 hybrid and eth location ԑijke = residual effect. the significance of gca and sca effects were performed computing the standard error for lines, testers and crosses and then tested against t-test by taking the degree of freedom of pooled error mean square [18, 19]. the proportional contributions of lines (gcal), testers (gcat), and their interaction (sca lxt) to the sum square of crosses were calculated as the ratio between sum of squares of each component and the cross sum of squares as given [19]. agriculture and food sciences research, 2020, 7(2): 113-124 116 © 2020 by the authors; licensee asian online journal publishing group 3. results and discussion the analysis of variance and general and specific combining ability effects were computed and the results are discussed below. 3.1. analysis of variance the analysis of variances for yield and yield related traits for individual and combined location are presented in appendix 1, 2 and table 2. significant differences were detected between the two locations for all of the studied traits except for ear length, indicating that the two locations differed in the environmental conditions to cause variation which agreed with the finding of aly and khalil [20]. entry difference were significant (p<0.01 or p<0.05) for all traits except for plant aspect table 2. entries differed in their performance from one location to another for variable like grain yield, plant and ear height, ear diameter and 1000 kernels as entry x location interaction was significant. beyene, et al. [21] and murtadha, et al. [22] also reported significant entry x location interaction effect for some yield and its components in maize and indicated presence of wide variability with regard to tested entry and locations. the result showed the location played significant role in the variation of these traits. if significant genotype x location interaction mean squares existed, different genes involved in controlling the traits show the inconsistency of the genes over locations [23]. the interaction of entry with location suggests further evaluation of the genotypes across more number of locations to minimize environmental effect from computation genetic variance. table-2. analysis of variance for yield and yield related traits of 48 testcross and two hybrid checks evaluated at holeta and ambo, 2017. 3.2. combining ability analysis significant differences (p<0.01, or p<0.05) were observed among the crosses for all traits except for anthesissilking interval at holetta and for number of kernels per row at ambo appendix 3 and 4. at ambo, gca mean squares due to lines were significant (p<0.01, or p<0.05) for most of the studied traits except for ear position and ear aspect, while tester gca mean squares were significant (p<0.01, or p<0.05) for days to 50% silking, anthesissilking interval, plant and ear height, ear aspect and 1000 kernels weight appendix 3. similarly, sca mean squares for line x tester were significant for grain yield, ear height, ear position and ear aspect. at holetta, gca mean squares of lines and testers were significant (p<0.01, p<0.05) for all traits except for ear length, anthesis silking interval and number of kernels per row and gca of tester for days to 50% anthesis, number of kernels per row and 1000 kernels weight. the mean squares of sca line x testers were also significant (p<0.01, or p<0.05) for seven traits and non-significant for other seven studied traits appendix 4. this indicates both additive and non-additive gene actions with different level from location to location were involved in the inheritance of these traits. the observed differences in the level of significance of gca and sca mean squares with changing locations were an indication of environmental effect on the preponderance of additive and non-additive gene action. zare, et al. [24] reported the different level of gca and sca mean square for different testing environment. in contrast, haddadi, et al. [25] found no environmental effect on the preponderance of additive and non-additive gene action. the significance of mean squares due to lines and testers indicated inbred lines variation among the lines and tester in their performance. meanwhile, significant line x tester interaction suggests that inbred lines performed differently according to the testers to which they were crossed. combined analysis of line x tester showed highly significant (p<0.01) difference among cross for all studied traits table 3a and 3b. cross x location interaction were significant (p<0.01, or p<0.05) for most traits except for days to 50% anthesis, anthesis silking interval and ear position. this indicates the presence of wide genetic variations among the studied materials. these findings are in agreement with those reported by aly and hassan [26]; aly and khalil [20] and mousa [27]. the mean squares of gca lines and testers and sca were significant (p<0.01, or p<0.05) for all traits except gca of testers for days to 50% anthesis, number of ear per plant and ears aspect, and sca for anthesis silking interval, number of ears per plant, number of kernel rows per ear and ear diameter, which confirms to the finding of mohammad, et al. [28] and dar, et al. [29]. significant gca and sca mean squares indicate that both additive and non-additive gene effects were involved for the variation of crosses through all studied traits. however, higher gca proportional contribution and gca/sca ratio greater than unit indicated predominance of additive gene action for all studied traits. this indicates that variations among crosses through all the studied traits were mainly due to additive gene effect which was in accordance with the finding of amare, et al. [30]. adebayo and menkir [31];satyanvesh [32];tolera, et al. [11] have earlier reported that additive gene actions were more important than non-additive gene actions for inheritance of grain yield and yield related traits. however, aminu, et al. [33] and arsode, et al. [34] reported that non-additive gene action was predominated in the inheritance of these traits. based on current findings, the inheritance of all traits was governed by additive gene effect, suggesting selection would be effective in improving yield and yield related traits. trait l, df=1 re(l)df=2 b(l*r) df=36 ent df=49 ent*l df=49 error df=62 mean±se(m) cv% r2 gy 8.38* 0.03 1.29 4.41* 2.63** 1.1 7.53± 0.52 13.9 0.86 ad 14162.4** 24.23** 2.96 13.33** 2.77 3.18 104.52±0.89 1.71 0.99 sd 18489.6** 19.34** 2.60 15.66** 2.51 3.31 105.15±0.91 1.73 0.99 asi 0.63** 0.001 0.005 0.007* 0.005 0.004 1.2± 0.03 5.52 0.86 ph 574.6** 779.0** 161.6 1631.89** 237.4* 139.1 251.07±5.9 4.70 0.93 eh 5724.5** 398.33** 45.04 943.11** 85.85* 54.64 136.66±3.7 5.41 0.95 epo 0.07** 0.0002 0.001 0.004** 0.0007 0.002 0.54±0.02 7.33 0.79 epp 1.49** 0.007 0.03 0.13** 0.05 0.03 1.70±0.09 10.18 0.86 ea 0.78* 0.91** 0.13 0.43** 0.19 0.13 3.12±0.18 11.56 0.84 pa 2.88** 0.75* 0.15 0.20 0.14 0.20 3.30±0.22 13.69 0.70 el 1.69 8.82** 0.98 3.61** 1.21 0.81 15.47±0.45 5.82 0.88 ed 1.62** 0.004 0.03 0.10** 0.03** 0.03 4.32±0.09 3.84 0.86 krpe 10.76** 0.58 0.63* 1.21** 0.47 0.37 12.86±0.3 4.74 0.86 kpr 19.22* 25.22** 7.43* 8.51** 6.50 4.22 32.3±1.03 6.37 0.83 tkw 193827.8** 27.26 743.1 3102.2** 1603.9* 947.3 305.0±15.39 10.09 0.90 agriculture and food sciences research, 2020, 7(2): 113-124 117 © 2020 by the authors; licensee asian online journal publishing group gca line x location interaction was significant (p<0.01, or p<0.05) for traits such as grain yield, ear height, number of ears per plant, ear length and number of kernels row/ear, while gca tester x location interaction effect showed significant for all studied traits except for days to 50% anthesis and silking, ear position and ear length table 3aand 3b. these suggest that the lines and testers were depended on the location in which they were grown and limited number of testing location would be insufficient. comparable results were reported aly [35]; fan, et al. [36] and noelle, et al. [37] for the materials they studied. on the contrary, non-significant gca x location interaction mean squares were reported by alemeshet [38]. significant gca x location interaction implies that the trend of variation of gca of lines and testers were different across location and selection for good combining lines would be effective if based on hybrid performance across a range of environment. this also suggests the need for selecting inbred lines at specific locations. however, mean squares of sca x location interaction were non-significant for all investigated traits which were in agreement with the finding of dagne, et al. [39]; adebayo and menkir [31] and fan, et al. [36]. but, tulu, et al. [40] for grain yield and ear per plant, noelle, et al. [37] for days to 50% silking and ear aspect and mosa, et al. [41] for ear length, number of rows per ear and number of kernels per row found significant sca x location interaction. table-3a. analysis of variance and heritability for yield and yield related traits of 48 testcrosses and two commercial hybrid checks evaluated at ambo and holeta, 2017. source df gy t/ha ad day sd day asi day ph cm eh cm epo % loc 1 7.76* 12661** 17787** 193** 351.72 5184 ** 0.067** rep(loc) 2 0.06 23.07** 18.89** 0.32 403.86 357.6 ** 0.000 cross 47 5.20** 13.92** 18.23** 3.03* 1952.07** 1208.2 ** 0.005** line(gca) 23 5.92** 21.48** 25.75** 2.30* 1019.01** 772.4** 0.006** tester(gca) 1 22.07** 10.45 105.02** 47.51** 59791.93** 33769 ** 0.033** line xtester 23 3.75** 6.50* 6.94** 1.82 370.08** 228** 0.002 cross x loc 47 2.57** 3.90 4.38* 1.90 221.59* 96.49* 0.001 line x loc 23 2.81** 4.08 4.60 1.39 205.39 93.50 * 0.001 tester x loc 1 20.90** 0.31 14.08* 17.06** 2665.16** 1307** 0.001 l x t x loc 23 1.54 3.87 3.75 1.75 131.55 46.84 0.001 gca/sca 1.83 3.38 4.37 2.40 9.78 9.82 4.67 error 58 1.07 3.14 3.16 1.65 129.31 52.26 0.001 pro. cont (%) l 55.66 75.55 69.11 37.20 25.55 31.29 67.33 t 9.02 1.60 12.25 33.38 65.17 59.47 15.04 lxt 35.32 22.85 18.64 29.42 9.28 9.24 17.63 h2 b 28.19 68.26 71.21 34.45 84.65 88.76 80 table-3b.analysis of variance and heritability for yield and yield related traits of 48 testcrosses and two commercial hybrid checks evaluated at ambo and holeta, 2017. source df epp no ea scale el cm ed cm krpe no kpr no tkw gm loc 1 1.35** 1.01** 1.88** 1.08** 4.20** 5.83 186083** rep(loc) 2 0.006 0.67* 6.69** 0.01 0.18 7.21 14.33 cross 47 0.147** 0.51** 4.74** 0.11** 1.30** 9.43** 3885.10** line(gca) 23 0.229** 0.80** 7.36** 0.18** 1.53** 11.68** 5356.10** tester(gca) 1 0.193* 0.05 0.88 0.34** 11.79** 10.46 34114.7** cross x loc 47 0.050* 0.21* 1.33* 0.04* 0.57* 6.31* 1909.8** line x loc 23 0.055* 0.18 1.55* 0.04 0.62* 6.84 1256.1 tester x loc 1 0.188* 1.94* 2.59 0.65** 1.87* 1.90 26660.1** lxt x loc 23 0.039 0.17 1.05 0.02 0.47 5.96 1486.45 gca/sca 3.45 3.28 3.23 5.09 3.37 1.70 6.21 error 58 0.031 0.13 0.85 0.02 0.36 3.96 820.01 pro.cont (%) l 74.55 76.39 76.027 77.16 57.76 60.65 67.46 t 2.99 0.22 0.003 6.42 19.32 2.36 18.68 lxt 22.45 23.39 23.64 16.42 22.89 37.04 13.86 h2 b 58.43 50.85 65.33 53.85 48.34 26.49 39.72 3.3. general combining ability effect estimates of gca effects of 24 inbred lines and two testers for combined data of yield and yield related traits are presented in table 4aand 4b. out of 24 lines, three of them had positive and highly significant gca effects and six lines exhibited negative and significant (p<0.01, or p<0.05) gca effects for grain yield. line l23 showed maximum gca effect (1.64t ha-1) followed by l11 (1.50t ha-1), whereas l13 revealed lowest gca effect (-1.57t ha-1) followed by l3 (-1.31t ha1). this depicts the presence of best and poorest general combiners in the group of the studied inbred lines. l23, l11 and l15 found to be good general combiners suggesting their ability to transmit additive genes in desirable direction for grain yield. l3, l7, l13, l21, l18 and l19 found to be poor general combiners as they had tendency to reduce grain yield. among the testers, fs59 revealed significantly positive gca effect. several researchers girma, et al. [9]; natol [42] reported both positive and negative significant gca effects for maize grain yield in different set of materials. high gca effects are attributed to additive or additive x additive gene effects which denote the fixable genetic components of variance [43]. the inbred lines with good general combining ability can make complementary single cross which can be used as seed parent for three-way or double cross hybrid development. for days to 50% anthesis and silking, gca effects of inbred lines ranged from -2.81 to 2.70 and -2.89 to 3.49 table 4a, respectively. l3, l7, l10, l13, l14 and l22 showed positive gca effects, while lines l1, l4, l5, l9 and l18 had negative gca effects for both traits. lines gca effect for anthesissilking interval ranged from -1.41 (l12) to 1.06 (l3) agriculture and food sciences research, 2020, 7(2): 113-124 118 © 2020 by the authors; licensee asian online journal publishing group and tester gca effects for anthesis-silking interval were -0.5 (t2) to 0.5 (t1). despite the importance of negative gca effects for anthesis-silking interval, only l12 was found to be good general combiner. negative and significant line gca affects for days to 50% anthesis and silking, and anthesis silking interval suggests the possibility of exploiting favorable genes for earliness to maturity and narrower anthesis silking interval in future breeding work. similarly, sundararajan and kumar [44] and demissew [13] suggested the importance of the negative gca effects for days to 50% anthesis and silking to develop early maturing varieties. for plant and ear height, l1, l5, l10, l12, and l14 revealed positive and significant lines gca effect, while l2, l8, l13, l18 and l22 showed negative and significant gca effects. inbred lines with negative gca effects are good general combiners for plant and ear height. girma, et al. [9] concluded that shorter plant height with lower ear placement is desirable. inbred lines with negative gca effect had tendency to reduce plant and ear height, hence genotypes with shorter plant height with lower ear placement can be good for lodging resistance. ji, et al. [45] also noted that cultivars with high ear positions are prone to root and stalk lodging and suggested that negative significant lines gca effects are desirable shorter stature and lower ear placement. significant gca effects of inbred lines ranged from -0.37 (l18) to 0.40 (l23) for number of ear per plant, -1.71 (l13) to 2.06 (l15) for ear length, -0.24 (l7) to 0.27 (l10) for ear diameter, -0.83 (l17) to 0.73 (l20) for number of kernel rows per ear and -3.02 (l1) to 2.27 (l4) for number of kernels per rows table 4b. l1, l9, l23 and l24 for number of ears per plant, l4, l7, l8, l10, l12, l14, l15 and l24 for ear length, l4, l10, l11, l12, l14, l20 and l21 for ear diameter and l8, l10, l11, l14, l20 for number of kernel rows per ear and l4 and l15 for number of kernels per rows showed positive and significant gca effects. on the other hand, l4, l8, l13, l18, l20 and l21 for number of ears per plant, l1, l9, l11, l13 and l19 for ear length, l1, l7, l16, l17, l19, l22, l23 and l24 for ear diameter, l2, l5, l16, l17, l21 and l22 for number of kernel rows per ear and l1, l3 and l9 for number of kernels per row revealed negative and significant line gca effect. ejigu, et al. [46] also reported certain inbred lines with good general combing ability and suggested the possibility of improving these traits through selection. the present study also identified inbred lines with good general combing ability which could be used for the improvement of traits of interest as these lines had potential to transfer favorable genes to their progenies. the range of lines gca effect for 1000 kernels weight varied from -45.84 (l7) to 77.36 (l21). inbred lines l11, l14, l21 and l22 had positive and significant line gca effects, while l2, l7, l9, l13, l15 and l19 showed negative and significant line gca effects. hence, l11, l14, l21 and l22 were good general combinersand had favorable allele frequency for developing varieties with heavy grain weight. concerning testers, fs59 was a good general combiner for grain yield, ear diameter, number of kernels row per ear and 1000 kernels weight. likewise, fs67 found to be good general combiner for days to 50% silking, anthesissilking interval, plant height, ear height and ear position. generally, inbred lines and testers with good general combiners could be utilized in the improvement of traits of interest either during hybrid or synthetic variety development. table-4a.estimates of general combining ability (gca) effect of lines and testers for yield and yield related traits across two locations, 2017. lines gy t/ha ad days sd days asi days ph cm eh cm epo ratio l1 0.45 -2.05** -2.26** -0.30 10.26* 7.72** 0.01 l2 -0.30 -1.06 -1.01 0.13 -15.83** -11.77** -0.01 l3 -1.31** 2.57** 3.49** 1.06* -5.15 -3.65 0.00 l4 -0.08 -2.05** -1.64* 0.85 10.30* 2.10 -0.02 l5 0.72 -2.81** -2.39** -0.08 27.22** 11.86** -0.01 l6 -0.64 -0.68 -1.51* -0.83 -2.32 1.71 0.01 l7 -0.86* 2.70** 2.99** 0.23 -5.81 4.72 0.04** l8 -0.07 -0.17 0.61 0.74 -10.37* -8.79** -0.02 l9 0.39 -2.30** -2.89** -0.57 1.82 -9.02** -0.04** l10 0.68 1.57* 1.61* 0.05 13.92** 8.72** 0.00 l11 1.50** 1.58* 1.24 -0.29 3.97 8.97** 0.03* l12 0.37 1.95** 0.61 -1.41** 9.59* 21.11** 0.06** l13 -1.57** 1.95** 2.11* 0.13 -14.11** -13.40** -0.02 l14 0.31 2.32** 2.74** 0.47 13.19** 5.97* 0.00 l15 1.26** 0.33 0.86 0.49 3.42 4.83 0.01 l16 0.05 -0.68 -0.89 -0.24 6.50 11.37** 0.03* l17 -0.08 -1.18 -1.39* -0.26 7.62 6.10* 0.01 l18 -0.78* -1.55* -1.64* -0.11 -16.52** -8.65** 0.00 l19 -1.13** -0.31 -0.26 0.09 -20.87** -2.90 0.05** l20 0.22 -0.30 -0.14 0.35 -4.47 -1.27 0.00 l21 -1.15** 0.20 -0.39 -0.64 5.51 5.22* 0.01 l22 -0.16 1.57* 1.74** 0.28 -10.03* -12.52** -0.03* l23 1.64** -1.05 -1.14 -0.16 -4.89 -12.52** -0.04** l24 0.57 -0.56 -0.51 0.06 -2.17 -15.91** -0.06** se 0.36 0.63 0.63 0.45 4.02 2.56 0.01 tester t1 0.34** 0.24 0.74** 0.51** 18.39** 13.28** 0.01** t2 -0.33** -0.24 -0.74* -0.51 ** -18.33** -13.28** -0.01** se 0.11 0.26 0.18 0.13 1.16 0.74 0.004 note: **significant (p<0.01), *significant (p<0.05), l=line, t=tester, se= standard error, gy=grain yield, ad=anthesis days, sd=silking days, asi=anthesissilking interval, ph=plant height, eh=ear height, epo=ear position. agriculture and food sciences research, 2020, 7(2): 113-124 119 © 2020 by the authors; licensee asian online journal publishing group table-4b. estimates of general combining ability (gca) effect of lines and testers for yield and yield related traits across two locations, 2017. lines epp no ea scale el cm ed cm krpe no kpr no tkw gm l1 0.24** -0.26* -1.54** -0.13** -0.21 -3.02** -12.62 l2 0.11 0.55** -0.58 -0.08 -0.44* 0.59 -24.07* l3 -0.05 0.06 -0.56 -0.11 0.38 -1.98** -4.06 l4 -0.14* -0.38** 0.81* 0.16** 0.36 2.27** 4.85 l5 0.06 -0.32* 0.29 0.02 -0.63** 0.38 1.95 l6 0.05 -0.56** -0.41 0.09 0.35 -0.38 -8.94 l7 -0.02 0.56** 1.05** -0.24** -0.07 0.51 -45.84** l8 -0.16* 0.00 0.98** 0.01 0.50* 0.91 18.04 l9 0.16* 0.32* -1.64** 0.03 0.37 -2.14** -26.96** l10 -0.05 -0.44** 0.92** 0.27** 0.48* 1.02 7.06 l11 0.05 0.62** -1.34** 0.19** 0.54* -0.30 31.41** l12 0.04 0.18 0.73* 0.11* -0.12 1.20 29.04** l13 -0.19** 0.06 -1.71** 0.10 -0.07 -0.75 -21.20* l14 0.02 -0.13 0.97** 0.11* 0.54* 0.57 -12.61 l15 0.01 -0.32* 2.06** 0.06 0.39 2.10** -21.46* l16 0.05 0.25 -0.44 -0.11* -0.66** -0.48 -15.47 l17 0.03 0.05 -0.08 -0.16** -0.83** -1.23 12.36 l18 -0.37** -0.38** 0.19 0.10 -0.01 0.50 -3.44 l19 -0.13 -0.01 -0.85* -0.21** 0.18 -0.02 -35.00** l20 -0.18** 0.06 0.42 0.20** 0.78** 0.73 14.49 l21 -0.19** 0.06 0.04 0.13** -0.64** -1.06 77.36** l22 0.07 0.12 0.16 -0.22** -0.65** -0.83 22.73* l23 0.40** 0.00 -0.27 -0.15** -0.27 0.64 3.06 l24 0.15* -0.12 0.82* -0.18** -0.31 1.19 9.30 se 0.06 0.13 0.32 0.05 0.21 0.70 10.12 tester t1 0.03 -0.02 0.06 0.04** 0.26** 0.25 -13.33** t2 -0.03 0.02 -0.06 -0.04** -0.26** -0.21 13.33** se 0.02 0.06 2.92 note: **significant (p<0.01), *significant (p<0.05), l=line, t=tester, se= standard error, epp=ear per plant ea=ear aspect el=ear length ed=ear diameter, krpe=kernel rows per ear, kpr=kernels per rows, tkw=thousand kernels weight. 3.4. specific combining ability effect estimates of specific combining ability (sca) effects for grain yield and yield related traits for all hybrids computed across locations are presented in table 5aand 5b. crosses l2 x t1 (1.03), l3 x t1 (1.04), l8 x t1 (1.10), l11 x t1 (1.14), l13 x t2 (1.64) and l23 x t1 (1.64) are exhibited significant positive sca effects for grain yield. crosses like l2 x t2 (-1.03), l3 x t2 (-1.04), l8 x t2 (-1.10), l11 x t2 (-1.14), l13 x t1 (-1.64) and l23 x t2 (-1.64) found to be poor specific combiners which could be due to the presence of unfavorable gene combinations in the parents. the positive sca effects indicate lines were from opposite heterotic group, while negative sca affects refer lines were from the same heterotic group. for instance, cross l2 x t1 showed significant positive sca effect, but had lower grain yield than standard checks. the high positive and significant sca effects were manifested by crosses of low x low (l13 x t2 and l3 x t2), low x high (l2 x t1 and l8 x t1) and high x high (l23 x t1, l11 x t1), indicating the presence of complementary gene action for grain yield. inbred lines l3, l2, l13 and t2 had poor general combining ability, but resulted in hybrids with higher sca effects for grain yield. this signifies that inbred lines with poor general combining ability might produce better hybrids depending on the other parent with which it combines. the sca effects of days to 50% anthesis (ad) and silking (ad) ranged from -1.98 (l16 x t1) to 1.98 (l16 x t1) and -2.14 (l21 x t2) to 2.14 (l22 x t1), respectively. crosses such as l13 x t2, l16 x t1 and l21 x t2 revealed negative and significant sca effects for days to 50% anthesis and silking, while l13 x t1, l16 x t2 and l21 x t1 showed positive sca effects. crosses with negative and significant sca effects for these traits are considered as good specific combiners, which indicate the earliness of the hybrids. on the other hand, crosses with positive and significant sca effects were found to be poor specific combiners, mainly for highland condition due to lateness. inbred lines with negative and significant gca effects did not necessarily produced crosses with negative and significant sca effects. but, tester fs59 had negative and significant gca effect and interacted well with l21 and l13 in cross combination thereby resulted in crosses with negative and significant sca effects. the interactions involved in the crosses were poor x good general combiners (l13 x t1 and l21 x t1) and poor x poor (t16 x t2). crosses l9 x t1, l16 x t2 and l21 x t1 had negative and significant sca effects for plant and ear height considered as good specific combinations. these crosses involved parents with general combiners of good x poor, poor x good and poor x poor gca effects. l13 x t2, l16 x t1 and l21 x t2 for plant height and l9 x t2, l10 x t1, l12 x t2, l16 x t2 and l21 x t1 for ear height showed positive and significant sca effects, indicating that these crosses were poor specific combination for these traits. accordingly, good specific combination could be utilized in developing lodging resistance hybrids. positive sca effects for plant and ear height indicates crosses had tendency to increase height in undesirable direction. crosses l8 x t1 for number of ears per plant, l9 x t1, l10 x t2 and l14 x t1 for ear length and l11 x t2 for number of kernels per row had significant and positive sca effects. the crosses involved parent interaction of poor x poor, good x poor and good x poor based on their gca effects. l8 x t2 for number of ears per plant, l9 x t2, l10 x t1 and l14 x t2 for ear length and l11 x t1 for number of kernels per row showed significant and negative sca effects. the crosses exhibiting significant and positive sca effects are considered as good specific combiners. the crosses that showed significant and negative sca effects found to be poor specific combiners. moreover, crosses with positive and significant sca effects had inbred lines with positive (l10 and l14) and negative (l9) gca effect. agriculture and food sciences research, 2020, 7(2): 113-124 120 © 2020 by the authors; licensee asian online journal publishing group finally, based on the gca effects of parents, and sca effects hybrids for different traits, it is evident that high specific combinations involved, inbred lines with high x high, high x low and low x low gca effects. the sca effects of crosses combinations exhibited no specific trends in cross combinations between parents having good and poor gca effects. this indicates high specific combiners were not only obtained from the combination of good x good general combiners, but also resulted from good x poor and poor x poor general combiners. the superiority of hybrids resulted from the crosses of high x low combiners could be due to interactions between positive alleles and negative alleles from good and poor combiners, respectively. the best performance of these combination may be caused by additive x additive (high x high), additive x dominance (high x low), or dominance x dominance (low x low) gene interactions dey, et al. [47]; talukder, et al. [48]. zhang, et al. [49] suggested that hybrids with high sca effects from parents with low gca effects might be primarily due to non-additive gene action which includes dominance and epistasis. for some of the studied traits, high gca effect parents resulted into hybrids with low sca effect which might be due to the lack of complementary of gene action. on the other hand, lowgca effect parents produced hybrids with high sca effects which might be caused by complementation of gene actions. generally, the crosses identified were best specific combiners for yield and yield related traits which can be used in the heterosis breeding. table-5a.estimates of specific combining abilities of line x tester for yield and yield related traits across two location, 2017. s/n code gy t/ha ad days sd days ph cm eh cm epp no 1 l1xt1 -0.14 -0.11 0.01 -0.91 2.22 -0.03 2 l1xt2 0.13 0.11 -0.01 0.85 -2.22 0.03 3 l2xt1 1.03* 1.12 0.26 1.86 -4.52 -0.02 4 l2xt2 -1.03* -1.12 -0.26 -1.93 4.52 0.02 5 l3xt1 -1.04* -0.12 -0.24 -4.16 -2.41 -0.01 6 l3xt2 1.04* 0.12 0.24 4.09 2.41 0.01 7 l4xt1 -0.17 0.03 -0.11 -4.46 -0.63 0.09 8 l4xt2 0.17 -0.03 0.11 4.40 0.63 -0.09 9 l5xt1 0.30 0.39 -0.11 -3.85 0.34 -0.07 10 l5xt2 -0.31 -0.39 0.11 3.78 -0.34 0.07 11 l6xt1 0.56 0.02 -0.74 -3.13 -4.54 0.00 12 l6xt2 -0.56 -0.02 0.74 3.06 4.54 0.00 13 l7xt1 -0.58 -0.85 -0.24 2.10 -2.03 -0.13 14 l7xt2 0.58 0.85 0.24 -2.17 2.02 0.13 15 l8xt1 1.10* 0.01 0.14 7.63 2.97 0.26** 16 l8xt2 -1.10* -0.01 -0.14 -7.70 -2.97 -0.26** 17 l9xt1 0.18 -0.86 -0.11 -10.57** -9.27** 0.05 18 l9xt2 -0.18 0.86 0.11 10.51** 9.27** -0.05 19 l10xt1 -0.72 0.76 0.89 5.60 7.72* -0.03 20 l10xt2 0.72 -0.76 -0.89 -5.66 -7.72* 0.03 21 l11xt1 1.14** 0.00 0.51 4.01 2.48 0.01 22 l11xt2 -1.14 0.00 -0.51 -4.07 -2.48 -0.01 23 l12xt1 0.34 0.64 0.89 2.29 12.86** 0.03 24 l12xt2 -0.34 -0.64 -0.89 -2.36 -12.86** -0.03 25 l13xt1 -1.64** 1.88* 1.89* -11.57** -5.67 -0.04 26 l13xt2 1.64** -1.88* -1.89* 11.51** 5.66 0.04 27 l14xt1 0.55 -0.23 -0.49 4.36 3.48 0.08 28 l14xt2 -0.55 0.23 0.49 -4.43 -3.48 -0.08 29 l15xt1 -0.04 0.76 0.14 1.56 2.61 -0.06 30 l15xt2 0.04 -0.76 -0.14 -1.63 -2.61 0.06 31 l16xt1 0.94 -1.98* -2.11* 14.87** 9.61** -0.05 32 l16xt2 -0.94 1.98* 2.11* -14.93** -9.61** 0.05 33 l17xt1 -0.73 -0.27 -0.11 4.03 0.09 -0.15 34 l17xt2 0.72 0.27 0.11 -4.09 -0.09 0.15 35 l18xt1 0.53 -0.87 0.39 9.55 2.35 0.00 36 l18xt2 -0.53 0.87 -0.39 -9.62 -2.35 0.00 37 l19xt1 0.03 -0.86 -1.24 -5.79 -4.90 0.04 38 l19xt2 -0.03 0.86 1.24 5.72 4.90 -0.04 39 l20xt1 0.29 -0.88 -1.11 7.64 1.97 0.10 40 l20xt2 -0.30 0.88 1.11 -7.70 -1.97 -0.10 41 l21xt1 -0.56 1.62* 2.14* -12.70** -8.02* -0.11 42 l21xt2 0.56 -1.62* -2.14* 12.63** 8.02* 0.11 43 l22xt1 -0.17 1.03 0.76 3.72 -5.26 0.09 44 l22xt2 0.17 -1.03 -0.76 -3.79 5.26 -0.09 45 l23xt1 1.64** -0.37 -0.86 -4.70 0.96 0.06 46 l23xt2 -1.64** 0.37 0.86 4.64 -0.97 -0.06 47 l24xt1 -0.87 -0.87 -0.49 -8.17 -2.41 -0.08 48 l24xt2 0.87 0.87 0.49 8.11 2.41 0.08 se(d) 0.52 0.89 0.89 5.69 3.23 0.09 note: **significant (p<0.01), *significant (p<0.05), se= standard error, gy=grain yield, ad=anthesis days, sd=silking days, ph=plant height, eh=ear height, agriculture and food sciences research, 2020, 7(2): 113-124 121 © 2020 by the authors; licensee asian online journal publishing group table-5b.estimates of specific combining abilities of line x tester for yield and yield related traits across two location, 2017. s/n code ea scale el cm kpr no s/n code ea scale el cm kpr no 1 l1xt1 0.02 -0.45 0.73 25 l13xt1 -0.05 -0.42 0.60 2 l1xt2 -0.01 0.45 0.73 26 l13xt2 0.05 0.42 -0.60 3 l2xt1 -0.04 0.27 -0.21 27 l14xt1 -0.10 1.03* 1.54 4 l2xt2 0.04 -0.27 0.21 28 l14xt2 0.10 -1.03* -1.54 5 l3xt1 -0.05 0.26 0.16 29 l15xt1 -0.05 -0.80 0.52 6 l3xt2 0.05 -0.26 -0.16 30 l15xt2 0.05 0.80 -0.52 7 l4xt1 0.02 -0.13 0.08 31 l16xt1 -0.11 0.27 0.42 8 l4xt2 -0.02 0.13 -0.8 32 l16xt2 0.11 -0.27 -0.42 9 l5xt1 -0.42* -0.03 0.22 33 l17xt1 -0.04 0.75 0.91 10 l5xt2 0.42* 0.03 -0.22 34 l17xt2 0.04 -0.75 -0.91 11 l6xt1 0.08 0.09 0.13 35 l18xt1 0.02 0.14 1.08 12 l6xt2 -0.08 -0.09 -0.13 36 l18xt2 -0.02 -0.14 -1.08 13 l7xt1 0.08 0.18 -1.08 37 l19xt1 0.02 0.03 1.48 14 l7xt2 -0.08 -0.18 1.08 38 l19xt2 -0.02 -0.03 -1.48 15 l8xt1 -0.23 -0.03 -0.01 39 l20xt1 -0.29 -0.22 0.65 16 l8xt2 0.23 0.03 -0.01 40 l20xt2 0.29 0.22 -0.65 17 l9xt1 0.21 1.25** 1.76 41 l21xt1 0.08 -0.55 -1.04 18 l9xt2 -0.21 -1.25** -1.76 42 l21xt2 -0.08 0.55 1.04 19 l10xt1 0.08 -0.99* -0.96 43 l22xt1 -0.10 -0.08 -0.99 20 l10xt2 -0.08 0.99* 0.96 44 l22xt2 0.10 0.08 0.999 21 l11xt1 0.27 -0.84 -2.39* 45 l23xt1 0.02 0.10 -1.13 22 l11xt2 -0.27 0.84 2.36* 46 l23xt2 -0.02 -0.10 1.13 23 l12xt1 0.45* -0.24 -0.50 47 l24xt1 0.15 0.39 -0.69 24 l12xt2 -0.45* 0.24 0.50 48 l24xt2 -0..15 -0.39 0.69 se 0.18 0.46 0.99 note: ** highly significant (p<0.01),*significant (p<0.05), ea=ear aspect, el=ear length, ed=ear diameter, krpe=kernel rows per ear, kpr=kernels per rows, tkw=thousand kernels weight. 4. conclusion analysis of variance revealed the presence of significant variation among the genotypes for all studied traits except for plant aspect. mean square of entry x location interaction for most of studied traits showed nonsignificant, indicating that hybrid performance was consistent across locations. nevertheless, genotype x location interaction was significant for some traits, suggesting further evaluation of selected genotypes over a number of locations. mean squares of gca and sca were significant for most of studied traits, implying that both additive and non-additive gene action were involved in the control of the inheritance of these traits. however, the proportional contribution of gca effect was higher and gca/sca ratio was greater than unit for all studied traits, suggesting that additive gene action was more important than non-additive gene action for these traits. estimation of gca and sca effects identified lines and crosses with good general and specific combiners for studied traits respectively. inbred lines l23, l11 and l15 for grain yield, l1, l9, l23 and l24 for number of ears per plant, l8, l14 and l15 for ear length, l8, l14 and l20 for number of kernels row per ear and l4 and l15 for number of kernels per row were the top good general combiners for the respective traits. crosses such as l11xt1, l13xt2 and l23xt1 for grain yield, l6xt2 for number of ear per plant, l9xt1, l10xt2 and l14xt1 for ear length and l11xt2 for number of kernels per row revealed good specific combining ability. finally, better performing testcrosses, inbred lines with desirable gca effects and cross combinations with desirable sca effects for grain yield and yield related traits could be used as source of useful genetic material for future maize breeding. references [1] b. prasanna, s. vasal, b. kasahun, and n. singh, "quality protein maize," current science, vol. 81, pp. 1308-1319, 2001. 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[49] x. zhang, l. liangjie, l. chao, g. baojian, and x. rugen, "combining ability of different agronomic traits and yield components in hybrid barley," plos one, vol. 10, pp. 1-9, 2015. agriculture and food sciences research, 2020, 7(2): 113-124 123 © 2020 by the authors; licensee asian online journal publishing group appendix appendix-1.analysis of variance for grain yield and yield related traits of 50 maize genotypes evaluated at ambo, 2017. traits rep(df=1) blk(rep)df=18 entry(df=49) error df=31 mean cv % gy 0.06 0.904 4.26** 1.19 7.74 14.08 ad 16.0* 1.01 6.23* 3.38 96.10 1.91 sd 13.69 2.75 9.63** 3.9 95.53 2.07 asi 0.0002 0.003 0.008** 0.002 1.1 4.24 ph 501.76 224.06 776.49** 207.71 207.71 5.70 eh 272.25* 32.20 399.64** 50.28 142.01 4.99 epo 0.00004 0.001 0.002 0.003 0.56 9.17 epp 0.01 0.027 0.094** 0.029 1.62 10.47 ea 0.12 0.059 0.20** 0.095 3.056 7.66 pa 0.49 0.16 0.19 0.18 3.42 12.52 el 8.82** 1.01 2.51** 1.09 15.37 6.78 ed 0.004 0.04 0.07** 0.03 4.41 4.05 krpe 0.37 0.93** 0.56 0.35 12.63 4.68 kpr 6.35 8.97 7.20** 5.68 32.56 7.32 tkw 22.37 987.20 3282.45** 1024.25 336.13 7.52 note: **significant (p<0.01),*significant (p<0.05), rep=replication, blk=block, cv=correlation variation. appendix i2. analysis of variance for grain yield and yield related traits of 50 maize genotypes evaluated at holetta, 2017. traits rep(df=1) blk(rep)df=18 entry(df=49) error df=31 mean cv% gy 0.006 1.67 3.12** 1.01 7.33 13.73 ad 32.49 ** 4.91 11.19** 2.98 112.93 1.53 sd 25.0** 2.43 10.23** 2.71 114.76 1.44 asi 0.001 0.006 0.004* 0.006 1.22 6.37 ph 1056.25** 99.13 1136.63** 70.54 249.37 3.37 eh 524.41** 57.88 650.52** 59.0 131.31 5.85 epo 0.0003 0.0004 0.002** 0.0005 0.52 4.35 epp 0.004 0.03 0.08** 0.04 1.79 10.5 ea 1.69** 0.20 0.44** 0.21 3.18 14.33 pa 1.0* 0.14 0.17 0.22 3.18 14.9 el 8.82** 0.95 2.53** 0.53 15.56 4.70 ed 0.004 0.01 0.08** 0.02 4.23 3.59 krpe 0.79 0.34 1.17** 0.39 13.08 4.79 kpr 44.08** 5.87* 7.92** 2.74 31.94 5.19 tkw 32.15 498.92 1742.34* 870.39 note: **significant (p<0.01),*significant (p<0.05), rep=replication, blk=block, cv=correlation variation. appendix-3.mean squares, and proportional contribution of lines, tester and line x tester of yield and yield related traits 48 maize testcross at ambo, 2017. source df gy ad sd asi ph eh epo rep 1 0.03 15.84* 12.76 0.11** 408.38 260.04* 0.0013 cross 47 4.76** 6.49** 10.89** 2.81(0.009)** 902.75** 511.62** 0.0025** line 23 6.23** 9.73** 13.11** 1.95(0.006) 687.48* 422.09** 0.003 tester 1 0.09 3.76 98.01** 59.32(0.18)** 19877.41** 10922.67** 0.0047 line x tester 23 3.48** 3.37 4.88 1.20(0.004) 293.04 148.49** 0.002* error 29 1.01 2.57 3.61 0.77(0.002) 211.96 44.85 0.0008 proportion contribution (%) line 64.12 73.36 58.91 34.07 37.27 40.37 58.44 tester 0.04 1.23 19.15 44.98 46.85 45.42 3.92 line x testers 35.84 25.41 21.93 20.95 15.89 14.2 37.65 ea ed el epp krpe kpr tkw rep 1 0.09 0.01 6.48** 0.01 0.06 0.68 7.23 cross 47 0.22** 0.07** 2.66** 0.11** 0.65* 7.18 3590.20** line 23 0.28 0.12** 4.48** 0.16** 0.73* 7.54 3535.40** tester 1 0.67** 0.01 0.44 0 2.62** 9.01 57934.13** line x tester 23 0.15* 0.02 0.93 0.06 0.27 6.73 1282.22 error 29 0.06 0.03 1.06 0.03 0.36 5.04 857.51 proportion contribution (%) line 60.68 88.45 82.47 74.35 65.43 51.45 48.19 tester 6.36 0.33 0.36 0 10.19 2.67 34.33 line x testers 32.97 11.22 17.18 25.65 24.38 45.88 17.48 note: **significant (p<0.01), *significant (p<0.05), gy=grain yield, ad=anthesis days, sd=silking days, asi=anthesissilking interval, ph=plant height, eh=ear height, epo =ear position, epp=ear per plant, ea=ear aspect, el=ear length, ed=ear diameter, krpe=kernel rows per ear, kpr=kernels per rows, tkw=thousand kernels weight. agriculture and food sciences research, 2020, 7(2): 113-124 124 © 2020 by the authors; licensee asian online journal publishing group appendix-4.mean squares, and proportional contribution of lines, tester and line x tester of yield and yield related traits of 48 maize testcross holetta, 2017. source df gy ad sd asi ph eh epo rep 1 0.05 15.66** 25.01** 0.55 247.00** 320.26** 0.0005 cross 47 3.05** 10.57** 11.73** 2.04 1226.24** 755.60** 0.0024* line(gca) 23 2.96** 15.71** 17.24** 1.67 587.61** 434.46** 0.0032** tester(gca) 1 37.79** 3.32 21.09** 3.43 39148.12** 22634.84** 0.0241** line x tester(sca) 23 1.63 5.74 5.81* 2.34 216.10** 125.48* 0.0007 error 29 0.97 3.1 2.71 2.50 46.94 55.52 0.0005 proportional contribution (%) line 47.5 72.75 71.93 40.11 23.45 28.14 64.69 tester 26.35 0.01 3.83 3.58 67.93 63.74 21.43 line x testers 26.15 26.58 24.25 56.31 8.62 8.13 13.88 ea ed el epp krpe kpr tkw rep 1 1.17** 0 4.70** 0.001 0.54 10.54 23.21 cross 47 0.50* 0.09** 3.17** 0.09* 1.48** 8.45** 2074.04** line(gca) 23 0.70** 0.09** 4.09 0.11* 1.55** 11 3015.38* tester(gca) 1 1.27** 1.02** 3.56 0.38** 12.98** 1.27 229.4 line x tester(gca) 23 0.26 0.04* 2.23** 0.05 0.91* 6.22* 1212.9 error 29 0.2 0.02 0.6 0.04 0.36 2.81 745.02 proportional contribution (%) line 69.15 51.46 63.17 62.95 51.22 63.69 71.15 tester 5.44 25.17 2.39 9.46 18.69 0.89 0.24 line x testers 25.41 23.36 34.44 27.59 30.1 36 28.62 note: **significant (p<0.01), *significant (p<0.05), gy=grain yield, ad=anthesis days, sd=silking days, asi=anthesissilking interval, ph=plant height, eh=ear height, epo =ear position, epp=ear per plant, ea=ear aspect, el=ear length, ed=ear diameter, krpe=kernel rows per ear, kpr=kernels per rows, tkw=thousand kernels weight. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 16 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 16-21, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.16.21 © 2020 by the authors; licensee asian online journal publishing group effect of credit on cassava productivity in kintampo south district using cobbdouglas production function model maxwell amedi1 moses dumayiri2 mohammed abdul-rashid suhuyini3 ( corresponding author) 1 food security & resilience technical program manager, world vision international, ghana. 2lecturer, department of banking and finance, school of business and law, university of development studies, ghana. 3phd candidate, department of agricultural economics, university of development studies, ghana. abstract the study estimated the effect of credit on cassava productivity in kintampo south district using cobb-douglas production function model. primary data was collected from 60 smallscaled cassava farmers while secondary data was gathered from literature. snowball sampling technique was used to obtain the sample size of 60 small-scaled cassava farmers. data obtained were analyzed using descriptive statistics and cobb-douglas production function model. results from the study showed that majority of the farmer’s accessed credit from vsla groups. the study found that, credit has significant effect on cassava output level with an elasticity of 1.83 implying that a percentage increase in credit supply will lead to 1.83 % increase in cassava output. the study concludes that, the output of cassava could be increased considerably by obtaining credit. the study recommends the ministry of agriculture to use vsla groups as vehicle of administering loans/credit to small-scaled farmers to increase productivity of farmers. secondly, small-scaled farmers in kintampo south district should be part of vsla and other co-operate groups to acquire credit which will enable them purchase necessary input required for cassava farming activities. keywords: cassava, productivity, credit, cobb-douglas production function model, kintampo south district. citation | maxwell amedi; moses dumayiri; mohammed abdulrashid suhuyini (2020). effect of credit on cassava productivity in kintampo south district using cobb-douglas production function model. agriculture and food sciences research, 7(1): 16-21. history: received: 11 november 2019 revised: 16 december 2019 accepted: 20 january 2020 published: 19 february 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 17 2. problem statement .......................................................................................................................................................................... 17 3. methodology ..................................................................................................................................................................................... 17 4. result and discussions ................................................................................................................................................................... 18 5. conclusion ......................................................................................................................................................................................... 20 6. recommendation.............................................................................................................................................................................. 20 references .............................................................................................................................................................................................. 21 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.16.21&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.16.21&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1337 https://orcid.org/0000-0001-7189-0774 agriculture and food sciences research, 2020, 7(1): 16-21 17 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by estimating the effect of credit on cassava productivity in kintampo south district using cobb-douglas production function model. 1. introduction cassava remains to be one of the most important crops grown in ghana because of its adaptability to different ecological zones and its economic values [1]. cassava farming creates employment for about 20% percent of the population in ghana in different forms of livelihood activities and serves as a major source of income for households. in spite of the relevance to the economy, most farmers’ find it difficult to access credit [2] to go into its production to ensure food security [3]. this is because generally agriculture cash flows are seasonal in nature (i.e. cash inflows and outflows seldom occur at the same time; therefore farmers are left with no option than getting external source of funding to meet their expenditures which they do not even get at all or record low yield [4]. agricultural credit therefore plays short-circuiting role in individual savings process to increase investment in agriculture [5]. in a research done by jan and khan [4] and sial, et al. [6] it came out clear that agricultural credit plays crucial role in procuring agricultural equipment’s and machineries, purchase of raw materials, payment of wages, acquisition of farm inputs (stem, weedicide, pesticides, herbicides) to increase agricultural productivity. in view of this, government’s strategy over the years to increase agriculture productivity and economic growth has been through agricultural programmes supported by the government or donors [7]. such programmes have overall goal of raising farmer-household income and reducing poverty through private sector-led and agribusiness development. recent research by isser [8] revealed that the volume of credit given by bank in in the ghana is on decline whiles the non-agricultural sector is higher. from literature, the few studies that exist include; factors that influence demand of formal savings and lending facilities in ghana by aryeetey and gockel [9] examination of credit supply from different sectors of the economy, agricultural credit and production response. in kintampo south district limited or no empirical studies exist on the effect of credit on cassava productivity. the study therefore seeks to ascertain the effect of credit on cassava productivity. this paper, therefore intends to fill the knowledge gap. the relevance of this study cannot be over emphasized. it is hoped that the facts and findings of the study will be relevant to the district assembly, policy makers and other government and non-governmental organizations. the study will help provide a better understanding of whether credit has any impact on cassava productivity. findings in the district will help in the design of policies that will alleviate poverty, reduce vulnerability, and improve household wellbeing. the study will also be very useful to the intellectual community as it adds to the available literature on the above subject matter. 2. problem statement cassava producers in the kintampo south district are mostly small-scaled farmers. most of these farmers cultivate land sizes not more than five acres. farmers, therefore, can barely have surplus on their generated income and thus, save little or none for investment in their farm operations. low savings among smallscaled farmers do not encourage major investment in agriculture in terms of mechanization and therefore contribute little output. percentage credit provided by the deposit money banks (dmb) to cassava producers in the kintampo south district has been on a persistent decline compared to the volume of credit to the non-agricultural sectors. district farmers who are fortunate to receive credit from these commercial banks receive low amount which scarcely have the propensity to increase production. the tedious and bureaucratic procedures encountered by the farmers when accessing such credits from financial institutions in the kintampo south district discourages farmers from taking credit. in spite of government efforts to facilitate credit deepening and credit widening among farmers through financial institutions, most farmers are restricted to accessing credit or shun completely from taking credit from formal financial institutions. they, therefore, rely primarily on informal credit sources that supply low amount. these concerns raise the following research questions; 1. what proportions of small -scaled cassava farmers in kintampo south district have access to credit from financial institutions? 2. does credit for small scale-scale cassava farmers in kintampo south district have any significant effect on their productivity? 2.1. objectives of the study the objective of the study is to assess the effect of credit on cassava productivity in kintampo south district of the brong ahafo region of ghana 2.2. specific objectives of the study 1. to estimate the proportion of small scale cassava farmers in kintampo south district those have access to credit from financial institutions. 2. to estimate the effect of credit on cassava productivity in the kintampo south district. 3. methodology 3.1. theoretical framework the cobb-douglas production function of the form y =adαhβ eɛ was also used to access the effect of credit on cassava output. this equation is widely used to represent the relationship of an output to inputs. the α, and β, are the output elasticity of the inputs and the ɛ is the error term. the effect of credit on cassava output. the cobb-douglas production function is employed to estimate the effect of credit on the production output by the small-scale cassava farmers in kintampo south district. the cobb-douglas production function is of the form: agriculture and food sciences research, 2020, 7(1): 16-21 18 © 2020 by the authors; licensee asian online journal publishing group y = adαhβlβfā the production function is linearized to obtain: iny = ina + αind + βinh + ᵞinl + ãinf + ý ………………………………….(eqn -1) where; y denotes total production of cassava (kg/acre). d denotes credit (amount in ghs). h denotes land (acres). l denotes labour input (ghs). f denotes fertilizer inputs (kg). a denotes total factor productivity. ý denotes error term which accounts for the random disturbances. α, β, ᵧ, and ã are the parameters to be estimated. the parameters α, β, ᵧ, and ã are the output elasticities of credit, land, labour and fertilizer respectively. these values are constants determined by available technology. output elasticity measures the responsiveness of output to a change in levels of credit, land, labour or fertilizer used in production, ceteris paribus. α, β, ᵧ, and ã > 0, implying that, each of credit, land, labour, and fertilizer has positive correlation with output. the efficiency or the total productivity factor of the available technology will be shown by the constant intercept, a. 3.2. methods of analyses objective 1 in order to estimate the proportion of small scale cassava farmers in kintampo south district that have access to credit from formal financial institutions, descriptive statistics was used to describe the proportion of farmers in the study area that are currently using credit from formal financial institutions. percentages were mainly used to describe results and then presented in tabular form. objective 2 in order to assess the effect of credit on cassava productivity in the kintampo south district cobb-douglas production function model was used. 3.3. sample size and sampling technique the snowball sampling technique was used to obtain the sample size of 60 smallscaled cassava farmers. one of the smallscaled cassava farmers who was available and ready to speak to was first contacted. after answering my questions, he directed me to the next farmer who then also directed me to another farmer after responding to my questions till i had a total of 60 small-scale cassava farmers. primary data was collected from the cassava farmers by administering structured questionnaires. 3.4. method of data collection interviews using structured questionnaire were the main techniques used in the gathering the requisite data. open and close-ended questions were used. some of the information collected from farmers includes age, sex, marital status, level of education, household size and farming experience e.t.c. the openended questions were to bring out understanding of the situation on the ground. 3.5. study area the study was conducted in kintampo south district of brong ahafo region of ghana. kintampo south district is a political and administrative entity which shares boundaries with kintampo north municipality to the north, to the south by nkoranza and techiman, to the east by atebubu and pru districts and to the west by wenchi municpal. it lies within longitude 10º 20º west and 20º 10º east and latitude 80º 15º north and 70º 45º south. the district which is one of the 27 in brong ahafo has jema as its capital with an estimated population of 6300 [10]. the district is mainly inhabited by the mo and bono together with a multiplicity of other ethnic groups such as the dagaabas, dagombas, frafra, ashantis, lobi and wala in the district. the district has 122 communities many of which are hard to reach especially in the wet or rainy season. the district has three (3) major weekly markets located at jema (tuesday), apesika (thursday) and anyima (monday). these markets are patronised mainly by farmers and middlemen from kintampo, techiman, tamale, wenchi and other areas. products sold at these markets are mainly food crops and household items. 4. result and discussions 4.1. sex distribution of respondents the sex distribution of the respondents as indicated in figure 1 revealed that, 67% of the farmers are males and 33% are females indicating that cassava farming in kintampo south district is dominated by men. this agrees with the findings of olaleye [11] that small scale farming are being carried out mostly by men while females involve in light farm operation such as weeding, processing and harvesting and marketing of cassava. agriculture and food sciences research, 2020, 7(1): 16-21 19 © 2020 by the authors; licensee asian online journal publishing group figure-1. sex distribution of respondents. source: field survey, 2018. 4.2. marital status and household size of respondents the result from table 1 shows that 58% of respondents are married and 20% are single whilst 22 % are widowed. about 23% of small scaled cassava farmers have household sizes between 0 and 5 whilst 13% of them have household size above 10 persons per family with majority of the farmers’ having household size between 5 and 10 representing 64%. this is consistent with adebayo and adeola [12] whose study also reveals that majority of the respondents have large family sizes which fall within the range of 6-10 persons per family. table-1. households marital status. marital status frequency percentage married 35 58.3 single 12 20.0 windowed 13 21.7 total 60 100.0 source: field survey, 2018. table-2. household sizes of cassava farmers. household size frequency percentage 1-5 14 23.3 6-10 38 63.3 ≥10 8 13.3 total 60 100.0 source: field survey, 2018. table-3. age distribution of respondents. age frequency percentage 18-30 13 21.7 31-40 18 30.0 41-50 23 38.3 ≥50 6 10.0 total 60 100.0 source: field survey, 2018. most of the small-scaled farmers are between the ages of 41-50 representing 38.3% as indicated in table 3. the farmers above 50 years represent 10% of small-scaled cassava farmers sampled. also 21.7% of the farmers are between 16-30 years whilst 30% are between 31-40 years. the economically active labour force engaged in smallscaled cassava farming at kintampo south district is in the range of 31-50 representing 68.3% which is appro. 68%. this agrees with finding from mofa [13] that 67% of similar age are engaged in small scaled cassava farming. table-4. educational level of respondents. level frequency percentage none 21 35 primary 18 30 jhs/mslc 15 25 shs/tech/voc 6 10 total 60 100 source: field survey, 2018. the result from table 4 shows that 35% of the farmers engaged in smallscaled cassava farming do not have any formal education. about 30% had primary education whiles 25% had junior high school and middle school certificate examination. it was only 10% that had senior secondary school education. agriculture and food sciences research, 2020, 7(1): 16-21 20 © 2020 by the authors; licensee asian online journal publishing group figure-2. proportion of small scale cassava farmers’ who access credit from formal financial institutions. source: field survey, 2018. from figure 2, the main source of funding for small-scaled cassava farmers in kintampo south district is through village savings and loans associations (vsla). the next available source of credit is from their personal savings. about 18% of farmers carry out their cassava farming activities using their own resources. about 5% of the respondents secure their credit from friends. the high percentage of farmers taking credit from the vsla was evident from the field because most of them were members of vsla groups and their participation was great. beyond this, the high figure recorded stems from the fact that, is quite easier to borrow from the group and the interest on the loans will be shared back among members during share out session by close of the year. 4.3. effect of credit on cassava output the cobb-douglas production function model was used to access the effect of credit on the output of cassava produced by the small-scaled cassava farmers. the estimated r-sqaure means that jointly, credit, labour; land fertilizer explained 70% of the total variation in the output of cassava produced. the estimated fvalue also suggested that the joint effect of the explanatory variables included in the model is significant at 1% percent. 4.4. results of the linear cobbdouglass production function table-5. results of effect of credit on cassava output. dependent variable: log (output) method least square sample (adjusted):660 included observation: 12 after adjustment variables coefficient robust std. err t p>|t| log (credit) 1.830853 .0708182 2.58 0.0362** log (fert) 0.250387 .721253 0.35 0.490 log (land) 0.341130 .725430 0.47 0.096 log (labour) 0.074659 .598665 0.125 0.722 c -4.518687 3.900769 -1.158 0.083 goodness of fit number of obs = 60 pseudo r2 = 0.807042 adjusted r-square = 0.696781 f (10, 110) = 2.83 prob. > f = 0.012070 note: ***, **and * are significant at 1%, 5% and 10% respectively. the output elasticity of credit was 1.83 implying that a percentage increase in credit supply will result in 1.83 percentage increase in cassava output. those of fertilizer, land and labour were 0.25, 0.34 and 0.07 respectively. the magnitude of output elasticity of credit shows that under the circumstances and the existing system of cassava farming at kintampo south district, credit supply will have a significant influence on output. that is, the production of cassava is considerably responsive to credit. this implies that the output of cassava could be increased considerably by obtaining credit without necessarily increasing the land size, fertilizer application and hiring more labour since they were not statistically different from zero. the credit supply will enable the farmers to acquire the necessary inputs such as fertilizer, weedicides, improved planting materials and modern implements required for production expansion. 5. conclusion it is worth concluding from the study that; • majority of the small scale cassava farmers need credit to enhance their productive farming activities. • the output of cassava is responsive to credit and the output could be increased significantly by obtaining credit without increasing land size, fertilizer and labour. 6. recommendation • based on the findings from the study, the following recommendations are therefore suggested: • the government through ministry of agriculture should use vsla groups as vehicle of administering loans/credit to small scaled farmers to achieve high recovery rate and increase productivity of farmers. agriculture and food sciences research, 2020, 7(1): 16-21 21 © 2020 by the authors; licensee asian online journal publishing group • the small scaled farmers in kintampo south district should be encouraged to be part of vsla and other cooperate groups to enable them acquire credit which will enable them purchase the necessary input required to for cassava farming activities. references [1] mofa, "ministry of food and agriculture, statistics, research and information directorate (srid), agriculture in ghana facts and figures, accra." retrieved from: http://agricinghana.com/wp-content/uploads/2017/07/agriculture-in-ghana-facts-and-figures2014.pdf, 2014. [2] bank of ghana (bog), "quarter 1 report." retrieved from: https://www.bog.gov.gh. [accessed may 28, 2017], 2017. [3] o. w. olowa and o. a. olowa, "issues, problems and policies in agricultural credit: a review of agricultural credit in nigeria," bangladesh e-journal of sociology, vol. 8, pp. 87-108, 2011. [4] i. jan and h. khan, "factors responsible for rural household participation in institutional credit programs in pakistan," african journal of business management, vol. 6, pp. 1186-1190, 2012. [5] m. amedi, "determinants of loan repayment performance among rice farmers in hohoe municipality," unpublished mphil. thesis, submitted to school of graduate studies of university of ghana, legon, 2012. [6] m. h. sial, m. s. awan, and m. waqas, "role of institutional credit on agricultural production: a time series analysis of pakistan," international journal of economics and finance, vol. 3, 2011. [7] mofa, "ministry of food and agriculture, statistics, research and information directorate (srid), agriculture in ghana facts and figures, accra." retrieved from: http://agricinghana.com/wp-content/uploads/2017/07/agriculture-in-ghana-facts-and-figures2016.pdf, 2014. [8] isser, "institute of statistical, social and economic research (isser), agricultural sector development: policies and options, working paper 1, accra, ghana," pp. 50-61, 2003. [9] e. aryeetey and gockel, "mobilising domestic resources for capital formation in ghana: the role of informal financial sector. aerc research paper no.3," 1991. [10] ghana statistical service (g.s.s), "population & housing census (phc) summary report of final results. accra ghana." retrieved from: https:/www.statsghana.gov.gh/[various, 2011. [11] r. olaleye, "effectiveness of development intervention for economic empowerment on rural women in ondo nigeria," an unpublished ph. d thesis submitted to the department of agricultural extension and rural development, university of ibadan, ibadan, 2000. [12] o. o. adebayo and r. g. adeola, "sources and uses of agricultural credit by small-scale farmers in surulere local government area of oyo state," anthropologist, vol. 10, pp. 313-314, 2008.available at: https://doi.org/10.1080/09720073.2008.11891069. [13] mofa, "ministry of food and agriculture, statistics, research and information directorate (srid), agriculture in ghana facts and figures, accra." retrieved from: http://agricinghana.com/wp-content/uploads/2017/07/agriculture-in-ghana-facts-and-figures2016.pdf, 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 2, no. 1, 19-24, 2015 http://www.asianonlinejournals.com/index.php/aesr 19 chemical composition and antioxidant activities of the essential oil from the clove buds (syzygium aromaticum) toward various oxidative stresses in vitro ke zhang 1 1 college of physical education, shanxi normal university, linfen, shanxi, china abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ............................................................................................................................................................................... 20 2. material and methods ............................................................................................................................................................... 20 3. results and discussion .............................................................................................................................................................. 21 4. conclusions ................................................................................................................................................................................ 22 references ...................................................................................................................................................................................... 22 the chemical composition and antioxidant activities of the essential oil from clove buds toward various oxidative stresses in vitro were investigated in this work. the results showed that 22 compositions of the essential oil were identified by gas chromatography (gc) and gc-mass spectrometry (ms), and engenol (76.23%) was found to be the main components of the essential oil, followed by βcaryophyllene (11.54%), caryophyllene oxide (4.29%), and eugenyl acetate (1.76%). the essential oil from clove buds had the better scavenging capacity against 2, 2’-azino-bis (3-ethylbenothiazoline-6sulfonic acid) diammonium salts (abts) and 2,2-diphenyl-1-picrylhydrazyl (dpph) radicals, high reducing power, and inhibiting lipid peroxidation reaction in lard emulsion, as well as exhibited different protection effects against dna damage caused by fe 2+ and 2,2'-azobis (2methylpropionamidine) dihydrochloride (aaph). and the antioxidant activities of essential oil showed a dose-response relationship to some degree. however, the essential oil did not have the protection against protein damage, but had a prooxidant effect on protein oxidative damage. keywords: chemical composition, antioxidant activity, essential oil, clove buds, dna, protein http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(1): 19-24 20 1. introduction reactive oxygen species (ros) and reactive nitrogen species (rns) are generated by many redox processes [1]. however, the excessive amounts of ros and rns can attack important biological molecules such as carbohydrates, proteins, lipids, dna and rna, which lead to cell death and tissue damage [2]. one of the principal causes of food quality deterioration is the oxidation of unsaturated lipids initiated by free radicals [3]. in addition, as it is known, the presence of high amounts of polyunsaturated fatty acids in oils and fats make them more susceptible to oxidation in the course of the time depending on external factors [4]. as a result of autoxidation especially which occurs with the effect of oxygen in the air, unpleasant taste and smell that are known as the signs of rancidity in oil occur [5]. in particular, the use of safer antioxidants from various natural materials is recommended because of the side effects of synthetic antioxidants. the flower bud of syzygium aromaticum (l.) merr. & perry. (family myrtaceae), commonly known as clove, is a well known food flavor and a popular remedy for dental disorders, respiratory disorders, headache and soar throat in traditional medicines of australia, and asian countries [6]. clove buds comprise around 20% of volatile oil rich in eugenol. clove oil has been listed as a ‘generally regarded as safe’ substance by the united states food and drug administration when administered at levels not exceeding 1500 ppm in all food categories [7]. clove bud essential oil still remain a research priority due to their wide range of pharmacological and biological activities such as antioxidant [8], antibacterial, antifungicidal, antiviral, and anesthetic effects [9]. however, these informations are still limited. in this study, the chemical composition and antioxidant activities of essential oil from the clove buds toward various oxidative stresses in vitro are investigated in order to provide a reference for the comprehensive development and utilization of clove buds. 2. material and methods 2.1. plant materials and reagents the dried buds of clove were purchased as commercial products from the local market. tptz were purchased from fluka. 2,2’-azino-bis abts, dpph and aaph were from sigma. other chemicals used were all of analytical grade. 2.2. extraction of essential oil the dried clove buds were ground and hydrodistilled for 4 h using a clevenger-type apparatus. the oil was separated from water and dried over anhydrous sodium sulfate and stored in tightly closed dark vials at 4 °c until use. 2.3. gc-ms analysis the analysis of the essential oil were performed using a hewlett-packard 5890 ii gc, equipped with a hp-5 ms capillary column (30 m×0.25 mm; film thickness, 25 μm) and a hp 5972 mass selective detector for the separation. the mass selective detector was operated in electron-impact ionization (ei) mode with a mass scan range from m/z 30 to 500 at 70 ev. helium was the carrier gas at a flow rate of 1 ml/min. injector and ms transfer line temperatures were set at 250 and 200 °c, respectively. the oven temperature was programmed from 40 °c for 2 min, raised to 180 °c at a rate of 3 °c/min, held isothermal for 2 min, and finally raised to 250 °c for 5 min. a sample of 0.1 μl of the essential oil and oleoresin were injected manually using a 1:50 split ratio. 2.4. dpph assay the dpph radical scavenging activity was determined according to the method of xu, et al. [10]. the scavenging rate on dpph radicals was calculated according to the formula, scavenging rate (%) = [(aoa1)/ao]×100, where ao is the absorbance of the control solution, a1 is the absorbance in the presence of samples in dpph solution. the scavenging activity of the sample on dpph radicals was expressed by ic50 value. ic50 value is the effective concentration at which dpph radicals are scavenged by 50% and is obtained by interpolation from regression analysis. 2.5. abts assay the abts cation radical scavenging activity was determined according to the method of xu, et al. [10]. the scavenging rate and ic50 value were calculated using the equation described for dpph assay. 2.6. ferric reducing antioxidant power (frap) assay the reducing ability was determined by using frap assay according to the method of xu, et al. [10]. the frap of the sample was expressed by ic50 value. the ic50 value is the effective concentration at which the absorbance is 0.5 and is obtained by the equation described for dpph assay. 2.7. antioxidant activities in lard emulsion the antioxidative effects of the essential oil on lipid peroxidation were evaluated according to the method of kamkar, et al. [11] with some modifications. each emulsion sample was transferred to a series of capped glass test tubes. then, essential oil (0.25, 1 and 4 g) and bht (0.01 g) were added to the test tubes and put in a dark oven at 70 °c respectively. the stability of emulsion to oxidation was evaluated 5 times over an 11-day period by analyzing the peroxide values (pvs) measured. 2.8. dna damage protective effect assay protection of dna from oxidative damage of fe 2+ the ability of samples to protect supercoiled pbr322 plasmid dna against fe 2+ and h2o2 was estimated with the dna nicking assay as described by xu, et al. [10]. agriculture and food sciences research, 2015, 2(1): 19-24 21 protection of dna from oxidative damage of aaph the ability of samples to protect supercoiled pbr322 plasmid dna against aaph was measured following the method described by zhang and omaye [12] with some modifications. to assay inhibition of dna damage induced by essential oil, 0.5µg of pbr322 dna was incubated with various concentrations of compounds and 2 µl of 25 mm aaph in pbs for 30 min at 37 o c. then the samples were electrophoresed on 0.8% agarose gel containing 0.5 μg/ml ethidium bromide and the dna was photographed under ultraviolet light. 2.9. determination of protein oxidation protein oxidation was assayed as described by xiang, et al. [13] with minor modifications. briefly, 2 µl 3mg/ml of bovine serum albumin (bsa) in pbs was mixed with 2µl of 0.5 mm cuso4 solution and 2 µl of 10 mm h2o2 solution and inhibited by various concentrations of test samples. after incubation for 2 h at 37 o c, the samples were then assayed with normal sds–page. 2.10. statistical analysis all experiments were conducted three times independently. one-way analysis of variance (anova) and duncan’s multiple range tests were carried out to determine significant differences (p < 0.05) between the means. 3. results and discussion 3.1. gc-ms analysis the chemical compositions of the essential oil from clove buds are analyzed by gc and gc-ms, and the results are presented in table 1. in total, 22 components in essential oil were identified, representing 95.80% of the total amount. the engenol (76.23%) was the major component of the essential oil, followed by β-caryophyllene (11.54%), caryophyllene oxide (4.29%), and eugenyl acetate (1.76%). similar to our finds, some studies reported that eugenol was the main component of clove essential oil, but its content was different. for example, tang, et al. [14] identified 19 components in the essential oil of clove buds and the content of engenol was 62. 21 %; guan, et al. [15] compared essential oils of clove buds extracted with supercritical carbon dioxide and other three traditional extraction methods, and found that there were differences in the components and content of essential oil of clove buds by various methods, but the content of engenol was more than 50%; jirovetz, et al. [16] identified 23 components of clove leaf essential oil and found that eugenol (76.8%) was the main component, followed by β-caryophyllene (17.4%), αhumulene (2.1%), and eugenyl acetate (1.2%). although we can find in the literature studies using the essential oil from clove buds, these differences in the content and components of essential oil are difficult to compare because the species, geographic regions, and extract methods used are different among studies. 3.2. dpph radicals scavenging capacity the scavenging capacity assayed herein on dpph radicals is shown in table 2. the ic50 values of essential oil on dpph radicals was 396.25 μg/ml, and the scavenging rates of essential oil on dpph radical increased by 3.89fold from 14.31% to 70.06% with the increase of its concentrations, showing a concentration-dependent scavenging of the dpph radicals at certain concentrations, which was also consistent with the previous reports [8, 16]. 3.3. abts radicals scavenging capacity the profile of scavenging capacity of essential oil on abts was similar to the result of the scavenging capacity on dpph radicals (table 2). somewhat differently, the ic50 value on scavenging abts radicals was 60.34 μg/ml. similarly, abts radicals scavenging capacity of essential oil increased dose-dependently at concentrations ranging from 10 to 100 μg/ml, which may be attributable to its hydrogen-donating ability. these differences in data between dpph and abts assays were likely due to different experimental conditions. 3.4. ferric reducing antioxidant power (frap) assay table 2 showed that the reducing power of essential oil from clove buds was in a concentration-dependent manner and increased with concentrations. the ic50 value was 683.4 μg/ml. these results suggested that essential oil from clove buds had a remarkable potency to donate electron to reactive free radicals, converting them into more stable non-reactive species and terminating the free radical chain reaction. 3.5. antioxidant activities in lard emulsion the pvs of lard emulsion increased gradually within 7 days after treating, but no obvious distinction among different treatments was found (fig. 1). thereafter, the pvs of control significantly and rapidly increased, and were much higher than the treated samples. these results indicated that the essential oil from clove buds had inhibiting lipid peroxidation reaction, and the antioxidative effects depend on its concentration and action period, even though far lower than that of the bht. 3.6. protection of dna damage to assay the potential of the essential oil to prevent dna damage, oxidative dna strand breakage induced by aaph and fe 2+ were measured with pbr322 dna. the damage of plasmid dna produces a relaxed open circular form and further a linear double-stranded dna molecule [17]. the plasmid dna was mainly of the supercoiled form in the absence of fe 2+ and h2o2 (fig. 2, lane 1). during the addition of fe 2+ and h2o2, the supercoiled form of dna converted into the open circular and linear forms (fig. 2, lane 2). from the gel analysis, the protection effects against dna damage offered against dna damage by essential oil (15.62–250 mg/ml) was concentration dependent, but a lower concentration of essential oil did not seem to have the protection effects (fig. 2, lane 3, 4). the results indicated that the essential oil might prevent the reaction of fe ions with h2o2, on the other hand, and it agriculture and food sciences research, 2015, 2(1): 19-24 22 probably quenched hydroxy radicals by donating hydrogen-atom or electron, and therefore protecting the supercoiled plasmid dna from hydroxy radicals dependent strand breaks [17]. as shown in fig. 3, similar results were found in protective effect assay of dna from oxidative damage of aaph. the protection offered against dna damage by essential oil (3.9–62.5 mg/ml) was also concentration dependent, and the essential oil exhibited a stronger protective effect against dna damage caused by aaph with a lower completely prevented concentration of 3.9 mg/ml (fig. 3, lane 3). the differences in protective effect of essential oil against dna damage caused by fe 2+ and aaph may come from different determination method [18]. nonetheless, these results showed that the essential oil from clove buds owned a higher potential to prevent dna damage. 3.7. inhibition of protein oxidation oxidation of cellular proteins has been described under many pathological conditions. the vulnerability of various amino acid residues of proteins to oxidation varies with reactive oxygen species [19]. the protection effect against protein oxidative damage was determined by the oxidation of bsa initiated by cu 2+ and the results are shown in fig. 4. the results showed not only non-protective effect against oxidation of bsa but also a prooxidant effect, which was augmented by increasing concentrations of the essential oil. similar to our findings, xiang, et al. [13] reported that carnosic acid had the potential to prevent bsa oxidant damage in low concentrations but showed a prooxidant effect against bsa when the concentrations of carnosic acid were higher and the effect was concentration related. some researches [20, 21] reported that polyphenol have prooxidant effect with the existence of mental ions. nakagawa, et al. [22] suggested that cu(ii) ions could convert (+)catechin from an antioxidant to a prooxidant in protein oxidation. according to our results of chemical composition analysis, engenol that was the major composition of the essential oil, as a phenolic compound, may be the most possible reason of producing prooxidant effect. however, the prooxidation mechanism of the essential oil from clove buds needs to be further studied. 4. conclusions in conclusion, this work showed that eugenol was the main component in essential oil from clove buds, and the essential oil exhibited different antioxidant activities toward various oxidative stresses in vitro. the essential oil from clove buds had the better scavenging capacity against dpph and abts radicals, high reducing power, and inhibiting lipid peroxidation reaction in lard emulsion during storage, as well as it exhibited different protection effects against dna damage caused by fe 2+ and aaph. and all the antioxidant activities of essential oil from clove buds showed a dose-response relationship to some degree. however, the essential oil did not show the protection against protein damage as we expected, but had a prooxidant effect on protein oxidative damage. references [1] m. valko, d. leibfritz, j. moncol, m. t. cronin, m. mazur, and j. telser, "free radicals and antioxidants in normal physiological functions and human disease," int. j. biochem. cell bio., vol. 39, pp. 44–84, 2007. [2] m. l. alexandrova and p. g. bochev, "oxidative stress during the chronic phase after stroke," free radical bio. med., vol. 39, pp. 297–316, 2005. [3] a. r. hras, m. hadolin, z. knez, and d. bauman, "comparison of antioxidative and synergistic effects of rosemary extract with atocopherol, ascorbyl palmitate and citric acid in sunflower oil," food chem., vol. 71, pp. 229–233, 2000. [4] f. e. sikwese and k. g. duodu, "antioxidant effect of a crude phenolic extract from sorghum bran in sunflower oil in the presence of ferric ions," food chem., vol. 104, pp. 324–331, 2007. [5] m. m. özcan and d. arslan, "antioxidant effect of essential oils of rosemary, clove and cinnamon on hazelnut and poppy oils," food chem., vol. 129, pp. 171–174, 2011. [6] m. domaracky, p. rehak, s. juhas, and j. koppel, "effects of selected plant essential oils on the growth and development of mouse preimplantation embryos in vivo," physiol. res., vol. 56, pp. 97–104, 2007. [7] m. a. kildeaa, g. l. allanb, and r. e. kearney, "accumulation and clearance of the anaesthetics clove oil and aqui-stm from the edible tissue of silver perch (bidyanus bidyanus)," aquaculture, vol. 232, pp. 265–277, 2004. [8] i̇. gülçin, m. elmastaş, and h. y. aboul-enein, "antioxidant activity of clove oil-a powerful antioxidant source," arab. j. chem., vol. 5, pp. 489–499, 2012. [9] k. chaieb, h. hajlaoui, t. zmantar, a. b. kahla-nakbi, m. rouabhia, k. mahdouani, and a. bakhrouf, "the chemical composition and biological activity of clove essential oil, eugenia caryophyllata (syzigium aromaticum l myrtaceae): a short review," phytother. res., vol. 21, pp. 501–506, 2007. [10] j.-g. xu, q.-p. hu, and y. liu, "antioxidant and dna-protective activities of chlorogenic acid isomers," j. agric. food chem., vol. 66, p. 11625−11630, 2012. [11] a. kamkar, a. j. javan, f. asadi, and m. kamalinejad, "the antioxidative effect of iranian mentha pulegium extracts and essential oil in sunflower oil," food chem. toxicol., vol. 48, pp. 1796–1800, 2010. [12] p. zhang and s. t. omaye, "dna strand breakage and oxygen tension: effects of β-carotene, α-tocopherol and ascorbic acid," food chem. toxicol., vol. 39, pp. 239–246, 2001. [13] q. xiang, x. meng, y. qiao, l. xu, and x. liu, "effect of carnosic acid on free radical-mediated oxidative damage to proteins," food sci., vol. 34, pp. 281–284, 2013. [14] y. tang, m. zhang, y. zhang, h. liao, j. ou yang, z. zeng, s. feng, and h. li, "composition and antimicrobial activity of the volatile oil from bud of syinga juliana c. k. schneid," natural science journal of xiangtan university. (in chinese), vol. 30, pp. 101–105, 2008. [15] w. guan, s. li, r. yan, s. tang, and c. quan, "comparison of essential oils of clove buds extracted with supercritical carbon dioxide and other three traditional extraction methods," food chem., vol. 101, pp. 1558–1564, 2007. [16] l. jirovetz, g. buchbauer, i. stoilova, a. stoyanova, a. krastanov, and e. schmidt, "chemical composition and antioxidant properties of clove leaf essential oil," j. agric. food chem., vol. 54, pp. 6303–6307, 2006. [17] b. n. singh, b. r. singh, r. l. singh, d. prakash, r. dhakarey, g. upadhyay, and h. b. singh, "oxidative dna damage protective activity, antioxidant and antiquorum sensing potentials of moringa oleifera," food chem. toxicol., vol. 47, pp. 1109–1116, 2009. [18] m. dizdaroglu, p. jaruga, m. birincioglu, and h. rodriguez, "free radical-induced damage to dna: mechanism and measurement," free radical bio. med., vol. 32, pp. 1102–1115, 2002. [19] q. yang, x. pan, w. kong, h. yang, y. su, l. zhang, y. zhang, y. yang, l. ding, and g. liu, "antioxidant activities of malt extract from barley (hordeum vulgare l.) toward various oxidative stress in vitro and in vivo," food chem., vol. 118, pp. 84–89, 2010. agriculture and food sciences research, 2015, 2(1): 19-24 23 [20] b. halliwell, "are polyphenols antioxidants or pro-oxidants? what do we learn from cell culture and in vivo studies?," arch. biochem. biophys., vol. 476, pp. 107–112, 2008. [21] q. wu, p. su, and y. liu, "the development of the antioxidant and prooxidant activities of flavonoids," science and technology of food industry. (in chinese), vol. 35, pp. 379–383, 2014. [22] k. nakagawa, m. kaku, t. abukawa, k. aratani, m. yamaguchi, and s. uesato, "copper (ii) ions convert catechins from antioxidants to prooxidants in protein carbonyl formation," j. health sci., vol. 53, pp. 591–595, 2007. table-1. chemical composition of essential oil from clove buds compound peak area (%) compound peak area (%) 2-pinene 0.02 engenol 76.23 α-pinene 0.03 β-caryophyllene 11.54 eucalyptol 0.14 α-caryophyllene 0.64 methyl salicylate 0.06 (-)-b-cadinene 0.12 chavicol 0.09 β-selinene 0.25 4-allylanisole 0.13 α-selinene 0.16 anethol 0.11 caryophyllene oxide 4.29 α-muurolene 0.01 jasmone 0.07 α-copaene 0.05 ledol 0.03 valencen 0.01 globulol 0.04 eugenyl acetate 1.76 cedrene 0.02 table-2. radicals scavenging capacity and frap of essential oil scavenging rate (%) and frap (abs) at different concentrations (μg/ml) regression equation ic50 (μg/ml) dpph 100 250 500 750 1000 y=24.73ln(x)-98.98 r 2 = 0.9974 396.3 14.3±1.2d 37.7±0.9c 56.1±2.1b 65.4±3.5a 70.2±4.3a abts 10 25 50 75 100 y= 0.7959x + 1.9747 r 2 = 0.9903 60.4 9.8±0.5e 22.9±2.4d 44.5±3.1c 61.6±2.8b 82.8±5.3a frap 100 250 500 750 1000 y = 0.0005x + 0.1583 r 2 = 0.9982 683.4 0.22±0.03e 0.28±0.05d 0.41±0.03c 0.54±0.07b 0.68±0.04a values represent means of three independent replicates ± sd. r2 refer to the regression coefficients. different letters within a row indicate statistically significant differences between the means (p < 0.05) for different concentrations essential oil. fig-1. effects of essential oil and bht on pvs of lard emulsion at 70 °c. data are expressed as the mean values of three independent replicates ± sd. different letters indicate statistically significant differences between the means (p < 0.05) for different treatments at the same treatment time. fig-2. the protective effect of the essential oil to dna damage caused by fe2+. lane 1, the native dna; lane 2, the dna treated with 1 mm feso4 and 1 mm agriculture and food sciences research, 2015, 2(1): 19-24 24 h2o2; lane 3-7, the dna treated with 15.62, 31.25, 62.5, 125, and 250mg/ml essential oil, respectively. fig-3. the protective effect of the essential oil to dna damage caused by aaph. lane 1, the native dna; lane 2, the dna treated with aaph; lane 3-7, the dna treated with 3.9, 7.81, 15.62, 31.25, and 62.5 mg/ml essential oil, respectively. fig-4. the inhibitions of the essential oil against bsa oxidative damage. lane1, the native bsa; lane 2, the bsa and ethanol; lane 3, the bsa treated with 0.5 mm cu2+; lane 4, the bsa and essential oil; lane 5-9, the bsa treated with 0.5 mm cu2+ and 0.625, 1.25, 2.5, 5, 10 mg/ml essential oil, respectively. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. 28 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 28-37, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.28.37 © 2020 by the authors; licensee asian online journal publishing group the anti-nutritional and proximate composition of rain tree (samanea saman) pod samples uzoukwu, a. e1 ubbaonu, c. n2 nwosu, j. n.3 ogueke, c. c.4 chukwu, m. n.5 ( corresponding author) 1,2,3,4department of food science and technology, federal university of technology, owerri, imo state, nigeria. 5department of food technology, abia state polytechnic, aba, abia state, nigeria. abstract the anti-nutritional and proximate composition of pods of samanea saman were determined. about 600g of wholesome, cleaned and dried pods were divided into two portions: one portion was ground as whole pod meal while the other portion was deseeded and ground as seedless meal. the removed seeds were ground separately as a sample to obtain three samples namely: whole pod, pulp and seed samples. triplicate data obtained were subjected to one-way anova using spss software of version 21. mean values and fisher’s least significant difference (lsd) were determined for the separation of the means at (p≤0.05). the whole pod had 1.49mg/100g oxalate, 1.97mg/100g hydrogen cyanide, 0.71mg/100g tannins, 27.07mg/100g saponins, 0.51mg/100g flavonoids and 1.71mg/100g alkaloids, which were significantly(p≤0.05) different from that of pulp (1.89mg/100g oxalate, 1.51mg/100g hydrogen cyanide, 0.86mg/100g tannins, 28.46mg/100g saponins, 1.00mg/100g flavonoids and 1.41mg/100g alkaloids) and seed samples (1.39mg/100g oxalate, 1.61mg100g hydrogen cyanide, 0.56mg/100g tannins, 26.51mg/100g saponins, 0.49mg/100g flavonoids and 1.86mg/100g alkaloids). the moisture contents were pulp (19.30%), whole pod (15.50%) and seed (9.20%). ash content were whole pod (4.70%), pulp (2.90%) and seed (2.60%); ether extract of whole pod, pulp and seed were 3.31%, 2.52% and 2.66% respectively. the crude proteins were whole pod (13.21%), pulp (10.98%) and seed (21.55%); crude fibre of the plant were whole pod (15.95%), pulp (6.77%) and seed (8.47%). the whole pod had 47.33% carbohydrate which was significantly (p≤0.05) lower than the carbohydrate of pulp (57.53%) and seed (55.52%). the utilization of the pods of s. samana is a significant source of bioactive that if harnessed in the formulation of nutraceutical beverage could offer a whole lot of health benefit to the users. keywords: saponins, tannin, alkaloid, oxalate, flavonoid, moisture, proteins, ash, fibre, carbohydrate. citation | uzoukwu, a. e; ubbaonu, c. n; nwosu, j. n.; ogueke, c. c.; chukwu, m. n. (2020). the anti-nutritional and proximate composition of rain tree (samanea saman) pod samples. agriculture and food sciences research, 7(1): 28-37. history: received: 2 january 2020 revised: 6 february 2020 accepted: 12 march 2020 published: 8 april 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 29 2. materials and methods ................................................................................................................................................................... 29 3. results and discussion ................................................................................................................................................................... 34 4. conclusion and recommendation ................................................................................................................................................ 36 references .............................................................................................................................................................................................. 36 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.28.37&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.28.37&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1484 https://orcid.org/0000-0002-0080-4231 https://orcid.org/0000-0002-0005-5840 https://orcid.org/0000-0001-7892-3655 https://orcid.org/0000-0002-7348-6326 https://orcid.org/0000-0003-3371-7983 agriculture and food sciences research, 2020, 7(1): 28-37 29 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study has provided information for the utilization of the rain tree (samanea saman) pod in food production. this study has also opened another area of research such as pod utilization in food formulation and animal feed development. 1. introduction the rain tree (samanea saman merr.) belongs to the family of leguminosae (pulse family) [1]. it is commonly called “saman”, “rain tree”, “monkey pod” and “cow tarmarind” in english, while it is called “acacia preta” and “cenizaro” in spain. the french call it “abre de pluie” and the philipines call it “mimosa”. the name “rain tree” was believed to have originated from the leaflets which are light-sensitive and close together on cloudy days or nights, allowing rain to fall through the leaves to the ground below [2]. it is native to northern south america and has naturalized throughout the tropics. the plant is easily recognized by its characteristic umbrella-shaped canopy giving it a domed shape [3]. samanea saman is a multipurpose tree and is the botanical name for rain tree, 5’o clock tree or monkey pod. s. saman is native to northern south america but currently naturalized and distributed throughout the tropics. s. saman often planted in parks and pastures, vacant lots, churches and school grounds, along roadsides and planned landscape. s. saman is moderately fast growing with a growth rate of 0.75–1.5 m/year. outdoors planting is done when the seedlings are 3–5 months old and 20–30 cm tall. it grows best in lowlands from sea level to 300 m high and rainfall of 600–3,000 mm [4, 5]. the pods of samanea saman (jack) merr tree is straight, some-what fleshy, indehiscent, 15 to 20 centimeters long and 2 centimeter wide. when the pod ripens, the pulp is sweet and sugary with a flavor like licorice that is much relished by children. in south america, the sweet pulp is cooked like a tamarind. the seeds are oblong and reddish in color. goats can eat the leaves and pods when feeds are scarce and the fruiting season of this tree coincides with the scarcity of good quality forages [6]. the nutritional composition of samanea saman pods and seeds as having crude protein values of 16.6% and 31.6%, crude fibre values of 12.0% and 14.0%, ether extract values of 1.4% and 4.3%, ash values of 3.5% and 4.3%, carbohydrate values of 18.5% and 20.3% respectively. the ripe fallen pod is commonly eaten by children and animals [7] while the sweet, sticky pulp is used in the production of fruit drink similar to tamarindo in latin america [2, 8]. the pods are edible and eagerly eaten by human and livestock (cattle, goat) both domesticated and wildlife. the seed of s. saman is already a good source of protein and energy. it contains 13.57% protein, 89.25% dry matter, 2.98%ether extract (ee), 2.19% crude fibre, 0.23% ash, and 6.44% nitrogen extract. apart from its nutrient content which is almost comparable to corn, the pod is easily available making it a cheap source of feed for chickens cruz [9]. staples and elevitch [2] reported that rain tree pods are nutritious with 12–18% protein, 40% digestibility and because of the sweet pulp are eagerly eaten by cattle, hogs, horses and goats. according to barcelo and barcelo [6] fresh samanea saman (jack) merr pods had dry matter content of 84.82%, crude protein of 9.45%, crude fiber of 8.34%, crude fat of 8.82% and ash of 5.12%. the samanea saman (jack) merrpod is rich in carbo-hydrates as reflected in its nitrogen-free extract of 53.09%. samanea saman (jack) merr pods are relished by livestock due to its sweet taste. the seeds are also surrounded by sticky, gummy substance. the whole pods are used as feed and nutrient supplements for mammals such as squirrels, cows, cattle and goats. okonkwo [10] reported that its seed has been used in the production of a local condiment called anyu, however, the few works done on this pod were directed towards determining their anti-microbial and phytochemical potentials, and the use of their seeds in the production of “anyu” a local condiment [10-12]. available literature shows there is little or no research work on the use of these pods in their utilization in other commercial food products such as bread, yet these pods are wasting in the gardens and road-sides where the trees are planted for ornamental purposes. even in the campus of federal university of technology, owerri (futo), the trees fruit heavily and animals (cows or others) at their will pick up the heavily littering pods within the fruiting season [6]. anti-nutrients are natural or synthetic compounds that interfere with the absorption of nutrients [13]. antinutritional factors are present in different food substances in varying amounts, depending on the kind of food, mode of its propagation, chemicals used in growing the crop as well as those chemicals used in storage and preservation of the food substances [14, 15]. some oil seeds and legumes used in the production of foods contain toxic substances and anti-nutritional factors; for example, tannin in castor oil bean, trypsin inhibitors in soybean, oxalates in locust beans, phytic acid in african oil bean, phytate in melon seeds [16, 17]. despite the protein quality in leguminous seeds, it does not however reach the same level as in animal products. this is due to unbalanced amino acids, presence of anti-nutritional factors and low digestibility of protein. processing methods, such as soaking, cooking or fermentation can improve the quality of legume protein [18]. during fermentation, micro flora may produce proteolytic enzymes which may be responsible for the increase in protein digestibility. also, the elimination of phytic acid contributes to the improvement in protein digestibility of fermented products [19]. the main objective of this research is to investigate the anti-nutritional and proximate composition of rain tree pod. the specific objectives are to analyze the anti-nutrients and proximate composition of samanea saman whole pods, pulp and seeds. the notable sweetness of the rain tree pulp when applied in food is hoped to provide an alternative source of sugar for food production, with a resultant reduction in the cost of production and consequently the price of food in the market. besides, it’s distinct but attractive flavour may introduce a variety in food products. a good result from this study will certainly introduce the underutilized pods to the food industry and add economic value to it. 2. materials and methods 2.1. collection of materials some ripe, fallen wholesome rain tree (samanea saman) pods were picked from the tree sites around the school of agriculture and agricultural technology (saat) of the federal university of technology, owerri (futo), agriculture and food sciences research, 2020, 7(1): 28-37 30 © 2020 by the authors; licensee asian online journal publishing group imo state, nigeria. some of the chemicals used for the analyses were obtained from the department of food science and technology laboratory, and school of agriculture and agricultural technology laboratory in futo. others were purchased from the finlab store owerri, imo state, nigeria. some of the analyses were conducted in the laboratories of the department of food science and technology and school of agriculture and agricultural technology, futo, while others were carried out in the department of zoology laboratory of university of jos and international institute for tropical agriculture (iita), ibadan, oyo state, nigeria. 2.2. methods 2.2.1. preparation of samples the pods were picked at various times from the tree site, sorted, washed and sun dried for about five days. six hundred grams (600g) of wholesome, cleaned and dried pods were divided into two portions; one portion was ground as whole pod meal (sample 1) shown in figure 1 and figure 2, while the other portion was deseeded and ground as seedless meal (sample 2) as shown in figure 2. then the removed seeds figure 3 were collected and ground separately as a sample (sample 3) to obtain three samples for the desired raw material analyses, namely whole pod, pulp and seed samples [20]. figure-1. a rain tree (samanea saman) plant in futo. figure-2. rain tree (samanea saman) pods. agriculture and food sciences research, 2020, 7(1): 28-37 31 © 2020 by the authors; licensee asian online journal publishing group figure-3. rain tree (samanea saman) seeds. 2.2.2. determination of anti-nutrients in the pods of rain tree 2.2.2.1. saponin content the method described by okwu [21] was used for the analyses of the anti-nutrients. twenty-five grams (25g) of the sample was placed in a two hundred and fifty milliliters (250ml) flask. one hundred milliliters (100ml) of 20% (v/v) aqueous solution of ethanol was poured into the flask; then the flask with the content was placed in a hot water bath at 55oc for 4 hours with occasional stirring. the mixture was filtered and the residue re-extracted with a fresh 100ml of 20% (v/v) ethanol solution as before. both extracts were combined and the volume reduced to about 50ml in a water bath set at 90oc. the mixture was allowed to cool, extracted with 20ml of diethyl ether in a 250ml separating funnel. the ether layer was discarded and 50ml of n-butanol was poured into the lower layer. the mixture was shaken and the two layers allowed separating. the organic layer was washed twice with 10ml of 5%(v/v) sodium chloride solution in a separating funnel. this was poured into a pre-weighed beaker and evaporated to dryness on a boiling water bath. the beaker was cooled and weighed again [22]. the percentage saponin content was determined using equation 1. %𝑆𝑎𝑝𝑜𝑛𝑖𝑛 = 𝑊3 − 𝑊2 𝑊1 × 100 1 (1) where w3 = weight of beaker and residue after evaporation to dryness. w2 = weight of beaker alone. w1 = weight of sample. 2.2.2.2. flavonoid content two grams (2g) of the ground sample was placed in a 250ml erlenmeyer flask and 100ml of 80% (v/v) methanol solution was added. the mixture was stirred with a magnetic stirrer for 3 hours and filtered with whatman filter paper (no. 42). the residue was extracted with a fresh 100ml of 80% (v/v) methanol as before and the mixture filtered again. both extracts were combined into a pre-weighed beaker and evaporated to dryness on a boiling water bath. this was cooled and weighed again [23]. the percentage flavonoid content was determined using equation 2. %𝐹𝑙𝑎𝑣𝑜𝑛𝑜𝑖𝑑 = 𝑊3 − 𝑊2 𝑊1 × 100 1 (2) where: w3 = weight of beaker and flavonoid. w2 = weight of beaker. w1 = weight of sample. 2.2.2.3. determination of alkaloid content two grams (2g) of the ground sample was placed in a 250ml flask and 100ml of 20% (v/v) acetic acid in ethanol was added to the flask. the flask was covered and allowed to stand for 4 hours while stirring with a magnetic stirrer. the mixture was filtered and the residue re-extracted with a fresh 100ml of 20% (v/v) ethanolic acetic acid solution. both extracts were combined in a beaker and concentrated on a water bath to about one quarter of the original volume. the concentrate was cooled and concentrated ammonium hydroxide added in drops until complete precipitation of the alkaloids had occurred. the resulting mixture was allowed to stand overnight. thereafter, it was filtered using a pre-weighed filter paper. the filter paper with the content was dried in an oven set at 70oc for 6 hours and cooled to ambient temperature in a desiccator. the weight of the filter paper and its contents were taken [24, 25]. the percentage alkaloid content was determined using equation 3. %𝐴𝑙𝑘𝑎𝑙𝑜𝑖𝑑𝑠 = 𝑊3 − 𝑊2 𝑊1 × 100 1 (3) where w3 = weight of filter and alkaloids after drying. w2 = weight of filter paper alone. w1 = weight of sample. agriculture and food sciences research, 2020, 7(1): 28-37 32 © 2020 by the authors; licensee asian online journal publishing group 2.2.2.4. determination of tannin content about 0.5grams portion of the ground sample was placed in a 100ml flask and fifty milliliters (50ml) of distilled water was added and the mixture was stirred for about one hour with a magnetic stirrer. the sample was filtered with a moist filter paper into a fifty milliliters (50ml) volumetric flask and made up to the mark with distilled water. five milliliters (5ml) of the filtrate was pipette into a tube and mixed with three milliliters (3ml) of 0.1ml of ferricchloride in 0.1n hydrochloric acid and 0.008m potassium ferrocyanide solution. the absorbance was measured in a spectrophotometer set at 120nm wavelength within ten (10) minutes of mixing. a blank sample was prepared and the colour was developed and read at the same wavelength. a standard was prepared using tannic acid to get 100ppm and measured [26]. the tannin content was calculated using the equation 4: 𝑀𝑔 𝑚𝑙 𝑇𝑎𝑛𝑛𝑖𝑛 =⁄ 𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑋 𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑 1 (4) 2.2.2.5. determination of cyanogenic glycoside content one gram (1g) of the ground sample was placed in a round bottom flask and 200ml of de-ionized distilled water was added and the flask allowed standing for 2 hours. a distillation unit was set up and about 150mls of the reaction mixture (sample and de-ionized distilled water) was distilled into a 250ml receiver flask containing 20ml of 2.5% (w/v) sodium hydroxide solution. one hundred milliliters (100ml) of the mixture was measured into a 250ml flask and 8ml of 6m ammonium hydroxide solution was added followed by 2ml of 5% potassium iodide solution. the content of the flask was mixed and titrated with 0.02n silver nitrate solution till there was no further turbidity [27]. the percentage cyanogenic glycoside content was determined using equation 5. %𝐶𝑦𝑎𝑛𝑜𝑔𝑒𝑛𝑖𝑐 𝑔𝑙𝑦𝑐𝑜𝑠𝑖𝑑𝑒 = 𝑇 × 108 × 𝑁 1000 × 𝑉𝑒 𝑉𝑎 × 100 𝑊 (5) where t = titre value. 108 = equivalent weight of silver nitrate. n = normality of silver nitrate solution. ve= extract volume. va= aliquot volume titrated. w= weight of sample in grams. 1000 = scaling factor to obtain silver nitrate content of 1.0ml of silver nitrate solution. 2.2.2.6. determination of oxalate content five grams (5g) of the ground sample was placed in a 100ml flask and 20ml of 0.3n hydrochloric acid solution was poured into the flask. it was stirred with a magnetic stirrer at 50oc for i hour. the mixture was filtered with filter paper (no. 42) and the extract stored and the process was repeated. both extracts were combined and made up to a volume of 100mlwith distilled water. twenty milliliters (20ml) of the filtrate was poured into a 100ml beaker and three drops of phenolphthalein indicator was added. then, 5n ammonium hydroxide solution was added in drops until the reaction mixture was alkaline (pinkish). glacial ethanoic acid was added in drops until the pink coloration disappeared and a few more drops were added to make the mixture acidic. next, 5ml of 5% calcium chloride solution was added to the solution and the mixture was allowed to stand for 3 hours. it was centrifuged at 300r.p.m for 15 minutes and then the residue washed 3 times with hot water using a centrifugation technique. the residue was dissolved in 3ml of 3n tetraoxosulphate (vi) acid solution. the resulting solution was titrated with freshly prepared 0.01n potassium permanganate solution until permanent pink coloration that lasted for about 30 seconds was obtained [28]. a blank titration was also conducted. the percentage oxalate content was determined using equation 6. %𝑂𝑥𝑎𝑙𝑎𝑡𝑒 = 𝑋 1 × 1 1000 × 100 1 𝑎 × 100 𝑆 𝑏 (6) where x = weight of oxalate obtained by multiplying molar mass and molarity of oxalate. 1000 = reference volume for molar concentration. 100a = total volume of extract. 100b = scaling factor to convert to percentage. s = weight of sample taken for analysis. 2.2.3. proximate composition of the samples of pods of rain tree 2.2.3.1. determination of moisture content the aoac [29] method was used. two gramme-portions of each of the freshly ground samples (1, 2 and 3) were weighed into previously weighed dry crucibles. the crucibles with samples were dried in an oven at 105oc, cooled in desiccators for ten minutes, reweighed and returned into the oven until a constant weight was attained [26]. the moisture content was calculated with the following equation 7: % moisture content = 𝑊1−𝑊2 𝑊1 × 100 1 (7) where: w1 = the weight of sample before drying. w2 = the weight of sample after drying. sample 1= whole pod, sample 2= pod without seed (pulp), sample 3= seed. agriculture and food sciences research, 2020, 7(1): 28-37 33 © 2020 by the authors; licensee asian online journal publishing group 2.2.3.2 determination of ash content the ash content was determined following the aoac [29] method. two grammes of the samples were weighed in triplicate into previously weighed silica crucibles. the samples inside the crucibles were charred on a heater inside a fume cupboard to drive off most of the smoke. the crucibles with the contents were transferred into a muffle furnace and heated for about four hours at 550oc. they were cooled in a desiccator and weighed. the heating was repeated until the samples turned greyish white and attained constant weight. the ash content was then calculated as shown in equation 8: %ash = 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑎𝑠ℎ 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑡𝑎𝑘𝑒𝑛 × 100 1 (8) 2.2.3.3. determination of crude lipid (ether extract) the total lipid content of the samples was determined using the aoac [29] soxhlet fat extraction method. five grammes (5g) of the sample were weighed into a pre-weighed fat free extraction thimble which was plugged tightly with cotton wool. the thimble was placed in the soxhlet extractor fitted up with reflux condenser, all connected to a boiling flask containing 200ml of petroleum ether (boiling point 60oc) on a heating mantle. as the flask and petroleum ether were heated, the solvent evaporated and condensed into the thimble extracting oil from the sample and refluxed into the boiling flask with the extracted oil. this was done for 4 hours. at the end of extraction, the solvent (petroleum ether) was evaporated by heating at 70oc on a hot plate leaving the lipid extract in the flask. the flask with its contents were placed in an oven and dried at 110oc for one hour, cooled in a desiccator and re-weighed [26]. the percentage lipid was calculated as shown in equation 9: %ether extract 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑜𝑖𝑙 𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 × 100 1 (9) 2.2.3.4. determination of crude protein the crude protein contents of the samples were determined using the micro-kjeldahl apparatus as described by aoac [29]. two grammes of each of the samples was placed in a kjeldahl flask and 30ml concentrated sulphuric acid (h2so4) added followed by the addition of 10g potassium sulphate and 1g copper sulphate. the mixture was gently heated for few minutes until frothing ceased; the heat was increased and the sample allowed digesting for three hours. the digest was allowed to cool, diluted with distilled water (washing the digestion flask) up to 100ml. ten milliliters(10ml) of the dilute digest was pipetted into a distillation flask and 10ml of 40% (w/v) sodium hydroxide added. the mixture was distilled and the liberated ammonia collected in 10ml of 2% boric acid containing indicator. this was titrated with 0.01n hydrochloric acid to grey coloured end point. a blank was also prepared without a sample and treated as above [26]. the amount of crude protein was then calculated by multiplying percentage nitrogen in the digest by the conversion factor (6.25) as shown in equation 10. %𝑁 = (𝑎 − 𝑏) × 0.01 × 14 × 𝑣 𝑊 × 𝐶 × 100 1 (10) where a = the titre value of the digested sample. b = titre value of the blank sample. v = volume after dilution. w = weight of dried sample (mg). c = aliquot of sample used. 14 = atomic weight of nitrogen. crude protein = 6.25 x %n. 2.2.3.5. determination of crude fibre aoac [29] method was employed. two grams of the ground samples were weighed in duplicate into a 600ml, long pyrex beaker and 200ml of 1.25% h2so4 solution was added. the beaker was covered with a watch glass and the content gently boiled on a hot plate for 30 minutes. the acid was removed by filtering through a muslin cloth over a buckner funnel and the sample washed three times with 50ml of boiling water to free it of acid, before putting it back to the beaker. then, 200ml of 1.25% naoh solution was added to the residue in the beaker, which was covered with a watch glass and gently boiled on a hot plate for 30 minutes and then filtered. the residue was washed into a weighed no. 2 sintered glass crucible with 50ml of boiling water and later washed twice with 30ml portions of petroleum spirit. the crucible was dried in the oven at 80oc to a constant weight and then ignited in a muffle furnace at 600oc until a light gray coloured ash was obtained. the crucible and content were cooled to ambient temperature in a desiccator and then weighed [26]. the crude fibre content was calculated as shown in equation 11: % crude fibre = 𝑙𝑜𝑠𝑠 𝑖𝑛 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑛 𝑖𝑔𝑛𝑖𝑡𝑖𝑜𝑛 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 × 100 1 (11) 2.2.3.6. determination of carbohydrate content the carbohydrate content was obtained by difference [26] as shown in equation 12. %carbohydrate = (100% %moisture, %crude protein %fat-%ash-%crude fibre) (12) 2.2.3.7. statistical analysis triplicate data obtained were subjected to statistical analysis using spss software of version 21. mean values were determined and one-way anova was done as well as fisher’s least significant difference [30] was used to determine for the separation of the means at (p≤0.05). agriculture and food sciences research, 2020, 7(1): 28-37 34 © 2020 by the authors; licensee asian online journal publishing group 3. results and discussion 3.1. anti-nutritional composition (mg/100g) of rain tree (samanea saman) pod samples oxalate, hydrogen cyanide, tannins, saponins, flavonoids and alkaloids were identified in all samples of samanea saman pod table 1. oxalic acid as an anti-nutrient interfere with mineral availability particularly calcium. it binds with calcium and forms insoluble calcium oxalate which cannot be absorbed in the body. this may lead to death due to hypercalcimia in the renal tubules murray, et al. [31]; ifemeje [32]. ladeji, et al. [33] reported that oxalates cause irritation and swelling in the month and throat. the oxalate of the pulp sample was 1.89mg/100g which is significantly (p<0.05) higher than the oxalate (1.49mg/100g) of whole pod and 1.39mg/100g oxalate of seed sample table 1. this quantity of oxalate is within safe limit because 2% or more soluble oxalate can lead to toxicosis. they are therefore considered harmless when present in small amounts as shown in table 1. the amount of oxalate in samanea saman whole pod, pulp and seed samples is thus not harmful, more so, when cooking has been reported [34] to affect a significant reduction in total oxalate contents of plants. an unacceptably high value of 27.34 mg/100 g oil was reported for tropical almond [35]. cyanogenic glycosides are hydrolysed by β-glucosidase producing sugars and a cyanohydrin which spontaneously decompose to cyanohydric acid (hcn) and a ketone or aldehyde [36]. hcn is extremely toxic to a wide spectrum of organisms due to its ability of linking with metals (fe2+, mn2+ and cu2+) that are functional groups of many enzymes, inhibiting processes like reduction of oxygen in the cytochrome respiratory chain, electron transport in the photosynthesis, and the activity of enzyme like catalase, oxidase [32]. hydrogen cyanide is toxic when ingested by monogastric animals in large quantity [37]. the whole pod had hcn of 1.97mg/100g which is significantly (p<0.05) higher than the hcn (1.51mg/100g) of pulp and 1.61mg/100g of seed samples table 1. the hydrogen cyanide contents of these samples were low when compared to the report (4.31-4.77mg/kg) of ogueke, et al. [38] and maximum limit (10 mg/kg) recommended for garri [39]. reduction in hcn might be due to the processing methods employed [40]. high level of hydrogen cyanide has been implicated for cerebral damage and lethargy in man and animals [34, 41]. tannins are plant polyphenols, which have ability to form complexes with metal ions and with macromolecules such as proteins and polysaccharides [42]. dietary tannins are said to reduce feed efficiency and weight gain in chicks [37]. tannins are polyphenols and have been reported to exhibit anti-microbial actions. the presence of high level of tannins could confer on the users chemoprotective benefits [43]. the pulp had tannins of 0.86mg/100g which is significantly (p<0.05) higher than the tannins (0.71mg/100g) of whole pod and 0.56mg/100g of seed samples table 1. tannins may form a less digestible complex with dietary proteins. table-1. mean values (mg/100g) of anti-nutrient composition of rain tree (samanea saman) pod samples samples (mg/100g) anti-nutrients whole pod pulp seed lsd oxalate 1.49±0.01b 1.89±0.01a 1.39±0.01c 0.02 hydrogen cyanide 1.97±0.01a 1.51±0.01c 1.61±0.01b 0.02 tannins 0.71±0.01b 0.86±0.01a 0.56±0.01c 0.02 saponins 27.07±0.01b 28.46±0.01a 26.51±0.01c 0.02 flavonoids 0.51±0.01b 1.00±0.01a 0.49±0.01c 0.02 alkaloids 1.71±0.01c 1.41±0.01a 1.86±0.01b 0.02 note: means with different superscripts on the same row are significantly (p< 0.05) different. these tannins-protein complexes are astringent and adversely affect feed intake. the pod contains 0.86mg/100g of tannin due to this result taking the pod may not lower the availability of protein in the body or clotting with red blood cell as cause by excess tannin in human body. this suggests that this pod may be safe for consumption [44]. saponins are glycosides, which include steroid saponins and triterpenoid saponins [42]. high levels of saponins in feed affect feed intake and growth rate in poultry. reduction in feed intake has been ascribed to the bitter taste of saponins [37] and due to the irritating taste. excess saponins causes hypocholestrolaemia because it binds cholesterol making it unavailable for absorption [45]. saponin-protein complex formation can reduce protein digestibility [37]. saponins were predominant in all the samanea pod samples. the highest level of saponin was found in samanea saman pulp (28.46mg/100g), followed by the whole pod (27.07mg/100g) and the seed (26.51mg/100g). high levels of saponins have haemolytic effect on red blood cells and also reduce growth rate in animals. saponins also possess some beneficial properties such as cholesterol lowering, anti-tumor and as antioxidant [33]. the concentration of the value of saponins from the analysis was 28.46mg/100g which is within the who permissible limit of (48.50mg/100 g) as recommended in this study. saponins possess a carbohydrate moiety attached to a triterpenoid or a steroidal aglycone [46]. saponins reduce the uptake of glucose and cholesterol at the gut through intra-lumenal physico-chemical interactions. this could confer a chemo-protection against heart diseases to users [32]. flavonoids, a group of natural substances with variable phenolic structures, are found in fruits, vegetables, grains, bark, roots, stems, flowers, tea and wine. the flavonoids are categorized in different classes as alkaloids, terpenoids and phenolics. flavonoids carry out a number of protective functions in the human body. many flavonoids have evolved as bioactive compounds that interfere with nucleic acid or proteins and show antimicrobial or insecticidal and pharmacological properties [47].the highest level of flavonoids was found in samanea saman pulp (1.00mg/100g), followed by the whole pod (0.51mg/100g) and the seed (0.49mg/100g) as shown in table 1. flavonoids possess anti-inflammatory, anti-oxidant, anti-allergic, hepato-protective, anti-thrombic, anti-viral and anti-carcinogenic activities. these properties may explain the use of samanea saman for prevention and treatment of diseases [47]. alkaloids are beneficial chemicals to plants with predator and parasite repelling effects. however, they inhibit certain mammalian enzymic activities such as those of phosphor-diesterase, prolonging the action of cyclic amp. they also affect glucagons and thyroid stimulating hormones, while some forms have been reported to be agriculture and food sciences research, 2020, 7(1): 28-37 35 © 2020 by the authors; licensee asian online journal publishing group carcinogenic [48]. alkaloids were relatively lower in the samanea saman pulp (1.41mg/100g) than the seed (1.86mg/100g) and whole pod (1.71mg/100g) as shown in table 1. it is noteworthy that at the concentration of alkaloids in edible plants, they are usually non-toxic because steaming or boiling reduces their levels in plant extracts [32]. 3.2. proximate composition of rain tree (samanea saman) pod samples the fresh mature and ripe (brown) rain tree whole pod had a moisture content of 15.50% and understandably its pulp and seed had moisture contents of 19.30% and 9.20% respectively table 2. the values were significantly (p≤0.05) different from each other. esuoso [8] found that samanea saman husks, whole pods and seeds had moisture contents of 4.11%, 58.42% and 6.19%respectively. the slight variation in moisture content could be attributed to the drying conditions which the samples were subjected to before the analysis. factors such as environmental conditions could also affect the moisture content of food materials. barcelo and barcelo [6] showed that the samanea saman pod had a moisture content of 15.18%. the moisture content of 15.50% for the whole pod obtained in this study is in line with the values 15.30% and 15.80% reported by barcelo and barcelo [6]. the moisture content of foods determines the water activity which influences the rates of chemical, microbial and enzymatic reactions in foods. by lowering the water activity through drying, microbial deterioration of foods can be delayed, reduced or eliminated. the whole pod had higher ash content (4.70%) than the pulp (2.90%) or the seed (2.60%), indicating that it has a higher concentration of minerals than pulp and seed. this is in line with esuoso [8] who reported ash contents of 4.16% and 2.10% for samanea saman whole pod and seed respectively. barcelo and barcelo [6] reported a relatively higher ash content of 5.12% for samanea saman whole pod. though, there were slight variations in the ash contents of the samples, the values fall within a close range of 2.4% to 5.12%. ash in food constitutes the residue remaining after all the moisture has been removed as well as the organic materials burnt away by igniting at temperature of about 500oc [49]. thus, ash content represents the total mineral content in foods. table-2. mean values of the proximate composition of rain tree (samanea saman) pod samples. samples no. proximate composition (%) whole pod pulp seed lsd 1 moisture 15.50±0.01b 19.30±0.01a 9.20±0.1c 0.02 2 ash 4.70±0.01a 2.90±0.01b 2.6±0.01c 0.02 3 ether extract 3.31±0.01a 2.52±0.01c 2.66±0.01b 0.02 4 crude protein 13.21±0.01b 10.98±0.01c 21.55±0.01 0.02 5 crude fibre 15.95±0.01a 6.77±0.11c 8.47±0.01b 0.13 6 carbohydrate 47.33±0.01c 57.53±0.01a 55.52±0.01b 0.02 note: means with different superscripts on the same row are significantly (p< 0.05) different. the lipid content (ether extract) results in table 2 shows that samanea saman pod had the highest lipid content(3.31%) among the samples with samanea saman seed having 2.66% and the samanea saman pulp having the lowest lipid content (2.52%).the lipid content of all the samples were significantly (p< 0.05) different from each other. similarly, esuoso [8] reported a lower lipid value of 0.82% for the whole pod and 1.20% lipid content for the seed. the variations observed in the lipid contents could be as a result of agronomical factors and analytical techniques. with ether extract (fat) values of the samples ranging from 2.52% (in pulp) to 3.31% (in whole pod), certainly the pod is not a good oil source, thus its seed could not be classified as an oil seed. among the three areas of samanea saman pod studied, the seed had the highest protein value (21.55%). the whole pod had protein value of 13.21% while its pulp had a value of 10.98%. the differences between these values were significant (p<0.05). the protein value of 13.21% obtained in this study for the whole pod fell within the range of 10-18% reported by dihigo, et al. [50] and staples and elevitch [2]. semae, et al. [51] reported that the pods of rain tree to be highly digestible, having protein (15.31–18.00%) content, is low in cost and is non-toxic [52, 53]. esuoso [8] reported a whole pod protein value of 28.48% and that was the highest value observed in accessible literature for the pod. there was no specific report on the pulp with regards to proximate composition, but reports on the protein content of seed had values ranging from 22-31.60% [8, 50]. notwithstanding, the variations observed in the various reports, samanea saman pods and seeds exhibited high protein contents which could be explored in animal feed as nutrient source to improve animal diet and possibly for humans. the samanea saman whole pod had a mean crude fibre content of 15.95% as compared to values of 8.47% and 6.77% for the seed and pulp respectively. the crude fibre content of the three samples were significantly (p<0.05) different. the crude fibre value of 15.95% obtained in this study for the whole pod is comparably much lower than a value of 33.4% reported by esuoso [8] for the same material. also, a crude fibre value of 8.47% seed obtained in this study is comparably lower than the value 16.15% reported by esuoso [8] respectively for the same material. though agronomical factors may have contributed to the variations in the results obtained, different analytical conditions may have also led to the variation. crude fibres play important roles in digestion processes and the lowering of gastric cholesterol [21]. table 2 showed that samanea saman pulp had the highest carbohydrate content (57.53%) as compared to the seed with a value of 55.52% and the whole pod with the least carbohydrate content of 47.33%. there were significant (p<0.05) differences among the carbohydrate contents of the three samples. since the carbohydrate content was obtained by difference, the lower carbohydrate content observed in the whole pod could be a reflection of the higher crude fibre content of the whole pod which was much higher than that of the samanea saman pulp and seed. the result obtained in this research showed some similarities with those previously reported by other researchers, where the seed had higher carbohydrate than the pod. esuoso [8] also reported lower carbohydrate values of 26.73% and 34.89% for the samanea pod and seed, respectively. the relatively high carbohydrate content obtained in this research suggests that the various parts of samanea saman pods could serve as energy source when included in animal feed. agriculture and food sciences research, 2020, 7(1): 28-37 36 © 2020 by the authors; licensee asian online journal publishing group 4. conclusion and recommendation 4.1. conclusion the levels of hydrogen cyanide, oxalate, tannins and flavonoid were relatively low in all the samples of samanea saman pods. the results show that all the anti-nutrients were significantly (p<0.05) different in all the samples. the relative low levels of these anti-nutrients confirm the safety of the fruit for industrial utilization, human and animal consumption. it was observed that saponins were the predominant anti-nutrient identified in all the pod samples ranging from 26.51mg/100g (seed) to 28.46mg/100g (pulp) while the rest were less than 2.0mg/100g. the result of the anti-nutritional composition of samanea saman pods is evidence that utilization of samanea saman might not have any negative impact in the body. though saponin was the most abundant antinutrient in all the samanea saman pod samples, the highest level (26.51mg/100g) identified in the pulp is still within the safe level of less than 146mg/100g. the result of proximate composition of pod of rain tree showed that pulp had higher moisture content and carbohydrate content than whole pod and seed samples. it was observed that the whole pod had higher ash, ether extract and crude fibre than the pulp and seed samples whereas the seed was richer in protein content than the whole pod and pulp samples. the relative high protein (10.98% to 21.55%) and carbohydrate (47.33% to 57.53%) contents of the rain tree (samanea saman) pod samples suggest that they can be used as nutrient supplement in animal feeds and the seeds can be utilized as a rich protein meal in human diets. 4.2. contributions to knowledge within the limits of this research work, the study has, no doubt, contributed immensely to knowledge, which is as follows: • this study has provided information for the utilization of the rain tree (samanea saman) pod in food production. • this study has also opened another area of research such as pod utilization in food flavor and animal feed development. • the study has revealed the nutrient (protein, carbohydrate, lipid, ash, fiber and moisture) potential of these pods which are presently littering as waste. • this study has also opened another area of research such as pod utilization in food formulation and animal feed development. 4.3. recommendations this study leads to the need of further detailed search on the immense potentials of rain tree. the results of the present study are promising thus indicating the utilization of the pods of s. saman as a significant source of bioactive that if harnessed in the formulation of nutraceutical beverage could offer a whole lot of health benefit to the users. investors should be encouraged to invest in machines that can facilitate the removal of seeds from the pods. references [1] u. mbanali, m. chukwu, and m. iwuagwu, "variation in the functional properties of blends of heat-treated local thickening seeds and cocoyam flour," research journal of food science and nutrition, vol. 3, pp. 74-83, 2018.available at: https://doi.org/10.31248/rjfsn2018.053. 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s. mossalem 1* --gehan l. elenain 2 1 department of environmental research and medical malacology, theodor bilharz research institute (tbri), giza, egypt 2 department of parasitology, tbri, giza, egypt; natural sciences, university college, abu dhabi university, uae abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ........................................................................................................................................................................... 6 2. materials and methods .......................................................................................................................................................... 6 3. statistical analysis ................................................................................................................................................................. 7 4. results .................................................................................................................................................................................... 7 5. discussion ............................................................................................................................................................................... 7 references .................................................................................................................................................................................. 8 the present study was carried out to evaluate the molluscicidal activity of the pesticide emamectin (5% aqueous solution) and the methanol extract from the wheat bran (mewb) against b alexandrina and their infection with schistosoma mansoni was studied. the lc90 and lc50 values for emamectin were 50.4 and 22.3 ppm, respectively. infection of snails with s. mansoni under the effect of the tested agents was evaluated via four experimental groups, each of 50 snails for three consecutive days, one group of snails was exposed to 9.08 ppm aqueous emamectin solution, another group received 100 ppm methanol extract of wheat bran (mewb), a third group was administered by a combination of 9.08 ppm emamectin and 100ppm mewb, the fourth group was control maintained under similar experimental conditions. after three days, all the experimental groups were infected with s. mansonimiracidia and observed till shedding of cercariae. the physiological and histological changes were assessed before and after the infection.the changes in hemocytes of infected snails after administration with lc25 of emamectin or wheat bran was significantly (20% and 30%, respectively) suppressed compared to 75% for the control snails. however, the snails treated with joint of emamectin and mewb were the least infected snails (10%). on the other hand, the biochemical test results showed a remarkable reduction of got (p<0.01) gpt (p<0.05) and total protein (0.05) in the heamolyph extracted from the snails treated with an aqueous emamectin solution. yet, the levels of got were significantly increased in the groups administered with the mewb alone (p<0.01) or in combination with the emamectin (p<0.001). as for the total protein levels, there were slightly declining (p<0.05) in the group exposed to the aqueous emamectin and to the contrary, theses levels were significantly increased (p<0.05) in the snails of the two other experimental groups when compared to the controls. moreover, the haemocytes cells showed a differentiation which varied in number when detected under the microscope. administration of the aqueous emamectin solution resulted in a significant increase (p<0.05) of the amaebocytes and a considerable decrease (p<0.05) in the number of granulocytes and hyalinocytes compared to the control snails. on the contrary, a remarkable surge (p<0.05) of granulocytes and decrease of the amaebocytes was detected in the haemolyph of snails treated with mewb. keywords: biomphalaria alexandrina, emamectin, wheat bran, molluscicide, antioxidant, schistosoma mansoni. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2014, 1(1): 5-10 6 1. introduction schistosomiasis is the second major parasitic disease in the world after malaria [1]. it poses not only a health problem, but economic crisis as well. relatively 200 million people are infected with schistosoma and around 800 millions are at risk of infection [2]. fresh water snails are the intermediate hosts that transmit the human schistosomiasis [3]. the intestinal schistosomiasis is caused by scistosomamansoni which is transmitted by the intermediate host biomophalaria alexandrina, a fresh water snail found in egypt [4]. it is well-known that control of the freshwater snails might play a key role in the transmission of schistosomiasis. snails are approved to be the most vulnerable link in the schistosome life cycle and consequently, targeting their populations is an important approach to prevent the transmission of shistosomes. still comprehensive information about snails is required [5] to assist in developing a harmless molluscicide. application of molluscicides (synthetic or of plant origin) can confer a rapid and efficient technique in tumbling the snail populations [6]. attempts to reduce or eliminate populations of freshwater snails in africa have been concentrated on the intermediate hosts of schistosomes with less attitude to use the chemicals for the purpose of reducing transmission of schistosomiasis [4]. the chemical molluscicides have not achieved the expected outcomes [7] and the synthetic ones were approved to be costly in relation to the restricted budgets [7] available for the control of communicable diseases in many countries [8]. the development of molluscicides from plant origin is currently of interest in measures for snail control because they are affordable, safe and useful in local community self-help projects [9]. there are some plants with antioxidant activity such as rice bran which play an important role in reducing the pollution of aquatic media and were proved to be safe for all manner of objects and the surrounding environment [10]. it possesses not only detect the molluscicidal activity of the aqueous emamectin solution to assist in developing a candidate molluscicude, alone and combined with methanol extract from the wheat bran, as antioxidant agent, against b. alexandrina. 2. materials and methods 2.1. snails snails used in this study were adult biomphalariaalexandrina (8-10mm) collected from the river nile and irrigation scheme near cairo and brought to the laboratory, washed with dechlorinated water and examined for larval trematodes. unhealthy and infected snails were excluded, the healthy snails were maintained in plastic aquaria under standard conditions (25 o c ± 1 o c, ph = 7.2) for three weeks before being used in molluscicidal tests. snails were daily fed boiled lettuce leaves and blue green algae. 2.2. miracidia schistosomamansonimiracidia were obtained from schistosome biological supply center (sbsc), tbri, egypt. 2.3. emamectin (vetoken) emamectin (vetoken) is non-systemic insecticide which penetrates leaf tissues by translaminar movement. paralyses the lepidoptera, which stop feeding within hours of ingestion, and die 2-4 days.uses for control of lepidoptera on vegetables, brassicas, cotton and pine trees. the purchase company of emamectin is shanghai agrochina, international, trade, co. ltd chin,the structural formula of emamectin is (4″r) -4″-deoxy-4″(methylamino) avermectin b1 and mode of action: chloride channel activator. acts by stimulating the release of gaminobutyric acid, an inhibitory neurotransmitter, thus causing paralysis 2.3.1. extraction of methanol extract of wheat bran the wheat bran was dried in shade, then in an oven at 50 o c. ten grams powder was extracted with 250 ml 95% methanol at room temperature for one week. methanol was evaporated under vacuum and the residues were used for the bioassay tests. 2.4. the molluscicidal tests using aqueous emamectin and methanol extract of wheat bran the potential molluscicidal activities of emamectin and methanol extract of wheat bran were tested against adult b. alexandrina. different concentrations in ppm were prepared on the basis of weight/volume. for each concentration three glass containers each containing one liter dechlorinated water to which the test material was added directly on the water surface and stirred properly then 10 snails were introduced. the glass containers were covered by porous plastic sheets and maintained for 24 hours of exposure at normal laboratory conditions (25 o c ± 1 o c). after that, the snails were washed thoroughly with dechlorinated water, transferred to jars containing fresh dechlorinated water for another 24 hours for recovery. three replicates of control snails (10snails/l) were prepared in dechlorinated water. dead snails were distinguished and counted. further experiments were carried out such that laboratory bred b. alexandrina snails (8-10 mm) were deployed into four groups, each of 50 snails. they were continuously treated for 3 days with the tested agents one group emamectin solution was treated with the lc50 of emamectin, another group was treated with mewb, the third was exposed to a combination of 9.08ppm emamectin and 100ppm wemb, whereas the fourth group was maintained at the same conditions in dechlorinated water as a control group. dead snails were removed daily from all experimental groups. snails were considered dead if they probed and remained motionless or if the shell looked discolored. after 3 days of treatment, the survived snails at all groups were subjected to to infection with s. mansonimiracidia and kept till shedding of cercaria. the extent of the survival of the experimental snails were estimated by relating them to their counterparts of the control snails. determination of total protein, haemocytes number, glutamic oxaloacetic transaminase (got) and glutamic pyruvic transminase (gpt) in snails' hemolymph. agriculture and food sciences research, 2014, 1(1): 5-10 7 2.5. heamatocyte count haemolymph samples were collected according to michelson [11] by removing a small portion of the shell and inserting a capillary tube into the heart. the haemolymph pooled from 10 snails were collected in a vial tube (1.5ml) and kept in an ice-box. 2.5.1. total protein, glutamic oxaloacetic transaminase (got) and glutamic pyruvic transaminase (gpt) colorimetric estimation of total protein content in haemolymph of b. alexandrina snails was estimated according to the principle of biuret-tartrate method [12]. that of got and gpt was detected as stated by reitman and frankel [13]. 3. statistical analysis the means of the different groups were compared globally using the student's ttest sokal and rohlf [14]. the molluscicidal activity of the tested chemical was calculated by probit analysis, spss computer program version19, window7, mw office 2007. 4. results 4.1. molluscicidal activity of aqueous emamectin and methanol extract of wheat bran the effect of various concentrations of the aqueous emamectin (5%) and mewb on adult b. alexandrina snails after 24hr exposure was evaluated (table 1 and fig. 1). all mewb concentrations up to 100ppm were harmless to the snails. the lc25, lc50 and lc90 obtained with the aqueous emamectin solution against b. alexandrina snails were 9.08 ppm, 22.3ppm and 50.4 ppm, respectively. surprisingly, administration of joint 100ppm mewb and aqueous emamectin solution changed the activity of the emamectin against b. alexanderia snails to be 22.916ppm at lc25, 31.8 ppm at lc50 and 48.816 ppm in lc90, as depicted in table 2. 4.2. moullscicidal efficacy on the b. alexandrina infectivity withs. mansoni the percent s. mansoni infection to the experimental snails was remarkably lower (p<0.001) than that detected in the infected negative control snails. the lowest infection (10%) was determined by the group of snails administered by a joint of aqueous emamectin and mewb. 4.3. the effect of emamectin and mewb on b. alexanderina physiologic and histologic activity the enzyme activity measured by the levels of got, gpt and total protein showed variable responses to the administered substances. in the 9.08ppm emamectin treated snails, there was a significant decline in the levels of the three types of protein, whereas administration of mewb alone or in combination with the aqueous emamectin solution to snails increased the levels of got, gpt and total proteins in the haemolymph, considerably (table 2). administration of the emamectin resulted in a significant increase (p<0.05) of the amaebocytes and a considerable decrease (p<0.05) in the number of granulocytes and hyalinocytes compared to the control snails. on the contrary, a remarkable surge (p<0.05) of granulocytes and decrease of the amaebocytes was detected in the haemolyph of snails treated with mewb (table 3 & fig. 1). 5. discussion the aqueous emamectin (5%) is one of the most chemical pesticides used in cotton agriculture and causes pollution in the aquatic environment. the molluscicidal activity of aqueous emamectin (5%) alone and joint with the methanol extract from wheat bran was evaluated against b. alexandrina snails. the lethal concentrations of this molluscicide obtained against b.alexandrina snails lc90 and lc50 values were 50.4ppm and 22.3ppm.the current data showed a severe decrease (20% and 10%) in the infection rate of snails infected with s.mansoni after being continuously exposed to the sublethal dose lc25 of emamectin (5%) alone and in combination with the mewb, respectively. this may be explained by the deteriorations of physiological parameters of snails making them incongruous for the parasite development [15]. mahmoud, et al. [16] found that exposure of b. alexandrina and b. glabrata snails to sublethal concentration of cryptostegiagrandiflora 3 days pre-miracidial exposure led to a significant reduction in the infection rate with s. mansoni by 55.47% and 58.9%, respectively. in another study [17] ,administration of lc10 of daturastramonium and sesbaniasesban during s. mansonimiracidial exposure reduced the infection rate of b. alexandrina snails by 41.7% and 52.2%, respectively compared to the control group. all the recorded results in the present study concerning total protein levels, got and gpt activities are mostly in agreement with with similar research work conducted by many authors [18-21]. the present investigation showed that haemocytes in the experimental snails were significant different in number when compared to the control ones. three types of haemocytes were detected unevenly tribute in the haemolymph of b. alexandrina snails. they ranged from granulocytes 60%, amoebocytes 25% to hyalinocytes 15% in the lymph of the control snails, which is supported by the results obtained in different studies [15, 22-26]. administration of emamectin (5%) reduces the number of granulocytes and, hyalinocytes (40% and 10%, respectivel) but increased the number of amoebocytes by 50%. this observed surge in the number and size of amoebocytes might be due to their main role in encountering any invaders/foe that attack the snails' tissue and being activated by engulfing such administered harmful pesticide. previous similar observations were detected when snails were subjected to the molluscicideartemether for 3 days and obvious abnormalities were detected in haemocytes morphology [10] in the present study, exposure to the wheat bran alone or in combination with the emamectin has increased the number of haematocytes which might be due to the highly effective role of all extracts of wheat bran in inhibiting the linoleic acid peroxidation as demonstrated by laokuldilok, et al. 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[28] m. e. norhaizan, a. k. khairul-kamilah, a. zulkhairi, and a. azrina, "improving the lipid profile in hypercholesterolemia-induced rabbit by supplementation of germinated brown rice," j agric food chem 2011 jul 27, vol. 59, pp. 7985-91, 2011. agriculture and food sciences research, 2014, 1(1): 5-10 9 table-1. mortality rate of vetoken in comination with methanol extract of wheat bran against biomophalaria alexandrina snails total no. of dead snails total no. of snails concentration (ppm) 40 48 45 35 48 40 30 48 35 22 48 30 11 48 25 8 48 15 1 48 10 table-2.molluscicidal activity of vetoken (5%) alone and joint with wheat bran against b. alexandrina probit aqueous vetoken (5%) vetoken (5%) combined with 100 ppm mewb 95% confidence limits for concentration lower bound upper bound lower bound upper bound lc90 50.4ppm 48.816ppm 39.480 79.538 45.275 53.865 lc50 22.3ppm 31.847 16.748 31.370 29.804 33.977 lc25 9.08ppm 22.916 -5.032 15.834 19.937 25.235 fig-1. dose/probit regression line of joint vetoken/mewb on biomophalaria alexandrina table-3. the average percentage of s. mansoni infection to b. alexandrina in the experimental groups concentrations % infection got (g/dl) gpt (g/dl) total protein (g/dl) mean ± sd 9.08ppm vetoken 20%*** 0.318667±0.006351** 0.348667±0.005508* 5.44±0.1* 100ppm wemb 30%*** 0.415667±0.005508** 0.392333±0.005508* 6.776667±0.01527* 9.08ppm vetoken + 100ppm wemb 10%*** 0.542667±0.009018*** 0.415667±0.005508* 5.95±0.01* control 75% 0.350667±0.000577 0.353±0.001 5.748± 0.001 ***high significant at p<0.001; ** moderate significant at <0.01; *slightly significant at p<0.05 table-4.the haematocytes countbefore and after three-consecutive days’ exposure to aqueous vetoken and mewb concentration granulocyte amoebocyte hyalinocyte control 60% 25% 15% vetoken 40%* 50%* 10%* mewb 70%* 15%* 15% *significant at p<0.05 agriculture and food sciences research, 2014, 1(1): 5-10 10 fig-2. haemocytes under light microscope showing g granulocyte, a amoebocyte, and h hyalinocyte a. control; b. lc25vetoken-treated snails showed decreasing in the number of granulocytes; c. haemocytes from 100ppm mewbtreated snails showing an increase in the number and size of granulocytes. a is denoted for amaebocytes, g stands for granulocytes and h refers to hylinocytes (light microscope, 200x) views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. agriculture and food sciences research issn: 2411-6653 vol. 2, no. 1, 6-11, 2015 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 6 mycelia growth of pink oyster (pleurotus djmour) mushroom in different culture media & environmental factors iftekhar ahmad 1* --imran fuad 2 --zobada kanak khan 3 1 department of food engineering and tea technology, shahjalal university of science and technology, sylhet 2, 3 department of genetic engineering and biotechnology, shahjalal university of science and technology, sylhet abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ........................................................................................................................................................................... 7 2. materials and methods .......................................................................................................................................................... 7 3. results .................................................................................................................................................................................... 8 4. discussion ............................................................................................................................................................................. 10 references ................................................................................................................................................................................ 11 the study was undertaken to determine the optimum culture media and different other factors viz: optimum culture media composition, ph and temperature for a well-known mushroompink oyster (pleurotus djmour). four different types of culture media as-potato dextrose agar (pda), malt extract agar (mea), potato yeast dextrose agar (pyda), and yeast extract agar (yea) was prepared as different media for pure culture growth. calculation was conducted on average mycelium growth & duration of complete mycelium growth. the highest mycelia growth rate (0.24 cm) was observed for potato dextrose agar media and the lowest mycelia growth (0.11 cm) was observed in yeast media. the highest mycelia growth rate (0.41 cm) was observed at 25 0 c and the lowest mycelia growth (0.20cm) was observed at 30 0 c temperature. among the nutrient media compositions the fastest mycelium growth rate was observed in pda media at ratio of 15:150 (dextrose: potato) and minimum mycelia growth rate was observed in pda media at ratio of 25:250 (dextrose: potato). the ph that gives the highest mycelia growth rate (0.45cm) was observed in the ph 6.5 & the lowest (0.11cm) was observed in the ph 4.5 which statistically similar with ph 9.0. keywords: pink oyster, mycelium growth, potato dextrose agar (pda), malt extract agar (mea), potato yeast dextrose agar (pyda), yeast extract agar (yea). http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(1): 6-11 7 1. introduction mushroom describes a variety of gilled fungi, with or without stems, and the term is used even to describe both the fleshy fruiting bodies of some ascomycota and the woody or leathery fruiting bodies of some basidiomycota. pleurotus djamor, commonly known as the pink oyster mushroom, is a species of fungus in the family pleurotaceae. the potential of mushroom as fungal protein and as a source of medicinal compounds make the production of mycelium an attractive prospect. mushroom mycelium is used for medicinal and therapeutic purposes; mycelial biomass powder can be used to formulate various types of health tablets and capsules [1]. the biological efficiency of mushroom depends on the development of mycelia in the first cultural stage. healthy and active mycelial growth plays crucial roles in protecting themselves against several stress factors [2]. mushrooms are the source of extra ordinary power and virility and are used in the preparation of many continental dishes and have medicinal properties like anti-cancerous, anti-cholesteral, anti-tumorous. mushrooms are useful against diabetes, ulcer and lungs diseases [3]. mushrooms are the good source of protein, vitamins and minerals [4]. mushrooms contain about 85-95% water, 3% protein, 4% carbohydrates, 0.1% fats, 1% minerals and vitamins [5]. mushrooms contain appreciable amount of potassium, phosphorous, copper and iron but low level of calcium [6]. mushroom protein is intermediate between that of animals and vegetables [7]. mushroom also contain appreciable amount of niacin, pantothenic acid and biotin [8]. it is also important in biotechnological researches which are dealing with large scale production of mycelium (biomass) to obtain anti-cancer and antimicrobial substances, volatile compounds, biomass for food industry and enzymes production [9]. mycelia growth is strongly influenced by in vitro conditions and they obtain their nutrients by absorbing soluble inorganic and organic materials from medium which assures the maximum and most vigorous germination [10]. it has also been reported that healthy and active mycelia growth in used medium play crucial role for protecting themselves against several stress factors [ 2 ] . beside media the factors like temperature, ph and light affect to the mycelia growth. mycelium growth depends on several factors such as growing media, different media concentration, ph, temperature, nutrient element & some environmental factors. mycelia stage of mushroom was significantly influenced by the carbohydrate source, nitrogen source, temperature, ph and light intensity. fasola, et al. [11] reported that potato dextrose agar (pda) is the best culture media for mycelia growth of volvariella speciosa. fasola, et al. [11] also reported that the maximum, optimum and minimum mycelia growth of volvariella speciosa at the ph of 9.0, 6.0 and 3.0, respectively while no growth was observed at ph 2.0 and 10.0. generally, the growth of this fungus reduced at very strong acidic and alkaline ph. jonathan and fasidi [12] observed very good mycelia growth of volvariella esculenta at ph 6.0. this study was attempted to investigate the effects of different media, ph level, temperature, and different nutrient media composition on vegetative growth of pink oyster (pleurotus djamor) mushroom. 2. materials and methods the experiment was conducted in the tissue culture laboratory of national mushroom development and extension centre (namdec), sub centre (strengthening mushroom development project), horticulture center, mehedibag, sylhet, bangladesh. 2.1. preparation of culture media for pure culture potato dextrose agar ( pda), malt extract agar ( mea), potato yeast dextrose agar ( pyda), and yeast extract agar (yea) was prepared as different media for pure culture growth according to standard methods. media was autoclaved at a pressure of 15 pound square inch (psi) for twenty minutes at 121ºc. calculation was conducted on average mycelium growth & duration of complete mycelium growth. 2.2. sterile culture technique a healthy, fresh, clean mushroom fruit body was selected for culture. a section of mushroom was broken off. pour the different media into sterile tubes. using the index finger and thumb of the left hand, a piece of the mushroom was firmly grasped with the clean surface facing upward or sideways. with the right hand, the scalpel was grasped firmly by the handle. starting with the tip of the scalpel, a very small triangular shape was cut from the clean inner tissue. the tissue was gently "teased" free, and placed it quickly just inside the lip of the tube. then the bottom of the tube was taped for a few times on the heel of hand to position the mushroom tissue on the surface of the agar. the tube was then placed in a second storage can. these steps were repeated for the remaining tubes and mushroom materials. it was important to make several tubes from each mushroom specimen, since even with the most careful technique contamination can sometimes occur. 2.3. preparation of different synthetic pda media different amount of potato & dextrose were used for pda media. the pda was prepared by 0g, 5g, 10g, 15g, 20g, 25g of dextrose & 0g, 50g, 100g, 150g, 200g, 250g potato mixed with20g agar at ph 6.5. the mixture was boiled in gas burner until the agar dissolved. the media was poured into petri plate at 20 ml/plates and sterilized and solidified. then the plates were inoculated with testing mushroom species. then plates were transferred in incubation room for mycelium running at 20-25 o c temperature. 2.4. study of the different temperature on mycelial growth of p. djamor the selected medium was employed to evaluate the suitable temperature for mycelium growth of p. djamor. some composition was adjusted at 6.5 ph value before autoclave the inoculated plates of p. djamor on pda (potato dextrose agar) were placed at six different temperature values such as 10 0 c, 15 0 c, 20 0 c, 250 c 30 0 cand 35 0 c, with the ph value of each medium being 6.5. agriculture and food sciences research, 2015, 2(1): 6-11 8 2.5. study of the different ph on mycelial growth of p. djamor the media were adjusted to different ph values at 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5 and 9.0 with the addition of 1n hcl or 1n naoh before pouring into petri plates and autoclaving. the petri plates were inoculated with the inocula of the mushroom. after autoclaving, the petri plates were inoculated and transferred into incubation room for mycelial growth at 21 0 to 25 0 c temperatures. average mycelia growth and duration of completion of mycelium running of each replication were calculated. 3. results 3.1. study of the different growth media on mycelial growth of pleurotus djamor 3.1.1. mycelium growth per day four different culture media such as potato dextrose agar (pda), malt extract agar (mea), yeast extract agar (yea), malt extract yeast agar (meya) and potato yeast dextrose agar (pyda) medium were used to investigate the mycelial growth of pleurotus djamor. the highest mycelial growth rate (0.24cm/day) was observed in the potato dextrose agar (pda) media where lowest mycelial growth was found in yea media (0.11cm/day) (table 1). the density of mycelium on pda was higher than other culture media. table-1. mycelial growth rate per day by different media media mycelial growth rate (cm/day) potato dextrose agar 0.24 a malt extract agar medium 0.20 a potato yeast dextrose agar 0.16 ab yeast agar medium 0.11 b c.v (%) 9.16 note: means followed by same letters are not significantly different from each other at 5% level 3.2. total time required for complete mycelium growth in case of duration of complete mycelium running, the minimum time (5.25 days) was recorded in pda medium and the maximum time (10.25 days) was found in yea medium (figure 1). figure-1. days required for mycelium growth in line diagram the diagram describes that pda media takes the least amount of time to complete mycelium growth and yem takes the highest amount of time. 3.3. study of the different ph on mycelial growth of pleurotus djamor to determine the suitable ph on pda media with the ph range of 4.5 to 9.0 were used for mycelial growth of pleurotus djamor. the best mycelia growth (0.45 cm/day) was observed in slightly acidic medium (ph 6.5) which was statistically different from other ph values (table 2). table-2. mycelial growth rate (cm/day) at different ph level ph mycelial growth rate (cm/day) 4.5 0.11 c 5.0 0.13 c 5.5 0.15 c 6.0 0.31 b 6.5 0.45 a 7.0 0.31 b 7.5 0.30 b 8.0 0.26 b 8.5 0.15 c 9.0 0.12 c c.v% 4.93 note: means followed by same letters are not significantly different from each other at 5% level by interpreting the results we find that means are not significantly different from each other for ph of 4.5, 5.0, 5.5, 8.5, 9.0 and 6.0, 7.0, 7.5, 8.0 are not significantly different but 6.5 is significantly different mean than others. agriculture and food sciences research, 2015, 2(1): 6-11 9 3.4. time required for complete mycelium growth depending on ph in case of duration of complete mycelium running, the minimum time (5.0 days) was recorded at ph 6.0 and 6.5 which was statically similar to ph 6.0 and ph 7.0 and the maximum time (9.25 days) was found at ph 4.5 (figure 2). figure-2. days required for mycelium growth in line diagram at different ph. figure-3. mycelium growth at ph 6.5, gives the highest mycelium growth rate 3.5. study of the different temperature on mycelial growth of pleurotus djamor the best mycelial growth (.41 cm/days) was obtained at 25 0 c which was significantly higher on compare to all the treatment. the lowest mycelial growth (0.20 cm/day) was recorded in 30 0 c. 10 0 & 35 0 has no mycelium growth at all (table 3). table-3. mycelial growth rate at different temperature temperature ( 0 c) mycelial growth rate (cm/day) 10 -- 15 0.28 b 20 0.26 bc 25 0.41 a 30 0.20 c 35 -- c.v% 4.01 note: means followed by same letters are not significantly different from each other at 5% level 3.6. time required for complete mycelium growth depending on tempereture in case of duration of complete mycelium running, the minimum time (5.25 days) was recorded at 20 0 c, which was statistically similar to 15 0 c. in case of 35 0 c temperature, the duration of complete mycelium running was more than 7.75 days. agriculture and food sciences research, 2015, 2(1): 6-11 10 table-4. days required for complete mycelial growth at different temperature temperature ( 0 c) days required for complete mycelial growth 10 -- 15 6.50 a 20 5.20 a 25 7.00 a 30 7.75 a 35 -- c.v% 9.24 note: means followed by same letters are not significantly different from each other at 5% level 3.7. mycelial growth rate for various pda media composition (cm/day) the result of mycelia growth rate in different ratio of potato & dextrose used in the culture media as well as variety is shown below. highest mycelial growth rate (.33cm/day) was observed from the treatment t4 when pleurotus djmour was inoculated in pda at ratio of dextrose: potato (g/liter) (15/150) (table 5). table-5. mycelial growth rate (cm/day) for various pda media composition treatment mycelial growth rate (cm/day) t1 -- t2 0.28 ab t3 0.30 ab t4 0.33 a t5 0.31 ab t6 0.26 a c.v% 9.57 note: means followed by same letters are not significantly different from each other at 5% level [here t1= dextrose (g/liter): potato(g/liter) (0:0), t2= dextrose (g/liter): potato(g/liter) (5:50), t1= dextrose (g/liter): potato(g/liter) (10:100), t1= dextrose (g/liter): potato(g/liter) (15:150), t1= dextrose (g/liter): potato(g/liter) (20:200), t1= dextrose (g/liter): potato(g/liter) (25:250)] 3.8. duration of mycelium completion for various pda media composition (days) duration of mycelia completion in different ratio of potato and dextrose used in culture media is shown in table below. maximum days (14.00 days) required for the completion of mycelium running was observed from t6 when p.djmour was inoculated in pda at ratio of dextrose (g/liter) (25:250) because excess nutrient delay the mycelial growth. minimum days required for completion of mycelim running was observed from the treatment t4 when p. djmour was inoculated in pda at ratio of dextrose (g/liter): potato (g/liter) (15/150). table-6. duration of mycelium completion for various pda media composition treatment days required to mycelium complete (days) t1 -- t2 13.75 a t3 13.00 ab t4 11.50 b t5 12.25 ab t6 14.00 a c.v% 9.83 note: means followed by same letters are not significantly different from each other at 5% level 4. discussion the growth of mycelium depends on different factors such as media, ph, temperature, nutrient element and some environmental factors [13]. medium is important because it supplies necessary nutrient for the growth of mushroom mycelium. culture media permit acceleration of mycelial growth; ensure quality and year round production [14]. the highest mycelial growth rate (0.24cm/day) was observed in the potato dextrose agar media which was similar to khan, et al. [15] who reported that malt extract potato extract & wheat extract were the best media for cultivation of auricularia politrica. fasola, et al. [11] reported that ph 6.0 is best for mycelial growth of v. speciosa. ibekwe, et al. [16] reported that the optimum mycelia growth of pleurotus ostreatus was recorded at ph 6.5 which supports our proposition. likewise, chandra and purkayastha [17] and jonathan and fasidi [12] obtained very good growths of agaricus campestris and v. esculenta at ph 6.0. this result is not similar with the findings of chang and miles [10] who reported 7.5 as the best ph level for mycelial growth of v. volvacea. the lowest mycelial growth (.11 cm/day) was recorded at ph 4.5. mushroom mycelium is significantly influenced by various environmental factors such as light, temperature, ph, carbon sources etc. ph has great effects on nutrition and morphological development of mushrooms. generally the growth of fungi is reduced at very strong acidic and alkaline ph. different agar media such as potato dextrose agar, malt extract agar, lamberts agar, compost extract agar are mostly used for the growth of mycelium [18]. the effect of mycelial growth rate in four different culture media were shown in the results. significantly the highest mycelium growth (0.24cm/day) of pleurotus djamor was observed in potato dextrose agar media which is statistically similar (0.20cm/day) to malt extract yeast agar media and the lowest mycelium growth rate (0.11 cm/day) was found in yeast extract medium. this result is similar to kalm and kalyoncu [9]. the highest mycelium growth of pleurotus geesternaus was observed in potato dextrose agar media (0.41 cm/day) and this medium has been demonstrated to be highly supportive of mycelial growth of mushrooms by several authors such as jeffers and martin [19]. the lowest mycelium growth rate (0.29 cm/day) was found in yeast extract agar media. agriculture and food sciences research, 2015, 2(1): 6-11 11 the days required to the completion of mycelium running in different media is shown in the results. the lowest time (5.25 days) required for the completion of mycelium running of pleurotus djamor was observed in potato dextrose. agar which was statically similar to malt extract agar medium (6.75days) and highest time (10.25 days) was observed in yeast extract agar media. kalm and kalyoncu [9] reported that mea and pda are more suitable media for mycelial growth of morchella spp. for pleurotus djamor the temperature range of 10-35 0 c was considered to find out the most suitable one. the highest mycelial growth (0.41 cm/day) was found in 25 0 c in the case of pleurotus djamor .this result is supported by sung, et al. [20]; they stated that the favorable temperature for mycelial growth was 30 0 c for macrolepiota procea and pleurotus ostreatus. this result is similar to the report of shim, et al. [21]; they stated that the mycelial growth of p. fumosoroseus was favorable at the temperature of 20 to 25 0 c. for pleurotus djamor, 10 0 c & 35 0 c was found to be not suitable for mycelial growth. the test was conducted also to determine the best ratio in pda media for mycelia growth for pleurotus djmour. highest mycelial growth rate (.33cm/day) was observed from the treatment t4 when pleurotus djmour was inoculated in pda at ratio of dextrose: potato (g/liter) (15/150) which was statistically similar to t5. similar findings were reported by khandaker, et al. [22], who reported that pda media gave highest (0.33cm/day) mycelial growth of grifola frondosa. references [1] t. chen and z. p. xin, "taurine in the spores and extract powder of log cultivated ganoderma lucidum," acta edulis fungi., vol. 8, pp. 45-46, 2001. [2] r. s. herderio, m. d. pereira, a. d. panek, and e. c. eleutherio, "trehalose protecs saccharomyces cerevisiae from lipid peroxidation during oxidative stress," biochemical biophysical. acta, vol. 17, pp. 340-346, 2006. [3] t. h. quimio, "cultivation ganoderma the “pleurotus-way” mushroom," newsletter for tropics, vol. 6, pp. 12-130, 1976. [4] s. m. khan, a. g. kausar, and m. a. ali, "yield performance of different strains of oyster mushroom (pleurotus spp.) on paddy straw in pakistan," mush. sci. x1 sydney, pp. 675-67, 1981. [5] r. p. tewari, mushroom cultivation. extension bulletin vol. 8. india: indian institute of horticulture research, banglore, 1986. [6] e. e. anderson and c. r. feller, "the food value of mushroom agaricus compestri," pool. am. soc. hort., vol. 41, pp. 3010-303, 1942. [7] r. h. j. kurtzman, "nutrition of pleurots sapidus effects of lipids," mycologia, vol. 68, pp. 268-295, 1976. [8] t. r. subramanian, nutritive value. mushroom extension bulletin vol. 8. india: indian institute of horticulture research, 1986. [9] e. kalm and f. kalyoncu, "mycelial growth rate of some morels (morchella spp.) in four different microbiological media," american-eurasian j. agric. & environ. sci., vol. 3, pp. 861-864, 2008. [10] s. t. chang and p. g. miles, edible mushrooms and their cultivation. boca raton, florida, usa: crc press inc, 1987. [11] t. r. fasola, j. s. gbolagade, and i. o. fasidi, "nutritional requirements of volvariella speciosa (fr. ex. fr.) singer, a nigerian edible mushroom," food chemistry, vol. 100, pp. 904-908, 2007. [12] s. g. jonathan and i. o. fasidi, "physico-chemical studies on volvariella esculenta mass (singer), a nigerian edible fungus," food chemistry, vol. 85, pp. 339-342, 2004. [13] c. t. calam, the evaluation of mycelial growth. in: norris j. r. and ribbons d. w. (eds). carbohydrate-peptide linkage of acidic heteroglycopeptide with immunostimulating. methods in microbiology vol. 1. new york: academic press, 1971. [14] s. t. chang, "a 40-years journey through bioconversion of lignocellulosic wastes tomushrooms and dietary supplements," int. j. med. mushrooms, vol. 3, pp. 299-310, 2001. [15] s. m. khan, j. h. mirza, and m. a. khan, "physiology and cultivation of wood’s ear mushroom (auricularia polytricha (mont.) sacc.)," isms, vol. 13, p. 21, 1991. [16] v. i. ibekwe, p. i. azubuike, e. u. ezeji, and e. c. chinakwe, "effects of nutrient source and environmental factors on the cultivation and yield of oyster mushroom (pleurotus ostreatus)," pak. j. nutr., vol. 7, pp. 349-351, 2008. [17] a. chandra and r. p. purkayastha, "physiological studies on indian mushroom," transactions of the british mycological society, vol. 69, pp. 63-70, 1977. [18] v. n. pathak, n. yadav, and m. gour, mushroom production and processing technology. jodhpur, india: agrobios, chopasani road, 1998. [19] s. n. jeffers and s. b. martin, "comparison of media selective for phytophythora and phytium species," plant diseases, vol. 70, pp. 1038-1043, 1996. [20] j. sung, h. w. moon, and d. h. park, "growth condition of liquid culture by pleurotus ostreatus," the korean journal of mycology, vol. 27, pp. 1-9, 1999. [21] m. j. shim, m. w. lee, h. s. ro, h. s. lee, and t. s. lee, "the characteristics of culture conditions for the mycelial growth of macrolepiota procea," mycoboiology, vol. 33, pp. 15-18, 2005. [22] j. khandaker, s. yesmin, and m. moonmoon, "mycelial growth of pleurotus citrinopileatus on different culture media and environmental conditions," bangladesh j. mushroom. loncoff, g. 1981. guide to mushrooms. simon and schuster, new york, vol. 2, pp. 55-61, 2008. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. 20 © 2021 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 8, no. 2, 20-29, 2021 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2021.82.20.29 © 2021 by the authors; licensee asian online journal publishing group characterisation of phenolic compounds, sterols and geographical fingerprint of çekişte extra-virgin olive oils according to their geographical locations by using lc ims qtof mass spectrometry ayca akca uckun republic of turkey ministry of agriculture and forestry, olive research institute, cultivation techniques, bornova, i̇zmir, turkey. email: ayca.akca@tarimorman.gov.tr abstract in this context, aim of this study is to determine the effect of çekişte olive oils in different locations and show differences on geographical locations while taking geographical indication label. çekişte olive oil variety which cultivated in six different locations (birgi, bademli, beyazit, yeniceköy, zeytinlik, uzumlu) were evaluated the effects of geographical locations on the chemical characterization of in the southwest of turkey. the agricultural ecological map of each location was created using gis. olive oil samples were analyzed fatty acid, sterol and phenolic. moreover, lc ims qtof spectrometer and progenesis qi software were used to determine the geographical fingerprints of olive oil samples in different locations. results showed that oil qualities of some locations differ significantly depending on olive growing area (p <0.05), some of them not. the principal component analysis of the different locations analyzed revealed that "geographical location" factor significantly affects the olive oil quality. keywords: olive oil, sterols, phenolic compounds, lc ims qtof spectrometer, çekişte variety, geographical indication label. citation | ayca akca uckun (2021). characterisation of phenolic compounds, sterols and geographical fingerprint of çekişte extravirgin olive oils according to their geographical locations by using lc ims qtof mass spectrometry. agriculture and food sciences research, 8(2): 20-29. history: received: 20 july 2021 revised: 25 october 2021 accepted: 23 november 2021 published: 13 december 2021 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: the author gratefully thanks to olive research institute, ankara food control laboratory directorate, origin determination laboratory unit, republic of turkey ministry of agriculture and forestry and olive farmers of karaburun region. funding: this work has been carried out project no 1513, ‘’investigation of olive and olive oil with regional characteristics with climatic and topographic conditions and determination of geographical indication standardization’’ by tagem (agricultural research policies directorate). competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 21 2. material and methods ..................................................................................................................................................................... 21 3. results and discussion ................................................................................................................................................................... 23 references .............................................................................................................................................................................................. 29 mailto:ayca.akca@tarimorman.gov.tr http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.doi.org/10.20448/journal.512.2021.82.20.29 https://orcid.org/0000-0002-5592-496x agriculture and food sciences research, 2021, 8(2): 20-29 21 © 2021 by the authors; licensee asian online journal publishing group contribution of this paper to the literature there are a lot of studies carried out to differ evoos from geographical regions according to various chemical characterizations in turkey. however, there is not any research on the effect of the geographical location on the chemical characterization of the çekişte variety. in this study, the analysis of sterol and polyphenolic compounds of evoos in the same region but under different ecological zone (climatic and topographic conditions) was performed for the first-time using lc ims qtof mass spectrometry via appropriate extraction method. 1. introduction extra virgin olive oil (evoo) is considered the healing oil due to its essential nutrients. evoo is unique compared to other oils (vegetable, animal) with its chemical composition (antioxidants, phenols, fatty acid, vitamins, etc.). there are more factors affecting chemical composition, physico-chemical quality, sensory properties of olive oil. these are environmental factors (topography, ecological zone, humidity, altitude, climate), agronomic factors (irrigation, pruning, pesticide application, fertilization, harvesting time, ripening index) and post harvest factors (oil extraction system, oil storage conditions) [1]. in addition, it is argued that quality of olive oil is effected by olive variety and geographical regions. polyphenols and antioxidant are the main parameters to be considered in geographic fingerprint according to geographical region and olive variety in evoo [2]. also, fatty acid plays important parameter in the chemical of evoo which has a high content of fatty acids, for example oleic acid ranging from 56 % to 84 %. another important parameter is the sterol profile which is considered as a fingerprint to examine its geographic originality in the evoo. olive oil leaders such as spain, italy and greece use some tools to separate evoos by geographical location. one of the these tools is lc ims qtof mass spectrometer and progenesis qi software [3]. çekişte olive variety, dominated by the southwestern part of turkey and the coastal part, has a long history. the oil properties of the memecik olive variety are quite high. various studies are carried out to differ evoos from geographical regions according to various chemical characterization in turkey. however, there is not any research on the effect of the geographical location on the chemical charecterization of the çekişte variety which cultivated in the southwest of turkey and has a high oil content. in addition, the analysis of sterol and polyphenolic compounds of evoos in the same region but under different ecological zone (climatic and topographic conditions) was performed for the first time using lc ims qtof mass spectrometry via appropriate extraction method. only a few researchers have focused their attention on geographical location of olive tree crops and how it may reveal its affect on physical, chemical properties of evoos. the characterization of geographical indication studies related to evoos have not been published on literature in turkey. in this context, propose of this study is to determine the effect of çekişte olive oils in different locations (birgi, bademli, beyazıt, yeniceköy, zeytinlik, üzümlü) and show differences on geographical locations while taking geographical indication label. 2. material and methods 2.1. fruit samples the research was conducted throughout 2020/2021 olive season. olive fruit samples of the çekişte variety (~ 2,5 kg for each sample) were collected by hand randomly from 6 different locations (birgi, beyazıt, bademli, yeniceköy, zeytinlik, üzümlü) and 9 trees in each location. olive fruits (2,04-4,05) were collected at the ripening index. figure-1. agricultural ecology map of ödemiş region. agriculture and food sciences research, 2021, 8(2): 20-29 22 © 2021 by the authors; licensee asian online journal publishing group 2.2. remote sensing methods (gis) and agricultural ecology map agricultural ecology maps of the locations were created with remote sensing methods (figure 1). agricultural ecoregion maps which is the creation of similar homogeneous areas by bringing together the climate, topographic, soil parameters that make up the land features by means of gis. the classification system is based on the system. table-1. agricultural ecological map information of ödemiş region. selçuk region ecological map altitude drought index warmest average coldest average slope birgi location 10721 500-900 m 0.75-1 (humid) 20-30 oc 0-10 oc %2-12 bademli location 10731 300-500 m 0.5-0.75 (semi humid) 20-30 oc 0-10 oc %12-30 beyazit location 10721 500-900 m 0.75-1 (humid) 20-30 oc 0-10 oc %2-12 yeniceköy location 10331 300-500 m 0.5-0.75 (semi humid) 20-30 oc 0-10 oc %2-12 zeytinlik location 10100 100-300 m 0.5-0.75 (semi humid) >20 oc 0-10 oc %2-12 uzumlu location 10221 100-300 m 0.5-0.75 (semi humid) 20-30 oc 0-10 oc %2-12 2.3. oil extraction following the harvest, olive fruites were brought in laboratory and olive oil was extracted within 24 hours. fruits were taken from 6 different locations (birgi, bademli, beyazıt, yeniceköy, zeytinlik, üzümlü) and olive oils were used with "abencor" system for extracted. evoo were stored in dark glass bottles at 4 oc. 2.4. determination of sterol composition and amount by capillary column gas chromatography sterol composition and amount of evoos were determined by capillary column gas chromatography and erythrodiol and uvaol of total sterols were made by using turkish food codex related to olive oil sampling and analysis methods communiqué 2014/53 [4]. 2.5. chemicals and extraction of sterols and phenolic compounds methanol of lc grade used for the extraction of sterols and phenolics from samples and preparing the mobile phase were supplied from isolab. deionized water was obtained by filtration using a milli-q-system (millipore, bedford, ms, usa). ammonium acetate used for preparing the mobile phase was purchased from sigma aldrich (st. louis, mo, usa). extraction of sterols and phenolic compounds from evoos was carried out using liquid extraction method. meoh:h2o (80:20, v/v) was used as the extraction solvent. three grams of olive oil samples weighted into centrifuge tube, and added with 3 ml of 80 % meoh solution. after homogenization via vortex, samples were centrifuged for 10 min at 7500 rpm. the supernatant was collected, the pellet was used again. the same procedure was repeated 3 times. all supernatant fractions were collected, combined and filtered through 0,22 µm ptfe syringe filters and stored in vial until lc ims qtof analysis. the extracts were mixed with an equal volume of water prior to analysis. also, a pool sample consisting of all oil samples was prepared in the same way to check the accuracy of our research. the extracts were mixed with an equal volume of water prior to analysis. also, a pool sample consisting of all oil samples was prepared in the same way to check the accuracy of our research. 2.6. fatty acid composition the fatty acid composition was determined by gas chromatography (gc) after saponification/methylation with methanolic koh via the official method (eec reg.2568/91). the fatty acids was detected by the comparison of retention time in standard compounds. 2.7. lc ims qtof screening the lc ims qtof system (ultra-high performance liquid chromatography with a acquity uhplc i-class system (waters, milford, ma, usa) was coupled to a vion® ims qtof (waters, manchester, uk), ion mobility quadrupole time-of-flight) was used. the lc separation was performed using an acquity uplc beh c18 (100x2,1 mm, id. 1,7 µm particle size, waters) analytical column. the oven was set at 30 °c. the solvents used consisted of (a) 90% h2o, 10% meoh, and 5 mm ammonium aceate and (b) 100% meoh and 5 mm ammonium aceate. the used flow gradient started with 1% solvent b with flow rate 0.35 ml/min during 1 min, increasing to 39 % fort he following two min and then incereasing to 99 % fort the next 11 min. organic conditions were kept for 2 min and then initial conditions were restored within 0.1 min and the column re-equilibrated for 2.5 min. the total elution programme was 18 min. the injection volume was 3 µl. the qtof system was operated in positive ionization mode, capillary voltage of 3.0 kv, mass range 50-1200 m/z. source temperature of 120 °c, desolvation temperature of 400 °c. external calibration was performed using a leu-enkephalin solution injected during the run 1 min intervals. data were collected under low collision energy of 6.0 ev and high collision energy of 15 to 45 ev table 2. olive oil samples were analyzed in waters brand vion lc ims qtof system (waters, milford, ma, usa) in order to determine the origin tables 2 and 3. all samples were analyzed in the same batch without any stopping. data acquisition and data analysis were carried out by unifi (waters, usa) software. then, the raw datas were subjected to principal component analysis (pca) using progenesis qi (nonlinear dynamics, waters, usa) software. unifi data format were converted to .uep format using the peak picking options figure 2. agriculture and food sciences research, 2021, 8(2): 20-29 23 © 2021 by the authors; licensee asian online journal publishing group figure-2. lc ims qtof screening use phase. table-2. the operating conditions of the vion lc ims qtof system. sample manager: ims qtof: column manager: scan settings: wash solvent: metanol:su (70:30, v/v) sample temperature: 8.0 ºc analyzer mode: sensitivity capillary voltage: 3.0 kv source temperature: 120 ºc desolvation temperature: 400 ºc cone gas: 50 l/h desolvation gas: 1000 l/h temperature: 30 ºc scan settings: 50 m/z – 1200m/z high deffinition mse low energy: 6.00ev high energy ramp: 1515 ev table-3. flow gradient of vion lc ims qtof system. time (dk) flow (ml/dk) % a % b curve 0.00 0.350 99.0 1.0 6 1.00 0.350 99.0 1.0 6 3.00 0.350 61.0 39.0 6 14.00 0.350 1.0 99.0 6 16.00 0.350 1.0 99.0 6 16.01 0.350 99.0 1.0 6 18.00 0.350 99.0 1.0 6 2.8. statistical analysis social sciences (spss) program release 16.0 was made for statistical analyses. the results are determined as mean ± standard deviation (sd) of five measurements for each analytical data. significant differences were determined at p < 0.05 between the values of all parameters according to the one-way anova: post hoc comparisons (duncan test). the principal component analysis (pca) was made using progenesis qi sofware. it was applied to separate evoos each geographical locations according to all parameters. 3. results and discussion 3.1. total phenolic content the total amount of phenolic was determined in evoos. statistical analysis are found significant differences (p <0.05) in total phenol contents among oil samples. total phenol content ranges from 107.319 to 180.892 mg/kg-1 in olive oils. figure-3. total phenol contents in olive oil samples from çekişte variety in six locations in the west of turkey. results are shown as means ± sd (n = 5). a-b different letters indicate significantly different values at p < 0.05 according to duncan test. agriculture and food sciences research, 2021, 8(2): 20-29 24 © 2021 by the authors; licensee asian online journal publishing group evoos of zeytinlik locations have highest phenolic content (180.892 mg/kg-1). zeytinlik location was showed that geographical location have a significant effect on the total phenolic content figure 3. del monaco, et al. [5] examined some of evoos from different locations and varieties in a study. he reported that characterization of evoos according to geographical regions and total phenol content differs greatly according to geographical location in italy. this study reveals that it is in line with our results. another study related to zarazi varieties in tunisia, higher phenolic content was detected in olive oil samples in the south of tunisia. result of study, it was stated that water scarcity increases the phenolic content due to drought creates a stress condition that triggers phenolic synthesis in olive fruits. there are significant differences among geographical regions in terms of different phenolic compounds in this study and also in previous studies. bajoub, et al. [6] found that qualitative and quantitative phenolic composition of evoos is strongly influenced by agricultural parameters, but effected by the genetic factors and environmental conditions particularly climate and topography. our datas showed that there are different phenolic contents of olive trees in same altitude due to soil structure, nutritional status. altitude effects phenolic content as positive in evoo. 3.2. fatty acid composition important fatty acid compositions were determined in çekişte olive oil samples. as shown in table 4, oleic (c 18: 1), palmitic (c 16: 0), linoleic (c 18: 2) and stearic acids (c 18: 0) are the main fatty acids in evoo. the contents of oleic acid is between 79.09 % (zeytinlik), 77.33 % (bademli). palmitic acid content is respectively 12.52 % (bademli), 11.80 % (üzümlü), 11.76% (zeytinlik). the highest linoleic acid content was in zeytinlik location (79.31 %). fatty acid composition was found different among geographical locations (table 4). morelló, et al. [7] studied that fatty acid changes due to genetic factors as well as environmental conditions. piravi-vanak, et al. [8] found that fatty acid composition of evoo is significantly affected by topographical and climatic conditions. table-4. fatty acid composition (%) of evoos in six different geographic locations in the southwest of turkey. note: each value represents the mean of five determinations (n = 5) ± standard deviation. nd not determined. a-f different letters in the same row indicate significantly different values (p < 0.05) according to duncan test. 3.3. effects of geographical location on phytosterol contents some sterols are the main sterols such as β-sitosterol, campesterol and δ-5-avenasterol. other sterols are minor sterols such as stigmasterol, clerosterol and-5-24-stigmastadienol in evoo (table 5). as shown in table 5, phytosterol are mostly depending on the geographical location of evoo. in the çekişte variety, the highest phytosterol content is β-sitosterol, followed by δ-5-avenasterol. significant differences were found in the contents of β-sitosterol and δ-5-avenasterol according to different geographical locations. (p <0.01). the highest β-sitosterol content was found in üzümlü location (86.70 %), then bademli (84.53 %) and birgi location (84.33 %). regarding the content of δ-5-avenasterol, zeytinlik location showed the highest value (10.87 %), while it was the lowest (7.83 %) in üzümlü location. table-5. phytosterol composition (%) of evoos in 6 different locations in turkey (birgi, bademli, beyazıt, yeniceköy, zeytinlik, üzümlü). locations campesterol stigmasterol δ5-24 stigmastadienol β -stosterol δ5avenasterol clerosterol apparent β -sterol bademli 2.38± 0.02ef 0.62± 0.00d 0.70± 0.00b 84.53± 0.05b 10.00± 0.02d 0.44± 0.00d 94.46± 0.00b beyazıt 2.39± 0.00e 0.53± 0.02e 0.52± 0.05e 84.30± 0.02b 10.54± 0.00b 0.68± 0.05b 93.26± 0.02c birgi 2.65± 0.01c 0.50± 0.00f 0.66± 0.03c 84.33± 0.00b 9.79± 0.14e 0.78± 0.03a 94.92± 0.01b üzümlü 2.85± 0.03a 0.77± 0.08a 0.43± 0.11f 86.70± 0.01a 7.83± 0.03f 0.38± 0.11e 93.43± 0.13c yeniceköy 2.59± 0.00d 0.64± 0.03c 0.87± 0.06a 83.82± 0.03c 10.14± 0.00e 0.55± 0.06c 93.18± 0.07cd zeytinlik 2.70± 0.04b 0.69± 0.01b 0.63± 0.02d 83.24± 0.00c 10.87± 0.05a 0.30± 0.02e 95.48± 0.03a note: apparent β-sitosterol (sum of clerosterol + β-sitosterol + δ-5-avenasterol + δ-5, 24-stigmastadienol). each value represents the mean of five determinations (n = 5) ± standard deviation. a-f different letters indicate significantly different values (p < 0.05) in the same row according to duncan test. ödemiş region birgi bademli beyazıt yeniceköy zeytinlik üzümlü 1 miristik acid (c14:0) 0.01 ± 0.03a 0.01 ± 0.04a 0.01 ± 0.04a 0.01 ± 0.05a 0.01 ± 0.02a 0.01 ± 0.04a 2 palmitik acid (c16:0) 11.58 ± 0.12e 12.52 ±0.08a 11.64 ± 0.07d 11.37 ± 0.05f 11.76 ± 0.12c 11.80 ± 0.06b 3 palmitoleik acid (c16:1) 0.81 ± 0.06d 0.87 ± 0.04b 0.86± 0.13c 1.01± 0.08a 0.88± 0.05a 0.78± 0.04e 4 heptadekanoik acid ( c17:0) 0.14± 0.07d 0.18± 0.12b 0.15± 0.11c 0.13± 0.12e 0.15± 0.13c 0.19± 0.09a 5 heptadesenoik acid (c17:1) 0.35± 0.07b 0.35± 0.02b 0.34± 0.04c 0.33± 0.07d 0.33± 0.06d 0.39± 0.04a 6 stearik acid (c18:0) 2.25± 0.11b 2.56± 0.08a 2.06± 0.13d 1.95± 0.12e 2.14± 0.09c 2.21± 0.08b 7 oleik acid (c18:1) 78.60± 0.06b 77.33± 0.03d 78.64± 0.03bc 79.09± 0.06b 79.31± 0.05a 78.45± 0.08c 8 linoleik acid (c18:2) 5.03± 0.04a 4.83± 0.09b 5.03± 0.09a 4.83± 0.05b 4.35± 0.04c 4.83± 0.06b 9 linolenik acid ( c18:3) 0.69± 0.02d 0.79± 0.05b 0.79± 0.03b 0.78± 0.07c 0.62± 0.05e 0.81± 0.03a 10 araşidik acid (c20:0) 0.31± 0.12a 0.29± 0.09b 0.28± 0.07c 0.26± 0.08e 0.26± 0.05e 0.27± 0.03d 11 gadoleik/eikosenoik acid (c 20:1) 0.22± 0.04c 0.25± 0.07b 0.17± 0.12e 0.22± 0.08c 0.19± 0.06d 0.26± 0.05a 12 behenik acid (c 22:0) nd nd nd 0.04± 0.06a nd nd 13 lignoserik acid ( c24:0) nd nd nd nd nd nd 14 trans oleik acid (c18:1t) nd nd nd nd nd nd 15 trans linoleik acid +trans linolenik acid (c18:2 t+c18:3 t) nd nd nd nd nd nd agriculture and food sciences research, 2021, 8(2): 20-29 25 © 2021 by the authors; licensee asian online journal publishing group çekişte evoos has low stigmastereol and campesterol content. in all evoos, the campesterol content was found below the limit between 2.85 % (üzümlü) and 2.70 % (zeytinlik) according to eu regulations (4%). there is significant difference in campesterol content according to the geographical locations. in addition to apparent β sitosterol, it was detected by the sum of β-sitosterol and other three sterols (δ5-24-stigmastadienol, clerosterol and δ-5-avenasterol). çekişte evoo are found approximately the limit of 94 %. the highest apparent β-sitosterol (95.48 %) was detected in zeytinlik location table 5. in evoos, the content of stigmasterol is lower than campesterol as previous research [9]. in this study, different phytosterol content revealed in çekişte evoos were found to be similar as chemlali varieties. many studies showed that various factors affect the sterol content such as olive variety, ecological zone, soil structure and harvest time [10]. this result of study is parallel with previous studies. 3.4. determination of marker ions in evoos by using lc ims qtof screening the determination of marker ions in olive oils carried out using lc ims qtof mass spectrometry system figure 4. bademli beyazıt birgi üzümlü agriculture and food sciences research, 2021, 8(2): 20-29 26 © 2021 by the authors; licensee asian online journal publishing group figure-4. peak intensity chromatograms of evoos (a-acarlar, b-gökçealan, c-havutculu, d-şirince, e-sultaniye, fzeytindağ, g-pool) from ultra-performance liquid chromatography–quadrupole time-of-flight ms in esi+ ionization mode. lc ims qtof system was used to determine the geographical indications of evoos. primarily, all samples extracted were injected into the lc system under the conditions specified in the method. the total ion chromatograms obtained in the positive ionization mode of ödemiş region in figure 4 are given. as can be seen in the total ion chromatograms, there are differences between the olive oil methanols: water extracts studied in the study. 3.5. statistical analysis using progenesis qi software progenesis qi software is widely used in metabolomics researches in recent years. in this study, progenesis qi was used for multivariate statistical analysis. the spectral regions before 1.0 min and after 13 min of analysis were excluded from data evaluation. yeniceköy zeytinlik ödemiş pool bademli agriculture and food sciences research, 2021, 8(2): 20-29 27 © 2021 by the authors; licensee asian online journal publishing group beyazıt birgi üzümlü yeniceköy zeytinlik agriculture and food sciences research, 2021, 8(2): 20-29 28 © 2021 by the authors; licensee asian online journal publishing group figure-5. esi (+) ms spectra of olive oils from ödemiş region. esi (+) ms spectra of selçuk region are given in figure 5. the ms spectrum of each olive oil sample is different from each other. while some masses gave higher intensity in some evoos, some showed lower intensity. in addition, some masses are found in some evoos but not in others. progenesis qi software was used to reveal these differences statistically. as can be seen in figure 5, there are some differences among the olive oil extracts. a distinct clustering among the evoos was detected, which suggest that the metabolites significantly changed between different region. figure-6. (a) the plot is based on the first 2 principal component for data obtained in esi+ mode (b) the plot is based on the first and third principal component for data obtained in esi+ mode. note: a: pca1 x pca2 b:pca1 x pca3. 3.6. principal component analysis (pca) pool samples were located in the centre of the pca pilot shows that the analytical system is reliable. the samples in the same group are clustered together. pca1, pca2, pca3 were used to show the clustering of the evoos. pca1, pca2 and pca3 were 47.97 %, 22.40 % and 13.05 %, respectively. the total variance was determined as 83.42 in the 95 % confidence level. as can be seen in the pca plot, it seems feasible to seperate the oil samples using this method (figure 6). the obtained data were filtered with anova p (p value≤0.05), not fragmented and max-fold change. the markers in each sample were identified using ez info software. in this study, we used a method for untargeted metabolomics in evoos. the detected masses were then subjected to library scanning. chemspider library, lipidblast library, and elemental composition (h, c, n, o, p and h, c, n, o, separately) were used for scanning. the compounds were summarized along peak number, retention time, observed m/z, empirical formula, adduct, mass error, and msigma, isotope similarity ratio (%) score and proposed compounds in table 1. during the data processing and compounds identification, same compounds have different rt but have the same m/z ratio. ödemiş pool a b agriculture and food sciences research, 2021, 8(2): 20-29 29 © 2021 by the authors; licensee asian online journal publishing group as a result, each olive oil samples belonging to the ödemiş region is clustered in different regions. it has been determined that especially evoos belonging to zeytinlik are clustered in quite different regions from the others, and a successful distinction can be made with this research. it has been determined that different climatic and topographical conditions cause differences in the geographical origin of evoos. in this respect, it is recommended that evoos to be labeled as geographical indication should be labeled on the basis of small local region. abbreviatıons and nomenclature gi :geographical indication; pca :principal component analysis; evoo : extra virgin olive oil; rt : retention time; lc: liquid cromatography; ims: ion mobility spectrometry; qtof: quadropole time-of-flight references [1] d. arslan, y. karabekir, and m. schreiner, "variations of phenolic compounds, fatty acids and some qualitative characteristics of sarıulak olive oil as induced by growing area," food research international, vol. 54, pp. 1897-1906, 2013.available at: https://doi.org/10.1016/j.foodres.2013.06.016. 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[10] j. pardo, m. cuesta, r. gómez, j. granell, and m. alvarez-ortí, "quality of virgin olive oil from the designation of origin (do)“aceite campo de montiel”(ciudad real, spain) from two crop seasons," italian magazine of fatty substances, vol. 89, pp. 103111, 2012. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 28 © 2023 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 10, no. 2, 28-31, 2023 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v10i2.5209 © 2023 by the author; licensee asian online journal publishing group participatory demonstration and evaluation of improved onion variety in east shewa zone of oromia regional state, ethiopia fistum miruts1 belay roba2 derara sori3 ( corresponding author) 1,2,3ethiopian institute of agricultural research, melkassa agricultural research center, ethiopia. 1email: fitsummiruts@gmail.com 2email: robabelay1984@gmail.com 3email: soriderara2016@gmail.com abstract the study was carried out in selected districts of east shewa zone oromia regional state to compare the performance of a recently released onion variety (nafis) with their recommended agronomic recommendations to a control (bombay red) through an on-farm demonstration. the demonstration was implemented on 9.3 hectares of 37 farmers’ farmland. the nafis variety outperformed the control, with a mean yield of 293.2 qt/ha. the mean technology gap was 106.8 qt/ha and the technology index recorded was observed to be 26.7%. farmers' preferences were used to evaluate onion varieties with criteria of yield, disease, bulb size, bulb color (deep red preferred), pungency, storability, and market preference. nafis variety is preferred over the bombay red variety. in addition, lack of improved seeds, high chemical costs, and insect and disease infestations are among the major constraints limiting onion production and productivity in the study area. trips and fungi (purple bloch) have also been identified by farmers as a major disease affecting onion production in the districts. in general, the demonstrated variety (nafis) was found to be feasible and preferred by the farmers to cultivate in the east shewa zone as compared to the check (bombay red). therefore, onion yield in farmers' fields can be improved largely by cultivating the nafis variety in the study area and similar environments. keywords: demonstration, extension gap, east-shewa, onion, technology gap, technology index. citation | miruts, f., roba, b., & sori, d. (2023). participatory demonstration and evaluation of improved onion variety in east shewa zone of oromia regional state, ethiopia. agriculture and food sciences research, 10(2), 28–31. 10.20448/aesr.v10i2.5209 history: received: 22 august 2023 revised: 13 november 2023 accepted: 27 november 2023 published: 6 december 2023 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by ethiopian institute of agricultural research (grant number: 62-05). institutional review board statement: the ethical committee of the ethiopian institute of agricultural research, ethiopia has granted approval for this study. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 29 2. materials and methods ................................................................................................................................................................... 29 3. result and discussion ..................................................................................................................................................................... 30 4. conclusion and recommendations ............................................................................................................................................... 31 references .............................................................................................................................................................................................. 31 mailto:fitsummiruts@gmail.com mailto:robabelay1984@gmail.com mailto:soriderara2016@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v10i2.5209 https://orcid.org/0000-0003-1390-4669 https://orcid.org/0009-0004-2835-8212 https://orcid.org/0009-0003-3233-0995 agriculture and food sciences research, 2023, 10(2): 28-31 29 © 2023 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this paper contributes to the existing literature in several ways. first, unlike most other studies, it focuses on the traits growers consider for sustainable onion production rather than evaluating the technology in yield performance only. second, the study is unique in that the technology is new and contributes significantly to the farming community's production and productivity. 1. introduction in ethiopia, onion is an important cash crop that is widely used to enhance the flavor of various foods. it is grown under both irrigation and rainfed conditions, by smallholder farmers and commercial growers, with, irrigation covering the majority of the land [1]. the study area has a high potential for onion production because of the abundance of irrigated farmland and the presence of relatively better market access [2]. according to a report from the central statistics agency [3], the area covered by onions in 2019/20 was approximately 28,185.11 hectares, with a total production of 2,624,782.85 quintals. there was a 29% increase in area coverage and a 20% increase in production when comparing the 2019/20 onion cropping season to the 2012/2013., crop productivity, however, is decreasing year after year. there was a 25 qt/ha yield difference between the 2012/13 (100.24 qt/ha) and the 2019/20 (75.29 qt/ha) cropping season table 1. these low yield results indicate the presence of a large productivity gap, which is attributed to several constraints, the most significant of which are the lack of an authorized body that produces and distributes improved onion seed varieties and the use of inappropriate agronomic practices [2, 4]. with this in mind, the purpose of the study was to compare the performance of a recently released onion variety (nafis) with the check (bombay red) through on-farm demonstration and to analyze farmers' preferences for the demonstrated varieties. table 1. area coverage, production, and productivity of onion in ethiopia. production year area coverage (ha) production (qt) productivity (qt/ha) productivity change (qt/ha) 2012/13 21,865.37 2,191,886.02 100.24 2013/14 24,375.70 2,197,352.67 90.15 -10.09 2014/15 22,771.88 2,307,451.89 101.35 11.2 2015/16 29,517.01 2,648,493.54 89.73 -11.62 2016/17 33,603.39 3,274,752.45 97.45 7.72 2017/18 31,673.21 2,938,875.85 92.79 -4.66 2019/20 28,185.11 2,624,782.85 75.29 -17.5 source: csa (2012/13 – 2019/20). 2. materials and methods 2.1. description of the study area the research was carried out in three districts of the east shewa zone (adama, lume, and adamitulu jido kombolcha) of the oromia regional state. the zone has ten districts and is divided into 17.2% slight, 41.3% moderate, and 41.5% severe agricultural drought risk subzones. the zone economy is characterized by a mixed farming system that relies on a large rain-fed farm [5]. east shewa zone has three agroecologies namely highland, midland, and lowland. this zone received an annual temperature range of 12-39°c. the zone has a total land area of 971,159.21 hectares [6]. figure 1 shows the distribution of the study areas. figure 1. map of the study area. agriculture and food sciences research, 2023, 10(2): 28-31 30 © 2023 by the authors; licensee asian online journal publishing group 2.2. farmer's selection and field establishment as adama, lume, and adamitulu jido kombolcha (atjk) districts are potential areas in onion production, farmers who have access to irrigation, interest in the technology, a willingness to allocate and manage fields of the demonstration, and a willingness to collaborate with researchers and extension agents were chosen to host the demonstration. the newly released variety (nafis) was demonstrated side by side with a locally popular variety (bombay red). a plot size of 0.25 hectares was used for each demonstration. in onion production, all recommended packages reported by moa [7] applied for instance, using 40 cm water furrows with 20 cm rows on beds and 5 cm between plants in the demonstration plots are employed in farmers’ fields. all varieties were planted on the same day. follow-ups and critical advice from respective researchers and agricultural experts have been provided to demonstration host farmers. the melkassa research center provided complete onion technology packages for the demonstration. 2.3. training of farmers since training is one of the extension methods playing a significant role in increasing the awareness of the farming community on a particular technology, host farmers (37) and development agents (10) in the study were given training on onion production and management practices. the training consists of both theoretical and practical training. researchers discovered gaps during their regular visits. to fill the observed gap, field-level training was provided to the farmers by grouping them into small groups and discussing issues raised by farmers. 2.4. data collection and analysis both qualitative and quantitative data were collected and analyzed through supervision and follow-up of the activity with the participation of all stakeholders. a data record sheet was created to collect all necessary data. thus, field observation and contact with the target farmer during the field visit were used to collect data. farmers' yield data and preferences for the variety were collected. finally, the collected data was analyzed using simple statistical tools. the following equation was used to calculate the gaps between the potential yield and the demonstration yield, the extension gap, and the technology index [8]. • technology gap = potential yield – demonstration yield (1) • extension gap = demonstration yield – yield under existing practice (2) • technology index = (potential yield demonstration yield / potential yield) x 100 (3) 3. result and discussion 3.1. yield performance the mean yield of the nafis variety obtained under the demonstration was 271.19, 322.09, and 286.16 q/ha in the adama, lume, and atjk districts respectively. the highest yield (346 qt/ha) was in the adama district during 2018/19 whereas the smallest yield (196.38 qt/ha) was in the same district during 2019/20 for the nafis variety. the difference in yield amount over location and demonstration year was observed, which might be due to management differences in climatic deviations, as well as the change in the location of the demonstration in each year. the result of the demonstration revealed that the recently released varieties (nafis) performed extremely well when compared with the check (bombay red) during all the years of demonstration table 2. accordingly, a mean yield advantage of 33.9, 127.3, and 48.9% of nafis variety was recorded over the check-in adama, lume, and atjk districts respectively. the pooled mean shows also a 41.4% yield advantage of the nafis variety over the check-in of all demonstrated districts. the yield improvement in onion and other crops under demonstration has adequately been documented by abady, et al. [9] and kinfe and tesfaye [10]. table 2. yield performances of nafis variety. location year no. of demo. area nafis yield (qt/ha) check (qt/ha) yield increment (%) adama 2018/19 8 2 346.00 232.37 48.90 2019/20 5 1.25 196.38 164.97 19.04 mean 13 3.25 271.19 198.67 33.97 lume 2018/19 8 2 311.27 101.34 207.14 2019/20 5 1.25 332.90 225.83 47.41 mean 13 3.25 322.09 163.59 127.28 atjk 2018/19 6 1.5 241.23 233.03 3.52 2019/20 5 1.25 331.08 286.86 15.41 mean 11 2.75 286.16 259.95 48.90 pooled mean 37 9.25 293.15 207.40 41.35 3.2. technology gap, extension gap, and technology index the newly released onion variety (nafis) outperformed the control (bombay red). table 3 shows that the percentage increase in yield over check is 36.5 in the adama district, 97 in the lume district, and 10 in the atjk district. the technology gap in adama, lume, and atjk districts was 128.8, 77.9, and 113.8 qt/ha, respectively. this may be attributed to differences in soil fertility status and weather conditions. as a result, it appears that location-specific recommendations are required to close the yield gap between different varieties. regarding the extension gap, the lume and adama districts had the greatest extension gap (158.5 qt/ha). the extension gap highlighted the importance of educating onion-producing farmers through various means for the adoption of improved high-yielding varieties with the recommended packages. by using the nafis variety, farmers can change the trend of a wide extension gap. the technology index in table 3 indicates the viability of the variety (nafis) in the farmers' field. the lower the value of the technology index, the higher the technology's feasibility [11, 12]. across districts, the technology index ranges from 19.48 to 32.2 percent. the nafis variety is found to be more feasible for farmers to produce in the study area than the bombay red variety (check). agriculture and food sciences research, 2023, 10(2): 28-31 31 © 2023 by the authors; licensee asian online journal publishing group table 3. productivity of onion (nafis variety), yield gaps, and technology index. location potential yield (qt/ha) demonstration (qt/ha) bombay red (qt/ha) % increase over check technology gap extension gap technology index (%) adama 400 271.19 198.67 36.50 128.81 72.52 32.20 lume 400 322.09 163.59 96.89 77.91 158.50 19.48 atjk 400 286.16 259.95 10.08 113.84 26.21 28.46 mean 400 293.15 207.40 47.82 106.85 85.74 26.71 3.3. farmers feedback farmers' preferences were used to evaluate onion varieties with criteria of yield, disease and insect resistance, bulb size, bulb color (deep red preferred), pungency, storability, and market preference. according to the ranking of onion varieties based on farmers' criteria, farmers preferred the nafis variety over the bombay red variety. table 4 presents below the onion producer farmers preference of onion variety based on their own criteria. as a result nafis variety was preferred by all listed criteria for production. the focus group discussion participants mentioned that a lack of improved seeds, high chemical costs, and insect and disease infestations are among the major constraints limiting onion production and productivity in the study areas as shown in table 5. trips and fungi (purple bloch) have also been reported by farmers as a major disease affecting onion production in the districts. table 4. preference ranking of onion varieties based on farmers' preference criteria. varieties market preference better yield good bulb color average bulb size resistance to insect pungency resistance to disease storability nafis 1st 1st 1st 1st 1st 1st 1st 1st bombay red 2nd 2nd 2nd 2nd 2nd 2nd 2nd 2nd table 5. major challenges of onion production. major challenges disease insect cost of pesticides unstable price uis* total scores rank disease disease disease disease uis 3 2nd insect insect insect uis 2 3rd cost of pesticides cost of pesticides uis 1 4th unstable price uis 0 5th uis* 4 1st note: *uis (unavailability of improved seed). 4. conclusion and recommendations the demonstration yielded an encouraging result and allowed stakeholders to demonstrate the new variety's productivity potential by comparing it to local variety on farmers' fields. this could alleviate some of the barriers to technology transfer and adoption. according to the findings of the study, conducting demonstrations of the improved variety (nafis) increased onion yield potential significantly. the crop's yield potential can be increased to a greater extent by cultivating the nafis variety of onions in the east shewa zone. as a result, the farming community's income and livelihood will improve. the study also emphasized the importance of educating farmers to improve their use of improved varieties. the demonstration also raised awareness and encouraged farmers to plant the newly demonstrated onion variety (nafis). as a result, a strategy to address the wide farming community should be developed to increase onion production and productivity. references [1] z. asfaw and d. eshetu, "production and management of major vegetable crops in ethiopia addis ababa, ethiopia ethiopian institute of agricultural research, addis ababa, ethiopia; kopia ethiopia centre," vol. 1. addis ababa, ethiopia: printed at ethcana printing press, 2015, p. 149. [2] a. teshome, b. feyissa, and d. abnet, "pepper and onion yield estimate parameters as affected by seed treatment options: in the selected districts of central rift valley; oromia regional state of ethiopia," open access library journal, vol. 2, p. e1243, 2015. http://dx.doi.org/10.4236/oalib.1101243 [3] csa, "agricultural sample survey report on area and production of major crops, meher season," vol. 1. addis ababa, ethiopia: statistical bulletin, 2020, p. 587. [4] y. mahilet, "adoption of onion production package by smallholder farmers in east shoa zone of oromia regional state, ethiopia," a research project submitted to van hall larenstein university of applied sciences in partial fulfillment of the requirements for the degree of master of development, specialization: rural development and food security, 2012. [5] g. legesse and k. v. suryabhagavan, "remote sensing and gis based agricultural drought assessment in east shewa zone, ethiopia," tropical ecology, vol. 55, no. 3, pp. 349–63, 2014. [6] n. asfaw, g. shimalis, e. yassin, and b. beriso, "crop production system and their constraints in east shewa zone, oromia national regional state, ethiopia," vol. 5, no. 2, pp. 30-39, 2020. https://doi.org/10.11648/j.ijees.20200502.11 [7] moa, horticultural crop packages, manual for agricultural extension experts. addis ababa, ethiopia: amharic version, 2018. [8] s. samui, s. maitra, d. roy, a. mandal, and d. saha, "evaluation of front line demonstration on groundnut," journal of the indian society costal agricultural research, vol. 18, no. 2, pp. 180-183, 2000. [9] s. abady, g. liku, and d. yadeta, "participatory varietal selection and evaluation of twelve sorghum (sorghum bicolor (l) moench) varieties for lowlands of eastern hararghe," international journal of plant breeding and crop science, vol. 4, no. 3, pp. 281285, 2017. [10] h. kinfe and a. tesfaye, "yield performance and adoption of released sorghum varieties in ethiopia," edelweiss applied science and technology, vol. 2, no. 1, pp. 46-55, 2018. https://doi.org/10.33805/2576.8484.115 [11] n. chauhan, "impact and yield fissure inspection of gram through trainings and flds by kvk tapi in gujarat," indian journal of agricultural research and extension, vol. 4, pp. 12-15, 2011. [12] k. jeengar, p. panwar, and o. pareek, "front line demonstration on maize in bhilwara district of rajasthan," current agriculture, vol. 30, no. 1/2, pp. 115-116, 2006. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://dx.doi.org/10.4236/oalib.1101243 https://doi.org/10.11648/j.ijees.20200502.11 https://doi.org/10.33805/2576.8484.115 198 © 2019 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 6, no. 2, 198-202, 2019 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2019.62.198.202 © 2019 by the authors; licensee asian online journal publishing group studies on the growth and yield of prostrate groundnut variety (arachis hypogaea l.) as affected by oil palm bunch ash in umudike southeastern nigeria akpan a. u.1 eka, m. j.2 ( corresponding author) 1,2department of agronomy, michael okpara university of agriculture, umudike, southeastern, nigeria. abstract an experiment on studies on the growth and yield of prostrate groundnut variety (arachis hypogaea l.), as affected by oil palm bunch ash in umudike, southeastern, nigeria, was conducted at the eastern farm of michael okpara university of agriculture, umudike, abia state. the research was laid out in a randomized complete block design (rcbd), with three replications. the treatments comprised the oil palm bunch ash levels of 0, 4, 6 and 8 tha-1. result, showed that number of leaves per plant increased at 4 and 8 weeks after planting (wap), and declined at 12 wap due to senescence. number of branches per plant increased at 4, 8 and 12 wap, while shoot dry weight increased at 4 and 8 wap, and declined at 12 due to senescence. dry matter content was higher at 4 wap, but dropped at 8 and 12 wap, due to the translocation of photoassimilates (biomass) as the crop developed in age. 8 tha-1 of oil palm bunch ash produced highest growth parameters compared with the control (0 tha -1). improved number of seeds per pod (18.27 pods); number of seeds per plant (27.7 seeds); weight of pods per plant (20.89g); weight of seeds per plant (15.27g), 100-seeds weight (57.8g); harvest index (0.17); shelling percentage (74.2%); pod yield (5.22 tha-1) and seed yield (3.82 tha-1) were obtained at 8 tha-1, although performances were statistically similar. consequently, 8 tha-1 of oil palm bunch ash is recommended for the cultivation of groundnut in umudike, southeastern, nigeria. keywords: growth, yield, groundnut, oil palm bunch ash. citation | akpan a. u.; eka, m. j. (2019). studies on the growth and yield of prostrate groundnut variety (arachis hypogaea l.) as affected by oil palm bunch ash in umudike southeastern nigeria. agriculture and food sciences research, 6(2): 198-202. history: received: 8 april 2019 revised: 17 may 2019 accepted: 28 june 2019 published: 22 august 2019 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: the authors appreciate all the workers of the university farms for their unalloyed supports, throughout the duration that the research lasted. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 199 2. materials and methods ................................................................................................................................................................. 199 3. results and discussions ................................................................................................................................................................ 200 4. conclusion ....................................................................................................................................................................................... 201 references ............................................................................................................................................................................................ 201 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.198.202&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2019.62.198.202&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://www.asianonlinejournals.com/index.php/aesr/article/view/936 http://orcid.org/0000-0003-3334-8399 agriculture and food sciences research, 2019, 6(2): 198-202 199 © 2019 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this research has further revealed that oil palm bunch ash is veritable organic manure, with dual purposes of ameliorating a declining soil fertility and acidity. equally, groundnut cultivation in southeastern nigeria can be carried out with the application of organic manures like the oil palm bunch ash. 1. introduction groundnut (arachis hypogaea l.), is a major crop grown in the arid and semi-arid zones of nigeria. it is either grown for its nut, oil or its vegetative residue (haulms) [1]. groundnut, also known as peanut, is an important food and cash crop grown across west africa. the crop is cultivated mainly by small-household and resources poor farmers. it is a self-pollinated, tropical annual legume [2]. it is an important crop in nigeria as it occupied 13-27% of the total area in some state of the country [3]. groundnut is a legume that ranks 4th among the oil seed crops and 13th among the food crops of the world. it provides high quality edible oil (48-50%), used in cooking, margarine, salads, easily digestible protein (26 to 28%), and about half of the 13 essential vitamins and more than a 2/3rd of the 20 essential minerals necessary for normal human growth and maintenance. it produces high quality fodder for livestock [4]. being a legume, it synthesizes atmospheric nitrogen into root nodule with a symbiotic association of rhizobium and thereby improves soil for fertility [5]. the main factors responsible for low yield in groundnut are the inadequate use of nutrients as well as nutrient deficiencies. organic manure such as oil palm bunch ash is one of the major sources of essential plant nutrients gaining importance in groundnut production. oil palm bunch ash is an organic residue derived from the incineration of oil palm bunch refuse after fruit extraction. according to akpan [6] the scarcity and high cost of inorganic fertilizer led to intensification of research into low-cost, internally sourced, cheap, affordable and adoptable organic materials that could serve as liming agents and fertilizers. akpan [7]; akpan and mbah [8] and akata [9] studied effect of liming materials such as plant derived ash and saw dust on cucumber and cassava yields, the materials were found to increase soil ph and crop yield. ntare, et al. [10]; obi [11] and moyin-jesu [12] found out that plant derived ash including those of wood and cocoa pod, increased soil chemical properties, ph and yield of vegetables, cassava and maize. mbah and akpan [13] reported that ash is an effective fertilizer and liming material for improving soil fertility, while ikeh, et al. [4] stressed that combined application of oil palm bunch ash and poultry manure at 2.5 tha-1 rate each resulted to high tuber yield of water yam. effiong, et al. [14] found that oil palm bunch ash and poultry manure treatments increased the soil ph, o.m, n, p, k, ca, mg by 6%, 13%, 19%, 28%, 32%, 33% and 21% respectively. akpan [6] reported that oil palm bunch ash increased soil nutrient contents and uptake of nutrients by maize and these effects led to significant increase in maize yield. essien [15] attributed the superiority of organic manure to the slow release of balanced nutrient during decomposition. this makes organic waste of greater residual beneficial effects distributed over longer period than inorganic fertilizers asiegbu [16]. gomez and gomez [17] reported high growth and yield of groundnuts at 4 tha1 of oil palm bunch ash manure. the objective of this research therefore, is to evaluate the growth and yield of prostrate groundnut (arachis hypogaea l.) as affected by oil palm bunch ash in umudike, southeastern, nigeria. 2. materials and methods the experiment was conducted at the teaching and research farm of michael okpara university of agriculture, umudike, during 2018 cropping season. umudike is located on latitude 050 29’n and longitude 07033’e, on the elevation of 122.0m above sea level. it has a maximum and minimum temperature of 33oc and 24oc, annual rainfall of 21, 690.8mm and relative humidity range of 50%-95% respectively. rainfall pattern is however bimodal, with rainfall commencing from march to october, with a short dry spell in august, nrcri [18]. the experiment was laid out in a randomized complete block design. (rcbd) with three replications. the treatments comprised the oil palm bunch ash levels of 0, 4, 6 and 8 tha-1. the prostrate groundnut variety (g153), used in this study was obtained from the institute for agricultural research (iar), ahmadu bello university, zaria, kaduna state. the experimental layout was 24. 5 x 7.4 m (181.3 m2), with each plot measuring 1.8 m x 2.0 m; 0.5 m spacing between plot and 1 m spacing between replicates. planting spacing was 50 cm x 30 cm, resulting in a plant population of 66,667 plants ha-1. before planting, the site was mechanically ploughed and subsequently harrowed. planting was done in may 5th on bed. two to three seeds were planted per hole, while thinning to one plant per stand was conducted at 14 days after planting (dap). oil palm bunch ash manure at the rates of 0,2,4, 6 and 8 tha-1, was applied by ring method after thinning at 14 dap. empty oil palm bunches were obtained from a local oil mill. they were dried properly before incineration to obtain the ash. the research plots were weeded once at 3 wap, with hoe, while insect pests were controlled by the use of cypermethrin-10 ec. at 2.5 ml litre-1 of water using a knapsack sprayer once at 4 wap [19]. harvesting was conducted at the end of august, precisely 4 months after planting. data collection on vegetative parameters such as: number of leaves per plant, number of branches per plant, shoot dry weight per plant and dry matter content were conducted at 4, 8 and 12 wap, while reproductive parameters including: number of pods per plant, number of seeds per plant, weight of pods per plant, weight seeds per plant, 100 – seed weight, harvest index, shelling percentage, pod yield and seed yield (tha-1), were obtained after harvest at maturity. before the commencement of the research, composite soil samples were randomly collected at the depth 0-20cm for both physical and chemical analyses. oil palm bunch ash manure was equally analyzed, while meteorological data bordering on rainfall (mm), relative humidity (%) and temperature (0c) were obtained from the meteorological unit of national root crops research institute (nrcri), umudike. both, vegetative and reproductive parameters were subjected to analysis of variance (anova) method [20] while fisher’s least significant difference (f-lsd) at 5% probability level was adopted to separate means. agriculture and food sciences research, 2019, 6(2): 198-202 200 © 2019 by the authors; licensee asian online journal publishing group 3. results and discussions 3.1. meteorological properties of the research site the meteorological properties of the research site table 1 revealed that monthly rainfall increased from the month of april, (337.5mm) and dropped in the month of november (45mm) preparatory to the commencement of dry season. both minimum and maximum temperatures were almost stable throughout the year, although with insignificant fluctuations. relative humidity on the other hand increased from march (80%), with obvious drop in november due to the beginning of dry season table 1. the fluctuation in relative humidity was tandem with that of monthly rainfall. the rainfall situation was in line with the recommendation of ibia and udo [21]; chude, et al. [22] for groundnut cultivation in southeastern nigeria. table-1. meteorological properties of the research site during 2018 cropping season. months jan. feb. mar. april may june july aug. sept. oct. nov. dec. rainfall (mm) 0 80.1 9.6 337.5 246.6 326.6 237.0 173.3 334.7 273.4 45.0 7.3 tempts. 0c min. 22.6 23.8 24.6 24.7 24.9 23.1 24.1 24.4 23.3 23.7 23.9 24.1 max 34.1 34.5 34.4 32.5 32.6 30.9 29.6 30.4 30.1 30.8 32.3 32.1 r/humility (%) 53 77 80 76 81 80 86 87 85 83 80 71 source: meteorological unit, nrcri, umudike. 3.2. physico-chemical properties of the research site the physico-chemical properties of the research site table 2 showed that the soil was sandy loam in texture, while the sand fraction of (67.2%) was capable of sustaining the growth of groundnut. the ph of 4.2 , showed that the site was strongly acidic, and not in conformity with the ph of 5.5 to 6.5 recommendation by ibia and udo [21] and chude, et al. [22]. organic matter (0.87%); organic carbon (0.5%) and potassium (0.079 cmokg-1) were very low, while nitrogen (0.182%) and phosphorus (67.8 cmokg-1) were moderate and high respectively. the site therefore, requires liming or addition of plant ash for the remediation of its acidic status. the properties of oil palm bunch ash showed that the acidity level was low due to the high ph of 10.8. it is very rich in almost all the analyzed parameters table 2. the rich status of oil palm bunch ash conforms with the reports of varying researchers such as: ntare, et al. [10]; effiong, et al. [14]; akpan [6] and ikeh, et al. [4]. plant ash including oil palm bunch ash are veritable liming and fertilizer materials. table-2. physico-chemical properties of the experimental site. soil physical properties oil palm bunch ash properties sand (%) 67.2 silt (%) 12.8 clay (%) 20 textural class sandy loam soil chemical properties ph (h2o) 4.2 10.8 organic matter (%) 0.87 8.02 organic carbon (%) 0.5 4.65 total nitrogen (%) 0.182 0.28 available p (mgkg-1) 67.8 8.20 calcium (cmolkg-1) 2.8 3.40 potassium (cmolkg-1) 0.079 11.50 magnesium (comolkg-1) 1.6 0.48 sodium 0.185 0.87 source: field data for soil and oil palm bunch ash samples. 3.3. growth of groundnut as affected by oil palm bunch ash the evaluated growth attributes showed that number of leaves per plant table 3 for all treatment increased at 4 and 8 wap, with a decline at 12 wap due to senescence. higher number of leaves per plant was recorded at 8 wap for the treatments, while 8 tha-1 of oil palm bunch ash produced highest number of leaves per plant of 73.7 (4wap); 156.2 (8 wap) and 146.0 (12wap), compared with the control. number of branches per plant increased at 4, 8 and 12 wap, with 8 tha-1 of oil palm bunch ash producing highest number of braches per plant of 6. 85, (4wap); 7.58 (8 wap) and 8.83 (12 wap) compared with 0 tha-1 (control), although performances were statistically similar. shoot dry weight table 3 also increased at 4, 8, and dropped at 12 wap due to the process of senescence, while 8 wap produced higher short dry weight for all treatments. 8 t ha-1 of oil palm bunch ash gave higher shoot dry weight of 21. 3% (4wap): 69.2% (8wap) and 45.8% (12wap), compared with the control. dry matter contents table 3 for all treatments was high at 4 wap, with a decline at both 8 and 12wap due to probably to the translocation of phyto-assimilate to sinks as the crop increased in age. 8 tha-1 of oil palm bunch ash produced highest dry matter content of 23.3% (4 wap); 19.6 (8wap) and 21.5 (wap), compared with the control which produced the least values, although performances were statistically similar table 3. both number of leaves per plant and number of branches per plant were significantly (p<0.05) affected at 12wap and 4wap respectively, while shoot dry weight and dry matter contents were not significantly (p> 0.05) affected at 4, 8 and 12wap. the increase in number of leaves per plant at 4 and 8 wap and a decline at 12 wap, was earlier reported by akpan [6]; akpan [7] and akpan and mbah [8]. uko, et al. [2]; awodun, et al. [20] and akata [9] all reported higher growth attributes at highest rates of oil palm bunch ash fertilizer, during their respective researches. table-3. effect of oil palm bunch ash on number of leaves per plant, number of branches per plant, shoot dry weight per plant and dry matter content of prostrate groundnut at 4, 8 and 12 wap during 2018 cropping season. agriculture and food sciences research, 2019, 6(2): 198-202 201 © 2019 by the authors; licensee asian online journal publishing group treatments no. of leaves per plant no. of branches per plant shoot dry weight per plant (%) dry matter content (%) 4wap 8wap 12wap 4wap 8wap 12wap 4wap 8wap 12wap 4wap 8wap 12wap otha-1 52.0 123.3 65.5 5.50 6.67 7.63 14.4 40.7 37.7 19.7 19.0 22.3 4tha-1 58.5 124.1 96.3 5.50 7.27 8.00 15.4 50.9 44.0 25.3 17.2 19.3 6tha-1 58.8 134.1 126.7 5.53 7.42 8.13 17.8 54.9 45.6 19.6 18.9 21.5 8tha-1 73.7 156.2 146.0 6.85 7.58 8.83 21.3 69.2 45.8 28.7 23.3 22.8 means 60.7 132.6 110.0 5.84 7.23 8.15 17.2 53.2 44.0 23.3 19.6 21.5 l sd (0.05) ns ns 57.9 1.05 ns ns ns ns ns ns ns ns source: field data for vegetative parameters. 3.4. yield of prostrate groundnut as affected by oil palm bunch ash fertilizer all the evaluated yield parameters table 4 including number of pods per plant, number of seeds per plant, weight of pods per plant, weight of seeps per plant, 100-seed weight, harvest index, shelling percentage (%), pod yield (tha-1) and seed yield (tha-1), all improved with increased levels of oil palm bunch ash, although performances were statistically similar. 8 tha-1 of oil palm bunch ash produced highest number of pods per plant (18.27 pods); number of seeds per plant (27.7 seeds); heavier pods (20.89g), heavier seeds (15.27g); 100-seed weight (57.8g); harvest index (0.17); shelling percentage (74.2%); pod yield (5.22 tha-1) and seed (3. 82 tha-1) compared with otha-1 (control) with lesser values. the evaluated yield parameters were however not significantly (p>0.05) affected by the treatments, suggesting that their performances were generally statistically similar. this suggests that the acidic status of the sites must have locked-up the release of macro-nutrients to the plant. various researchers, including uko, et al. [2]; awodun, et al. [20] and akata [9] all reported improvement in yield parameters with increased rates of oil palm bunch ash fertilizer. the pod yield of 5.22 tha-1 obtained at 8 tha-1 was higher than the pod yield range of 3,000 4,500 kgha-1 reported by olorunju [23] while the seed yield of 3.82 tha-1 obtained at 8 tha-1 was higher than 2. 58 tha-1 reported by gomez and gomez [17]. the seed yield of 3. 82 tha1 at 8 tha-1 of oil palm bunch ash was however lower than the 4.0 tha-1 reported by uguru [24]. table-4. effect of oil palm bunch ash on some selected yield attributes of prostrate groundnut during 2018 cropping season. treatments no. of pods per plant no. of seeds per plant wt. of pods per plant (g) wt. of seeds per plant (g) 100 seeds weight (g) harvest index shelling percentage (%) pod yield tha-1 seed yield (tha-1) otha-1 12.00 22.3 16.54 13.69 50.9 0.09 72.2 4.13 3.42 4tha-1 12.83 23.8 18.71 13.93 51.7 0.13 73.0 4.68 3.48 6tha-1 14.75 24.2 19.58 14.35 53.1 0.15 74.2 4.90 3.59 8tha-1 18.27 27.7 20.89 15.27 57.8 0.17 74.2 5.22 3.82 means 14.46 24.5 18.93 14.31 53.4 0.13 73.4 4.73 3.58 l sd(0.05) ns ns ns ns ns ns ns ns ns source: field data for reproductive parameters. 4. conclusion the research has revealed that the values of both the growth and yield parameters increased with corresponding increases in the rates of oil palm bunch ash, although their performances were generally statistically similar. 8tha-1 of oil palm bunch ash equally produced highest growth and yield attributes, compared with 0tha-1 (control). in view of the above therefore, 8 tha-1 of oil palm bunch ash is hereby recommended for the cultivation of groundnut in umudike, southeastern, nigeria. the application of lime or plant ash for the remediation of the acidic status of the site is strongly recommended. this will ensure the release of nutrients to the crops. references [1] a. a. makinde, k. o. oluwasemire, o. a. akintola, and h. a. ademiyi, "seasonal variability and row arrangement effect on the performance of groundnut (arachis hypogaea l.) and okra (abelmoschus esculentus) intercrop in a forest-savanna transition zone of nigeria," journal of applied agricultural research, vol. 4, pp. 111-120, 2012. [2] a. e. uko, e. b. effa, and a. u. akpan, "effect of oil palm bunch ash and poultry manure on the growth and yield of groundnut (arachis hypogaea l) in an ultisol of uyo, akwa ibom state, nigeria," in proceedings of the 3rd national annual conference of crops science society of nigeria, held at the university of nigeria, nsukka, between september 27-30, 2016, 2016, pp. 158-163. [3] a. akinmutimi and l. agwu, "comparative effects of commercial lime (caco 3) and ground eggshell on the uptake of calcium and dry matter yield of maize in an ultisol of southeastern nigeria," nigeria agricultural journal, vol. 45, pp. 13-20, 2014. [4] a. ikeh, n. ndaeyo, e. akpan, e. udoh, and o. akata, "evaluation of complementary use of organic manure for sustainable water yam production in uyo, southeastern nigeria," american journal of research communication, vol. 1, pp. 33-48, 2013. [5] a. o. ikeh, o. r. akata, and e. a. akpan, "effect of oil palm bunch refuse ash on sustainable production of egusi melon (colocynthis citrullus) in acid costal plain soil of uyo southeastern, nigeria," in proceedings of the third national annual conference of crop science society of nigeria. held at the university of nigeria, nsukka, from september 27-30, 2016, 2016, p. 96. [6] a. u. akpan, "effect of planting density and variety on the vegetative growth and fresh pod yield of two vegetable cowpea (vigna unguiculata (l) walp) in southeastern, nigeria," journal of science of agricultural food technology and the environment, vol. 13, pp. 74-78, 2014. [7] a. u. akpan, "effect of sowing dates on growth and fresh pod yield of three improved cowpea varieties (vigna unguiculata (l) walp.) in umudike, southeastern nigeria," journal of the science of agriculture, food technology the environment, vol. 14, pp. 57-62, 2015. [8] a. akpan and e. mbah, "growth and yield of vegetable cowpea varieties (vigna unguiculata (l) walp) as influenced by potassium fertilizer rates in umudike, nigeria," nigerian journal of agriculture, food and environment, vol. 12, pp. 121-126, 2016. [9] o. r. akata, "effects of fertilization on a cassava based intercropping system in uyo. southeastern, nigeria," m.sc thesis of the department of crop science, university of uyo, akwa ibom state, nigeria, 2006. [10] b. r. ntare, a. t. diallo, j. ndjeunga, and f. waliyar, "groundnut seed production manual," ed. niamey, niger: international crops research institute for the semi-arid tropics, (icrisat), 2008, pp. 6-10. [11] e. m. obi, "growing of native vegetable in nigeria," food and agricultural organization handbook, vol. 12, pp. 66-69, 2001. [12] e. moyin-jesu, "determination of soil nutrient levels for maximum yield of okra (abelmoschus esculentum) using sole and amended plant residues," pertanika journal of tropical agricultural science, vol. 31, pp. 233-245, 2008. agriculture and food sciences research, 2019, 6(2): 198-202 202 © 2019 by the authors; licensee asian online journal publishing group [13] e. u. mbah and a. u. akpan, "effect of organic manure sources on the growth and yield of groundnut in lowland derived savanna of ishiagu, ebonyi state, nigeria," journal of agricultural food and environment, vol. 13, pp. 233-245, 2017. [14] g. s. effiong, t. o. ibia, p. i. ogban, and n. d. inyang, "evaluation of locallysourced liming material for acid soils of akwa ibom state, southern nigeria," american-eurasian journal of agronomy, vol. 2, pp. 144-151, 2009. [15] i. e. essien, "effect of organic fertilizer on soil micro-organism, growth, yield and nutritional contents of cucumber (cucumis sativum l.) in uyo, akwa ibom state, nigeria," unpublished m. sc thesis of the department of soil science and land resources management, university of uyo, 2015. [16] j. asiegbu, "effects of organic matter substrate sources and time of photosynthate—sink removal on flower and pod production in okra, abelmoschus esculentus (l) moench," east african agricultural and forestry journal, vol. 52, pp. 293-297, 1987. available at: https://doi.org/10.1080/00128325.1987.11663530. [17] k. a. gomez and a. a. gomez, statistical procedure for agricultural research, 2nd ed. new york: john wiley and son inc, 1984. [18] nrcri, "national root crops research insitutute," handbook, vol. 6, pp. 5-7, 2007. [19] k. rajgopal, k. chandran, j. misra, p. bhalodia, and r. mathur, "evaluation of bold-seeded groundnut accessions for confectionery attributes," international arachis newsletter, vol. 20, pp. 18-20, 2000. [20] m. awodun, s. ojeniyi, a. adeboye, and s. odedina, "effect of oil palm bunch refuse ash on soil and plant nutrient composition and yield of maize," american-eurasian journal of sustainable agriculture, vol. 1, pp. 50-54, 2007. [21] t. o. ibia and e. j. udo, guide to fertilizer use for crops in akwa ibom state, nigeria (teaching, research and extension guide): sibon books limited, 2009. [22] v. o. chude, w. b. malgui, i. y. amapu, and a. o. ano, "manual on soil fertilizer assessment," pp. 34-35, 2004. [23] p. e. olorunju, "evaluation of production, multiplication and distribution system of improved groundnut germplasm in nigeria," atcp/raf/7823(a) report. federal ministry of agriculture, abuja, nigeria2000. [24] m. i. uguru, crop production: tools, techniques and practices. nsukka: ephrata press, 2011. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 65 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 65-68, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.65.68 © 2020 by the authors; licensee asian online journal publishing group physical characteristics of the essential oil extracted from released and improved lemongrass varieties, palmarosa and citronella grass beriso mieso1 abdela befa2 ( corresponding author) 1analtytical chemistry ethiopia institute of agricultural research, wondo genet agricultural research center shashamene, ethiopia. 2food science and technology, ethiopia institute of agricultural research, wondo genet agricultural research center shashamene, ethiopia. abstract the grass species lemongrass, palmarosa, and citronella/nardus grass are one of the aromatic and medicinal plants which know days wondo genet agriculture center is working on their essential oil yield increment and releasing different varieties. the essential oils were extracted using hydro distillation methods. this study investigated the analysis of physical characteristics of lemongrass varieties, palmarosa, and citronella grass essential oils. from the released three lemongrass varieties wg-lomisar-java has the highest oil content (1.25 %± 0.02) and the other two varieties namely lomisar-i and wg-lomisar-ua (upper awash) have the same oil content (1.2%± 0.02). wg-lomisar-ua (upper awash) has the largest relative density/specific gravity (0.9015± 0.0002) and wg-lomisar-java has the smallest value (0.9007± 0.0002). in the case of the refractive index, lomisar-i has a large value (1.4890± 0.0002) and wg-lomisar-java has a small value (1.4870± 0.0002). the oil content of palmarosa was 1.2%± 0.03, the specific gravity was 0.8815± 0.0002 and the refractive index was 1.4720± 0.0002, whereas citronella grass contains 1.3%± 0.01 oil content, 0.9108± 0.0002 specific gravity, and 1.4800± 0.0002 refractive indexes. keywords: citronella grass, essential oil, lemongrass, palmarosa, physicochemical. citation | beriso mieso; abdela befa (2020). physical characteristics of the essential oil extracted from released and improved lemongrass varieties, palmarosa and citronella grass. agriculture and food sciences research, 7(1): 65-68. history: received: 30 january 2020 revised: 3 march 2020 accepted: 8 april 2020 published: 13 may 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: authors are greatly acknowledged wondo genet agricultural research center for critically supporting on the released fund when we work in this research and also the authors would like to thanks all wondo genet researcher those stand beside as during this work and writing of manuscript from start to end. funding: the authors were gratefully acknowledged ethiopia institute of agricultural research for financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 66 2. materials and methods ................................................................................................................................................................... 66 3. results and discussion ................................................................................................................................................................... 67 4. conclusion ......................................................................................................................................................................................... 67 references .............................................................................................................................................................................................. 67 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.65.68&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.65.68&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1627 agriculture and food sciences research, 2020, 7(1): 65-68 66 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study investigated the analysis of physical characteristics of lemongrass varieties, palmarosa, and citronella grass essential oils. 1. introduction essential oils are concentrated essences extracted from different parts of plants, containing hundreds of substances, but typically with the prevalence of one, two or three of them that characterize the fragrance [1]. essential oils are highly concentrated secondary metabolites of diverse functions in the plant system. ancient romans, greeks, egyptians, the middle, and the far east used regularly essential oils as perfumes, food flavors, deodorants, pharmaceuticals, and embalming antiseptics [2]. in spain and france from the early 1300s, distillation was developed to produce more concentrated essences of aromatic grasses and herbs like mints, citronella, lemongrass, lavender and sage [3]. in ethiopia wondo genet agricultural research center is among research centers found in the ethiopian institute of agricultural research center (eiar) which coordinates research activities on lemongrass, palmarosa, and citronella grass. lemongrass, cymbopogon citratus, is a perennial medicinal plant belonging to family geramineae, and it is distributed worldwide especially in tropical and subtropical areas of africa, asia, and america [4]. the chemical composition of lemongrass essential oil depends on many factors such as genetic diversity, temperature, light intensity, maturity stage, and agricultural practice and main effective compounds of lemongrass are citral and essential oil [5-10]. cymbopogon martini (roxb.) wats. var. motia, known as palmarosa, is a perennial herb, widely distributed in tropical and subtropical regions [11]. it contains essential oil, whose main components are geraniol and geranyl acetate [12]. the essential oil from cymbopogon martini is widely used as a flavoring for food, beverages, and snuff products, as a valuable component for perfumes, cosmetic and pharmaceutical products, and against the action of various bacteria, fungi and microorganisms [11]. cymbopogon nardus, common name citronella grass, is a perennial aromatic plant from the poaceae grass family, originating in tropical asia and based oil production, citronella grass is also used for culinary purposes, as a fragrance, aroma and flavor flavoring [12, 13]. this study aims to evaluate the physical properties of the essential oil extracted from released and improved grass species namely lemongrass varieties, palmarosa, and citronella grass. 2. materials and methods 2.1. sample collection and preparation the experiment was carried out in the wondo genet natural product laboratory.the samples of three released lemongrass varieties namely lomisar-i, wg lomisar-java and wg lomisar-upper awash (ua), palmarosa and citronella grass at optimum harvesting age was collected from wondo genet agricultural research center experimental field. the sampling site was located at an altitude of 1800 m a.s.l., and latitude and longitude of n 39° 1' 44" e 8° 25' 59". the collected samples were weighed and transported to wondo genet natural product laboratory for extraction. 2.2. essential oil extraction the essential oil was extracted from fresh leaves using the clevenger type apparatus for 3 hours by hydrodistillation methods and dried with anhydrous na2so4 according to tepe, et al. [13]. 2.3. physical quality parameter of essential oil 2.3.1. essential oil content from the essential oil extracted essential oil contents of grass species samples were calculated based on the formula described below oil content (w/w (%) = mass of extracted oil (g) mass of extracted sample (g) × 100 2.3.2. the specific gravity of the essential oil the specific gravity was determined according to the method described by chophi, et al. [14]. the 5ml of distilled water was added to the cleaned pycnometer. the distilled water was weighed (mwater) (make sure that there is no bubble or air inside the pycnometer while weighing). the distilled water was removed and the pycnometer was dried. then the same volume of oil was added into the pycnometer and was weighed (moil). finally, specific gravity or relative density was calculated using the following formula specific gravity/relative density = 𝑀𝑎𝑠𝑠 𝑜𝑓𝑜𝑖𝑙 𝑀𝑎𝑠𝑠 𝑜𝑓𝑤𝑎𝑡𝑒𝑟 x 100 2.3.3. refractive index determination the refractive indexes of the essential oils were measured by refractometer (reichert, ar200) according to the method described by chophi, et al. [14]; pirbalouti, et al. [15]. the prism of the digital spectrophotometer was cleaned and the read button was pressed first to make sure that it is cleaned well. the sample was applied to the prism of a digital spectrophotometer using a micropipette. finally, the result of the refractive index was read and recorded. the triplicate analysis was taken place and the result was the average of triplicate value in all cases. 2.4. data analysis significance differences in physical properties of the essential oil extracted from released and improved grass species were subjected to analysis of variance (anova) using the microsoft excel software. https://en.wikipedia.org/wiki/perennial https://en.wikipedia.org/wiki/poaceae https://en.wikipedia.org/wiki/asia agriculture and food sciences research, 2020, 7(1): 65-68 67 © 2020 by the authors; licensee asian online journal publishing group 3. results and discussion the physical quality parameter of aromatic plants in this study like essential oil content, relative density or specific gravity, refractive index, appearance, color, and odor were analyzed and the results were shown in the following tables table 1 and table 2. table-1. physicochemical parameters of lemongrass varieties (n=3). variety type eoc% (w/w) specific gravity refractive index appearance color odor lomisar-i 1.2±0.01 0.9012± 0.0002 1.4890± 0.0002 clear liquid yellowish sweet smell, lemony wg-lomisar-java 1.25±0.02 0.9007± 0.0002 1.4870± 0.0002 clear liquid yellowish sweet smell wg-lomisar-ua (upper awash) 1.2± 0.02 0.9015± 0.0002 1.4883± 0.0002 clear liquid yellowish sweet smell note: where eoc= is essential oil content. from table 1, it was shown that wg-lomisar-java has the highest oil content with a value of 1.25 % ± 0.02 from the three released varieties. the remaining two varieties namely lomisar-i and wg-lomisar-ua (upper awash) have the same oil content (1.2%± 0.02). the value of essential oil content of all lemongrass varieties of this research finding was less than the finding of pirbalouti, et al. [15]. the difference was maybe because of the difference in sample location and agroecology between two samples. wg-lomisar-ua (upper awash) has the largest relative density/specific gravity (0.9015± 0.0002) followed by lomisar-i (0.9012± 0.0002) and wg-lomisar-java (0.9007± 0.0002). the value of the relative density/specific gravity of lemongrass varieties in this study was greater than the values expressed in taweechaisupapong, et al. [16]. the reason for the difference in values between two findings was maybe because of the difference in the sample location climatic condition and agroecological difference including soil type. in the case of the refractive index, lomisar-i has a large value (1.4890± 0.0002) and wg-lomisar-java has a small value (1.4870± 0.0002). the values of the refractive index of this research finding were found within the range of values observed in taweechaisupapong, et al. [16]. the appearance of all three varieties was clear liquid and their color was yellowish. the odor of lomisar-i was a sweet smell, lemony and that of the remaining two varieties was a sweet smell. analysis of variance (anova) oil content, specific gravity, and refractive index were significantly different from variety to variety because of fcalculated > fcritical. table-2. physical parameters of palmarosa and citronella grass (n=3). plant name eoc% (w/w) specific gravity refractive index appearance color odor palmarosa 1.2± 0.03 0.8815± 0.0002 1.4720± 0.0002 clear liquid pale yellow pleasant odor with rose-like smell citronella 1.3± 0.01 0.9108± 0.0002 1.4800± 0.0002 clear liquid pale yellow sweet smell note: from where eoc= is essential oil content. table 2, it was shown that palmarosa has 1.2%± 0.03 oil content, 0.8815± 0.0002 specific gravity and 1.4720± 0.0002 refractive indexes. the value of essential oil content of this study was less than the value of essential oil reported by rajeswara, et al. [11]. again, the refractive index and specific gravity of palmarosa oil were less than values reported by taweechaisupapong, et al. [16]. citronella grass contains 1.3%± 0.01 oil content,0.9108± 0.0002 specific gravity, and 1.4800± 0.0002 refractive index. the values of the refractive index of this study were within the range of results reported baker, et al. [12]; phovisay, et al. [17] and the values of specific gravity were greater than the report of baker, et al. [12]; phovisay, et al. [17]. the difference maybe because of the difference in the sample location and agroecology. both of them are clear liquid and have a pale-yellow color. the odor of palmarosa was pleasant odor with the rose-like smell but the odor of citronella grass was a sweet smell. 4. conclusion in this study physicochemical quality parameters analysis of three lemongrass varieties, palmarosa and citronella grass were analyzed. the parameters were essential oil content, refractive index, specific gravity, appearance, color, and odor. the values of these physicochemical quality parameters were significantly different from one variety to the other in lemongrass. references [1] m. f. mendes, extraction modes. handbook of food products manufacturing. hoboken, new jersey: john wiley & sons, inc, 2007. [2] a. mohammed, z. r. hail, c. demelza, and p. f. michael, "the growth and development of sweet basil (ocimum basilicum) and bush basil (ocimum minimum) grown under three light regimes in a controlled environment," agronomy, mdpi, vol. 9, pp. 743757, 2019. [3] t. t. joseph-adekunle and a. s. adelola., "growth response, herbage yield and proximate contents of peppermint (mentha piperita) as influenced by organic fertilizer types. in ecological and organic agriculture strategies for viable continental and national development in the context of the african union's agenda 2063," in scientific track proceedings of the 4th african organic conference. november 5-8, 2018. saly portudal, senegal, 2018. [4] w. s. soliman, "chemical composition evaluation of egyptian lemongrass, cymbopogon citratus, essential oil," international journal of scientific and engineering research, vol. 8, pp. 630-634, 2017. [5] a. akhila, essential oil-bearing grasses: the genus cymbopogon: crc press, 2019. [6] j. samy, m. sugumaran, and k. lee, herbs of malaysia. malaysia: federal publication sdn berhad, 2005. [7] i. b. jaganath, herbs: the green pharmacy of malaysia. serdang: mardi, 2000. agriculture and food sciences research, 2020, 7(1): 65-68 68 © 2020 by the authors; licensee asian online journal publishing group [8] n. tajidin, "growth performance and nutrient concentration of ‘hijau’lemongrass (cymbopogon citratus) as affected by maturity stages at harvest," the role of plant physiology the role of plant physiology in climate change adaptation in climate change adaptation and mitigation and mitigation, p. 35, 2010. [9] k. devi, transcriptome sequencing of cymbopogon winterianus and characterization of metabolic pathway genes. jorhat: amateur athletic union, 2015. [10] s. p. khanuja, a. k. shasany, a. pawar, r. k. lal, m. p. darokar, a. a. naqvi, and s. kumar, "essential oil constituents and rapd markers to establish species relationship in cymbopogon spreng.(poaceae)," biochemical systematics and ecology, vol. 33, pp. 171186, 2005. [11] r. b. rajeswara, d. rajput, and r. patel, "essential oil profiles of different parts of palmarosa (cymbopogon martinii (roxb.) wats. var. motia burk.)," journal of essential oil research, vol. 21, pp. 519-521, 2009. [12] b. p. baker, j. a. grant, and r. malakar-kuenen, "citronella & citronella oil profile," journal of essential oil research, vol. 24, pp. 350-353, 2018. [13] b. tepe, d. daferera, a. sokmen, m. sokmen, and m. polissiou, "antimicrobial and antioxidant activities of the essential oil and various extracts of salvia tomentosa miller (lamiaceae)," food chemistry, vol. 90, pp. 333-340, 2005. available at: https://doi.org/10.1016/j.foodchem.2003.09.013. [14] r. chophi, s. sharma, s. sharma, and r. singh, "trends in the forensic analysis of cosmetic evidence," forensic chemistry, vol. 14, p. 100165, 2019. [15] a. g. pirbalouti, m. oraie, m. pouriamehr, and e. s. babadi, "effects of drying methods on qualitative and quantitative of the essential oil of bakhtiari savory (satureja bachtiarica bunge.)," industrial crops and products, vol. 46, pp. 324-327, 2013. available at: https://doi.org/10.1016/j.indcrop.2013.02.014. [16] s. taweechaisupapong, t. boonmars, p. satthanakul, v. chatchanayuenyong, s. nitinon, u. soodtoetong, w. thipsongkroh, and w. khunkitti, "effect of lemongrass gel against candida albicans in rat model of oral candidiasis," sjst, vol. 42, pp. 299-304, 2020. [17] s. phovisay, x. briatia, v. chanthakoun, and s. savathvong, "effect of distillation methods on citronella oil (cymbopogon nardus) content," in iop conference series: materials science and engineering, vol. 639, pp. 1-6, 2019. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 175 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 175-181, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6249 © 2024 by the authors; licensee asian online journal publishing group banknotes residues and thermoplastic starch as sustainable reinforcements for ldpe-based agricultural composites martins, marcela fialho1 trindade, julio gabriel2 boaventura, tulio pacheco3 gil, camila silva brey4 ( corresponding author) 1,4department of chemical and materials engineering, federal university of lavras, lavras, minas gerais, brazil. 1email: marcela.martins1@estudante.ufla.br 4email: camilabreygil@gmail.com 2,3federal university of lavras, lavras, minas gerais, brazil. 2email: jgabriel0499@gmail.com 3email: tuliopb1@hotmail.com abstract this study examines the development and characterization of composites comprising low-density polyethylene (ldpe), thermoplastic starch (tps), and cellulose fibers sourced from recycled banknotes (br). the composites were fabricated using extrusion and injection molding techniques, followed by an evaluation of their morphological, thermal, mechanical, and water absorption properties. the inclusion of br fibers enhanced the mechanical performance of ldpe, notably improving tensile strength and elastic modulus, while the hydrophobic treatment of the fibers ensured minimal water absorption. the incorporation of tps increased the composites' biodegradability potential but reduced tensile strength and increased water absorption due to its hydrophilic nature. scanning electron microscopy revealed strong interfacial adhesion between the fibers and matrix and confirmed the complete dispersion of starch in tps-containing samples. thermogravimetric analysis highlighted the thermal stability of the composites and identified distinct degradation stages corresponding to the matrix, tps, and fibers. these results indicate that the developed composites offer a viable solution for agricultural applications, balancing mechanical performance and environmental sustainability. keywords: ldpe composites, recycled banknote fibers, thermoplastic starch, sustainable materials, polymer reinforcement. citation | fialho, m. m., gabriel, t. j., pacheco, b. t., & brey, g. c. s. (2024). banknotes residues and thermoplastic starch as sustainable reinforcements for ldpe-based agricultural composites. agriculture and food sciences research, 11(2), 175–181. 10.20448/aesr.v11i2.6249 history: received: 4 november 2024 revised: 5 december 2024 accepted: 11 december 2024 published: 26 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by conselho nacional de desenvolvimento científico e tecnológico-cnpq brazil (grant number: cnpq-brasil– 403724/2023-6) and fundação de amparo à pesquisa de minas gerais–fapemig (grant number: fapemig brasil– apq-03923-23, apq00413-22). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: characterization, data analysis and writing, m.f.m., j.b.t. and c.s.b.g.; conceptualization, methodology, materials processing, reviewing and editing, t.p.b. and c.s.b.g.; project administration and funding acquisition, c.s.b.g. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 176 2. materials and methods ................................................................................................................................................................. 176 3. results and discussion ................................................................................................................................................................. 177 4. conclusion ....................................................................................................................................................................................... 180 references ............................................................................................................................................................................................ 180 mailto:marcela.martins1@estudante.ufla.br mailto:camilabreygil@gmail.com mailto:jgabriel0499@gmail.com mailto:tuliopb1@hotmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6249 https://orcid.org/0009-0005-4707-4239 https://orcid.org/0009-0002-3284-0111 https://orcid.org/0000-0002-6143-5417 https://orcid.org/0000-0002-0767-3123 agriculture and food sciences research, 2024, 11(2): 175-181 176 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study uniquely integrates recycled banknote fibers into ldpe composites, combining hydrophobic fiber treatment with thermoplastic starch to enhance mechanical properties and biodegradability. unlike previous works, it explores the interplay of these components, demonstrating the feasibility of upcycling cellulose waste while achieving a balance between performance and environmental sustainability. 1. introduction synthetic polymers are widely used across industries due to their versatile and desirable properties. lowdensity polyethylene (ldpe) is widely used in agriculture for various applications such as mulching films and packages for fertilizers due to its flexibility, durability, and resistance to moisture and chemicals [1, 2]. however, these polymers are derived from non-renewable sources, which are both limited and environmentally harmful due to improper disposal and long degradation times [2, 3]. to reduce the environmental impact associated with synthetic polymers in agriculture, biodegradable natural materials are increasingly being explored [1, 4-6]. starch stands out as a promising candidate due to its low cost and large-scale production. however, despite being a promising biodegradable material, starch has several limitations that drive its incorporation into other polymers like ldpe to improve performance. those limitations include poor mechanical properties, high moisture absorption and low thermal stability [7]. to be compatible with ldpe, starch must undergo a destructuring process, transforming its granular structure into an amorphous material called thermoplastic starch (tps) [8, 9]. despite the potential of combining ldpe and tps, poor compatibility between the materials leads to reduced mechanical performance. to improve the properties of this mixture, compatibilizing agents, typically organic acids, are often employed [10-12]. since the main objective of this paper is to seek more sustainable alternatives, we proposed the use of banknotes residues (br) from bills unfit for circulation. such waste was made available by the central bank of brazil, which states that approximately 800 tons of banknotes waste were discarded in 2017 [13]. banknotes are composed of high-quality cellulose fibers, among other components, which guarantee properties such as mechanical and water resistance, which are essential for the blend of ldpe and tps. in a previous study gil, et al. [13] br were incorporated on ldpe matrix at concentrations between 1-20% resulting on physical interactions between the residues and polymer matrix, improving mechanical properties. these improvements include increased tensile strength and reduced elongation percentage, which are valuable for agricultural applications where durability and strength are required. considering these promising outcomes, the addition of tps into a ldpe/br composite is intended to enhance the biodegradability, aiming to minimize the environmental impact. in this paper, we developed a composite of ldpe, tps, and br, with the goal of creating a sustainable alternative to conventional agricultural polymers. this composite material not only demonstrates performance comparable to synthetic polymers but also incorporates renewable and recycled resources, making it a viable, ecofriendly substitute in agricultural settings. 2. materials and methods the ldpe, code ei-1630, was supplied in pellet form by quattor.low-density. the corn starch with 70% amylose, and pure glycerol were used to produce thermoplastic starch (tps). the banknote residue samples were provided by the central bank of brazil (banco central do brasil). the preparation of the composite material using recycled banknote waste as reinforcement involved several steps. first, the thermoplastic starch (tps) was produced from a mixture of corn starch, glycerol, and water in a ratio of 60:24:14% by weight. this mixture was stirred at 60°c for 30 minutes. next, the banknote waste was shredded into small fragments, which were then mixed with the ldpe to form the primary matrix material. this mixture underwent two passes through a single-screw extruder to ensure sample homogeneity. tps was then added to this mixture, followed by a third extrusion, according to the formulation specified for each test specimen as outlined in table 1. following the mixing and extrusion steps, samples were injected into a mold with dimensions according to american society for testing and materials (astm) d 638 – 03 standards. the produced materials were subsequently characterized to determine the composite’s final properties and to evaluate the influence of fiber and tps addition on the polymer matrix. table 1. composites composition. sample ldpe (%w/w) banknote residues (%w/w) tps (%w/w) ldpe 100 0 0 ldpe/br 85 15 0 ldpe/br/tps10 76.5 13.5 10 ldpe/br/tps20 68 12 20 ldpe/br/tps30 59.5 10.5 30 2.1. mechanical testing the samples were characterized by tensile tests according to astm d638 standards. testing was performed on a universal testing machine (emic dl 3000) using a 2 kn load cell. results were analyzed through one-way analysis of variance (anova) followed by tukey’s multiple comparison test. statistical significance was set at p < 0.05. all analyses were conducted using past software (version 2.17c, 1999–2013). agriculture and food sciences research, 2024, 11(2): 175-181 177 © 2024 by the authors; licensee asian online journal publishing group 2.2. thermogravimetric analysis (tga) thermogravimetric analysis curves were recorded on a seiko sii nanotechnology inc. exstar 7200 instrument under nitrogen with a heating rate of 10 °c/min over a temperature range of 20 to 800 °c. nitrogen was used as the purge gas at a flow rate of 30 ml/min. 2.3. morphological analysis the morphological analyses of the composites were conducted using images obtained with a scanning electron microscope (sem) (zeiss, leo, evo 40) equipped with extended variable pressure (xvp). samples were mounted on carbon tape with the cross-sectional area positioned upward. all samples were coated with gold by sputtering. 2.4. fourier transform infrared spectroscopy (ftir) ftir analysis in the 4000 to 650 cm⁻¹ range was performed using a nicolet 8700 spectrometer equipped with fourier transform and an atr accessory. znse crystal was used with 84 scans at a resolution of 4 cm⁻¹. 2.5. water absorption test the water absorption test was conducted in accordance with astm d570. prior to immersion, samples were dried in an oven at 60°c for 48 hours to determine the initial mass. specimens were then immersed in water at room temperature (23 ± 1°c). after 48 hours, the samples were removed from the water, quickly dried with paper, and weighed. this process was repeated until the mass stabilized. at the end of the test, a water absorption curve (%) versus immersion time (hours) was obtained as per astm d570 – 98. the percentage of water absorption in the composites was calculated using equation 1: ∆𝑀(%) = 𝑀𝑓−𝑀𝑖 𝑀𝑖 𝑥100 (1) where δm is the water absorption percentage, and mi and mf represent the sample mass before and after immersion, respectively. 3. results and discussion the tensile properties of composites and pure ldpe are shown in table 2. as previously shown by gil, et al. [13] the incorporation of banknote fibers into ldpe matrix introduces desirable reinforcing characteristics without the need for additional compatibilizing agents. similarly, in this study, the ldpe/br sample demonstrated a comparable performance, as shown in table 2, with higher elastic modulus, lower tensile strain, and increased tensile strength compared to pure ldpe and samples containing tps. as expected, the addition of cellulose fibers increased the elastic modulus of ldpe. studies indicate that enhanced strength between the fiber and polymer matrix signifies good adhesion in the composite, which aligns with findings from other authors [13, 14]. the behavior of the samples with the addition of tps showed, based on the analysis of table 2, that the addition of starch reduces the maximum stress supported by the material. it results in an increase in the elastic modulus and a reduction in the percentage of deformation. the incorporation of starch in the composite, as expected, causes embrittlement, due to the replacement of a resistant matrix with a fragile material [15]. table 2. mechanical properties (young’s modulus, tensile strength and elongation at break) of ldpe and ldpe composites. sample young’s modulus (mpa) tensile strength (mpa) elongation at break (%) ldpe 47.7 ± 6.6a 9.85 ± 0.22a 133 ± 4a ldpe/br 73.8 ± 6.7b 10.8 ± 0.15b 19.7 ± 1.9b ldpe/br/tps10 66.2 ± 8.4b 8.16 ± 0.14c 28.1 ± 3.3c ldpe/br/tps20 62.4 ± 8.4b 8.25 ± 0.59c 31.0 ± 11c.d ldpe/br/tps30 65.2 ± 10.4b 8.35 ± 0.29c 24.0 ± 3d thermal analysis of the samples can be observed in figure 1. for the banknotes residues a minor weight loss can be observed at temperatures below 100ºc due to moisture present in the material. in the range of 200 to 370ºc, a more significant weight loss in the br was observed, primarily due to the degradation of hemicellulose and cellulose, resulting in approximately 64% mass loss at 305ºc. beyond this range, the weight continues to decrease, attributed to the degradation of lignin and subsequent decomposition of the br components. after reaching 600ºc, the br retained a significant percentage of its weight compared to the other samples, which can be attributed to the hydrophobic components added to banknotes, providing greater resistance to high temperatures. the thermal characteristics of the br align with findings reported in other studies [13, 16]. in ldpe, a substantial weight loss is noted between 340 and 510°c, occurring in a single decomposition step. the ldpe/br sample, however, displays two mass-loss intervals: the first between 270 and 370ºc, and the second between 400 and 510ºc, corresponding to br and ldpe degradation, respectively. as noted in previous studies, compared to ldpe the thermal stability of the composite decreased, due to the decomposition temperature of hemicellulose, cellulose, and lignin present in the br [13]. note: identical letters indicate statistically similar means (within the same column) according to tukey’s test at a 5% significance level. agriculture and food sciences research, 2024, 11(2): 175-181 178 © 2024 by the authors; licensee asian online journal publishing group figure 1. tga of ldpe and ldpe composites. the tga curve for thermoplastic starch presents three stages of mass loss. the first, between 25 and 125°c, occurs due to water loss; the second, between 290 and 384°c, is considered as the main degradation stage; and the third stage, above 480°c, refers to the decomposition of residual matter [17]. the samples containing tps present mass loss in the intervals of 200 to 270 °c, 270 to 370 °c and 370 to 510 °c, as shown in table 3, which suggests the influence of fibers, tps and ldpe, respectively. studies have shown that the higher the starch content, the higher the storage modulus of ldpe/tps blend [15]. this can be attributed to the rigid character of starch compared to the ldpe structure. table 3. percentage of weight loss. sample 30-200°c 200-270°c 270-370°c 370-510°c residues at 510°c ldpe/br 0.29 0.81 7.67 85.86 5.37 ldpe/br/tps10 2.24 2.78 13.07 77.3 4.61 ldpe/br/tps20 4.06 4.6 16.09 70.27 4.98 ldpe/br/tps30 4.97 6.7 19.62 60.05 8.66 figure 2 presents the ftir analysis data. in the br spectrum, absorption bands are observed between 3334 and 3274 cm⁻¹, corresponding to the hydroxyl groups present in cellulose and hemicellulose; between 2916 and 2848 cm⁻¹, associated with the stretching vibrations of methylene groups; and at 1032 cm⁻¹, linked to c–o stretching. the pronounced peak at 1032 cm⁻¹ indicates the significant presence of c–o stretching, consistent with findings in related literature, a characteristic of both hemicellulose and cellulose fibers [16]. in the ldpe spectrum, distinct peaks appear at 2914, 2846, 1463, and 719 cm⁻¹, which correspond to asymmetric ch₂ stretching, symmetric ch₂ stretching, bending deformation, and rocking deformation, respectively [13]. in the ldpe/br samples, the absorption bands characteristic of pure ldpe was predominantly observed, with no clear indication of br presence in the spectrum. this can be attributed to the low fiber content relative to the percentage of ldpe, which results in absorption bands of ldpe being intense enough to mask the characteristic bands of the fiber [13]. figure 2 also shows the ftir spectra of samples containing ldpe/br/tps. mendes et al. analyzed the ftir spectrum of tps and identified absorption bands characteristic of starch and glycerol at 920, 1022, and 1140 cm⁻¹ due to c–o stretching; at 1648 cm⁻¹, related to ester group formation; at 3277 and 2914 cm⁻¹, associated with o–h and c–h stretching, respectively; and at 1423 cm⁻¹ due to glycerol [17]. the presence of ester bonding at regions around 1642 cm⁻¹ and 1000 to 1200 cm⁻¹ indicates an anhydride ring opening and bonding between starch and ldpe. some of these tps characteristic bands are present in the spectra of the samples, as highlighted in figure 2, confirming the presence of tps. additionally, these bands are more pronounced in the spectra with higher tps content, specifically in the ldpe/br/tps20 and ldpe/br/tps30, compared to the ldpe/br/tps10 sample. agriculture and food sciences research, 2024, 11(2): 175-181 179 © 2024 by the authors; licensee asian online journal publishing group figure 2. ftir spectra of the banknotes residues, pure ldpe, and ldpe composites. water absorption over time for the samples is shown in figure 3. as observed, pure ldpe does not absorb water (absent blue bar on graph), maintaining a stable weight throughout the process due to its hydrophobic nature [18]. the addition of fiber resulted in a quick absorption of 0.82%, with the weight stabilizing after the first measurement, indicating a fast yet minimal uptake by the composite. although fibers are generally hydrophilic, banknote fibers contain components that provide hydrophobic properties (water resistance) compared to other cellulose fibers, explaining the low absorption observed [13]. with the addition of tps, which has hydrophilic properties, an increase in water absorption over time was noted [18]. the sample showing the greatest mass variation throughout the experiment was ldpe/br/tps20. figure 3. percentage of water absorption as a function of time. the morphology of the produced samples was examined using sem, as shown in figure 4. for comparison, images of the pure ldpe matrix and the fiber were also obtained (figures 4 a-b). in polymeric materials, this analysis is crucial, as the physical-mechanical properties of the composites are directly influenced by the morphology of their constituents and their interfacial interactions. the images of the fibers (figure 4a) show loose fibers and also fibers bound together by the resin that forms the banknotes. as observed in figure 4b, the sem image of pure ldpe displays a smooth and homogeneous surface, characteristic of its amorphous structure. the absence of significant microstructural features, such as fibers or particles, highlights its uniform composition. the ldpe/br samples, on the other hand, demonstrate a notable difference in morphology, with fibers embedded within the polymer matrix (figure 4 c). unlike the smooth surface of pure ldpe, the composite samples exhibit a more fibrous and heterogeneous appearance, indicative of the incorporation of the fiber component. as already analyzed by gil et al., the ldpe/br samples show an absence of removed or empty fibers, suggesting a certain degree of interfacial bonding between the fiber and the ldpe matrix [13]. furthermore, when tps is added to the composite, as seen in figures 4 d-f, the fracture surface becomes even more irregular and heterogeneous. studies have previously revealed low interfacial adhesion between ldpe and tps [14] however, the introduction of fibers appears to significantly enhance these interactions. another critical observation is the absence of starch granules in the sem images, indicating the complete destruction of the original corn starch structure during tps preparation [19]. the differences in morphology between the pure agriculture and food sciences research, 2024, 11(2): 175-181 180 © 2024 by the authors; licensee asian online journal publishing group ldpe and the composite samples underscore the successful incorporation of fibers and tps, as well as the potential for these modifications to influence the material's mechanical properties. figure 4. sem images: (a) banknotes residues, (b) ldpe, (c) ldpe/br, (d) ldpe/br/tps10, (e) ldpe/br/tps20 and (f) ldpe/br/tps30. 4. conclusion this study successfully demonstrated the potential of developing composites using ldpe, cellulose fibers from recycled banknotes, and tps for agricultural applications. the inclusion of cellulose fibers notably enhanced the mechanical properties of the polymer matrix, improving its tensile strength and stiffness without the need for compatibilizing agents. these findings highlight the efficiency of reusing cellulose-rich materials like banknotes as reinforcement in polymer composites. the addition of tps introduced a biodegradable component into the material, but also brought challenges such as increased water absorption and reduced mechanical strength due to its hydrophilic nature. despite this, the proper dispersion of tps, evidenced by the absence of starch granules in the sem images, reflects the effectiveness of the processing method used. thermal analysis confirmed the distinct decomposition stages for the matrix, fibers, and tps, emphasizing the complex interactions within the composite. the stability and interfacial bonding observed are promising indicators of the material's suitability for practical applications, especially in environments where moderate mechanical demands and sustainability are key factors. the findings underscore the feasibility of developing eco-friendly composites with balanced mechanical and environmental properties. future work could explore alternative fiber treatments or compatibilization strategies to further enhance the integration of tps and fibers, aiming to expand the usability of these materials in diverse sectors. references [1] k. lewicka et al., "current trends of polymer materials’ application in agriculture," sustainability (2071-1050), vol. 16, no. 19, p. 8439, 2024. https://doi.org/10.3390/su16198439 [2] k. n. palansooriya et al., "low-density polyethylene microplastics alter chemical properties and microbial communities in agricultural soil," scientific reports, vol. 13, no. 1, p. 16276, 2023. https://doi.org/10.1038/s41598-023-42285-w [3] p. nayanathara thathsarani pilapitiya and a. s. ratnayake, "the world of plastic waste: a review," cleaner materials, vol. 11, p. 100220, 2024. https://doi.org/10.1016/j.clema.2024.100220 [4] a. samir, f. h. ashour, a. a. hakim, and m. bassyouni, "recent advances in biodegradable polymers for sustainable applications," npj materials degradation, vol. 6, no. 1, p. 68, 2022. https://doi.org/10.1038/s41529-022-00277-7 https://doi.org/10.3390/su16198439 https://doi.org/10.1038/s41598-023-42285-w https://doi.org/10.1016/j.clema.2024.100220 https://doi.org/10.1038/s41529-022-00277-7 agriculture and food sciences research, 2024, 11(2): 175-181 181 © 2024 by the authors; 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licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 89-96, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.89.96 © 2020 by the authors; licensee asian online journal publishing group physicochemical and antioxidant properties of oils used by local fried food vendors in d/line-port harcourt, rivers state emelike, n.j.t1 ujong a. e.2 achinewu, s. c.3 ( corresponding author) 1,2,3department of food science and technology, rivers state university, nkpoluoroworukwo, port harcourt, rivers state, nigeria. abstract frying initiates different physical and chemical changes which can cause degradation of oil and quality of the fried food. this study was conducted to investigate and monitor the physicochemical and antioxidant properties of frying oils used by local fried food vendors in d/line-port harcourt, rivers state. a total of five frying oils were collected randomly during frying operations from the study area and analyzed at weekly intervals (3 weeks) for physicochemical and antioxidant properties. the results showed that all the frying oils collected were above the permissible limit for free fatty acid and saponfication value while peroxide values were within the codex regulatory limit after three weeks of collection. the results also revealed a decrease in iodine values of the frying oils except for oil samples collected from agudama street and railway close. moisture content of all the frying oils was below the 0.3% maximum limit while smoke points were not in line with the recommended standard as all the oils had smoke points <170oc even up to the third week of collection. increased usage of the oils during frying also resulted to a decrease in total phenolic content except for frying oil collected from kaduna street while a reverse was observed for total flavonoid content. the degradation in the quality of oils used by local fried food vendors in d/line, port harcourt during frying operations is an important health issue which could cause damaging health effects due to the toxic substances produced. keywords: antioxidant, physicochemical, oil, frying, d/line, collection. citation | emelike, n.j.t; ujong a. e; achinewu, s. c (2020). physicochemical and antioxidant properties of oils used by local fried food vendors in d/line-port harcourt, rivers state. agriculture and food sciences research, 7(1): 89-96. history: received: 18 march 2020 revised: 22 april 2020 accepted: 25 may 2020 published: 15 june 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 90 2. materials and methods ................................................................................................................................................................... 90 3. results and discussion ................................................................................................................................................................... 91 4. conclusion ......................................................................................................................................................................................... 95 references .............................................................................................................................................................................................. 95 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.89.96&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.89.96&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1735 https://orcid.org/0000-0002-8440-7072 https://orcid.org/0000-0001-6572-2328 https://orcid.org/0000-0001-7161-8148 agriculture and food sciences research, 2020, 7(1): 89-96 90 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to literature by evaluating the level of degradation in the quality of frying oils used by local fried food vendors in d/line, port harcourt during frying operations. 1. introduction frying is one of the most common and longstanding culinary techniques used for preparation of foods throughout the world. it involves submerging the food in hot, liquid fat at high temperature of 150oc-190oc [1]. during this process, the fried food absorbs some amount of oil and as a result, certain proportions of degraded products from the oil are accumulated by the fried food [2]. frying also initiates different physical and chemical changes which results in extensive degradation. the frying process is also open to atmospheric oxygen and high temperature which can however cause degradation in the oil leading to unpleasant odour and flavor [3]. several studies have shown that continuous use of vegetable oils for frying results in degradation which affects the physical and chemical properties of the oil quality idun-acquah, et al. [4]. codex alimentarius commission/fao/who food standards [5] reported that frequent frying of vegetable oils leads to deterioration and lipid oxidation results in the formation of peroxides which are responsible for primary oxidation in the oil [6]. degradation of vegetable oils during frying result in loss of nutritive value and causes damaging health effects due to the toxic substances produced [6]. according to codex alimentarius commission/fao/who food standards [5] continuous deep fat frying decreases the unsaturated fatty acids of oil and increases foaming, colour, viscosity, density, specific heat and free fatty acid contents. there has been an increase in the consumption of street fried foods in port harcourt metropolis resulting from high demand for such foods and the changing lifestyle and growth in the number of working women. food items that are often cooked by frying include yam, potato, fish, meat, plantain etc. quality control measures are lacking specifically as related to the quality of oil used by these fried foods street vendors in frying. most of them use vegetable cooking oil numerous times for frying before discarding so as to reduce cost. the frying oil is also infrequently discarded, with these vendors instead simply adding more as oil is absorbed by the fried food [7]. according to flores, et al. [8] this practice decreases the rate of hydrolytic alterations and therefore mask or slow down this type of deterioration. mensah and obeng [9] reported that vegetable oils for cooking are to be used 3-6 times before being discarded as waste; however most of these fried food vendors use these oils even more than the supposed number of times. some use the oils for frying until the colour changes to dark while others use it until the flavor of the product is unacceptable. good [10] also reported that repeated usage of vegetable oil for frying lowers the smoke point which makes the oil to smoke on heating at a lower temperature. the consumption of repeatedly heated vegetable oil during frying has been linked with increasing the risk of developing atherosclerosis, total serum lipid and low density lipoprotein (ldl) levels [11, 12]. there is need to investigate the effect of repeated frying on the quality of oils used by these fried food vendors in order to provide basic data and knowledge of the quality of oils used for frying. therefore, this study was aimed at determining the physicochemical and antioxidant of vegetable oils used by local fried food vendors in d/line, port harcourt, rivers state. 2. materials and methods 2.1. sample collection a total of five (5) oil samples (100 ml each) were collected at weekly intervals for period of three weeks from five streets, all situated in d/line, port harcourt. the streets covered were agudama, emekuku, kaduna, wogu, and railway close. these oil samples were repeatedly used vegetable oils collected during frying operations. fresh unused branded oil (kings refined vegetable oil) and unbranded (local vegetable oil) were also collected and used as control samples. the oil samples were transported to the biochemistry laboratory of the department of food science and technology, rivers state university for analysis within 1 hr after collection and checked for physicochemical and antioxidant properties. all chemicals used for this study were of analytical grade and obtained from the department of food science and technology laboratory, rivers state university, port harcourt. 2.2. methods 2.2.1. physico-chemical analysis iodine value, peroxide value, free fatty acid content, saponfication value, smoke point and moisture content of the oils collected were all determined using the aocs method [13]. 2.2.2. antioxidant analysis 2.2.2.1. the total phenolic content total phenolic of the oils was determined by the folin–ciocalteu reagent (fcr) according to the procedure reported by emelike, et al. [14]. the oil sample (0.3 ml) was weighed into a conical flask and 3.0ml of folin– ciocalteu reagent added into it. after 5min, 6% sodium carbonate (3.0 ml) was added and the mixture allowed standing at room temperature for 90mins. the absorbance of the mixture was measured at 725nm using uv/vis spectrophotometer (usa). the total phenolic content was calculated from the calibration curve and the results were expressed as mg of gallic acid equivalent per gram of oil (mggae/g oil). 2.2.2.2. total flavonoid content total flavonoid content was determined using the method of boham and kocipai-abyazan [15]. the oil samples (0.5 ml) was weighed into a conical flask and 20 ml of 80% aqueous methanol added and shake using orbital shaker for 3 hrs. this was followed by filtering using whatman filter paper thereafter the filterate was transferred into a moisture can and evaporated for 1 hr at 105oc, cooled and weighed. agriculture and food sciences research, 2020, 7(1): 89-96 91 © 2020 by the authors; licensee asian online journal publishing group 2.2.3. statistical analysis all analysis was performed in duplicate and data was subjected to analysis of variance (anova) using spss version 23 for windows (ibm corporation, new york, usa) according to the method of wahua [16]. variability within the means was separated by using duncan multiple range test (dmrt) which was defined at (p<0.05). 3. results and discussion 3.1. physicochemical properties of the frying oils 3.1.2. free fatty acid composition the result of free fatty acid values (%) of the frying oils collected is shown in figure 1. the chief composition of oils is fatty acids and the degree of unsaturation is the very first factor influencing the oxidative stability of frying oils. in this study, the lowest ffa of 0.01% and 0.03% was recorded in samples collected at week 1 from wogu and kaduna street respectively with no significant difference (p<0.05) from each other. the highest ffa of 6.90% was recorded in oil samples collected from wogu street at week 3. an increase in the ffa was observed after third week of collection for railway street (1.46-2.92%), kaduna street (0.03-1.02%), wogu street (0.016.90%) and emekuku street (2.34-3.77%) while that agudama street decreased from 2.78-1.01%. oil samples collected from wogu and kaduna street at first week as well as the branded oil used as control had ffa values (0.01-0.33%) within the maximum permissible ffa of 0.3% for edible oils while others were above the range. at week 2 and 3, all the frying oils (except for branded) collected had ffa values above the specification. a study conducted by idun-acquah, et al. [4] also revealed similar results with %ffa ranging from 0.62-2.41% after 5 days of frying. the differences observed in the ffa values of the frying oils are a consequence of new oil added during frying process which dilutes these values. blending of vegetable oils alter their fatty acid profiles [17] and can steeply retard oxidation of oils during frying. idun-acquah, et al. [4] also stated that deviation in the ffa values may be attributed to the high temperatures attained by the oil as well as the water from the fresh food product causing the natural fatty acids in the oil to get hydrolyzed into free fatty. according to emelike, et al. [14], low moisture in oil decreases the fatty acid content while high moisture leads to its increase. increase in ffa from this study can also be attributed to thermal and oxidative decomposition resulting to breakdown of long carbon chains into shorter chains. free fatty acids are also formed as a result of the cleavage and oxidation of double bonds to form carbonyl compounds at elevated temperature heating lalas [18]. mcwilliams [19] reported that the release of free fatty acid results to the formation of acreolin and smoking of the oil. acreolin is visible as bluish and acrid smoke and its vapour may cause eye, nasal and respiratory tract irritations in low level exposure. acreolin also induces the respiratory, ocular and gastrointestinal irritations by inducing the release of peptides in nerve terminals innervating these systems [20]. therefore, the oils collected at the second and third week may contain some amount of acreolin since their ffa exceeded the maximum specification. this study agrees well with other studies that the ffa content of oils increases with the number of frying cycles [21] as well as with the frying time [22]. according to thomas [23], smoke point of oils depends greatly on free fatty acid. the more ffa an oil contains, the quicker it will breakdown and start smoking. this result also indicates that smoke points of the frying oils will be very low. figure-1. free fatty acid values (%) of the oils collected at weekly intervals. 3.1.3. peroxide value of frying oils the result of the peroxide values of frying oils is shown in figure 2. for all the samples collected at week 1, the unbranded oil used as control had the highest peroxide value (pv) of 9.47meqo2/kg while the lowest pv of 0.80meqo2/kg was observed for the branded vegetable oil used as control. the pv of the oils were significantly (p<0.05) different from each other for all the weeks. it was observed that samples collected from agudama, emekuku and wogu streets had decreasing pv of 7.79-2.20meqo2/kg, 5.79-2.29meqo2/kg and 6.573.90meqo2/kg after the 3 weeks of collection while all other samples (except the controls) increased. the decrease in pv of these frying oils may be that the oils were changed to fresh oils at point of collection at week 2 and 3. the increase in the pv of the other samples following frying is because of the oxidation of carbon atoms adjacent to the double bonds in the triacylglyceride structure leading to the formation of hydroperoxides. pv is a useful biomarker of the preliminary stages of rancidity occurring under mild conditions and the freshness of the lipid matrix thus the agriculture and food sciences research, 2020, 7(1): 89-96 92 © 2020 by the authors; licensee asian online journal publishing group greater the pv, the faster the oxidation of the oil occurring [24]. the observed increase in pv during heating of oils has been reported by other authors [25, 26]. a study conducted by idun-acquah, et al. [4] obtained pv ranging 10.00-25.250 meq o2/kg after 5 days of frying. they reported the deviations of pv from the standard value to be due to the continuous exposure of the oil to light, high temperatures and atmospheric oxygen, which reacts with the oil to form peroxides. according to codex regulatory, oils to be utilized in deep frying should have a codex regulatory maximum pv of 10meqo2/kg [27]. in this study, pvs increased but all the oils were still concordant with the maximum codex standard. figure-2. peroxide values (meq02/kg) of the oils collected at weekly intervals. 3.1.4. iodine value of frying oils the result of the iodine values of the frying oils collected and analyzed is presented in figure 3. iodine value (iv) is chemically the mass of iodine in grams that is consumed by 100gram of a chemical substance by mass as oleic acid. iodine value is often used to determine the amount of unsaturation in fatty acids. the higher the iodine index, the faster is the tendency of oil oxidation during heating at elevated temperatures as in deep frying [28]. the maximum iodine value observed was the control (branded) sample with 44.69gi2 while the control (unbranded) sample had the least iv with gi2. iv for all the oil samples at weekly intervals were significantly (p<0.05) different from one another. samples collected from agudama and railway streets showed an increase in the iv after three weeks of collection and this could probably be due to a change in the used oil to fresh oil or the addition of fresh oil into the used oil in the frying medium. on the other hand, oil samples collected from emekuku, wogu and kaduna streets showed a decrement in iv. the observed decrement of the iodine value is concordant with the decrement in double bonds attributed to oxidation and thermal decomposition and has been reported by omara and kigenyi [29]. chebet, et al. [30] reported that a decrease in iodine value is an indication of deterioration in the vegetable oils. iodine value is of major interest in regard to oxidative stability of oils after frying of food. it is therefore established from the study that the decrease in the iodine value of the oils after frying shows relatively higher oxidation. figure-3. iodine values (gi2) of the oils collected at weekly intervals. 3.1.5. saponification value of frying oils result of the saponification values of the frying oils collected is shown in figure 4. saponification value (sv) is used to determine the saponification number of a fat or oil which is an index of the average molecular weight of the triaclyglceride in the sample. saponification number is a very important factor in soap production. from the results agriculture and food sciences research, 2020, 7(1): 89-96 93 © 2020 by the authors; licensee asian online journal publishing group obtained, the reference sample (unbranded oil) and samples collected from railway at second week had sv within the standard 196-206mgkoh/g of oil by national standardization body [31]. there was an increase in the saponification values of all the oils (except for samples collected from wogu and railway) even after second and third weeks of collection with values ranging between 238.99-353.37mgkoh/g, 231.13-360.96mgkoh/g, and 208.27-293.89mgkoh/g for samples collected from agudama, emekuku and kaduna streets, respectively. this increase above the specification for vegetable oils indicates that the oil to be discarded after frying can be used for soap production. this result correlates with the findings of alajtal, et al. [32] who reported an increase in the saponification values of sunflower, corn and olive oils after frying from 21.09-22.44mgkoh/g, 19.4121.15mgkoh/g and 21.54-22.27mgkoh/g oil respectively. figure-4. saponification values (mgkoh/g) of the frying oils collected at weekly intervals. 3.1.6. smoke point of frying oils result of the smoke points of the frying oils collected is presented in figure 5. the smoke point is the temperature at which a fat or oil produces a continuous wisp of smoke when heated. the maximum smoke point observed was for the reference samples (170oc and 160oc for branded and unbranded vegetable oils respectively). at week 1, the smoke point of all the frying oils ranged between 123-155oc, week 2, 130-165oc and week 3, 110150oc. a decline in the smoke point of frying oils collected from agudama, emekuku and wogu street for all the three weeks studied. for kaduna and railway, an increase was observed only at week 2, following this was a decrease. this was consistent with the findings of choudhary and grover [33] that a decrease in smoke point was observed after first, second and third frying of rice bran oil blends. according to sarwar, et al. [34] the smoke point of cooking oil must be at least 170oc. the findings of the present study were not in line with this statement as all the frying oil samples collected (except for the branded control) had smoke point <170oc even up to the third week of collection. this may be due to the level of impurities in the oil from the frying process. figure-5. smoke points (oc) of the frying oils collected at weekly intervals. 3.1.7. moisture content of frying oils result of the moisture content of the frying oils collected is shown in figure 6. for all the samples collected at week 1, the moisture content of the frying oils were found to be in the range of 0.13-0.20% with samples collected from agudama street recording lowest while samples from railway and the unbranded sample were highest. after three weeks of collection, the moisture was found to be in a range of 0.10-0.20% with frying oils from railway recording lowest and unbranded samples recording highest. however, there was no significant (p>0.05) difference in the moisture content of the frying oils studied. the frying oils collected had values below the 0.2% maximal limit for volatile matters at 105oc in oil and fats [35]. agriculture and food sciences research, 2020, 7(1): 89-96 94 © 2020 by the authors; licensee asian online journal publishing group figure-6. moisture content (%) of the oils collected at weekly intervals. 3.2. antioxidant properties of the frying oils 3.2.1. total phenolic content (tpc) result of the total phenolic content of the frying oils collected is presented in figure 7. total phenolic content of the frying oils at week 1, 2 and 3 ranged from 0.01-0.37mggae/g, 0.02-0.37mggae/g and 0.000.37mggae/g, respectively. there was a decrease in the tpc of frying oils collected from agudama, worgu, emekuku and railway streets at 3rd week of collection while those from kaduna recorded an increase. total phenolic content of control (branded oil) was significantly higher (p<0.05) than all other frying oils while total phenolic content of unbranded oil did not differ with oil collected from kaduna at 3rd week of collection and from railway, emekuku and agudama at first week of collection. the decrease in total phenolic content of oil during frying operations is as a result of the exposure of the frying oil to high temperature and long time during deep frying. the total phenolic content of the oils are close to that of güzel, et al. [36] for vegetable oils in turkey (0.133-1.596mggae/ml). the results are higher than that of xuan, et al. [37] for commercial vegetable edible oils marketed in japan (1.76-39.16mggae/g). results presented here are in correlation with that of gómez-alonso, et al. [38] where the reduction of antioxidant activity of olive oil correlated well with the number of deep-frying cycles. phenolics are known for their role in the oxidative stability of oils. their presence may also prevent deterioration of oils through quenching of radical reactions responsible for rancidity [39, 40]. . figure-7.total phenolic content (mggae/g) of the oils collected at weekly interval. 3.2.2. flavonoid content of the oils result of the flavonoid content of the frying oils collected is shown in figure 8. initial flavonoid content of the frying oils collected ranged from 0.77-2.20% with oil collected from wogu street observed to be the lowest while the branded oil was highest. at week 2 and 3, oils collected from agudama and kaduna streets were lowest in flavonoid content. flavonoid content of branded oil was significantly higher (p<0.05) than the frying oils collected while at week 3, it showed no significant difference (p>0.05) except for oil collected from kaduna street and unbranded oil. flavonoids are known for their antioxidant, anti-inflammatory, anti-atherosclerotic and anticarcinogenic properties [41]. they are among antioxidant defensive systems protecting vegetable oils against oxidative damage. agriculture and food sciences research, 2020, 7(1): 89-96 95 © 2020 by the authors; licensee asian online journal publishing group figure-8.flavonoid content (%) of the oils collected at weekly interval. 4. conclusion the study results showed degradation in the physicochemical and antioxidant properties of the vegetable cooking oil after repetitive use of frying. all oil samples collected at first week from wogu and kaduna streets were within an acceptable range for percentage free fatty acid and as the weeks progressed, they were above the permissible limit. there was also an increase in peroxide value of some oils while others showed a decrease, however all the frying oils collected were within the codex regulatory limit. all the oils collected showed a decrease in iodine value except for samples collected from agudama and railway close while increase in saponification value above the specification for the oils was observed. smoke point of the oils not in line with standard limits even up to third week of collection while moisture content was below the 0.3% maximum limit. total phenolic content of the oils were relatively low and this decreased as collection weeks progressed except for oils from kaduna street while a reverse was observed for flavonoid content. this study therefore shows the level of degradation in the quality of oils used by these fried food vendors during frying operations. safety measure should therefore be put in place as these degradations in oil quality could cause damaging health effects due to the toxic substances produced. degraded oils are not suitable for human consumption but can be used for alternate uses including biodiesel and soap production. references [1] f. a. rehab and a. m. el anany, "physicochemical studies on sunflower oil blended with cold pressed tiger nut oil during deep frying process," grasas y aceites, vol. 63, pp. 455-465, 2012. available at: https://doi.org/10.3989/gya.057612. 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[41] k. s. mala and a. e. kurian, "nutritional composition and antioxidant activity of pumpkin wastes," international journal of pharmaceutical, chemical and biological sciences, vol. 6, pp. 336-344, 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 50 © 2022 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 9, no. 2, 50-58, 2022 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v9i2.4110 © 2022 by the authors; licensee asian online journal publishing group valorization of garlic peel as a potential ingredient for the development of valueadded rice based snack product pukhelein jesparwell lyngdoh1 subhajit ray2 ( corresponding author) 1,2department of food engineering &technology, central institute of technology kokrajhar, kokrajhar, india. 1email: jesparwelltongkhar@gmail.com tel: +919501884237 2email: subhajit@cit.ac.in tel: +919330054980 abstract the present study was designed to develop and evaluate the nutritional, functional, sensory properties and texture profile data of garlic peel powder fortified viz. 2%, 5% and 10% value added pukhelein and compared with control sample. garlic peel showed crude fiber, carbohydrate and ash content viz. 60.57%, 21.71% and 16.34%, total phenolic content (tpc) of 108.79 µg/ml gae, total flavonoid content (tfc) of 33.27 mg quercetin/ml and antioxidant activity (33.38 mg/ml) respectively. 5% formulation exhibited the highest crude fiber, protein content, tpc (326.6 µg/ml gae), tfc (91.38 mg quercetin/ml) and total antioxidant content (54.68 µg/ml).sensory evaluation data showed good result in 5% formulation in terms of appearance, texture and taste. texture profile analysis showed that a considerable decrease in hardness, springiness, cohesiveness, gumminess and chewiness and resilience. the outcome of the study revealed that 5% garlic peel powder fortified pukhelein exhibited the best result among all formulations. keywords: garlic peel, utilization, functional components, sensory properties, value addition, pukhelein. citation | jesparwell lyngdoh; subhajit ray (2022). valorization of garlic peel as a potential ingredient for the development of valueadded rice based snack product pukhelein. agriculture and food sciences research, 9(2): 50-58. history: received: 18 may 2022 revised: 12 july 2022 accepted: 27 july 2022 published: 16 august 2022 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. authors’ contributions: both authors contributed equally to the conception and design of the study. competing interests: the authors declare that they have no conflict of interest. acknowledgment: authors are grateful to the department of food engineering &technology, central institute of technology kokrajhar, kokrajhar, assam, india for providing the necessary laboratory facilities to carry out this research work. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study followed all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 51 2. materials and methods ................................................................................................................................................................... 52 3. result and discussion .................................................................................................................................................................... 54 4. conclusion ......................................................................................................................................................................................... 57 references .............................................................................................................................................................................................. 57 mailto:jesparwelltongkhar@gmail.com mailto:subhajit@cit.ac.in https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v9i2.4110 https://orcid.org/0000-0003-0081-9836 agriculture and food sciences research, 2022, 9(2): 50-58 51 © 2022by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by emphasizing the development and evaluation of garlic peel powder fortified rice based innovative value-added snack product pukhelein with respect to nutrition, functional, sensorial and textural properties. 1. introduction garlic(allium sativum) is a species ofbulbous flowering plant in the genus allium and has been known since the ancient times to have many benefits. it is also used as a food ingredient all around the world because of its characteristic flavor and aroma. garlic is considered as a medicine in many countries as it is believed to treat and prevent diseases like high blood pressure, coronary heart disease, high attack and cholesterol. it is also used in cancer treatment like breast cancer, prostate cancer colon and bladder cancer. it contains allicin and has high antifungal and anti-viral properties [1]. through various studies, different parts of the plants have been shown to possess many properties which are valuable for functional ingredients for food and promoting health-beneficial effects [2, 3]. however, behind the well-known usefulness of garlic plant, there is one part of it which is still widely considered as waste. the garlic peel/skin is usually discarded directly without any further use [4].garlic peel could be used as an alternative for more costly waste water treatment processes. due to the high consumption of garlic high amount of peels were discard, in the community. in regards to environment this waste was used as an adsorbent to remove methylene blue from aqueous solution [5].garlic peels also has high antimicrobial and antioxidant properties and its possibility of using it as a preservative in cooked beef [6].there have been several efforts to study the potential of garlic peel even though they are still very limited. rahmawati, et al. [7] was explored the phytochemical activities of garlic peel extract and its potential utilization as a natural food additive and functional ingredient in the future. there studies are of two phases; in the first phase they identify and compares the phytochemical content and their activities between aqueous and ethanolic extract. in the later phase, they focus on their extraction method and it was found that the highest amount of antioxidant activity, flavonoid and phenolic content could be obtained in treatment of raw material-solvent ratio20gm/100ml. ifesan, et al. [6] carried out research on the properties of antioxidant and antimicrobial garlic peel extract and its possible way of using it as a natural food additive and preservative in cooked beef. chitsaz, et al. [8] did a study on the effects of garlic peel/peel in biochemical, haematological and digestive enzyme parameters of juvenile belunga species and its uses as feed formulation and there are also studies of which further agrees the effectiveness of garlic peel in marine organism and biochemical digestive enzyme, which was selected as a fish model due to its importance in commercial sturgeon aquaculture. hameed and ahmad [5] studied on the potential use of garlic peel for the removal of methylene blue from aqueous solution. garlic peels a cheap and waste material, was found to be very effective to remove methylene blue from aqueous solution. kallel, et al. [9] were reported on the phenolic extraction from the disposed garlic husk (gh) using different solvents (water, methanol and ethanol) and the result obtained stated that garlic husk is a potential source of natural antioxidant that can be used in different medicinal, biological and in food industry application. based on the evidence provided by the studies above, there might be a possibility to utilize garlic peel and garlic peel extract as a natural food additive or even as a functional food or a value addition to any product in the future. this study was to determine the proximate composition, flavonoid content, phenolic content, antioxidant capacity and to investigate the nutritional, functional, textural and sensory qualities of value added garlic peel powder fortified pukhelein. table 1 represents the availability of phytochemical components in garlic peel with respect to the ethanol and water extraction. table 1. list of phytochemical/functional components in garlic peel. secondary metabolites ethanol extract garlic peel water extract garlic peel alkaloid saponin + + agi antimicrobial activity(s. cerevisiae) + flavonoid + + antioxidant activity + + phenolics + + note: +indicates presence, indicates absences [10]. table 2 represents the applicability of different categories of garlic waste in various food products and their beneficial activities. table 2. summary table of potential application of different forms of garlic waste. form of garlic waste application matrix/ final product function/ benefits reference garlic peel powder cooking rice add vitamins and nutrients [11] garlic peel powder wheat bread bread with enhance antioxidative properties [11] ginger peel powder wheat flour extrudate enhance vitamin and antioxidant [12] garlic peel extract soup enhance soup with flavor and nutritional component [11] garlic peel powder beef use as a food additive [13] garlic peel ash hair strengthen the hair [12] garlic extract face treat against itchy skin [11] agriculture and food sciences research, 2022, 9(2): 50-58 52 © 2022by the authors; licensee asian online journal publishing group 2. materials and methods 2.1 collection of samples garlic samples were purchased from the local market adjacent to the institute, kokrajhar, assam, india. figure 1 illustrates the manufacturing flow diagram of garlic peel powder. figure 1. processing of garlic peel powder. 2.2. determination of proximate composition 2.2.1. determination of moisture content moisture content of garlic peel was determined by the following formula according to the description of aoac [14]. moisture (%) = (w1w2) x 100 w where, w1 = weight of the sample and the dish before drying. w2 = weight of the dish after drying. w = weight of the sample. 2.2.2. determination of ash content ash content of garlic peel was determined by using muffle furnace as described by aoac [14]. ash (%) = w2-w1 x 100/ ws where, w2 = wt. of the crucible with ash. ws = wt. of the sample. w1 = weight of the crucible. 2.2.3. determination of protein content protein content of garlic peel was determined as described by lowry, et al. [15]. 2.2.4. determination of crude fiber crude fiber of garlic peel was determined by the following formula as described by aoac [14]. crude fiber % by wt. = (w 1 – w2) x100 w where, w1 = wt. after drying. w2 = wt. after ashing w = wt. of the sample 2.2.5. determination of crude fat content crude fat of garlic peel was determined by the following formula as described by aoac [14]. % crude fat = (w2 – w1) x 100 w where, w1weight of the flask. w2weight of the flask and extracted fat, wweight of the sample 2.2.6. determination of carbohydrate content carbohydrate content of garlic peel was determined by using the formula as described in anthrone method [16, 17]. concentration of unknown= [(absorbance of unknown*concentration of standard) /absorbance of standard] agriculture and food sciences research, 2022, 9(2): 50-58 53 © 2022by the authors; licensee asian online journal publishing group 2.3. extraction of sample extraction of samples was done by taking 1 g of garlic peel powder suspended in 35 ml of methanol were mixed and kept for 1 hour in a sonication bath. the extract was mixed rapidly by centrifuge for 10minutes. the extract was filtered using whatman filter paper and is store in the dark prior further analysis. 2.3.1. determination of total phenolic content (tpc) total phenolic content of garlic peel was estimated spectrophotometrically [18]. 2.3.2. determination of total flavonoid content total flavonoids content of garlic peel was estimated by the method described by malik and singh [19]. 2.3.3. estimation of total antioxidant capacity by dpph (radical scavenging activity) total antioxidant capacity (radical scavenging activity) was measured by using the free radical,dpph (2,2diphenyl-1picrylhydrazyl) [20]. radical scavenging activity (%) = [(ao-as)/ao] x 100 where, ao is absorbance of control (blank) and as is absorbance of sample extract 2.4. utilization of garlic peel powder in traditional food for the development of value-added product garlic peel powder contributes a significant role for the development of value added pukhelein. figure 2 illustrates the manufacturing flow diagram of garlic peel powder fortified value added pukhelein. figure 2. development of garlic peel powder fortified traditional rice based product (pukhelein). figure 3. garlic peel powder fortified pukhelein. figure 3 illustrates the development of value added garlic peel powder fortified rice based snack product pukhelein. 2.5. texture profile analysis texture profile analysis (tpa) was followed by instrumental method in terms of the different textural parameters e.g. hardness, chewiness, cohesiveness, gumminess, resilience and springiness [21]. 2.6. sensory evaluation evaluation of sensory parameters was performed by nine point hedonic rating test as described by ray [22]. 2.7. development of valueadded rice based snack pukhelein ricebased snack pukhlein were prepared by incorporating of garlic peel powder in various proportions in rice flour. the product was shown in figure 4. agriculture and food sciences research, 2022, 9(2): 50-58 54 © 2022by the authors; licensee asian online journal publishing group figure 4. pukhlein (control and fortified with different proportions of garlic peel powder). table 3. proximate composition of garlic peel. parameters (%) garlic peel ash 16.34 carbohydrates 21.71 crude fiber 60.57 fat 0.83 moisture content 3.36 protein 8.24 3. result and discussion 3.1. proximate composition experimental result obtained from table 3 showed that garlic peel was a very rich source of ash, crude fiber and protein content. however, a very negligible amount of lipid exists. less moisture content established the profound by product stability. the result was significantly comparable with the result obtained by kallel, et al. [9]. significant mineral content of garlic peel indicates its potentiality of fortification and at the same time higher level of crude fiber directs its digestibility. table 4. total phenolic, flavonoid and antioxidant content. functional parameters result total phenolic content (gallic acid equivalent µg/ml) 108.7961351 total flavonoid content (mg quercetin/ml) 33.27 antioxidant activity (mg/ml) 33.38 3.2. phytochemical / functional composition in garlic peel extract experimental result obtained from table 4 showed that the total phenolic as 108.79 µg/ml gallic acid equivalent, total flavonoid33.27mg/ml quercetin and antioxidant activity of 33.38 mg/ml respectively. tpc value showed higher value as compared with the result of syeda, et al. [23]. similarly, total flavonoids content (tfc) exhibited significantly enhanced value than the result obtained from syeda, et al. [23]. similarly, antioxidant activity by dpph radical scavenging assay method exhibited the higher value than the experimental investigation of syeda, et al. [23]. moreover, the result revealed that garlic peel has a potential to be regard as an anticancer as phenolic was used in cancer treatment [1]. decades of research suggest people who eat more generous amount of antioxidant-rich food have increased protection against disease. protection of food product was related to antioxidant ability responsible for providing fight against free radicals and reduces oxidative stress. the result of the study showed better result in terms of methanol extracted total antioxidant capacity (tac) or antioxidant activity than kotenkova and kupaeva [24]. 3.3. proximate composition of garlic peel powder fortified pukhelein the proximate composition of value-added rice-based snack (pukhlein) will be represented in table 5. table 5. proximate composition of value added pukhelein. parameters control 2% 5% 10% ash 0.39 0.51 0.56 0.53 carbohydrates 54.7 55.4 55 55.5 fats 16.1 15.8 15.3 14.6 crude fibre 2.39 2.53 5.1 3.8 protein 4.07 5.9 6.05 5.8 moisture 20.2 19.8 19.3 18.4 experimental result revealed that the incorporation of garlic peel powder increases the ash and crude fiber in the value-added product (pukhlein). higher ash content means high in mineral content in the garlic peel. similarly, agriculture and food sciences research, 2022, 9(2): 50-58 55 © 2022by the authors; licensee asian online journal publishing group the crude fibre is increased as the garlic peel is rich in crude fibre. the incorporation of garlic peels exhibited increasing protein content but decreasing fat content in the valuedadded product. therefore, while comparing all formulations of garlic peel powder fortified value added product, 5% formulation showed better result in terms of increased level of ash, crude fiber and protein content. 3.4. functional potential of valueadded pukhlein figure 5. functional potential of value-added rice based snack (pukhlein) with different proportion of garlic peel powder. figure 5 showed that 5% garlic peel powder fortified value added pukhelein contains maximum amount of total phenolic content (tpc), total flavonoids content (tfc) and total antioxidant capacity (tac). in general garlic peel powder addition to pukhelein enhanced the functional potential due to the occurrence of function components in significant level. moreover, 5% formulation showed the better result due to possibly the release of potential functional components in comparison to other. 3.5. sensory evaluation figure 6. sensory evaluation of value-added rice snack (pukhlein) with different proportion of garlic peel powder figure 6 illustrates the evaluation of sensory parameters of formulated garlic peel powder value added pukhelein.sensory evaluation was done by using nine point hedonic rating test. in this investigation, the sensory panel was constituted by twenty five panellist including faculty members, pg and ph.d.scholars of the institute. based on the evaluation of panellist, it was found that 5% garlic peel powder fortified rice based snack pukhelein exhibited the best result in terms of appearance, taste, aroma, texture and overall acceptability in comparison to other formulations. no such comparative data of other researchers were found. 3.6. textural characteristics of value-added rice snack (pukhlein) with different proportion of garlic peel powder figure 7. hardness of fortified pukhelein. agriculture and food sciences research, 2022, 9(2): 50-58 56 © 2022by the authors; licensee asian online journal publishing group figure 7 represents the hardness as one of the important textural attribute of value added garlic peel powder rice based snack pukhelein. figure 8. springiness of fortified pukhelein. figure 8 represents the springiness as one of the important textural attribute of value added garlic peel powder rice based snack pukhelein. figure 9. cohesiveness of fortified pukhelein. figure 9 represents the cohesiveness as one of the important textural attribute of value added garlic peel powder rice based snack pukhelein. figure 10. gumminess of fortified pukhelein. figure 10 illustrates the gumminess as one of the important textural attribute of value added garlic peel powder rice based snack pukhelein. agriculture and food sciences research, 2022, 9(2): 50-58 57 © 2022by the authors; licensee asian online journal publishing group figure 11. chewiness of fortified pukhelein. figure 11 illustrates the chewiness as one of the important textural attribute of value added garlic peel powder rice based snack pukhelein. figure 12. resilience of fortified pukhelein. figure 12 illustrates the resilience as one of the important textural attribute of value added garlic peel powder rice based snack pukhelein. experimental investigation based upon various figures viz. hardness, springiness, cohesiveness, chewiness, gumminess and resilience obtained from texture profile analysis (tpa) revealed the significance of 5% garlic peel powder in comparison to other formulations e.g.2%, 10% and even control i.e.0% fortified pukhelein due to decreased value. these characteristics established it as a novel product. no such comparative data of other researchers were found in this endeavour. 4. conclusion in this study, garlic peel as a valuable by product was considered for the development of value added product pukhelein. different properties viz. nutritional, functional, sensory and textural attributes were analyzed for conrol,2%,5% and 10% garlic peel powder fortified value added product. finally, after considering all analytical datas and sensory perception it was established that 5% formulation was considered to be the best one. references [1] s. ejaz, i. chekarova, j. w. cho, s. y. lee, s. ashraf, and c. w. lim, "effect of aged garlic extract on wound healing: a new frontier in wound management," drug and chemical toxicology, vol. 32, pp. 191-203, 2009.available at: https://doi.org/10.1080/01480540902862236. [2] s. durairaj, s. srinivasan, and p. lakshmanaperumalsamy, "in vitro antibacterial activity and stability of garlic extract at different ph and temperature," electronic journal of biology, vol. 1, pp. 5–10, 2009. [3] h. amagase, b. l. petesch, h. matsuura, s. kasuga, and y. itakura, "intake of garlic and its bioactive components," the journal of nutrition, vol. 131, pp. 955s-962s, 2001.available at: https://doi.org/10.1093/jn/131.3.955s. [4] l. natalie, "10 good reason to save onion and garlic peel. retrieved from farmer’s almanac: https://www.farmersalmanac.com," 2021. [5] b. hameed and a. ahmad, "batch adsorption of methylene blue from aqueous solution by garlic peel, an agricultural waste biomass," journal of hazardous materials, vol. 164, pp. 870-875, 2009.available at: https://doi.org/10.1016/j.jhazmat.2008.08.084. [6] b. o. t. ifesan, e. a. fadipe, and b. t. ifesan, "investigation of antioxidant and antimicrobial properties of garlic peel extract (allium sativum) and its use as natural food additive in cooked beef," international journal of scientific reports, vol. 3, pp. 711–721, 2014. [7] d. rahmawati, d. andika, and s. a. fortunata, "gaelic peel extract phytochemical evaluation and extraction optimization," journal of functional food and nutraceutical, vol. 1, pp. 41-46, 2019. http://www.farmersalmanac.com,/ agriculture and food sciences research, 2022, 9(2): 50-58 58 © 2022by the authors; licensee asian online journal publishing group [8] h. chitsaz, h. oraji, a. keramat amirkolaei, and r. akrami, "effect of garlic peel on haematological, biochemical and digestive enzyme activity in beluga juvenile (huso huso)," iranian journal of aquatic animal health, vol. 4, pp. 13-28, 2018.available at: https://doi.org/10.29252/ijaah.4.1.13. [9] f. kallel, d. driss, f. chaari, l. belghith, f. bouaziz, r. ghorbel, and s. e. chaabouni, "garlic (allium sativum l.) husk waste as a potential source of phenolic compounds: influence of extracting solvents on its antimicrobial and antioxidant properties," industrial crops and products, vol. 62, pp. 34-41, 2014.available at: https://doi.org/10.1016/j.indcrop.2014.07.047. [10] b. divya, b. suman, m. venkataswamy, and k. thyagaraju, "a study on phytochemicals, functional groups and mineral composition of allium sativum (garlic) cloves," international journal of current pharmaceutical research, vol. 9, pp. 42-45, 2017.available at: https://doi.org/10.22159/ijcpr.2017.v9i3.18888. [11] a. toppo, good reason to use garlic and onion peels: kj krishi jagran, 2020. [12] a. s. longhurst, "garlic and health, healthline.com. retrieved: https://www.healthline.com," 2019. [13] b. o. t. ifesan, s. siripongvutikorn, n. hutadilok-towatana, and s. p. voravuthikunchai, "evaluation of the ability of eleutherine americana crude extract as natural food additive in cooked pork," journal of food science, vol. 74, pp. 353-57, 2009. [14] aoac, official method of analysis. washington dc: association of official analytical chemists, 2000. [15] c. o. lowry, n. rosebrough, a. farr, and r. randall, "protein measurement with the folin phenol reagent," journal of biological chemistry, vol. 193, pp. 265-75, 1951.available at: https://doi.org/10.1016/s0021-9258(19)52451-6. [16] e. e. layne, "methods in enzymology," vol. 3, p. 447, 1975. [17] d. t. plummer, an introduction to practical biochemistry, 3rd ed. uk: mcgraw hill, 1990. [18] aoac, . official method of analysis. washington dc: association of official analytical chemists, 2017. [19] e. p. malik and m. b. singh, plant enzymology and hittoenzymology, 1st ed. new delhi: kalyani publishers, 1980. [20] w. brand-williams, m. cuvelier, and c. berset, "use of free radical method to evaluate antioxidant activity," lebensmittelwissenschaft & technologie, vol. 28, pp. 25-30, 1990. [21] t. p. guinee, cheese rheology. in: fuquat, j.w. &fox,p.f.(eds.): encyclopedia of dairy sciences, 2003. [22] s. ray, sensory properties of foods and their measurement methods. m.s.khan and m.shafiur rahman (eds).techniques to measure food safety and quality. switzerland: springer nature, 2021. [23] a. z. n. syeda, a. irfan, f. z. ameer, z. muhammad, m. anum, j. c. amar, and a. shomaila, "determination of antimicrobial and antioxidant potential of agro-waste peels," proceedings of the brazilian academy of sciences, vol. 92, pp. 1-12, 2020.available at: https://doi.org/10.1590/0001-3765202220200515. [24] e. a. kotenkova and n. v. kupaeva, "comparative antioxidant study of onion and garlic waste and bulbs," iop conference series: earth and environmental science, vol. 333, p. 012031, 2019.available at: https://doi.org/10.1088/1755-1315/333/1/012031. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.healthline.com,/ 136 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 136-145, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6227 © 2024 by the authors; licensee asian online journal publishing group encapsulation of vegetable oils in polylactic acid nanofibers to improve oil retencion in feed aquaculture aline moreira de alcantara1 tulio pacheco boaventura2 renan rosa paulino3 priscila vieira e rosa4 julio cesar ugucioni5 juliano elvis oliveira6 ( corresponding author) 1,2federal university of lavras, lavras, minas gerais, brazil. 1email: alcântara@ufla.com.br 2email: tuliopb1@hotmail.com 3,4department of animal science, federal university of lavras, lavras, minas gerais, brazil. 3email: renan.paulino@ufla.br 4email: priscila@ufla.br 5department of physics, federal university of lavras, lavras, minas gerais, brazil. 5email: julio.ugucioni@ufla.br 6department of engineering, federal university of lavras, lavras, minas gerais, brazil. 6email: juliano.oliveira@ufla.br abstract the feed used in fish farming contains lipids that erode and disperse in water, adversely affecting fish nutrition and contributing to water pollution. this study evaluated the encapsulation of corn and gold linseed oils in polylactic acid (pla) nanofibers produced via solution blow spinning. the nanofibers were characterized using fourier-transform infrared (ftir) spectroscopy, scanning electron microscopy (sem), contact angle analysis, and encapsulation efficiency tests. additionally, the release profiles of the oils were studied under simulated tank water and gastrointestinal conditions. the results demonstrated successful encapsulation of the oils within the pla matrix. ftir analysis confirmed the presence of oil in the nanofibers (wavenumbers 2925 and 2853 cm-1), with encapsulation efficiencies ranging from 81.54±10.36% to 99.14±4.55%. sem revealed uniform nanofiber morphology with smooth surfaces and average diameters between 157±49 and 385±133 nm. all nanofibers exhibited contact angles above 90°, indicating hydrophobic behavior. feed samples containing free oils showed significantly higher lipid release into water compared to feed incorporating encapsulated oils. the findings suggest that encapsulating vegetable oils in pla nanofibers reduces lipid dispersion in water while maintaining nutrient availability. this polymeric system offers a sustainable alternative to enhance aquaculture feed efficiency and reduce environmental impacts, contributing to cleaner water and healthier fish. keywords: farming, formulation, nutricion, oil leakage rate, pellet, solution blow, spinning. citation: |alcantara, a. m. de, boaventura, t. p., paulino, r. r., rosa, p. v. e, ugucioni, j. c., & oliveira, j. e. (2024). encapsulation of vegetable oils in polylactic acid nanofibers to improve oil retencion in feed aquaculture. agriculture and food sciences research, 11(2), 136–145. 10.20448/aesr.v11i2.6227 history: received: 31 october 2024 revised: 6 december 2024 accepted: 10 december 2024 published: 20 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the conselho nacional de desenvolvimento científico e tecnológico-cnpq brazil (grant number: cnpq-brasil –150480/2023-7) and fundação de amparo a pesquisa de minas gerais–fapemig (grant number: fapemig-brasil –apq-05593-24, bpd-00406-22, apq-02146-21), and coordenação de aperfeiçoamento de pessoal de nível superior – capes-brazil. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: conceptualization, methodology, validation, formal analysis, investigation and writing – original draft, a.m.d.a., t.p.b. and r.r.p.; methodology, validation, formal analysis, investigation, resources, data curation, investigation and writing, p.v.e.r. and j.c.u.; methodology, validation, formal analysis, investigation, resources, data curation, investigation, reviewing and editing, visualization, supervision, project administration and funding acquisition, j.e.d.o. all authors have read and agreed to the published version of the manuscript. mailto:alcântara@ufla.com.br mailto:tuliopb1@hotmail.com mailto:renan.paulino@ufla.br mailto:priscila@ufla.br mailto:julio.ugucioni@ufla.br mailto:juliano.oliveira@ufla.br https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6227 https://orcid.org/0000-0002-7343-2280 https://orcid.org/0000-0002-6143-5417 https://orcid.org/0000-0001-7915-5694 https://orcid.org/0000-0002-0052-079x https://orcid.org/0000-0002-6362-5919 https://orcid.org/0000-0001-9570-8308 agriculture and food sciences research, 2024, 11(2): 136-145 137 © 2024 by the authors; licensee asian online journal publishing group contents 1. introduction .................................................................................................................................................................................... 137 2. materials and methods ................................................................................................................................................................. 138 3. results .............................................................................................................................................................................................. 139 4 discussions ....................................................................................................................................................................................... 142 5. conclusion ....................................................................................................................................................................................... 143 references ............................................................................................................................................................................................ 144 contribution of this paper to the literature this article proposes the use of nanotechnology as a sustainable solution for incorporating vegetable oil into fish feed, enhancing fish nutrition while reducing nutrient waste and water contamination. 1. introduction the fish feed should contain proteins, lipids, vitamins, and carbohydrates for efficient maintenance of adequate animal growth within the shortest time [1]. it is important to give special attention to the balance between intake, retention, and excretion, because excessive intake of certain nutrients is responsible for intoxication events and, consequently, water pollution [2, 3]. water contamination from fish farms can occur due to therapeutic residues, fish pathogens, and feed‐derived wastes, and the loss of these ingredients is also related to economic disadvantages [4]. depending on production systems in fish farming and particularly the kind of effluent treatment, the waste load per ton of fish production can become a serious environmental challenge [3]. monitoring and treating this feed waste in effluent directly is an inaccurate and costly process. the remainder of the undigested feed is dispersed on water (fat, vitamins, carbohydrates, and proteins) or excreted in the feces as solid waste, and the byproducts of fish metabolism are excreted as dissolved waste (ammonia, urea, phosphate). concerns about the oils used in feed fortification also include how their waste will affect water quality since they are highly related to aquatic pollution. oil leaching can lead to the formation of a mechanical barrier in the water surface, preventing light and oxygen from penetrates the water column and consequently reducing photosynthesis and phytoplankton formation. the decrease of phytoplankton in the aquatic medium is also responsible the reducing oxygen supply to the water column [5]. therefore, the formulation of diets with low environmental impact is necessary to avoid damage to water quality in aquaculture production systems [6]. in this context, the use of controlled release systems containing natural products in feed formulation is an alternative to reduce water contamination from fish farming. fish meal and oil play an important role as lipid source in the fish diet. because of their rich content in highly unsaturated fatty acids, particularly eicosapentaenoic (epa) and docosahexaenoic (dha) acids, they act as nutritional supplements [2]. essential unsaturated fatty acids promote fish growth by regulating metabolism and improving immunity. the problem is that the overuse of fish meal and oil in farmed fish diets results in higher nitrogen and phosphorus discharges into the water systems, which causes eutrophication [7, 8]. moreover, these products are very expensive and obtained from finite sources, therefore their use represents not only an environmental but also an economic problem. focused on developing more sustainable aquaculture processes several authors have been studying alternative lipid sources to replace fish oil [9, 10]. gold linseed oil (lo) and corn oil (co) represent a good alternative to fulfill this role, since they are a sustainable source of fatty acids that are metabolized by the fish organism, leading to the formation of long-chain polyunsaturated essential fatty acids of the n-3 (dha and epa) and n-6 (arachidonic acid) series, respectively. also, both corn and gold linseed oils are highly digested by all fish species. another advantage of their use is that they do not contain dioxins and pollutants [11] and some studies have demonstrated that these oils would improve the diet conversion rate of fish as well as fish health [12]. in recent years, there has been an increasing number of authors in the literature that studying the possibility of improving the composition and quality of animal feed using nanostructured materials in feed formulations [13-17]. oral administration of nutrients to farmed fish brings inherent challenges that must be considered when designing a nanostructured carrier. furthermore, each compartment of the gastrointestinal tract has a unique environment that includes its complement of enzymes and specific ph levels. nanofibers can overcome these challenges and deliver their nutrients to the intestine, where most of the absorption occurs, then traveling to the bloodstream and finally being distributed through all the organism [18]. techniques for obtaining polymeric nanofibers have been developed in the last decade. solution blow spinning (sbs) is a fiber manufacturing process that uses two parallel concentric nozzles. nanofiber production by sbs depends on several factors such as concentration and viscosity of the polymeric solution, ejection rate, air pressure, and working distance. these variables have a direct influence on the fiber production rate and fiber morphology [19-21]. in comparison with other techniques, such as electrospinning, the sbs technique has a higher ejection rate, fiber production, and versatility regarding the use of solvents, polymers, and applications, in addition to its safety and low cost [22, 23]. nanofibers obtained by the sbs technique have several applications in animal science like as controlled delivery of progesterone to control the estrus cycle in livestock animals [24] and encapsulation of diclofenac sodium to create in situ devices for pododermatitis treatment [25]. thus, it is notable that immediate improvements in fish feed formulations are needed for a more efficient administration of nutrients, which involves reducing waste and water contamination from fish farming and increasing the biodisponibility of nutrients in the intestinal tract. with this aim, we now report the development and physicochemical characterization of hydrophobic polylactic acid (pla) nanofibers containing different ratios of gold linseed and corn oil, produced by the solution blow spinning technique. we posit that biodegradable and hydrophobic nanofibers could protect fatty acid against release in the aquatic environment and stomach chamber, improving fatty acid release effectiveness and, thus, creating the potential for a smart fish feed with the lowest environmental impact. agriculture and food sciences research, 2024, 11(2): 136-145 138 © 2024 by the authors; licensee asian online journal publishing group 2. materials and methods 2.1. materials polylactic acid (3251 d, 66kda, chemical abstracts service number 9051-89-2) was obtained from nature works (usa). corn oil (brand lizza), gold linseed oil (brand pazze), and the materials used in feed production were purchased locally. chloroform (chemical abstracts service number 67-66-3) was obtained from synth (brazil). the lipophilic dye sudan iii (chemical abstracts service number 85-86-9) was purchased from êxodo científica (brazil). 2.2 solution and fiber production the polylactic acid solution was prepared as follows: 12% (w/v) pla was dissolved in chloroform under continuous stirring for 40 minutes. after the complete polymer dissolved, 50% (w/w) of vegetable oils were added to the solution [26, 27]. all the different fractions of corn oil (co) and gold linseed oil (lo) used are shown in table 1. the solution was kept under stirring and at room temperature for another 15 minutes. finally, 0.125% (w/w) of the lipophilic dye sudan iii was added, and it were kept under stirring for 15 minutes. table 1. different fractions of corn oil and gold linseed oil used to formulate the polylactic acid solutions that will pass through the solution blow spinning process. sample code corn oil (%v/v) linseed oil (%v/v) co:lo – 100:0 100 0 co:lo – 75:25 75 25 co:lo – 50:50 50 50 co:lo – 25:75 25 75 co:lo – 0:100 0 100 pla nanofibers 0 0 the fibers were obtained using the solution blow spinning technique. the spinning system was formed consisted of a source of compressed air (chiaperini, model mc 12 bpv,150 liters), a polymer injection pump (new era pump systems, model syringe pump al1000), and a glass syringe (20 ml and 19.33 mm). a rotating horizontal cylinder covered with aluminum paper was used to collect the nanofibers. the experimental apparatus was isolated inside a glass front-opening wooden box to maintain moisture (40-50%) and temperature control (approximately 30°c, using a reflective incandescent light bulb – tovalight e.27) throughout the process. the process took 3 hours for each sample and the spinning parameters used were adjusted based on the methodologies nepomuceno, et al. [27] and bonan, et al. [26]: feed rate – 6.00 ml/h; working distance – 20 cm; protrusion between the two concentric nozzles – 0.4 mm; air pressure – 172.37 kpa. the fibers were collected and stored for subsequent characterization. 2.3. fiber characterization the absorption spectrum of the samples was obtained by fourier-transform infrared spectroscopy (ftir) with the attenuated total reflection (atr) sampling technique. a total of 32 scans were considered for a wavenumber range of 4000-400 cm-1 with a resolution of 4 cm-1 [28]. nanofibers were coated with a gold layer by sputtering (balzers, sputter coating device 050) for the analysis of their morphology using a scanning electron microscope (sem). the sem system used was a leo evo 40 model (carl zeiss) equipped with extended variable pressure (xvp), bruker’s quantax eds for x-ray microanalysis, and a cryosystem from gatan. for each experiment, the average fiber diameter was determined with the image analysis software image j, using approximately 100 measurements randomly taken from the samples [29]. the hydrophobicity/hydrophilicity of the nanofibers was evaluated through contact angle measurement immediately, 5 seconds, and 60 seconds after the injection of a sessile drop on the nanofiber. the experiment was performed with a kruss drop shape analyzer goniometer – dsa25 (hamburg, germany). to determine encapsulation efficiency, 30 mg of the nanofibers were dissolved in 10 ml of chloroform and the solution was quantified in a uv-vis (ultraviolet-visible spectroscopy) spectrophotometer (shimadzu, model uv2601) at a wavelength of 513 nm. this process was performed in 5 different regions of each sample and the encapsulation efficiency was determined through equation 1 (designation). 𝐸𝐸 (%) = 𝐶𝐸 𝐶𝐼 ⁄ 𝑥 100% (1) where: ce = concentration of sudan iii encapsulated in nanofibers (average calculated using the 5 regions); ci = concentration of sudan iii initially added to the nanofibers. 2.4. feed production a formulation simulating a conventional feed for omnivorous fish in the growth phase (the largest group of fish cultivated in the world) was produced: 45% soybean meal, 34.8% cornflour, 11.5% corn gluten meal 60, 5% corn oil/gold linseed oil, 1.5% dicalcium phosphate, 1.5% mineral and vitamin supplement, 0.5% sodium chloride, 0.19% dl-methionine, 0.125% sudan iii (a chemical marker used to track the release of vegetable oils), and water was added until the mixture obtain the proper consistency. the mixture was pelletized in an extruder (adamo) and the product was dehydrated in a laboratory oven at 50°c for 24 hours [30]. six feed samples were produced, containing contained the same corn oil/gold linseed oil fractions proposed in table 1. 2.5. assessment of nutrient liberation in an aqueous medium (in vitro) approximately 30 mg of feed and feed containing nanofibers were placed into a falcon tube containing 3 ml of water, 1% (v/v) tween 80, sodium bicarbonate (40 mg/l), and a ph equal to 7.0 [31]. the tubes were kept in an note: polylactic acid (pla), corn oil (co) and gold linseed oil (lo). agriculture and food sciences research, 2024, 11(2): 136-145 139 © 2024 by the authors; licensee asian online journal publishing group incubator under at 25°c and 120 rpm [32]. this process represents the preparation of one aliquot that was subsequently analyzed in a uv-vis spectrophotometer (shimadzu, model uv-2601) around at wavelength of 534 nm. for each sample, 10 aliquots were prepared and collected after 10 minutes. 2.6. assessment of in vitro digestibility the uv-vis spectrophotometer (shimadzu, model uv-2601) was used to measure absorbance around a wavelength of 534 nm. tests of in vitro digestibility were based the methodology wang, et al. [32]. the simulated gastric fluid of fish was prepared as follows: 20 mg of feed and feed containing nanofibers were added to an erlenmeyer containing 7 ml of a sodium chloride (0.15 m) solution, 5% (w/v) of pepsin, and 1% (v/v) of tween 80; the solution was posteriorly acidified with hcl until the ph of 2.0. the solution was maintained in an incubator for 1 hour in the dark (the recipients were covered with aluminum paper to prevent the passage of ambient light), at 25°c, and 100 rpm. the simulated intestinal fluid of fish was prepared as follows: 20 mg of feed and feed containing nanofibers were added to an erlenmeyer flask containing 7 ml of a sodium chloride (0,15 m) solution, 1.5% (w/v) of pancreatin, 0.5% (w/v) of amylase, 0.3% (w/v) of bile salts, and 1% (v/v) of tween 80; the solution ph was posteriorly adjusted to 6.0. the solution was maintained in an incubator for 5 hours in the dark (the containers were covered with aluminum foil paper to prevent the passage of ambient light), at 25°c, and 40 rpm. the tests described in the previous paragraphs were also performed for the 6 feed samples containing different fractions of the free vegetable oils in its formulations. the presence of lipids in the solutions was evaluated using uv-vis spectrophotometer. 2.7. statistical analysis all data were subjected to the shapiro-wilk normality test and levene's test for homoscedasticity. parametric data were analyzed by variance analysis (anova) (one-way or twoway) followed by tukey's test at 5% probability. past software were used for data analysis. 3. results fiber characterization. 3.1. scanning electron microscopy (sem) figure 1 shows the sem images obtained for all the nanofibers produced in this paper, demonstrating that they exhibited uniform morphology and smooth surface. table 2 shows average diameter values for each sample, calculated using the software image j. table 2. average fiber diameter and standard deviation of pla nanofibers containing different fractions of vegetable oils. sample co: lo 100:0 co: lo 75:25 co: lo 50:50 co: lo 25:75 co: lo 0:100 pla nanofibers average diameter (nm) 168±52d 209±56c 385±133a 257±83b 157±49d 167±46d note: the letters a, b, c, and d in the table indicate statistical differences between results. results followed by different letters differ significantly between treatment (p<0.05). ony-way anova and tukey’s test were used to determine statistical significance. figure 1. scanning electron microscopy of nanofibers containing: (a) co: lo – 100:0, (b) co: lo 75:25, (c) co: lo 50:50, (d) co: lo 25:75, (e) co: lo – 0:100, (f) pla nanofibers. according to statistical analysis, higher values of average diameter were obtained for pla nanofibers containing mixtures of corn oil and gold linseed oil (cl: lo – 50:50). the incorporation of pure oils did not affect agriculture and food sciences research, 2024, 11(2): 136-145 140 © 2024 by the authors; licensee asian online journal publishing group the samples average diameter and the minor values of average diameter were obtained for pla nanofibers, co: lo 0:100 and co: lo – 100:0. 3.2 fourier transform infrared spectroscopy (ftir) figure 2 shows the ftir spectra obtained from the different samples. the infrared analysis reveals peaks in the spectra of the various samples produced. notable peaks were observed in the regions around 3000, 2925, and 2853 cm-1. figure 2. the infrared spectrum of pla nanofibers containing corn oil and gold linseed oil in different proportions and of the pure vegetable oils in the wavenumber region from 4000 to 400 cm-1. this region corroborates the encapsulation of the vegetable oils. 3.3. contact angle table 3 shows the average contact angle values 60 seconds after the injection of water drop onto the surface of the nanofibers. all samples are hydrophobic since the contact angle was higher than 90° for all nanofibers. the highest contact angle were observed for the samples co: lo – 50:150 and co: lo – 25:75. the minors contact angle value were found for the samples pla nanofibres and co: lo – 0:100. table 3. contact angles of pla nanofiber mat and pla/vegetable oils blend nanofiber mats with their standard deviations. sample average contact angle (°) co: lo – 100:0 109.9±2.7 ac co: lo – 75:25 116.7±7.4ab co: lo – 50:50 106.3±1.8 c co: lo – 25:75 107.8±3.4 c co: lo – 0:100 114.5±2.7a pla nanofibers 114.6±4.8a note: the letters a, b and c in the table indicate statistical differences between results. results followed by different letters differ significantly between treatment (p<0.05). ony-way anova and tukey’s test were used to determine statistical significance. 3.4. encapsulation efficiency table 4 shows the encapsulation efficiency for each formulation. the results reveal that the co: lo – 50:50 sample had the lowest encapsulation efficiency (81.54±10.36%). the highest encapsulation efficiency was obtained for samples co: lo –100:0 (99.14±4.55 %) and co: lo – 0:100 (97.32±11.09 %). agriculture and food sciences research, 2024, 11(2): 136-145 141 © 2024 by the authors; licensee asian online journal publishing group table 4. encapsulation efficiency of polylactic acid nanofibers containing different fractions of vegetable oils. sample encapsulation efficiency (%) co:lo – 100:0 99.14±4.55a co:lo – 75:25 94.06±9.47abc co:lo – 50:50 81.54±10.36b co:lo – 25:75 85.97±4.72 abc co:lo – 0:100 97.32±11.09 ac note: the letters a, b and c in the table indicate statistical differences between results. results followed by different letters differ significantly between treatment (p<0.05). ony-way anova and tukey’s test were used to determine statistical significance. 3.5. assessment of nutrient liberation in an aqueous medium (in vitro) in figure 3 it is possible to observe the fraction of oil released into the water during 10 minutes of exposure of the different materials. when comparing the composition of the feed samples with and without oil encapsulated in nanofibers, it is observed that the co: lo – 75:50 and co: lo – 50:50 samples exhibited statistically similar oil release. the co: lo – 100:0, co: lo – 25:75, and co: lo – 0:100 samples, containing oil encapsulated in nanofibers, showed lower oil release compared to the samples with free oil in the feed. when comparing oil release across different feed compositions, the co: lo – 0:100 sample showed the highest release. for feeds with oil encapsulated in nanofibers, the co: lo – 50:50 sample recorded the highest oil release. figure 3. comparison between the maximum quantity of vegetable oils released by the fish feed and feed containing nanofibers after 10 minutes in contact with an aqueous medium simulating fish tank condition. note: results followed by different capital letters differ significantly between the different treatment (fish feed a fndeed containing nanofibers) of the same compositions (p<0.05). results followed by different lowercase letters differ significantly between the different compositions of the same treatment (p<0.05). two-way anova and tukey’s test were used to determine statistical significance. 3.6. assesment of in vitro digestibility figure 4a shows the results of the digestibility test of the feeds in gastric fluid over a period of 1 hour. when comparing the same feed composition with and without oil encapsulated in nanofibers, it is observed that all feed samples without encapsulated oil exhibited higher digestibility compared to those containing encapsulated oil. when comparing the different feed compositions, the co – 50:50 sample showed the highest digestibility. a similar pattern was observed for feeds with encapsulated oil, where the co – 50:50 sample also presented the highest digestibility. figure 4b presents the results of the digestibility test of the feeds in intestinal fluid over a period of 5 hours. when comparing the same feed composition with and without oil encapsulated in nanofibers, it was found that the co – 50:50 sample with encapsulated oil showed higher digestibility than the other feed samples. for the other compositions, all feed samples without encapsulated oil exhibited higher digestibility compared to the formulations with encapsulated oil. when comparing the different feed compositions in intestinal fluid, the highest digestibility was observed for the co – 100:0 sample, while the lowest digestibility was recorded for the co – 50:50 sample. among the feeds with encapsulated oil, the highest digestibility in intestinal fluid was also observed for the co – 50:50 sample. agriculture and food sciences research, 2024, 11(2): 136-145 142 © 2024 by the authors; licensee asian online journal publishing group figure 4. comparison between fish feed and feed containing nanofibers in vitro digestibility. the test was performed maintaining the samples in simulated gastric fluid for 1 hour (a) the test was performed maintaining the samples in simulated intestinal fluid for 5 hours (b). note: results followed by different capital letters differ significantly between the different treatment (fish feed a fndeed containing nanofibers) of the same compositions (p<0.05). results followed by different lowercase letters differ significantly between the different compositions of the same treatment (p<0.05). two-way anova and tukey’s test were used to determine statistical significance. 4 discussions 4.1. scanning electron microscopy (sem) the study of nanofibers morphology can understand the encapsulation process. the addition of oil mixtures led to an increase in the average diameter of nanofibers (table 2), which can be attributed to an increase in the polymeric solution viscosity. the relationship between viscosity and the average diameter of nanofibers was already noted in several papers [19, 21]. this behavior indicates that solutions containing different fractions of vegetable oils are more viscous, creating fibers with higher diameter values. 4.2. fourier-transform infrared spectroscopy (ftir) infrared characterization enables the analysis of intermolecular interactions between the polymer and the active compounds through the identification of their chemical groups. the peaks observed for the polylactic acid nanofibers, corn oil, and gold linseed oil in the region from 3100 to 2800 cm-1 of the infrared spectrum agree with what is described in the literature [26, 33, 34]. the peaks show in the infrared spectrum region that confirm the encapsulation of vegetable oils in the polylactic acid matrices because it shows two signature peaks of corn and gold linseed oils (at 2925 cm-1, relative to ch3 symmetric stretch and at 2853 cm-1, relative to stretching vibration of bonds between carbon (c) and hydrogen (h) (ch stretch) in the nanofibers [33-36]. the ftir results (figure 2) suggest that weak interaction occurs between the gold linseed oil, corn oil, and the polylactic acid macromolecules occurs since there is no evidence of the formation of new primary bonds that can be associated with the shift or formation of new peaks in the infrared spectrum. rahmani, et al. [33] observed alterations in the infrared spectrum of silicon nanowires (sinws) after the deposition of poly(3-hexylthiophene) on its surface. the formation of new peaks related to stretching vibration of bonds between carbon (c) and hydrogen (h) (c-h) and silicon (si) and carbon (c) vibrations (si-c) and the absence of the carbon (c) and carbon (c) (c=c) bond peak evidenced the poly(3-hexylthiophene) adsorption in the sinws surface through the formation of si-c bonds after the rupture of the double bond between carbons. another example is the paper of yu, et al. [34], in which a shift of characteristic peaks of polyacrylonitrile nanofibers to a lower wavenumber after the encapsulation of different concentrations of acyclovir in the polymeric matrix, indicating the formation of hydrogen bonds between the carbon (c) and nitrogen (n) (c≡n group) from polyacrylonitrile and the oxygen (o) and hydrogen (h) (–oh group) from acyclovir. the presence of characteristic peaks of polylactic acid and vegetable oils in the nanofibers is an indication that the solution blow spinning process was efficient to encapsulate these nutrients. 4.3. contact angle it was expected that nanofibers would exhibit a hydrophobic behavior, with contact angle greater than 90° (table 3), because both polylactic acid and vegetable oils are nonpolar molecules and, therefore, are incapable of forming intermolecular interactions with water. moreover, the interaction between a polar liquid and a nonpolar surface is weak since the interactions inside the liquid are favored, resulting in a decrease of contact with the surface [36]. paragkumar, et al. [37] showed that pla films had a contact angle equal to 96°. this shows that the production of materials in nanoscale can improve properties such as hydrophobicity [38]. therefore, the results align with following what is described in the literature and it can be affirmed that all nanofibers produced in this paper have great potential to retain the diffusion of the oil fraction in aquatic environments since there will be no significant interaction between the polymeric matrix and water. agriculture and food sciences research, 2024, 11(2): 136-145 143 © 2024 by the authors; licensee asian online journal publishing group 4.4. encapsulation efficiency knowing that the encapsulation process was successful, it is necessary to quantify the number of active compounds that were trapped in nanofibers. the co: lo – 50:50 nanofiber had the one with the lowest encapsulation efficiency, which indicates that this sample will present an initial burst more evident than the other formulations, as shown by wang, et al. [39]. in general, these results agree with what is reported in the literature about encapsulation in nanofibers. in the paper of wang, et al. [39] the encapsulation efficiency of rosemary essential oil in polylactic acid nanofibers obtained by electrospinning was 90.18±1.77%, whereas the same encapsulation in pellets obtained by the extrusion technique led to an encapsulation efficiency of 68.77±1.41%. this behavior was also observed when comparing these two projects: pérez-masiá, et al. [40] obtained an encapsulation efficiency of 44.0±5.5% when encapsulating folic acid in starch microcapsules (electrospraying technique), and fonseca, et al. [41] obtained an encapsulation efficiency of 95.6±0.0% when encapsulating folic acid in starch nanofibers (electrospinning technique). the higher values of encapsulation efficiency observed to nanofibers when compared with other polymeric matrices is due to properties like high surface-to-volume ratio, which reduce the number of interest compounds that can be present in the material surface. through the comparison between the values demonstrated in table 2 and table 4, it can be assumed that the diameters of the nanofibers do not interfere directly in the encapsulation process, since large diameters are not necessarily associated with higher encapsulation efficiency values. 4.5. assessment of nutrient liberation in an aqueous medium (in vitro) in fish farming, the feed does not stay in contact with water for more than 10 minutes [42] and through analysis of figure 3, it is notable that in this period all nanofibers released between 14-21% of the nutrients encapsulated, while all feed samples released between 14-51% of nutrients during the same time. these results show that nanofibers are more effective in protecting nutrients against premature release. the nutrient release from nanofibers in the first 10 minutes of contact with an aqueous medium (figure 3) has a direct connection with the high amount of vegetable oils encapsulated (50% w/w). this behavior was already explained by karuppuswamy, et al. [43] who noticed that greater active agent concentrations led to faster burst release because there was a higher quantity of the compounds deposited in the polymeric matrix surface that could be easily dispersed in the medium [44]. the higher percentage of nutrients release observed for the feed can be explained by the fact that the feed commonly used in fish farming has approximately 40 essential nutrients that have to be present in specific quantities to ensure a balanced diet [45]. since the formulation is extremely complex and composed of several hydrophilic and hygroscopic fractions, like soybean meal, cornflour, and dicalcium phosphate, a fast erosion of the feed granules occurs when in contact with water. this rapid erosion leads to an accelerated migration of the apolar fraction of nutrients, for example, vegetable oils. therefore, they are dispersed in the aqueous medium before ingestion by fish. notably, nanofibers were capable of retain the action compounds much more efficiency than the feed containing free oils. these results prove that the encapsulation of active agents was successful and confirms that the proposal to apply nanofibers in fish farming to feed fortification is promising since lower release in water is directly associated with greater availability of nutrients that can be absorbed in fish metabolism. 4.6. assessment of in vitro digestibility the fish feeding process starts when the feed comes into contact with water. at this moment, part of the nutrients added in the formulation is lost to the aquatic medium. then, the pellets are consumed by the fish and the fraction of the active agents that were not released in water are digested in the stomach and, subsequently, in the intestine. the analysis of figure 3 and 4 show the absence of a release standard between the feed samples, which indicates that the formulation presents certain heterogeneity and that the chemical composition of the oil fraction added in the feed has a direct impact on the apolar nutrients release since there was a big difference between the percentage of nutrients released in the simulated mediums for each feed sample. therefore, considering the heterogeneity of the formulation and the loss of feed nutrients that occurs in water, each pellet has different quantities of active compounds when they reach the gastrointestinal tract. this leads to consequences in the growth, development, and nutritional value of fishes since each one will consume different concentrations of nutrients. the final weight variation of farmed fishes that are fed with the conventional feed has been already observed by several authors, such as [46-48]. nanofibers showed a more significant release in the simulated intestinal fluid than in simulated gastric fluid, which is a promising result since nutrients stay longer in the intestine, where most of its absorption occurs [16]. furthermore, statistical analysis shows that even with different encapsulation efficiency and average diameter values, nanofibers can have similar effectiveness. the sample with the lowest encapsulation, efficiency (co: lo – 50:50) was the one that released the greatest number of vegetable oils in water (figure 3), which makes sense if we think that the lower the quantity of nutrients encapsulated the greater the fraction that is free in the polymeric matrix surface. the sample with the highest encapsulation efficiency (co: lo – 100:0) was the one that released the greatest amount of vegetable oils in the intestinal fluid (figure 4b), indicating that the polymeric matrix effectively protects the active agents, ensuring a ph-induced release. since the feed components are not wrapped by a protective layer, nutrients are free and released indiscriminately both in the aqueous medium and in the simulated gastrointestinal fluids. thus, the feed contains a small number of nutrients when reaches the intestinal fluid, which affects the growth and development of fishe. therefore, vegetable oil encapsulation in nanofibers can be extremely advantageous, not only environmentally but also economically and nutritionally. 5. conclusion the results showed that the encapsulation of vegetable oils in polylactic acid nanofibers obtained by the solution blow spinning technique was efficient, and the product can be applied as a controlled release system to agriculture and food sciences research, 2024, 11(2): 136-145 144 © 2024 by the authors; 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licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 78-83, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6906 © 2025 by the authors; licensee asian online journal publishing group yield and fruit quality of barhee date palms as affected by gibberellic acid, boron and active dry yeast sprays mostafa, e.a.m.1 rasha, s. abdel-hak2 n.e. ashour3 m.m.s. saleh4 ( corresponding author) 1,2,3,4pomology department, national research centre, dokki, cairo, egypt. 1email: ea.mostafa@nrc.sci.eg 2email: rashaabdelhak777@gmail.com 3email: nashour2008@yahoo.com 4email: mmsssa2000@yahoo.com abstract the present investigation was conducted over two consecutive seasons on 15-year-old barhee date palms (phoenix dactylifera l.) cultivated in an exclusive orchard 63 kilometers along the cairoalexandria desert road in sandy soil under a drip irrigation system. for three times, bunches of the tested palms were misted with ga3, boric acid, or active dry yeast: once a day before pollination, once a month after fruit set, and once a month before harvest. ga3 was sprayed at the previous dates with 50, 100, and 50 ppm, respectively, while boric acid was sprayed at the same dates with 2000, 1500, and 1000 ppm. similarly, active dry yeast was sprayed at the same dates with 250, 500, and 250 ppm. the investigated treatments were compared with untreated palms. the findings indicate that all spraying treatments improved fruit properties, both chemical and physical, and boosted fruit set and yield compared to the control. in general, the most effective treatment to increase fruit set, production, and the physical and chemical properties in both seasons was the procedure of spraying inflorescences with active dry yeast three times (250, 500, and 250 ppm). keywords: active dry yeast, boron, barhee date palm, fruit quality, gibberellic acid, yield. citation | e a m, m., abdel-hak, r. s., ashour, n. e., & saleh, m. m. s. (2025). yield and fruit quality of barhee date palms as affected by gibberellic acid, boron and active dry yeast sprays. agriculture and food sciences research, 12(1), 78–83. 10.20448/aesr.v12i1.6906 history: received: 7 may 2025 revised: 2 july 2025 accepted: 11 july 2025 published: 15 july 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 79 2. materials and methods ................................................................................................................................................................... 79 3. results ................................................................................................................................................................................................ 80 4. discussion .......................................................................................................................................................................................... 82 5. conclusion ......................................................................................................................................................................................... 82 references .............................................................................................................................................................................................. 82 mailto:ea.mostafa@nrc.sci.eg mailto:rashaabdelhak777@gmail.com mailto:nashour2008@yahoo.com mailto:mmsssa2000@yahoo.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6906 https://orcid.org/0000-0002-2978-190x agriculture and food sciences research, 2025, 12(1): 78-83 79 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature comparing active dry yeast as a bioactive material with two chemical agents (ga3 and boron) may provide an opportunity for dry yeast to be used as an alternative treatment, especially if it produces distinctive results concerning fruit set, yield, and fruit properties of one of the most economical date palm cultivars. 1. introduction the oldest fruit crop grown domestically in the middle east is the date palm, and many people rely heavily on its fruits for their nutritional needs. the ministry of agriculture and land reclamation has released statistics showing that egypt is one of the top ten countries in the world for date production. there are approximately fifteen million fruitful female palms (14,955,331) planted on 115,610 feddans, yielding 1,684,917 metric tonnes of dried, semi-dry, and fresh dates [1]. in all agricultural systems, increasing crop yield through the effective use of fertilizers is a key objective. researchers are motivated to develop more effective methods of fertilizer application because excessive nutrient loss from agricultural land causes public concern. the benefits of foliar fertilization include homogeneous fertilizer distribution, rapid nutritional response, and lawful application rates. furthermore, it is simple to control concealed hunger [2, 3]. when applied topically, plant growth regulators are the most effective means for controlling growth, flowering, yield, and quality of fruit, particularly its size, as well as the maturation of the fruit. the growth regulators' capacity to expand date cell size and promote fruit development makes them well-known [4]. with ga3 spray at 100 or 150 ppm after 50 days of pollination, sakkoty dates' fruit weight, flesh percentage, fruit dimensions, and moisture percentage all increased significantly compared to untreated ones. total sugars were somewhat lower with ga3 than in the control [5]. when compared to the control, the chemical and physical characteristics of the barhee cultivar were enhanced by spraying salicylic acid (sa), benzyl adenine (ba), gibberellic acid (ga3), and naphthalene acetic acid (naa). growth regulator treatments generally improved the shelf life of barhee date palms [6]. it has been demonstrated that date palms benefit from boron spray application in terms of fruit set and quality because it affects a variety of plant functions, including hormone transition, flowering, pollen tube growth, and cell membrane functions. it also regulates the production of proteins, carbohydrates, and phenols, as well as the transportation of sugars from areas of production to places for growth and storage [7]. fruit growth, setting, retention, yield, and quality are all positively impacted by micronutrients [8-10]. a number of functions, including protein synthesis, glucose metabolism, and sugar transport, depend on boron [11]. certain microelements, such as boron, appear to significantly influence date palm yield and fruit quality, aiding in achieving proper fruit set and quality [12, 13]. additionally, bio-fertilization is recommended as a safe fertilization technique to boost the productivity and quality of numerous fruit varieties. certain strains, such as fixing bacteria and yeasts, are used as biological fertilizers to dissolve silicate, phosphorus, and nitrogen, making bio-fertilizers commercially accessible. many advantages and benefits of employing active dry yeast as a bio-fertilizer are attributed mainly to the stimulation of photosynthesis, increased levels of vitamins, and cytokinin (a naturally occurring plant hormone) [14]. active bread yeast was sprayed on valencia orange trees once in march or august, or twice, to improve fruit set, growth, number, weight, and volume, as well as the yield and other fruit quality parameters [15]. when sukary date palms were sprayed with yeast at 4% and 8%, the amount of soluble solids and total sugars was reduced [16]. the aim of this research is to investigate the impact of applying gibberellic acid, boron, and yeast to the inflorescences of barhee date palms on fruit yield and quality. 2. materials and methods this study examined the results of foliar spraying with ga3, boric acid, and active dry yeast on fruit set, yield, and quality over two consecutive seasons on 15-year-old, uniform barhee date palms (phoenix dactylifera l.) cultivated in a private orchard located 63 kilometers along the cairo-alexandria desert road. with the exception of the tested treatments, the experimental palms were given the standard horticultural procedures used in the commercial orchard after being chosen as healthy, uniform, and almost identical in vigor. in both seasons, the same male pollen was used to pollinate every palm. a drip irrigation system was employed to irrigate the palms. the palms underwent four spraying treatments, each consisting of three replicates and three bunches (4 replicates × 3 replicates × 3 bunches = 36 bunches on 12 palms). the treatments were arranged in a randomized complete block design, and the experimental bunches were sprayed with ga3, boric acid, or active dry yeast three times: once a day before pollination, once a month after fruit set, and once a month before harvest. ga3 was sprayed at the specified dates with 50, 100, and 50 ppm, respectively, while boric acid was applied at the same dates with 2000, 1500, and 1000 ppm. similarly, active dry yeast was sprayed at the same dates with 250, 500, and 250 ppm, respectively. in this manner, the treatments were sprayed with: 1. water only (control). 2. ga3 at 50, 100, and 50 ppm, respectively, at the three dates. 3. boric acid at 2000, 1500, and 1000 ppm, respectively, on the three dates. 4. active dry yeast at 250, 500, and 250 ppm, respectively, on the three dates. a 6:1 carbon to nitrogen supply was used to activate the dry, pure yeast powder, according to barnett et al. [17]. urea at 0.25% was added to the spraying solution for each treatment. a small handheld sprayer was used to apply the spray until runoff. the spraying solution was mixed with tween 20 (1%) as a wetting agent. fruits were harvested in early october when they were fully ripe, indicated by the color of their skin, which should turn yellow entirely. agriculture and food sciences research, 2025, 12(1): 78-83 80 © 2025 by the authors; licensee asian online journal publishing group the following measurements were carried out: fruit set percentage: ten strands per spath were measured for both abnormal and normal fruit set. the following calculation was used to determine the percentage of fruit setting: fruit set % = (total number of normal fruit set number of abnormal fruit set) / total number of fruits × 100. 2.1. yield /palm (kg) weighting bunches at harvest (early october in both seasons) allowed us to determine the overall production per palm. 2.2. average bunch weight the number of bunches per palm was divided by the total yield per palm to determine the average bunch weight. 2.3. average fruit number/strand the number of fruits on ten randomly chosen strands was counted to establish the average number of fruits per strand. 2.4. fruit physical properties ten fruits were randomly selected from each bunch (as a replication), and thirty fruits per treatment were used to assess the fruit weight, flesh, and seed weight (g), as well as the fruit length and diameter (cm). 2.5. fruit chemical properties fruit chemical characteristics were assessed at harvest by measuring the percentage of total soluble solids and calculating the percentage of acidity by titrating against 0.1 naoh, in accordance with the association of official agricultural chemists [18]. using the welbur method, the amounts of carotene and total chlorophyll (mg/100g peel fresh weight) were determined [19]. 2.6. statistical analysis according to snedecor and cochran [20], analysis of variance was conducted on the data gathered for both seasons, and the means were separated at the 5% level using the duncan multiple range test [21]. 3. results 3.1. yield both bunch weight and the earliest fruit set are considered indicators of yield. the results in table 1 demonstrate how ga3, boron, and active dry yeast sprays affected the percentage of fruit set and bunch weight of barhee date palms during the two seasons. throughout the two seasons under study, it was found that the outcomes followed a similar pattern. compared to the untreated palms (control), it was evident from the statistics that all treatments significantly increased the fruit set percentage, bunch weight, and yield per palm. the application of active dry yeast produced the highest yield per palm (149 & 155 kg), the heaviest bunch weight (14.96 & 15.50 kg), and the highest percentage of fruit set (70.30% & 71.56%) during the two seasons studied. in terms of the average number of fruits per strand, table 1 demonstrates that the ga3 treatment produced the maximum number of fruits (22.6 & 23.1 fruits/strand), followed by the active dry yeast treatment (22.0 & 22.53 fruits/strand). on the other hand, the control treatment gained the lowest fruit set (58% & 59%), bunch weight (11.8 & 12.2 kg), yield per palm (118 & 122 kg), and fruit number per strand (19.2 & 19.6 fruit/strand) in both seasons of the study, respectively. table 1. effect of spraying ga3, boron, and active dry yeast on fruit set, bunch weight, yield per palm, and fruit number per strand of barhee date palms during two seasons. treatments fruit set% bunch weight (kg) yield/palm (kg) fruit number/strand 1st season 2nd season 1st season 2nd season 1st season 2nd season 1st season 2nd season t1 control 58.00d 59.53c 11.80b 12.20d 118.00c 122.00d 19.20d 19.63c t2 ga3 65.00c 68.00b 14.00a 13.43c 140.00b 134.30c 22.00b 22.53a t3 boric acid 67.60b 69.00ab 14.16a 14.50b 141.66b 145.40b 21.06c 21.63b t4 active dry yeast 70.30a 71.56a 14.96a 15.50a 149.66a 155.00a 22.60a 23.06a note: means having the same letters within a column are not significantly different at 5% level. 3.2. fruit quality 3.2.1. physical properties results from both seasons indicate that spraying inflorescences with different treatments had a substantial impact on the fruit dimensions, fruit weight, flesh weight, and seed weight of barhee date palms when compared to the control table 2. in the first and second seasons, respectively, foliar spraying with active dry yeast produced the largest fruit length (3.6 and 3.7 cm) and fruit diameter (2.76 and 2.73 cm). spraying ga3 produced fruit lengths of 3.57 and 3.56 cm and fruit diameters of 2.7 and 2.8 cm in both study seasons. throughout the two study seasons, the control treatment produced the smallest fruit dimensions, measuring 3.1 and 3.2 cm for fruit length and 2.3 and 2.36 cm for fruit diameter. agriculture and food sciences research, 2025, 12(1): 78-83 81 © 2025 by the authors; licensee asian online journal publishing group the highest fruit weight (22.4 & 22.9 g) and flesh weight (20.96 & 21.33 g) were recorded in the first and second seasons, respectively, by foliar spray with active dry yeast. this was followed by foliar spray with boron, which produced fruit weights of 21.2 & 21.9 g and flesh weights of 19.60 & 20.26 g in both study seasons. the control treatment produced the lowest fruit and flesh weights, weighing 18.06 and 19.0 g for fruit and 16.66 and 17.46 g for flesh throughout the two experimental seasons. in terms of seed weight, the foliar spray containing boron produced the maximum seed weight (1.60 & 1.63 g), followed by the foliar spray containing active dry yeast (1.46 & 1.56 g) across the two study seasons. the foliar spray with ga3 yielded the lowest seed weight, weighing 1.16 and 1.26 g in both study seasons. table 2. effect of spraying ga3, boron, and active dry yeast on fruit dimensions, fruit weight, flesh weight, and seed weight of barhee date palms during two seasons. treatments fruit length (cm) fruit diameter (cm) fruit weight (g) flesh weight (g) seed weight (g) 1st season 2nd season 1st season 2nd season 1st season 2nd season 1st season 2nd season 1st season 2nd season t1 control 3.1d 3.20c 2.30b 2.36b 18.06d 19.0d 16.66d 17.46d 1.40b 1.56a t2 ga3 3.5b 3.56b 2.70a 2.80a 20.03c 20.53c 18.86c 19.26c 1.16c 1.26b t3 boric acid 3.3c 3.50b 2.73a 2.86a 21.20b 21.90b 19.60b 20.26b 1.60a 1.63a t4 active dry yeast 3.6a 3.73a 2.76a 2.73a 22.43a 22.90a 20.96a 21.33a 1.46ab 1.56a note: means having the same letters within a column are not significantly different at 5% level. 3.2.2. chemical properties table 3 presents the results of ga3, boron, and active dry yeast sprays on various chemical characteristics of barhee fruits over two seasons. these qualities include total soluble solids, acidity percentage, total sugars percentage, and tss/acid ratio. ga3, boron, and active dry yeast sprays significantly enhanced the dates' chemical composition by increasing the percentages of total soluble solids, total sugars, and tss/acid ratio, while decreasing the percentages of total acidity compared to untreated samples. the results might simply show that the total soluble solids were equal to the amount of sugar, and the overall acidity content was the opposite. in general, spraying active dry yeast resulted in the highest total soluble solids (32.53 & 32.90%), tss/acid ratio (89.56 & 88.83), and total sugars (53.06 & 55.06%) during the two seasons of the study, respectively, followed by foliar spray with ga3, which recorded 31.2 & 32% for tss and 86.66 & 84.93 for tss/acid ratio. the second highest for total sugars (50.50 & 52.66%) was observed when boric acid was sprayed. the lowest percentages of tss, tss/acid ratio, and total sugars were obtained with the control treatment, which recorded 28.83 & 29.3% for tss, 58.03 & 57.43 for tss/acid ratio, and 42.56 & 42.80% for total sugars in the first and second seasons, respectively. according to the results in table 3 over the two study seasons, the water spray treatment (control) had the highest fruit acidity percentages (0.49% and 0.51%, respectively). however, both ga3 and active dry yeast had the lowest overall acidity percentages (0.36% and 0.37%) for both seasons, respectively. table 3. effect of spraying ga3, boron and active dry yeast on tss, acidity, tss/acid ratio and total sugars of barhee date palms during the two seasons. treatments tss% acidity% tss/acid total sugars% 1st season 2nd season 1st season 2nd season 1st season 2nd season 1st season 2nd season t1 control 28.83c 29.30c 0.49a 0.51a 58.03c 57.43d 42.56d 42.80d t2 ga3 31.20b 32.00b 0.36c 0.37c 86.66a 84.93b 46.46c 48.56c t3 boric acid 28.73c 28.70c 0.43b 0.41b 65.16b 70.00c 50.50b 52.66b t4 active dry yeast 32.53a 32.90a 0.36c 0.37c 89.56a 88.83a 53.06a 55.06a note: means having the same letters within a column are not significantly different at 5% level. all treatments significantly increased both reducing and non-reducing sugars throughout the duration of the two study seasons when compared to the control, as shown by the results in table 4. the highest percentages of reducing and non-reducing sugars, 40.86% and 42.66%, and 12.03% and 12.4%, respectively, were produced by the palms sprayed with active dry yeast in both seasons. the control yielded the lowest results, with reducing sugars at 32.3% and 32.36%, and non-reducing sugars at 9.36% and 10.1% during the two study seasons. the other treatments recorded intermediate results. according to the results in table 4, the total chlorophyll content in the fruit peel was significantly increased by spraying inflorescences with active dry yeast. the highest content (6.44 and 6.53 mg/100g in fresh peel) was recorded in the first and second seasons, respectively, followed by foliar spraying with boric acid, which produced 6.27 and 5.80 mg/100g in the fresh peel across the two seasons. during both study seasons, the control treatment's fresh peel had the lowest total chlorophyll concentration, registering 2.44 and 2.64 mg/100g. the findings in table 4 show that the total carotene content in the fruit peel was significantly increased by spraying water only (control), which produced the highest content (8.31 & 8.49 mg/100g) in the first and second seasons, respectively, while the ga3 treatment resulted in the lowest content (3.29 & 4.06 mg/100g) in the two seasons. the other treatments yielded intermediate results. agriculture and food sciences research, 2025, 12(1): 78-83 82 © 2025 by the authors; licensee asian online journal publishing group table 4. effect of spraying ga3, boron and active dry yeast on reducing, non-reducing sugars, total chlorophyll and carotene of barhee date palms during the two seasons. treatments reducing sugars% non-reducing sugars% total chlorophyll mg/100g carotene mg/100g 1st season 2nd season 1st season 2nd season 1st season 2nd season 1st season 2nd season t1 control 33.20c 32.36d 9.36d 10.1c 2.44c 2.64c 8.31a 8.49a t2 ga3 36.03b 37.56c 10.43c 11.00b 2.47c 4.20b 3.29d 4.06d t3 boric acid 39.50a 40.66b 11.00b 12.00a 6.27b 5.80a 6.43c 7.05c t4 active dry yeast 40.86a 42.66a 12.03a 12.40a 6.44a 6.53a 6.49b 7.14b note: means having the same letters within a column are not significantly different at 5% level. 4. discussion the benefits of foliar fertilization include rapid reactions to applied nutrients, consistent fertilizer distribution, and low application rates [2]. applying zinc and boron to date palms increased their productivity, fruit set percentage, and bunch weight [9]. another theory is that boron enhances fruit removal force because it is involved in numerous enzymatic processes that control the metabolism of proteins and carbohydrates, which are essential for growth and development [8]. by applying boron and bio-fertilization, date palm fruit quality had improved. many processes, including the transport of sugars and the metabolism of carbohydrates and proteins, depend on boron to enhance the weight and dimensions of fruits and to speed up their maturity [11]. numerous researchers [8-10, 16] found that spraying micronutrients increased fruit yield and quality, and the obtained results were consistent with those findings. in addition to increasing productivity, proper boron nutrition is essential for fruit quality [22]. the obtained results are in line with numerous studies sarrwy, et al. [8]; omar, et al. [9]; mostafa [10]; khayyat, et al. [13]; osman [23]; ashour, et al. [24]; harhash and abdel-nasser [25]; soliman and al-obeed [26]; zaen el–daen, et al. [27] and jabbar and hassan [28] who found that foliar spray of boron on date palms significantly affected fruit setting. on the other hand, the ga3 application has a noticeable impact on fruit volume, diameter, and length. this effect may be explained by gibberellic acid's ability to increase cell elongation, as well as the elasticity and permeability of the cell wall, which increases the amount of water and nutrients entering the cells and causes them to swell and enlarge [29, 30]. our obtained results agree with those observed in many experiments [24, 28, 31-33]. the presence of yeast is advantageous for fruit development, since applying 0.2% yeast solution to keitte mango trees once during full bloom resulted in the development of fruits, increasing their weight and quantity per tree, as well as fruit length, width, and weight. additionally, it improved the pulp-to-fruit ratio and increased total soluble solids [34]. with its abundance of cytokinins and nutritional components, yeast is a natural (safe and non-polluting) component. additionally, it is rich in reducing sugars, enzymes, minerals, amino acids, and vitamins b1, b2, b3, and b12, all of which improve the chemical and physical properties of the fruits [35, 36]. this may be the reason for the effects of yeast extract. additionally, yeast contains proteins and cytokinins that promote cell growth and division [17]. regarding the impact of active dry yeast on the fruit's physical and chemical properties, the results obtained are consistent with those found in other studies [10, 16, 37, 38]. 5. conclusion overall, it can be said that applying ga3, boric acid, and active dry yeast to the inflorescences of barhee date palms one day prior to pollination, one month after fruit set, and one month before harvest, respectively, has a favorable impact on fruit set and fruiting. the active dry yeast spray outperformed the boric acid and ga3 sprays in both study seasons. references [1] ministry of agriculture and land reclamation agricultural statistics, ministry of agriculture and land reclamation statistics central administration for agricultural economy. egypt: ministry of agriculture and land reclamation agricultural statistics, 2016. 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[38] m. jafarpour and b. eghbal, "effect of active dry yeast (saccharomyces cerevisiae) on the quality and storage life of fresh-cut apple," journal of food processing and preservation, vol. 40, no. 6, pp. 1171–1179, 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/s0176-1617(11)81192-2 https://doi.org/10.2307/3001478 1 © 2022 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 9, no. 1, 1-8, 2022 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v9i1.3758 © 2022 by the authors; licensee asian online journal publishing group agriculture adaptation strategies of tunisian oasis households to climate change amira abdelhamid1 houcine jeder2 ahmed salah3 ( corresponding author) 1,3faculty of economics and management, university of tunis el-manar, tunisia. 1email: amira.abdelhamid83@gmail.com 3email: ahmedtun@yahoo.fr 2regional researches center on horticulture and organic agriculture, sousse, and laboratory of economy and rural communities, arid regions institute, university of gabes, medenine, tunisia. 2email: dejderhoucine@yahoo.fr abstract climate change should have impacts on tunisian arid regions oasis households are likely to bear the most significant negative impacts, the case of the oasis of "metouia" in the governorate of "gabes". this research is based on an econometric analysis through cross-sectional probit models involving 50 oasis households. the binary probit models showed that certain factors contribute significantly to the adaptation strategies identified, such as: the age of the agricultural household head, agriculture as a main activity, the agricultural production system adopted the mode of ownership of agricultural land, extension for farmers. all actions aimed at improving the resilience of tunisian oasis agricultural households to climate change focus mainly on the strategies adopted by farmers in terms of water management, the technical choices and the production systems adopted combined with the experience and local know-how. keywords: climate change, adaptation, agricultural, households, econometric analyses, tunisia. citation | amira abdelhamid; houcine jeder; ahmed salah (2022). agriculture adaptation strategies of tunisian oasis households to climate change. agriculture and food sciences research, 9(1): 1-8. history: received: 29 december 2021 revised: 15 february 2022 accepted: 1 march 2022 published: 8 march 2022 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. authors’ contributions: all authors contributed equally to the conception and design of the study. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study followed all ethical practices during writing. contents 1. introduction ......................................................................................................................................................................................... 2 2. theoretical and methodological framework ............................................................................................................................... 2 3. results and discussions ..................................................................................................................................................................... 5 4. conclusion and recommendations .................................................................................................................................................. 7 references ................................................................................................................................................................................................. 7 mailto:amira.abdelhamid83@gmail.com mailto:ahmedtun@yahoo.fr mailto:dejderhoucine@yahoo.fr https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v9i1.3758 https://orcid.org/0000-0001-7958-4894 https://orcid.org/0000-0003-1953-8209 https://orcid.org/0000-0002-3735-7092 agriculture and food sciences research, 2022, 9(1): 1-8 2 © 2022by the authors; licensee asian online journal publishing group 1. introduction agriculture lays a heavy burden on the environment in the process of providing humanity with food and fibers [1]. however, agriculture and food systems as well as the rural economies in the maghreb and north africa regions have been experiencing major drastically reduced agricultural production through extreme weather events, such as recurrent droughts and floods in these recent decades [2, 3].in these regions, the climate variability causes severe impacts on agriculture through long drought periods. recurrent droughts often affect entire countries over multiyear periods and can result in serious social problems caused by water scarcity and the intensive demand of water for agriculture. impacts anticipated under projected climate change such as increasing rainfall variability, increasing temperature, increasing evaporation rate and water deficit pose a significant challenge to the maghreb region. mean temperatures of morocco, algeria and tunisia are expected to rise by between 2 and 4°c until 2100. already by 2020, rainfall is expected to drop by between 5 and 20% [4]. tunisia is one of the maghreb countries and is very vulnerable to the water shortage. most of the water resources have medium to poor quality, and the salinity is often high. water deficits and droughts are ongoing risks for tunisian agriculture. the agricultural sector that pro vides approximately 13% of the annual gross domestic product to tunisia is very vulnerable to climatic changes also due to poor soils, limited ground and surface water, low rainfall and recurrent droughts particularly in the arid regions of tunisia in these regions, the consumption of irrigation water in irrigated areas and oases continues to increase to ensure the sustainability of the farming activity and guarantee the income of several households agricultural [5]. for example, in the arid regions of south-eastern tunisia, the gabes oases is known since antiquity for their vegetable crops that historically conserved the seeds, their greatest source of resilience facing difficult climate conditions (dry land). howewer, since the past few decades, the oases are getting very damaged (over-exploitation of resources, urbanisation…) as well as multiple factors have been behind this degradation [6]. the majority of young people and women are not interested in agriculture and the older populations that are, are turning to more profitable field crops. producing local seeds is forsaken for the benefit of improved hybride seeds that are often combined with the use of chemical fertilizers (highly polluting the soil). the lack of valorisation of local indigenous seeds, the absence of a seed market or even laws restricting the commercialisation of local seeds. this all explains why farmers are discouraged and have difficulties guaranteeing their own production. the risk is that future generations that loses their capacity to adapt to climate change and no longer is able to guarantee food security for the population surrounding the oases [7]. faced with climatic risks on agricultural activity in oases of irrigated perimeters, reviewing the concept of adaptation through a multidimensional vision touching different economic, social, agronomic, hydrological and political aspects is important to know and detect the main intervening factors in adaptation strategies. the adoption of a bottom-up approach that focuses on autonomous adaptation behavior seems to me more adequate to draw recommendations for adaptation strategies appropriate to oasis production systems in south-eastern tunisia, case of the metouia oasis in the governorate of gabes. therefore, this article aims to study the perception and choice of appropriate measures among smallholders in the oases of south-eastern tunisia for adaptation to climate change. the rest of the article is structured as follows: section two describes the theoretical framework. section three presents the methodology. section four discusses the results and section five provides conclusions and policy recommendations. 2. theoretical and methodological framework farmers' behavior towards adaptation to climate change is shaped by socio-economic, physical and behavioral factors [8]. institutional arrangements for the farmers and their working environment, development for access to markets and climatic factors are useful in shaping the behavior of smallholders [3, 9, 10]. the study of behavior adapting to climate change has many angles of analysis. there is both theoretical and empirical analysis. some questions are positive, many others are prescriptive. there are microeconomic issues and more macroeconomic problems. within this diversity, several approaches have been developed in particular: integrated assessment approaches, empirical (econometric) analysis, economy-wide simulation with models and decision support tools. each of these approaches can help shed light on different aspects of the adaptation problem [11]. for the problem of adopting certain strategies or measures to adapt to climate change in agriculture, the behavioral study of the perception or the choice of strategies requires understanding the reaction of economic agents to current climate and weather events. much of this evidence is provided by often interdisciplinary studies [12, 13]. however, more and more researchers are using large data sets at the household or farm level to explore how economic agents adapt fully and rationally. the evidence is particularly rich for the agricultural sector. such an economic approach in relation to adaptive behavior such as "the econometric approach to climate" has been reviewed by dell, et al. [14] and hsiang [15]. these two works mainly focus on assessing the impact and effect of climatic factors on economic variables such as labor productivity, production and growth, rather than on the benefits, costs or extent of the economy adaptation. however, many of the ideas of hsiang and dell et al also apply to the econometrics of adaptation. several researchers have sought to identify climatic effects both in transversely and chronologically, by comparing the impacts and / or adaptation behaviors across different climatic regimes by measuring the impact of particular meteorological events, such as floods, over time. increasingly, they have access to panel data. crosssectional studies are closely associated with the "ricardian approach" developed by mendelsohn, et al. [16]; kurukulasuriya, et al. [17]; seo and mendelsohn [18]; wang, et al. [19]. given the great diversity of climates around the world, these studies provide ample evidence of adaptive behavior. the approach of nominal and ordered econometric models can simultaneously model the influence of all the explanatory variables on each of the different contribution of this paper to the literature research has shown that certain cultural practices can contribute to the resilience of agriculture in arid regions of tunisia to climate change through adequate water management, technical choices and production systems adopted combined with the experience and local know-how of farmers. agriculture and food sciences research, 2022, 9(1): 1-8 3 © 2022by the authors; licensee asian online journal publishing group adaptation practices, while allowing the potential correlation between the unobserved disturbances as well as the relationship between the adoptions of different adaptation practices [20]. consequently, an agricultural household is confronted with the decision to adopt or not an adaptation strategy taking into account the parameters perceptions of climate change. necessarily, this decision to adopt or not depend on the characteristics of agricultural households explained using an "ad-hoc" approach "through several factors: socio-economic, climatic, endowment of available resources (land, water and labor) and policies related to extension services as well as access to information at optimal time. principal component analysis method and the ordered and binary probit econometric model were mobilized in this work to study the behavior of small farmers in the methaoui oasis in south-eastern tunisia in the face of the challenges of climate change. 2.1. ordered econometric models several studies have used various methodological approaches to analyze the determinants of adaptations to climate change and the choice of adaptation strategies. most commonly used analytical approaches in the literature include discrete choice regression models like binary probit or logit [21, 22], multinomial probit or logit and multivariate probit [2, 3, 23-26]. other empirical studies used principal component analysis and the ricardian model [27]. thus, the decisions of perception and adaptation to climate change are intrinsically multivariate and the attempt at univariate modeling excludes the useful economic information contained in the interdependent and simultaneous adoption decisions. on the basis of this argument, the study adopted the econometric technique of ordered and binary probit models to simultaneously model the perception and the influence of the set of explanatory variables on the main adaptation strategies [20, 28]. 2.1.1. ordered and binary probit models ordered probit model is widely used approach to estimate models of ordered types. the ordered probit model is built around a latent regression in the same manner as the binomial probit model [29]: yi ∗ = β′xi + εi (1) equation 1 presents the latent variable (farmers' perceptions) in this study exhibits itself in ordinal categories which were coded as 0, 1, 2…j. the response of category j is thus observed when the underlying continuous response falls in the j-th interval as: 𝑦 (𝑓𝑎𝑟𝑚𝑒𝑟𝑠′𝑝𝑒𝑟𝑐𝑒𝑝𝑡𝑖𝑜𝑛𝑠) = { 0 𝑖𝑓 𝑦𝑖 ∗ ≤ 𝛿0 ∶ 𝑙𝑜𝑤 𝑝𝑒𝑟𝑐𝑒𝑝𝑡𝑖𝑜𝑛 1 𝑖𝑓 𝛿0 ≤ 𝑦𝑖 ∗ ≤ 𝛿1 ∶ 𝑚𝑜𝑑𝑒𝑟𝑎𝑡𝑒 𝑝𝑒𝑟𝑐𝑒𝑝𝑡𝑖𝑜𝑛 2 𝑖𝑓 𝛿1 ≤ 𝑦𝑖 ∗ ≤ 𝛿2 ∶ ℎ𝑖𝑔ℎ 𝑝𝑒𝑟𝑐𝑒𝑝𝑡𝑖𝑜𝑛 (2) where in equation 2, the variable y* (i = 0, 1, 2) is the unobservable threshold parameters that were estimated together with other parameters in the model. when an intercept coefficient is included in the model, yo* is normalized to a zero value and hence only j-1 additional parameters are estimated with xs. as binary data models adopt or not adopt (0/1) adaptation strategies, the probabilities for each of the observed ordinal response, that is, farmer’s perception to climate change in this study had 3 responses which could be low, moderate and high with ordinal values of 0, 1, 2. for adaptation of such a strategy j, the latent variable in this case is a binary dependent variable with yi = 1 to adopt strategies j or yi = 0 not adopt strategies j. binary probit models can also be motivated by an underlying continuous latent variable yi * which depends on β‘xi and an error term εi (for i = 1,…, n) as in the case of equation 3. if the latent variables would be observable, this would lead to linear regression models. however, latent variables are not observable. but they can be related to the observed binary dependent variables yi: 𝑦 (𝐹𝑎𝑟𝑚𝑒𝑟′𝑠 𝑠𝑡𝑟𝑎𝑡𝑒𝑔𝑖𝑒𝑠) = { 1 𝑖𝑓 𝑦𝑖 ∗ ≥ 0 , 𝑎𝑑𝑜𝑝𝑡 𝑠𝑡𝑟𝑎𝑡𝑒𝑔𝑖𝑒𝑠 𝑗 0 𝑖𝑓 𝑦𝑖 ∗ < 0 , 𝑛𝑜𝑡 𝑎𝑑𝑜𝑝𝑡 𝑠𝑡𝑟𝑎𝑡𝑒𝑔𝑖𝑒𝑠 𝑗 (3) if follows for the probability 𝑝(𝑥𝑖 , 𝛽) that 𝑦𝑖 takes the value one in the following equation 4: 𝑝𝑖(𝑥𝑖, 𝛽) = 𝑃(𝑦𝑖 = 1|𝑥𝑖, 𝛽) = 𝑃(𝑦𝑖 ∗ ≥ 0|𝑥𝑖, 𝛽) = 𝑃(𝛽′𝑥𝑖 + 휀𝑖 ≥ 0) = 𝑃(휀𝑖 ≥ −𝛽′𝑥𝑖) (4) different binary response models can be derived by different distribution assumptions for휀𝑖 .if the error term 휀𝑖 has a standard normal distribution (with expected value of zero and variance one), this leads to binary probit models. in this case 휀𝑖 is symmetrically distributed around zero so that it follows in binary probit model equation 5: 𝑝𝑖(𝑥𝑖, 𝛽) = 𝑃(𝑦𝑖 = 1|𝑥𝑖 , 𝛽) = φi(𝛽′𝑥𝑖) = 1 − φi(−𝛽′𝑥𝑖) (5) the assumption of a known variance of 휀𝑖 is not problematic since this variance is not identified in binary response models and thus cannot be estimated besides 𝛽 so that it has to be normalized[29]. interpretation of a parameter 𝛽ℎ in binary response models with respect to the (partial) effect of the corresponding explanatory variable 𝑥𝑖ℎ (ℎ = 2, … , 𝑘) on the probability 𝑝𝑖(𝑥𝑖 , 𝛽). the parameter 𝛽ℎ cannot be interpreted as simply as in the linear probability model, i.e. it cannot be interpreted as the change in𝑝𝑖(𝑥𝑖 , 𝛽). if 𝑥𝑖ℎ increases by one unit (for a quantitative explanatory variable). instead, the (partial) marginal probability effects of 𝑥𝑖ℎ in binary response models are as follows (for i = 1… n) in the following equation 6: 𝜕𝑝𝑖(𝑥𝑖, 𝛽) 𝜕𝑥𝑖ℎ = 𝜕𝐹𝑖(𝛽′𝑥𝑖) 𝜕𝑥𝑖ℎ = 𝑑𝐹𝑖(𝛽′𝑥𝑖) 𝑑(𝛽′𝑥𝑖) 𝜕(𝛽′𝑥𝑖) 𝜕𝑥𝑖ℎ = 𝑓𝑖(𝛽′𝑥𝑖)𝛽ℎ (6) agriculture and food sciences research, 2022, 9(1): 1-8 4 © 2022by the authors; licensee asian online journal publishing group while 𝐹𝑖(𝛽′𝑥𝑖) is the distribution function and 𝑓𝑖(𝛽′𝑥𝑖) is the density function both of the standard normal distribution in binary probit models. partial marginal probability effects of 𝑥𝑖ℎ in binary probit models with 𝜑𝑖(𝛽′𝑥𝑖) as the density function of the standard normal distribution in the following equation 7: 𝜕𝑝𝑖(𝑥𝑖, 𝛽) 𝜕𝑥𝑖ℎ = 𝜕φi(𝛽′𝑥𝑖) 𝜕𝑥𝑖ℎ = 𝑑φi(𝛽′𝑥𝑖) 𝑑(𝛽′𝑥𝑖) 𝜕(𝛽′𝑥𝑖) 𝜕𝑥𝑖ℎ = 𝜑𝑖(𝛽′𝑥𝑖)𝛽ℎ (7) the discussion so far has implicitly assumed quantitative explanatory variables. however, if explanatory variables are discrete or even binary, to calculate of the (partial) effect of an infinitesimal change of an explanatory variable 𝑥𝑖ℎ can be very inaccurate. therefore, a discrete change of𝑝𝑖(𝑥𝑖, 𝛽)due to a discrete change ∆𝑥𝑖ℎ is (for i = 1…, n) for the following equation 8: ∆𝑝𝑖(𝑥𝑖, 𝛽) = φi(𝛽′𝑥𝑖 + 𝛽ℎ∆𝑥𝑖ℎ) − φi(𝛽′𝑥𝑖) (8) if 𝑥𝑖ℎ is a dummy variable,∆𝑥𝑖ℎ = 1 implies the change from zero to one. again it is possible to calculate (partial) average (discrete) effects and (discrete) effects at the mean �̅� = 1 𝑛 ∑ 𝑥𝑖 𝑛 𝑖 of the explanatory variables. where, φ is the normal density function, h the threshold parameter and 𝑥𝑖ℎ the j the explanatory variables. the farmer's perceptions of climate change and the adoption of an adaptation strategy are specified as follows equation 9: 𝑌 ∗ = 𝛽0 + 𝛽1𝑥1 + + 𝛽2𝑥2 + 𝛽3𝑥3 + + 𝛽3𝑥3 + 𝛽4𝑥4 + + 𝛽5𝑥5 + + 𝛽6𝑥6 + 𝛽7𝑥7 + 𝛽8𝑥8 (9) y*(farmer’s perceptions) =1 (no perception), 2 (average perception) or 3 (good perception ( ordered probit model). y* (farmer's strategies) = 1 (adopt strategies j), 0 (not adopt strategies j). (binary probit models). x1= ages of farmer (years), continuous (in number) . x2 = level of education (ordered), 1 (literate), 2 (primary), 3 (secondary), 4 (university). x3 = main agricultural activity, binary (1 if agriculture, 0 other). x4 = place of residence, binary (1 if on the farm, 0 outside) . x5 = farm size, continuous (in hectare). x6 = type of agricultural production system, ordered (3-stage system, 2 classic system, 1 otherwise). x7 = agricultural land owner, binary (1 if farm owner, 0 other). x8 = membership of the agricultural development group, binary (1 if yes, 0 no). 2.2. the study area the métouia oasis is one of the coastal oases of the governorate of gabès. it is located 12 km north of the city of gabès (south-eastern tunisia) and covers an area of approximately 270 ha figure 1. the métouia oasis is characterized by low rainfall. the monthly distribution of precipitation is characterized by a period without rain (june, july and august) and a period with rainfall irregularly distributed over the other months and an annual water balance which is highly deficient. the oasis farms cover very small areas of around 1.5 ha on average. the crops are staged there in height, the palm trees are on the first floor, the pomegranate trees are on the second floor and alfalfa and market gardening are on the third floor. the drainage network is ineffective and moderately maintained. the métouia oasis is characterized by the presence of a very shallow water table which closely conditions the evolution of the soil throughout the oasis [30, 31]. figure 1. location map of the métouia oasis, south-eastern tunisia. agriculture and food sciences research, 2022, 9(1): 1-8 5 © 2022by the authors; licensee asian online journal publishing group 2.3. data type and sources data used in this analysis were collected from a household survey conducted in métouia oasis of the governorate of gabes on south-eastern tunisia. the zones have been chosen to take a representative sample of 50 farms for this case study. data were gathered at the household level on socio-economic characteristics, agricultural production system characteristics, extension institutions and climate change perceptions table 1. table 1. summary of household characteristics. variable (type variable) description number (%) means perception (ordered variable) climate change perceptions 50(100%) no perception average perception good perception 6(20%) 5(17%) 19(63%) age (discrete variable) ages of farmer 50(100%) 50 education (ordered variable) level of education 50(100%) literate =1 primary =2 secondary =3 university=4 3(6%) 37(73%) 0(0%) 10(20%) main activity (binary variable) main agricultural activity 50(100%) if agriculture = 1 other = 0 25(50%) 25(50%) residence (binary variable) place of residence 50(100%) if on the farm = 1 outside = 0 45(90%) 5(10%) farm size (continuous variable) farm size 50(100%) 4.46 agricultural system (binary variable) type of agricultural production system 50(100%) if stage system = 1 if other system = 0 14(26%) 36(74%) land owner (binary variable) agricultural land owner 50(100%) if farm owner = 1 if other =0 18(26%) 32(74%) membership (binary variable) membership of (gda) 50(100%) if membership = 1 if no =0 23(26%) 37(74%) source: data survey. 3. results and discussions 3.1. perception of climate change for oasis farming households the objective of this section is to identify the determining factors of perception of climate change for the oasis farming households. the ordered probit regression model was used to find out the contributing factors implicitly (ad-hoc) to the perception of the phenomenon of climate change and which can play in the development of adaptation strategies. the results of the ordered probit regression model are presented in table 2. table 2. results of ordered probit regression model for perception of climate change. perception of climate change via ordered probit regression model coef. std. err. p>|z| x1: age 0.036 0.038 0.349 x2: educ 0.808 * 0.437 0.064* x3: agr 0.692 0.773 0.371 x4: resid 3.364 *** 0.732 0.000*** x5: size 0.611** 0.193 0.002** x6: system 0.467 0.544 0.391 x7: owner 1.512 * 0.788 0.055* x7: membership 1.991*** 0.723 0.006*** number of obs. = 50 wald chi2(8) = 504.42 prob > chi2 = 0.000 pseudo r2 = 0.353 log pseudo likelihood = -17.636 notes: *** significant at 1%, ** significant at 5%, * significant at 10%. source: model results. the results of the probit regression model ordered in table 2 show an overall significance of the model at the 1% level (prob> chi2). the positive contribution and the level of significance of the independent variables also determine the importance of these variables in the functioning of the oasis production system. indeed, a significance level of 1% for the residence variable (resid) clearly reflects that the farmer, who is installed in the oasis, felt the change of the environment and the deterioration of the oasis agricultural systems from day to day other than a farmer who is outside the oasis. the other variable that is significant at the 1% level is membership of an agricultural development group (membership) which is responsible for the activity of agriculture in the oasis, among other things the distribution of irrigation hours. being a member of this group means ease of access to information and extension and staffing of main irrigation hours or additional hours. therefore, we can deduce that these two variables are of the first order from the point of view of perception of climate change among oasis farming households. then, in second order, we find the variable area of the agricultural holding (size) significance at the 5% level. this variable shows that the perception can be perceived at the level of large farms whose activities require a lot of production inputs such as water, labor, etc. in third order, at the level of positive significance of agriculture and food sciences research, 2022, 9(1): 1-8 6 © 2022by the authors; licensee asian online journal publishing group 10%, we find variables like the level of education (educ) as well as the property of the land (owner) which plays an important role in the understanding of the phenomenon of climate change and their impact on oasis systems when the farmer owns their agricultural land. the land ownership variable (owner) also reflects a socio-cultural aspect among some farmers, beyond that oasis activity is a source of agricultural income, but it is also a natural heritage characterizing the region which must be preserved for biodiversity, sustainable development and for future generations. 3.2. adaptation strategies on oasis farming households the results of the ordered probit regression model showed the positive and significant contribution of certain key variables that can be used firstly to understand the behaviors of oasis households with respect to the phenomenon of climate change and secondly, avenues for reflection to develop adequate strategies adaptation. in the table 3, the principal component analysis method (pca) was used according to a set of classification criteria in relation to the variables of positive perception of climate change identified by the ordered probit model. among these criteria are: csd: change of sowing date; add_irrig: additional irrigation; purch_irrig: purchase of additional irrigation hours; sal_an: sale of animals to finance agricultural activity; adopt_crop: adoption of other crops; political_instr: state intervention through subsidies and encouragement; acces_credit: access to credit to invest and finance agricultural activity. table 3. component matrix of principal component analysis (apc). classification criteria components 1 2 3 csd -0.394 -0.244 0.729 add_irrig 0.799 0.404 0.254 sal_an -0.593 0.542 -0.051 adopt_crop -0.630 -0.392 0.577 political_instr -0.206 0.628 0.534 acces_credit 0.714 0.175 -0.315 purch_irrig 0.887 0.305 0.160 note: extraction method: principal component analysis. a. 3 components extracted. component 1 characterized by the positive contribution of criteria (add_irrig, purch_irrig and acces_credit) can be interpreted as strategy 1: adaptation strategy in terms of water saving policy. component 2 characterized by the positive contribution of criteria (sal_an and political_instr) can be interpreted as strategy 2: autonomous adaptation incentive strategy and component 3 characterized by the positive contribution of the criteria (csd and adopt_crop) can be interpreted as strategy 3: technical strategy and production system. these three adaptation strategies reflecting the behaviors of oasis farming households which are deduced by the main component method on the data from the survey questionnaires are also confirmed by local actors, whether they are experts or agents of the service agricultural extension in the region. 3.3. determinants factors of adaptation strategies of oasis farming households to climate change the results of ordered binary probit models in the table 4 show that the overall significance of a level 5% (prob> chi2) for the strategy 1 and strategy 2; at level 1% (prob> chi2) for the strategy 3 reflecting acceptance of the choice of adaptation variables to describe the strategies identified. the results also show the significant contribution of certain variables to express adaptive behavior among farmers in the oasis of methouia. table 4. binary probit models results. explanatory variables strategy 1 strategy 2 strategy 3 coef. p>|z| coef. p>|z| coef. p>|z| x1: age 0.259*** 0.003 0.065 0.250 0.155** 0.027 x2: educ 0.583 0.147 0.504 0.114 0.395 0.380 x3: agr 1.863** 0.045 1.299** 0.047 1.747*** 0.008 x4: resid 1.828 0.191 -1.177 0.430 -2.879 0.060 x5: size 0.050 0.795 0.044 0.854 0.075 0.755 x6: system -1.033 0.186 1.767*** 0.003 1.731*** 0.001 x7: owner 2.345*** 0.001 1.366* 0.069 0.841 0.249 x7: membership 2.472** 0.016 1.617* 0.079 2.051** 0.034 constante -19.642 0.002 -8.700 0.052 -13.740 0.015 number of obs. = 50 wald chi2(8) = 16.15 prob > chi2 = 0.040 pseudo r2 = 0.388 number of obs. = 50 wald chi2(8)= 23.02 prob > chi2 = 0.003 pseudo r2 = 0.413 number of obs. = 50 wald chi2(8)= 30.83 prob > chi2 = 0.000 pseudo r2 = 0.468 notes: *** significant at 1%, ** significant at 5%, * significant at 10%. the positive and significant contribution at the 1% level for variables such as age (age) and land ownership (owner) and also other variables that are significant at the 5% level as the main activity variable which is agricultural activity (agr) and the variable member of an agricultural development group (membership). these variables explain and justify the adoption of strategy 1 (adaptation strategy in terms of water saving policy) for some farmers in the oasis of methouia. indeed, an oasis household head who owns a farm, his main activity is agriculture and member of an agricultural development group where their access to water is possible, all these conditions allow him to access credit to invest in water saving for the purchase of drip irrigation or to build a water basin to store rainwater or additional irrigation water. for strategy 2 (incentive strategy for autonomous adaptation), the results of the binary probit models in table 3 show the positive and significant contribution at the 1% level for the classification variable of the production agriculture and food sciences research, 2022, 9(1): 1-8 7 © 2022by the authors; licensee asian online journal publishing group system (system), at the level of 5% for the variable (agr) and at the level of 10% for the two variables (owner and membership). this strategy explains that when agriculture is the main activity for the oasis household head who is also the owner of a farm and member of an agricultural development group for access to irrigation water, contribute to the choice of orientation of oasis agricultural production system. this orientation towards a new oasis production system can be interpreted as a kind of strategy of adaptation to climate change which is carried out for the benefit of the activity of the breeding in particular the cattle breeding which has known a significant deterioration due to of their significant cost. this autonomous adaptation strategy is achieved through the sale of heads of cattle to finance agricultural activity and also through the incentive procedure of public actors to encourage investment in profitable crops with high added value such as pomegranate trees. this strategy implicitly reflects the transformation and the dynamics of the functioning of oasis agricultural households in the south-eastern region, in particular in the oasis of methouia. for strategy 3 (technical strategy and production system), the results of the binary probit models in table 3 show the positive and significant contribution at the 1% level of the principal active variable (agr) and the production system orientation (system), and at the 5% level for age (age) and member of an agricultural development group (membership) variables. in fact, in recent years, we have noticed a change in the classic oasis production system in three stages (palm, arboriculture, market garden and fodder crops) associated with the activity of cattle and goat breeding towards a new production system oasis in two stages (arboriculture, vegetable and fodder crops) associated only with goat farming. this strategy is imposed by the phenomenon of urbanization, the change in lifestyle of oasis households and also the degradation of natural resources due to climate change. this positive contribution explains that these variables together play an important role in the strategy of technical adaptation and orientation of the production system, whether through the change of the date for certain vegetable crops thanks to the experience of older farmers although the orientation towards less costly agricultural production systems that consume less water. the perception of oasis households of climate change in the study area was consistent with the findings of other researchers around the world. indeed, the regression analysis of the ordered probit model revealed that certain variables such as education, agricultural area, residence, owner of agricultural land and membership of an agricultural development group have influenced the perception of climate change by farmers. the same interpretations for these variables are justified by the work developed in the central agricultural zone of the state of delta, nigeria [32]. for the different adaptation strategies and their determining factors which are identified by the binary probit regression model, for example, the variables: age, agriculture is the main activity, owner of agricultural land and membership of a development group agricultural, reflect the adaptive behavior of oasis households in methouia. the motivation for this adaptive behavior is based almost on four key terms: experience for autonomous adaptation and orientation of production systems, the owner of agricultural land for access to credit and membership in a group of agricultural development for access to water and information and an extension service. these same key adaptation terms also summarize the adaptation of farmers' livelihoods to environmental changes in the case of the minqin oasis, northwest china [33]. 4. conclusion and recommendations the study aimed to assess the perceptions and adaptation strategies of farmers to climate change in the oasis of methouia in south-eastern tunisia. it was found that the perception was raised among the majority of oasis farmers who were well aware that the climate was changing. the majority of farmers noted that there was an increase in temperature, decrease in rainfall, changes in the timing of rains and an increase in the frequency of droughts. the most common adaptation strategies among farm households were: crop diversification, change of production system, increase in water conservation practices, adjustment and management of livestock, the abundance of cattle breeding for the benefit of oasis agriculture and the increased use of irrigation technology through access to credit. the results of the study also show that certain variables such as level of education, residence on the farm, agricultural area, land owner and membership in the agricultural development group are crucial factors in influencing the probability oasis farmers to perceive climate change. likewise, factors such as the age of the head of household, education, the system of the land owner and membership in the agricultural development group, facilitate access to credit and also to extension and information on change climate. these variables can be considered as factors to trace the most adequate adaptation strategies for oasis farmers of methouia to climate change. any policy aimed at strengthening the adaptive capacity of farmers in the study area should consider the use of the factors mentioned above in developing adaptation strategies. the importance of these socio-economic and technical factors of production in the perception and strategies of adaptation to climate change in the case of agriculture is justified by several studies in the world [24, 33-39]. indeed, this study was an example to show that the bottom-up approach going from the individual scale for the case of the farmer to the global (community or society) to forecast the perception and ideas of the autonomous adaptation. this approach can be interpreted as the most effective methodological process in the design of adequate adaptation strategies which takes into account all the economic, social and ecological characteristics of a given region. today, it is time to rethink the development of adaptation strategies to climate change by strengthening the adoption of the bottomup approach on scientific and participatory bases with the actors concerned, first and foremost the farmer and their concerns for 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[39] j. b. smith, t. dickinson, j. d. donahue, i. burton, e. haites, r. j. klein, and a. patwardhan, "development and climate change adaptation funding: coordination and integration," climate policy, vol. 11, pp. 987-1000, 2011.available at: https://doi.org/10.1080/14693062.2011.582385. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. agriculture and food sciences research issn: 2411-6653 vol. 2, no. 2, 43-50, 2015 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 43 tillage methods to determine soil infiltration rates: a case study in uyo, nigeria ime etim 1 --i.i. ahuchaogu 2* --a. i. etuk 3 1,2,3 department of agricultural and food engineering university of uyo, akwa ibom state, nigeria abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ............................................................................................................................................................................... 44 2. results and discussion .............................................................................................................................................................. 46 3. conclusion .................................................................................................................................................................................. 47 references ...................................................................................................................................................................................... 49 bibliography .................................................................................................................................................................................... 7 this study evaluated the effect of tillage methods on the infiltration rate of a sandy loam soil in uyo. nigeria. tillage treatments adopted include zero –tillage, crude tillage, plough alone and plough harrow tillage. the result of the study revealed that the basic infiltration rates under zero crude, plough and plough harrow tillage in a sandy loam soil in uyo were 18mm/hr, 20mm/hr, 21mm/hr and 24mm/hr respectively. soil moisture content was averaged at 8.32% with a mean deviation of 0.372.the bulk density of the soil ranges from 0.611kg/m 3 to 0.613kg/m 3 . the mean percentages of sand, silt and clay from the analyzed soil sample were 74.43%, 13.64% and 16.69% respectively. a 2-way analysis of variance (anova) was used to test the significance difference between the average infiltration rates under the different tillage treatments. the result showed that there is a significant effect of tillage method on soil infiltration rate and soil infiltration rate under different tillage treatment followed the order plough + harrow > plough alone > crude tillage > zero tillage. the study recommends that farmers in uyo should endeavour to till their farm before planting at least with crude tillage implement as it will help to pulverize the soil surface and make it easier for irrigation and root penetration as well as root development. keywords: tillage, soil, infiltration, pulverize, bulk density. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(2): 43-50 44 1. introduction the yield of crops on a particular farmland depends majorly on the soil – plant water relationship. good soil manipulation enhances better penetration of water into the plant root zone, enabling root absorption of soil moisture for better plant growth and development. proper soil management world over has resulted in increased yield of crops, fibers and wood leading to food sufficiency and wealth creation. however where the soil is mismanaged vast areas of land has been eroded away leading to food scarcity, hunger and poverty. maltreatment of the soil in various parts of the world has led to soil erosion on a vast scale. the damaging aspect of soil erosion which is the physical removal of the upper layer of the topsoil leads to the loss of vital nutrient. loss of soil nutrient through erosion is a global phenomenon which can be eliminated by proper soil conservation methods. in the early stages of the development of crop culture, vegetation that competed with plants that provide food was removed by hands. food production was generally limited under these conditions. adewoyin and ajav [1]; ajav and adewoyin [2] stated that soil tillage is considered to be one of the biggest farm operations as it requires more energy on the farm. besides, it is usually used to reduce the effects of erosive force of the wind and water on the farm land. the practice of tillage include ploughing, harrowing, ridging is adopted for controlling erosion and conservation of moisture in the root zone. this in turn produces cloddy soil surface, reduces wind velocity and change the status of the moisture content and other soil properties. ogban, et al. [3] reported increase of organic matter as a result of tillage practice. this act of soil manipulation is majorly aimed at presenting the crop root zone of the soil for proper absorption for crop germination, growth and maturity. as the soil becomes wet, the water is held in pores and the capillary potential is increased, [4]. proper tillage practices can be used to improve soil related constraints while improper tillage operation may cause a range of undesirable processes such as destruction of soil structure, accelerated erosion, depletion of organic matter, disruption of water cycle, organic and plant nutrient. tillage practice which is majorly divided into two, namely primary and secondary tillage ultimately influences the soil physical properties for better aeration, moisture penetration and plants performance. kameníčková, et al. [5] stated that the treatment of the top layer of the soil plays a key role in the changes of the hydro-physical properties, mainly saturated hydraulic conductivity of the surface layer, soil perforation through tillage enhances soil water catchment and increases the infiltration of water into the soil surface, raising the hydraulic conductivity sorptivity values, [6]. there are about seven methods of tillage practices as important as they are, all of them cannot be practiced on the same kind of soil, under the same environmental condition to give the same desired positive result. ahaneku and dada [7] observed that desirable tillage methods are those that positively impact soil physical properties and result in good till. this enhances soil aeration, infiltration rate and water holding capacity of the soil. it also encourages root penetration, nutrient uptake and vigorous growth of crops through reduced penetration resistance of the soil to plant root and farm implement, [7] . the application of conservation tillage has the advantage of cultivating the leaves of the previous year’s crop residue on field before and after planting the next crop to reduce soil erosion and run-off. conservation tillage methods include no – till, strip-till, ridge-till and mulch-till. each methods requires different types of specialized or modified equipment and adaptation in management and has been found to reduce erosion by up to 60%90%.the depth of tillage may turn out to be of disadvantages to the exercise. conventional tillage practice including mouldboard ploughing to a 150 mm depth and other tillage operations could lead to loss of 50% of the soil organic matter and about the same amount of organic nutrient. after rainfall, tillage may be necessary to reduce evaporation loss and maintain adequate water in the root zone. though excessive tillage could lead to loss of organic matter but if the number of tillage operation is minimized and timed primarily to maximize water conservation the advance effect on organic matter can be minimized. the soil is essential for the survival of the human race on the earth as it provides most of the food required, fibers for clothing and wood for building materials. but in many parts of the world the soil has been so mismanaged that it will never be able to produce at least not in the near future. the soil serve as the foundation upon which plant stand and equally serve as a major water catchment and retention trough for the crops thereby making water readily available for the plant roots for increase yield. the physical properties of the soil and their various interactions are influenced largely by soil structure (as characterized by the size, shape and strength of aggregates or the size, shape and stability of pores) and crop residues or organic amendments (placement, quantity and type of organic materials). the performances of crops are equally affected by the physical properties of the soil including water retention capacity, strength, porosity, aeration and temperature. the soil water characteristics play a leading role in determining the nature and extent of variations in the other soil physical properties. tillage generally affects the structural properties of a soil by breaking down clods and thus altering the pour size distributions, and hence the stability of the pores is very dependent on soil organic matter bonds. therefore the basic of all good management is to encourage the development of a good stable soil structure with very active root development and organic activity. water enters the soil surface due to combined influence of gravity and capillary forces. both forces act in the vertical direction to cause percolation. this process of infiltration involves both transmission and storage of soil moisture. according to wuest, et al. [8] the most important factor determining whether water will soak in or run off is the ability of the soil surface to resist slacking and reconsolidation or crushing of the soil surface – slaking which occurs when soil aggregates (cluster or clumps of soil) break apart in water into separate soil particles. this thus makes soil aggregation a very important property of most soil as it controls water infiltration to a greater extent than the amount of sand, silt and clay and is a function of the surface maneuvering of the soil due to disturbance that be generated by farm implement. soil moisture acts as a modular between the land surface and atmosphere, thereby influencing climate and weather [9]. it influences various processes related to plant growth and hence ecological patterns, [10] and agricultural production, as well as a range of soil processes, [11]. the means of conserving agriculture and food sciences research, 2015, 2(2): 43-50 45 moisture falls into three categories namely increasing infiltration, reducing evaporation and preventing unnecessary plant growth. water infiltrates easily on sandy soils thus reducing water retention. however on heavy soils in most environments, water retention is high, residue levels could delay planting resulting in poor early seedling vigor. in many localities, infiltration is limited by surface crushing rather than by properties of the deeper soil. improvement of infiltration reduces run off and thus increase available soil water. the main methods use to increase infiltration are the use of soil amendment, soil management by tillage and conservation farming. these methods may be used separately or together. deep ploughing enhances weed control and the loosing of compacted soil layers. deep ploughing distorts farmland surfaces increase infiltration and water retention in the land holes and depressions. climatic factors like the rainfall makes soil moisture level rise, balances water loss through evapo-transpiration drainage and percolation to the ground water zone – loss of soil moisture is usually higher during the dry season of the year leading to higher evapo-transpiration rate. the moisture status of the soil is expressed in volumetric moisture content and the capillary potential of the water held in the soil pores. as the soil becomes wet, the water is held in large pores and the capillary potential increase, [4]. rainfall leads in increasing the soil moisture, the rate of infiltration depends on the intensity of the input and the initial moisture condition of the surface soil layer and hydraulic characteristics of the soil. small-scale effects such as the presence of a surface seal of low permeability (due to the management of surface soil particles by rain splash) or the presence of large channels and cracks in the surface soil may be important in controlling infiltration rates. if rainfall intensity is greater than the infiltration rate, water will accumulate on the surface and run off will begin, [12]. movement of water into the soil is controlled by gravity capillary action and soil porosity. of these factors, soil porosity is most important. soil porosity is controlled by its texture, structure, and organic content. coarse textured soils have larger pores and fissures than fine-grained soils and therefore allows for more water flow. pores and fissures found in soils can be made larger through a number of factors that enhance internal soil structure. the burrowing of worms and other organisms and penetration of plants roots can increase the size and number of macro and micro-channels within the soil. the amount of decayed organic matter found at the soil surface can also enhance infiltration. organic matters are generally more porous than mineral soil particles and can hold much greater quantities of water, [12]. suresh [13] views infiltration as the movement of water into the immediate soil surface. it is an important component in watershed modeling for the prediction of surface run off. in a study carried out at oregon state university, [14] identified soil organic matter as a soil factor that can affect the aggregation of soil particles and infiltration. the result also showed that a small increase in soil organic matter can have a substantial effect on soil aggregation and a very important effect on water infiltration even in a highly filled cropping system, where increased organic matter levels are very difficult to achieve or maintained near the soil surface. meek, et al. [15] evaluated factors that affect soil water infiltration to include traffic, tillage between crops and the formation of channels by roots of perennial crops. according to kooistra, et al. [16] the infiltration rate will be increased when sandy loam soil is filled because of the lower bulk density, infiltration rate will also decrease because large-pore continuity will be disrupted, and the importance of these two factors will depend on the level of compaction of the soil. the proper use of tillage, control and timing of traffic and selection of crops will allow a grower to maintain adequate infiltration levels so that adequate irrigation water can be applied. tillage may increase or decrease the infiltration rate depending on the degree of soil compaction. unsaturated hydraulic conductivity (k) values that were increased at least four fold by chiseling 0.43 m deep compared with an untilled check. infiltration of simulated rain when the level of tillage disturbance was reduces and suggested that the increase may have been caused by changes in the surface seal. lai [17] measured infiltration rates of a field where maize (zea may l) was cultivated for five years to be 480 mm/hr for no-till and 150mm/hr for the ploughed treatment. it was revealed that surface residue prevented surface seal in the notill treatment. meek, et al. [18] measured a 17% increase in infiltration rate in the field when soil was packed lightly before the first flood irrigation compared with no packing. patel and singh [19] reported that if the bulk density in a coarse textured soil was increased from 1.7 to 1.9 mg m3 hydraulic conductivity decrease by a factor of 260. meek, et al. [15] using the same soil also measured a decrease on infiltration rate of four times when traffic compacted soil from a bulk density of 1.7 to 1.89mg/m. tillage disturbs natural channels that have formed in soil the increase in porosity when soil is tilled may not result in an increase in filtration rate because of disruption of the vertical continuity of the pores [16]. plant roots are important in forming new channels. root growing initially may decrease infiltration rates, but later decomposition of roots leaves channels that result in increased infiltration rates. 1.1. materials and methods 1.1.1. site location and selection figure-1. (a) map of nigeria showing akwa ibom state (b) map of akwa ibom state showing uyo agriculture and food sciences research, 2015, 2(2): 43-50 46 the experiment was carried out in uyo, akwa ibom state, nigeria, figure 1. uyo geographically lies in the niger –delta and is a key member of the south-south zone, which account for more than 90% of nigeria wealth. rich in oil and natural gas with fertile land mass measuring 8,412sqm. it has a coastline of 129km suitable for investment and tourism. lying between latitude 4 0 33’ north longitude 7 0 36 and 8 0 25 east. uyo falls within the tropical rainforest of nigeria with dominant vegetation of green foliage of trees, shrubs the state has two distinct seasons – the rainy season lasts from april to october while the dry season is from november to march. in the coastal region rainfall is almost all the year round. the rainfall varies from 3,000mm along the coast to 2000mm in land; the mean temperature varies between 25 – 28 0 c. with a population of about 2,385,756 people whose major occupations include farming, fishing and petty trading. 1.2. method soil samples were collected at the experimental sites and preliminary test and measurement of soil properties were made before the commencement of tillage operations. the tested properties include moisture content, bulk density, particle size distribution and soil class. 1.3. materials the materials used for the tillage and infiltration experiment included shovel, hammer, spirit level, stop watch, 200 liters plastic drum, timber 75mm x 100mm x 400mm, 600 liters of water, swarag tractor, 3 disc plough, tandem harrow and machete. determination of soil moisture content were done using moisture can (crucible) hot and oven dried and desiccators. core cylinders, weighing balance, oven, desiccators, hammer, rubber band spade, spatula among others were used to determine the bulk density of the soil sample. hydrometer method was used to determine the particle size distribution of the six soil samples, mechanical stirrers, stirrer’s cups soil hydrometer, thermometer, 1000ml measuring cylinder and weighing balance was the apparatus use while the dispersing reagent was sodium hexametaphosphate. the study considered four tillage treatment base on the type and combination of tillage implements used viz zero tillage, crude tillage, plough tillage and plough + harrow tillage. tillage depth was maintained at 16-18cm for all treatments except zero tillage and crude tillage. the soil porosity, bulk density, organic matter content and moisture content were determined using standard laboratory procedures. at the commencement of the experiment immediately following the tillage operations tillage depth was measured at 18cm randomly on the ploughed plot. a steel rude was inserted down into the tillage soil until a characteristic hard pan was encountered. the tillage depth was measured from the corresponding reading on the steel rule under the zero tillage, the soil was not tilled. the soil was left bare for infiltration measurement to be taken. under the crude tillage treatment, the soil was tilled using a steel spade. the tilling was made manually by the researcher. however, the foot-press method was adopted in order to maintain a uniform tillage depth. the plough tillage involved the use of a disc plough of diameter 50cm and effective plough width 0.8m. the plough was hinged on the tractor for the tillage operation. tillage was done at a single run. the average speed of the tillage operation was 200mph and uniform tillage depth range of 16 to 18cm. the plough +harrow method was obtained by a combined use of plough and harrow each for a single run operation. the plough was used first and followed by the harrow to break the tillage lump. the plough was hinged to the tractor for the tillage operation. tillage was done at a single run. the average speed of the operation was 200mph and a uniform tillage depth of 16 to 18cm. the infiltration rates and basic infiltration rates were determined using a double ring cylinder infiltrometer with each ring being 3mm thick, with diameter 60cm for outer ring and 30cm for inner ring, both were 30cm high. the concentric rings were driven with a mallet into the ground uniformly without till and unduly disturbing the soil to at least 15cm, the side of the rings were kept vertically and the measuring rod was driven into the soil so that approximately 12cm is left above the ground. spirit level was then used to achieve uniform level. ponding depth method was used and the test began by pouring water into the rings until the depth were approximately 100mm. the level of water in the inner ring was recorded at the beginning of the test and the water level was also noted after 2 minutes; the drop in water level in the inner ring was recorded using the graduated measuring rod and the water was added immediately to level back to approximately the original level at the start of the test. the tests were continued until the drop in water level is the same over the same time interval. readings were made at intervals of 2,3,5,10,20, and 20 minutes thereby giving a cumulative time of 80 minutes. thus infiltration rates were computed using equation (1) below for each tillage method. the cumulative infiltration rate was plotted against cumulative time for the four different tests under different soil tillage treatment. 2. results and discussion some engineering properties of the soil other than infiltration carried out at the commencement of the tillage operations included moisture content, bulk density, particle size distribution and soil classification. the initial soil moisture content of the soil had values ranging from 8.09% to 8.75%. soil moisture content was averaged at 8.32% with a mean deviation of 0.372. the initial bulk density of the soil had values ranging from 0.611kg/m 3 to 0.613kg/m 3 . the particles size distribution of the soil particles namely: sand, silt and clay were measured. the mean percentages of sand, silt, clay and organic matter from the analyzed soil sample were 74.43%, agriculture and food sciences research, 2015, 2(2): 43-50 47 13.64% and 16.69% respectively. these mean values of the soil particles size were used in the reading of the textural class of the soil samples from the textural triangle gives a sandy loam soil classification. observation and measurements obtained from the measurement of soil infiltration for the plot under zero tillage are presented in table1. infiltration measurements were taken at 1 day and 2 days after tillage respectively. the average cumulative infiltration (mm) under the different tillage treatment was computed as presented in table 3 & 4. these average values were used to plot infiltration rate curves for the tillage treatments as shown in figures 2, 3 and 4 respectively. this shows that infiltration rate under the different soil tillage treatments increased in the order plough +harrow > plough > crude > zero. this is evidenced in the curves as the infiltration curve for plough +harrow is posited above other curve. the infiltration curve for zero tillage was positioned below other curves. this implies that the rate of infiltration increased with the advancement of tillage method. to further verify the result, a 2-way analysis of variance (anova) was used to test the significance difference between the average infiltration rates (table 3) under the different tillage treatment. the anova result computed is shown in table 5 and the test of significance of table 6 show that there is a significant difference between infiltration rates under different tillage treatment (f cal = 15.25; f crit = 9.28. n =4 p = <0.05). however, there was no significant difference between the infiltration rates at different stages of till (f cal = 3.99; f crit = 10.13 n = 2, p < 0.05) this implies that tillage method affects the infiltration rate of the soil. 3. conclusion the study which evaluated the effect of tillage methods on the infiltration rate of the sandy loam soil of uyo. akwa ibom state, nigeria shown that under zero tillage infiltration rate was 18mmhr, crude tillage 20mmht while that under plough alone and plough + harrow tillage was 21mmht and 24mmht respectively. there is a significant effect of tillage method on the infiltration rate and the soil infiltration rate under different tillage treatment followed the order plough +harrow > plough alone> crude tillage >zero tillage. base on the result of the study, it is recommended that farmers in uyo should endeavour to till their farms land before planting as it help to pulverize the soil surface and make it easier for irrigation and easy root penetration as well as root development. table-1. computations of infiltration rate for plough tillage s/n time infiltration (1-d tilth) infiltration (2-d tilth) infiltration rate (1-d tilth) infiltration rate (2-d tilth) minutes mm (mm) mm/minute mm/minute 1 2 11 9 5.5 4.5 2 3 9 10 3 3.33 3 5 12 12 2.4 2.4 4 10 15 13 1.5 1.3 5 10 14 10 1.4 1 6 10 9 9 0.9 0.9 7 20 7 6 0.35 0.3 8 20 7 6 0.35 0.3 total 80 84 75 average** 1.05 0.94 **average infiltration rate = (total infiltration/total time) 1-d tilth = one day after tillage; 2-d tilth = two days after tillage **basic infiltration rate = 21mm/hr. figure-2. infiltration curves for plough tillage agriculture and food sciences research, 2015, 2(2): 43-50 48 table-2. computations of infiltration rate for plough + harrow tillage s/n time infiltration (1-d tilth) infiltration (2-d tilth) infiltration rate (1-d tilth) infiltration rate (2-d tilth) minutes mm (mm) mm/minute mm/minute 1 2 12 9 6 6 2 3 11 6 3.66 3.67 3 5 13 9 2.6 2.6 4 10 16 14 1.6 1.4 5 10 14 12 1.4 1.2 6 10 9 7 0.9 0.8 7 20 8 6 0.4 0.35 8 20 8 6 0.4 0.35 total 80 91 84 average** 1.14 1.05 **average infiltration rate = (total infiltration/total time) 1-d tilth = one day after tillage; 2-d tilth = two days after tillage **basic infiltration rate = 24mm/hr. figure-3. infiltration curve for plough + harrow tillage table-3. average infiltration depths under different tillage treatments cumulative time (minutes) average infiltration depth (mm) zero-tillage crude tillage plough tillage plough + harrow tillage 2 8.5 8 10 12 5 15 16 19.5 23 10 24.5 26.5 31.5 36 20 38 39.5 45.5 51 30 49.5 49.5 57.5 64 40 56.5 57.5 66.5 72.5 60 62.5 64.5 73 80 80 68.5 71.5 79.5 87.5 figure-4. comparative infiltration curves for different tillage treatments agriculture and food sciences research, 2015, 2(2): 43-50 49 table-4. average infiltration rate under different tillage treatments tillage method 1 day old tilth 3 days old tilth average zero tillage 0.85 0.86 0.855 crude tillage 0.91 0.88 0.895 plough tillage 1.05 0.94 0.995 plough + harrow tillage 1.14 1.05 1.095 table-5. analysis of variance (2-way classification) summary summary observations sum average variance zero tillage 2 1.71 0.855 5e-05 crude tillage 2 1.79 0.895 0.00045 plough tillage 2 1.99 0.995 0.00605 plough + harrow tillage 2 2.19 1.095 0.00405 1 day old tilth 4 3.95 0.9875 0.017358 2 days old tilth 4 3.73 0.9325 0.007292 table-6. anova test of significance source of variation ss df ms f p-value f crit rows 0.0694 3 0.023133 15.25 0.025 9.28 columns 0.00605 1 0.00605 3.99 0.140 10.13 error 0.00455 3 0.001517 total 0.08 7 references [1] a. o. adewoyin and e. a. ajav, "appraisal of the utilization and performance of farm tractors for ploughing operations in selected states of south-western nigeria," in proceedings of the 11th international conference & 32nd annual general meeting, nigeria institution of agricultural engineering, 2011, p. 20. 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[6] j. m. abrisqueta, v. plana, j. a. franco, and m. c. ruiz-sánchez, "effect of tillage and water pressure head on the hydraulic properties of a loamy soil surface," spanish journal of agricultural research, vol. 4, pp. 180-186, 2006. [7] i. e. ahaneku and o. a. dada, "effect of different tillage methods and temporal factor on soil physical properties," journal of agricultural engineering and technology, vol. 21, pp. 1-10, 2013. [8] s. wuest, j. williams, and t. johlke, "effects of tillage on water infiltration," columbia basin agricultural research annual report, no. 1068, 2006. [9] d. entekhabi and k. l. brubaker, "an analytic approach to modelling land-atmosphere interaction: stochastic extension," water resources research, vol. 31, pp. 633-644, 1995. [10] i. rodriguez-iturbe, "ecohydrology: a hydrologic perspective of climate-soil-vegetation dynamics," water resources research, vol. 36, pp. 3-10, 2000. [11] r. e. white, principles and practice of soil science.the soil as a natural resource. oxford, uk: blackwell science, 1997. [12] m. pidwirny. infiltration and soil storage. fundamentals of physical geography. 2nd. available http://www.physicalgeography.net/fundamentals/8l.html [15 august 2012], 2006. [13] d. suresh, land and water management principles. new delhi: shansi publishers, 2008. [14] s. machado, k. rhinhart, s. petrie, s. wuest, r. correa, and t. johlke, "long-term experiments at cbarc-pendleton, 2004. in s. petrie and d. long (eds.). agricultural experiment station," oregon state university, special report 1061, 2005. [15] b. d. meek, e. r. rechel, l. m. carter, w. r. detar, and a. l. urie, "infiltration rate of a sandy loam soil: effects of traffic, tillage, and plant roots," soil science society american journal, vol. 56, pp. 908-913, 1992. [16] m. j. kooistra, j. bouma, o. h. boersma, and a. jager, "physical and morphological characterization of undisturbed and disturbed ploughpans in a sandy loam soil," soil tillage research, vol. 4, pp. 405-417, 1984. [17] r. lai, influence of tillage methods and residue mulches on soil structure and infiltration rate. in w.w. emerson et al. (ed). modification of soil structure. new york: john wiley & sons, 1978. [18] b. d. meek, e. a. rechel, l. m. carter, and w. r. detar, "changes in infiltration under alfalfa as influenced by time and wheel traffic," soil science society american journal, vol. 53, pp. 238-241, 1989. [19] m. s. patel and n. t. singh, "changes in bulk density and water intake rate of a coarse textured soil in relation to different levels of compaction," journal of indian society of soil science, vol. 29, pp. 110-112, 1981. bibliography [1] a. w. western, r. b. grayson, and g. b. oschl, "scaling of soil moisture: a hydrologic perspective," annual rev. earth planet science, vol. 30, pp. 149-180, 2002. http://www.physicalgeography.net/fundamentals/8l.html agriculture and food sciences research, 2015, 2(2): 43-50 50 appendix-a. test result for zero tillage s/n time cumulative time infiltration cumulative infiltration infiltration rate infiltration rate minutes minutes mm (mm) mm/minute mm/hr 1 2 2 8 8 4 240 2 3 5 7 15 2.33 140 3 5 10 10 25 2 120 4 10 20 13 38 1.3 78 5 10 30 11 49 1.1 66 6 10 40 7 56 0.7 42 7 20 60 6 62 0.3 18 8 20 80 6 68 0.3 18 views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. 36 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 36-42, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.5699 © 2024 by the authors; licensee asian online journal publishing group effect of different additives on nutrient parameter and palatability of ensiled water hyacinth (eichhornia crassipes) netra prasad pokharel1 keshav dhakal2 pravin panth3 rajan koirala4 ( corresponding author) 1,2,3,4institute of agriculture and animal science, paklihawa campus, tribhuvan university, nepal. 1email: netrapokharel73@gmail.com 2email: dhakalkeshav786@gmail.com 3email: prabinpant92@gmail.com 4email: rajankoirala567@gmail.com abstract water hyacinth, eichhornia crassipes, is an invasive aquatic weed that covers major water bodies in nepal, and its silage has great potential to be used as ruminant’s feed. an experiment was conducted with an aim to explore its ensiling attributes along with palatability test for cattle was conducted at institute of agriculture and animal science, rupandehi in a completely randomized experimental design with three replications. the quality and palatability of water hyacinth silage prepared with additives such as rice straw, molasses, wheat flour, and rice bran were assessed. the treatments included: water hyacinth with rice straw(t1); water hyacinth with rice straw and rice bran(t2); water hyacinth with rice straw and wheat flour(t3); water hyacinth with rice straw and molasses(t4); water hyacinth with rice straw, wheat flour and molasses(t5); water hyacinth with rice straw, wheat flour and rice bran(t6) and water hyacinth with rice straw, wheat flour, molasses and rice bran(t7). the results showed, crude protein (cp), crude fiber (cf), ether extract (ee), total ash (ta), ph, and palatability had significant differences across all treatments (p<0.05). cp (15.13) and cf (23.73) were found to be highest under control whereas rice straw, wheat flour and rice bran had the highest ee (12.74), ta (15.13), ph (3.87), palatability (100%), and considerably high cp (12.40) and cf (19.75). hence, silage of water hyacinth with rice straw, wheat flour, and rice bran has high nutrient content, palatability and can be used as a feed alternative to solve the problem of feed scarcity. keywords: crude fiber, crude protein, ether extract, feed alternatives, nutrient content, palatability, silage, water hyacinth. citation | pokharel, n. p., dhakal, k., panth, p., & koirala, r. (2024). effect of different additives on nutrient parameter and palatability of ensiled water hyacinth (eichhornia crassipes). agriculture and food sciences research, 11(2), 36–42. 10.20448/aesr.v11i2.5699 history: received: 25 march 2024 revised: 29 april 2024 accepted: 13 may 2024 published: 5 june 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: the ethical committee of the institute of agriculture and animal science, paklihawa, tribhuvan university, nepal has granted approval for this study on 23 april 2023 (ref. no. 3114). transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 37 2. materials and methods ................................................................................................................................................................... 37 3. results and discussion ................................................................................................................................................................... 39 4. conclusion ......................................................................................................................................................................................... 41 references .............................................................................................................................................................................................. 41 mailto:netrapokharel73@gmail.com mailto:dhakalkeshav786@gmail.com mailto:prabinpant92@gmail.com mailto:rajankoirala567@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.5699 https://orcid.org/0009-0001-9237-3992 https://orcid.org/0000-0002-0422-084x https://orcid.org/0009-0004-9591-2236 https://orcid.org/0009-0004-0663-2380 agriculture and food sciences research, 2024, 11(2): 36-42 37 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the research provides information that differs in palatability and chemical quality of silage from other similar research and is a unique study against ensiled water hyacinth. the paper adds the knowledge of how different additives can be utilized as feed alternative to solve the problem of feed scarcity. 1. introduction water hyacinth (eichhornia crassipes martius) is one of the most common weed species among floating water weeds in tropical and subtropical locations around the world due to its rapid development [1]. it originated in the amazon basin and quickly spread over latin america, africa, southeast asia, and the pacific region by 1950 [2]. water hyacinth appeared for the first time in ethiopia in 1965, near koka reservoir as well as the awash river ayana [3]. fessehaie [4] and taye, et al. [5] reported water hyacinth infestations in gambela regional state, the blue nile from lake tana to sudan, and lake ellen near alem tena. according to the international union for conservation of nature, these plants are among the 100 most harmful invasive species and the top ten worst weeds worldwide [6]. of the 182 foreign flowering plants found in nepal, 27 are regarded as invasive [7]. there is an immediate rise in livestock production to fulfill the requirements for animal protein for the world’s growing population [8]. the need for animal protein in the tropics has been escalating. there was a 31% deficit in dry matter content required for the well-being of existing cattle and poultry in nepal. scarce feed resources, particularly during the colder months from october to may, lack of understanding regarding feed nutrient composition, absence of expertise in feed formulation, and improper utilization of feed resources based on physiological requirements collectively impede the growth of nepal's livestock sector [9]. this scarcity hampers the timely production of high-quality research due to inadequate research infrastructure and facilities. the aquatic nuisance known as water hyacinth (eichhornia crassipes) poses a significant threat due to its tendency to obstruct water bodies such as rivers, lakes, marine routes, and sewage lagoons. this is particularly prominent in tropical and subtropical regions worldwide [10]. water hyacinth has recently received a lot of interest because of its potential uses as animal feed, aqua feed, water filtration, fertilizer, biogas generation, and potentially human nourishment, among other things [11, 12]. fresh water hyacinth contains 2.38% crude protein, 0.27% crude fat, 0.91% crude fiber, and 3.7% nitrogen-free extract. water hyacinth's dry matter, which is mineral-rich and contains 10-20% crude protein, can be used to substitute some protein in feed and roughage sharma [13]. biswas and mandal [14] discovered that water hyacinth comprised 15.58% crude protein, 19.97% crude fat, 1.33% crude fiber, and 1.85% crude ash after drying and chopping, whereas the leaves contained 16.04% crude protein and 14.97% crude fat. abdelhamid and gabr [15] demonstrated that water hyacinth boasts a crude protein value of 20% on a dry basis, thereby making it a suitable option for animal feed. impressively, water hyacinth can produce 6–10 times more protein per unit area compared to soybeans, hinting at its potential as ruminant feed in regions abundant with water hyacinth. the research site has an abundance of rivers and ponds, which leads to the creation of enormous numbers of water hyacinths. a sizable section of the pond is covered with water hyacinth. once the chemical makeup and nutritional value of water hyacinth are known, farmers may utilize it as an unusual feed for their animals, which will save feed costs and boost output. water hyacinth has high quantities of cellulose and hemicellulose, which may be utilized as ruminant energy sources [16]. because of the low amount of dry matter in water hyacinth, yellowing is often preferred to avoid silage losses [17]. water hyacinth may be effectively ensiled using feed additives and organic acids, and ruminants have indicated that the silages are enjoyable. dairy calves have been fed an ensiled combination of water hyacinth, grain straw, urea, and molasses. this has enhanced milk output [18]. therefore, the purpose of this study was to determine the chemical makeup, nutritional value, and use of water hyacinth during a feed shortage. 2. materials and methods 2.1. site selection the research was carried out at the horticulture farm of paklihawa campus, nepal, started in may 2023. the study area is located at 27.4829°n and 83.4457°e. figure 1 illustrates study location which was chosen owing to the existence of suitable ponds, water reservoirs, and lowlands for the collection of eichhornia crassipes. figure 1. gis map showing the study area. agriculture and food sciences research, 2024, 11(2): 36-42 38 © 2024 by the authors; licensee asian online journal publishing group 2.2. research design and layout research was carried out in a complete randomize design (crd) with seven treatments and three replications. wilted water hyacinth along with additives were used as a treatment. table 1 presents the details of treatments used during experimentation. table 1. details of treatments used in research. treatments treatment details t1 wilted water hyacinth + rice straw (10%) (control) t2 wilted water hyacinth + rice straw (10%) + rice bran (10%) t3 wilted water hyacinth + rice straw (10%) + wheat flour (10%) t4 wilted water hyacinth + rice straw (10%) + molasses (10%) t5 wilted water hyacinth+ rice straw (10%) + wheat flour (10%) + molasses (10%) t6 wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%) t7 wilted water hyacinth+ rice straw (10%) + wheat flour (10%) + rice bran (10%) + molasses (10%) 2.3. silage preparation • water hyacinth was harvested from the dande river of paklihawa. after removing the roots, it was thoroughly washed and allowed to wilt in the shade for about 2 days. • both the petiole and leaf fractions were cut into 4-5 cm pieces and mixed with additives according to the above-mentioned protocols in the polythene bags, each containing hyacinth with different additives of 3 kg, and vacuumed before it was left to ensile. • prepared bags were kept in an isolated place (free from rodents), maintaining temperature for 45 days. • regular monitoring was done to check for undesirable conditions. • after 45 days of preparation, representative silage samples were taken for a laboratory test. a palatability test was also done on the farm. 2.4. preparation of a plant sample a sample from each treatment of silage was taken. the sample was dried immediately in an oven at 70˚c (6080˚c) and kept for 24 hours. while drying, the samples should be thinly spread in the oven. the dried sample was ground in a mixture, and 0.20 g of plant sample was weighted, taken in a filter paper, and dropped as a package into a 100-ml digestion flask or a tube, adding 2 g of digestion mixture along with 10 ml of concentrated h2so4. the material was digested at low temperatures until foaming ceased. the temperature was raised to 400˚c, causing the acid to condense about one-third of the way into the center of the digesting flask. the flask was spun often, and digestion proceeded until carbonaceous particles formed and the color changed to green-blue. before the solution solidified, the flask was refrigerated and 40 ml of filtered water was added. the solution was transferred to a 100ml volumetric flask by rinsing the digestion flask with 3-4 batches of tiny volumes of distilled water and making up the volume. four drops of mixed indicator solution were added to 20 milliliters of 4-percent boric acid solution in 125 milliliters, and the flask was then put under the condenser. a 20-ml aliquot of the digested solution was transferred to a distillation flask, and 100 ml of distilled water was added. keeping the flask at a 45-degree angle, 20 milliliters of a solution containing 40 percent of sodium hydroxide were poured down the neck until it reached the bottom without mixing. the flask was quickly affixed to the distillation equipment and stirred to mix. the boiling temperature of the distillation flask prevented the boric acid from being drawn back. the distillate was created after about 75 milliliters of distillation. the nitrogen content was determined by titrating the distillation product with 0.05n hcl. at the end, the mixed indicator's hue simply changes from blue to reddish. for every batch of 21 samples, run the blank using all of the ingredients and procedures, excluding the plant sample. % n = (s−b) × n × 7 𝑊 where, s= volume of standard acid (ml) used up by sample. b= the volume of standard acid (ml) used up by blank. n = normality of the standard acid. w = oven dry weight of sample. 14= equivalent weight of nitrogen. 20= aliquot. 2.5. ether extract (crude fat) first, a filter paper thimble was created, put on a weighing scale, and its weight was recorded. a silage sample was ground. 2 g of sample was taken into the thimble, and cotton was placed to cover the sample. a thimble was folded to enclose the sample. a cellulose thimble was taken and labeled with a sample number. a clean and dry flat bottom flask was taken. the weight of the flask was also taken. a soxhlet extraction unit was setup placing, the sample was placed in it, and a sufficient amount of n-hexane was added. through the condenser of the soxhlet extractor, water was run. the soxhlet extractor was run for 6 hours. after 6 hours, thimble was taken out, and wastage n-hexane was collected. a sample was taken out of the thimble, and wastage n-hexane was collected from the thimble. by condensation, the rest of n-hexane was collected from the flask. the flask was rotated to evaporate the excess n-hexane from it. inside the flask, extracted oil was seen. to remove moisture and hexane, a flask was placed inside the oven. the temperature was set at 110˚c and dried for 30 minutes. after 30 minutes, dried flask was taken out, and placed, and cooled in the desiccator. after cooling, the final weight of the flask with fat was taken. agriculture and food sciences research, 2024, 11(2): 36-42 39 © 2024 by the authors; licensee asian online journal publishing group weight of sample = ws. weight of flask = w1. weight of flask with fat = w2. now, crude fat (%) = (w2− w1) ws × 100 2.6. crude fiber 2.6.1. boiling in acid a 2 g oven-dried sample was weighed before adding 200 ml (0.158m) of sulfuric acid to the cylinder and pouring it into a conical flask. the sample was added to the flask shown above. following 30 minutes of boiling, a flask was put on a hot plate and shook by hand. the trash flask was placed inside a funnel lined with cotton fabric. the boiling sample was filtered to remove the acid solution. the flask was thoroughly cleaned with hot water to eliminate any remaining acid residue. a 200 ml 0.313 m naoh solution was measured. 2.6.2. boiling in base the naoh solution was put into the conical flask to clean the filtrate. the flask was shaken to mix before being put on a heated plate. after 30 minutes of boiling, the flask was shook with one hand. the trash flask was placed next to a funnel lined with cotton fabric. the base solution was drained from the boiling sample using filtration. the flask was thoroughly cleaned with hot water to eliminate the base residue, and fiber was collected in the crucible. 2.7. drying of fiber the crucible was put on a heated plate to evaporate any extra water. the obtained fiber was dried in an oven at 105℃ for 2 hours. after drying, the crucible was weighed and the result was recorded. 2.8. incineration of fiber the crucible was placed in a muffle furnace and fiber was burnt at 550℃ for 2-4 hours. after burning, the crucible was cooled using a dessicator. after 10-20 minutes, the crucible was taken from a dessicator and weighed. wt. of sample = ws. wt. of crucible with fiber = w1. wt. of crucible with ash = w2. now, crude fiber = (w1−w2) ws × 100 2.9. ash content the crucible was weighted, and its weight was noted to 4 decimal places. 2 g of sample was weighted in a crucible, and the weight was recorded to 4 decimal places. the sample was placed in muffle furnance at 600˚c for 2 hours. after that, it was cooled in the dessicator and weighed within 1 hour after reaching room temperature. the sample was now weighed and recorded at 4 decimal points. % ash = ashed wt. – crucible wt. crucible and sample wt.−crucible wt. ×100 2.10. palatability test representative 12 cattle of the same breed were fed 2 kg of silage from each treatment for 3 consecutive days from each replication, and feed intake was noted. the first day of the trial was done by feeding 1.5 kg of silage from each treatment. the cattle were allowed to feed on a given silage weight (w1). after 15 minutes, the left-over feed (w2) was recorded. percentage left over feed = 𝑊2 𝑊1 × 100% 2.11. ph test representative samples were taken to the lab and grinded into fine dust, then weighed at 5 g each. then each sample was dissolved in 20 ml of distilled water and left for 10 minutes. data were collected by using a ph meter in the lab. 2.12. data analysis and statistical tools the tabulation and processing of data were done using ms excel 2016, and the data were statistically analyzed following an analysis of variance (anova) using r-studio (r 4.3.2). the significant differences between treatment means were compared by the least significant difference (lsd) at the 5% level of probability. arc gis was used for making a topographic map of the study area. 3. results and discussion 3.1. physical quality parameter after 45 days, the silage in each treatment was ready for use. ph is one of the primary quality parameters for silage. silage is often categorized as very good (ph 3.8 to 4.2), excellent (ph 4.2 to 4.5), and fair (ph >4.5) depending on ph [19]. in the experiment, wilted water hyacinth + rice straw (10%) + rice bran (10%) and wilted water hyacinth + rice straw (10%) revealed very good quality with phs of 4.51 and 4.21, respectively (table 2). the odor of wilted water hyacinth + rice straw (10%) + molasses (10%) was rated excellent with a brownish green color. nearly all treatments have a ph of less than 4, with wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%) having the best ph. agriculture and food sciences research, 2024, 11(2): 36-42 40 © 2024 by the authors; licensee asian online journal publishing group table 2. effects of different feed additives on ph, color, and odor of silage after 45 days. treatments ph color odor wilted water hyacinth + rice straw (10%) (control) 4.51±0.06a dark brown bad wilted water hyacinth + rice straw (10%) + rice bran (10%) 4.21±0.09b brown bad wilted water hyacinth + rice straw (10%) +wheat flour (10%) 3.67±0.27cd brown good wilted water hyacinth + rice straw (10%) + molasses (10%) 3.16±0.05e brownish green very good wilted water hyacinth + rice straw (10%) + wheat flour (10%) + molasses (10%) 3.39±0.04de golden brown good wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%) 3.87±0.03bc brownish green good wilted water hyacinth+ rice straw (10%) + wheat flour (10%) + rice bran (10%) + molasses (10%) 3.70±0.07cd golden brown good lsd (p<0.05) 0.36*** cv% 5.32 grand mean 3.80 note: cv: coefficient of variation; lsd: least significant difference followed by the same letter in a column are significantly different. ***= treatment means are significantly different at a 0.1% level of significance. in a column, the different letters represents data differs significantly at 5% level of probability. a presented: highest significant mean among treatments. b presented: second highest significant mean among treatments. c presented: significant mean after a and b. d presented: lowest significant means among treatments. the water-soluble carbohydrates included in food additives influence silage fermentation [20]. molasses has 700 grams of carbs per kilogram of dry matter, whereas rice bran has just 53 grams [21, 22]. molasses increased the fermentation process more than rice bran. as ph is a valid metric of fermentation, increasing the ph of the rice bran-added compounds may imply a slower pace of fermentation [23]. good silage tends to be a yellow-green to brownish green hue [24] having an agreeable sweet, sour, and smelling [19]. 3.2. chemical quality of the silage the effect of different additives on the chemical quality of silage is presented in table 4. crude protein is the measure of approximate protein content in silage. wilted water hyacinth + rice straw (10%) has the highest crude protein content (15.13%), followed by wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%) + molasses (10%). the crude protein content of wilted water hyacinth, rice bran, and molasses was found to be 174, 53, and 33 gram per kg respectively [25]. our findings contradicted the findings of indulekha, et al. [19], who found a reduction in the nutrient composition of silage with the addition of straw. the silages supplemented with rice straw had the greatest crude fiber content. additionally, the quantity of crude fiber rose with the inclusion of rice bran. the combination of wilted water hyacinth + rice straw (10%) and wilted water hyacinth + rice straw (10%) + rice bran (10%) has the greatest fiber content. with a crude fiber content of 23.73 and 21.88 percent respectively (table 3). the addition of straw and bran enhanced the crude fiber content of silage. when wheat bran is added to silage, the amount of crude fiber and crude fat increases, as stated by [26]. the maximum ether extract was recorded in wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%), followed by wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%) + molasses (10%). the wheat flour and rice bran have enhancing effects on the ether extract. the maximum total ash content was recorded in wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%), followed by wilted water hyacinth + rice straw (10%) + rice bran (10%). total ash content is an index for measuring total inorganic matter in food samples, which reflects the quantity of minerals present [27]. the higher ash content may be because the root sections used in the ensiling procedure have the ability to absorb different minerals from the water component. although the effects of giving high ash diets to ruminant animals are not fully known, high ash levels in dairy cow diets or forages may operate as a covert antagonist against the effectiveness of dairy nutrition programs [28]. table 3. effects of different feed additives on crude protein, crude fiber, ether extract, and the total ash content of silage after 45 days. treatments crude protein (%) crude fiber (%) ether extract (%) total ash (%) wilted water hyacinth + rice straw (10%) (control) 15.13±0.20a 23.73±0.84a 11.83±0.09cd 12.11±0.05c wilted water hyacinth + rice straw (10%) + rice bran (10%) 10.10±0.19d 21.88±0.76b 11.39±0.22d 14.22±0.16b wilted water hyacinth + rice straw (10%) +wheat flour (10%) 11.06±0.11cd 21.04±0.03bc 12.12±0.05bc 11.69±0.03cd wilted water hyacinth + rice straw (10%) + molasses (10%) 8.09±0.13e 21.01±0.01bc 12.38±0.23ab 11.52±0.29d wilted water hyacinth + rice straw (10%) + wheat flour (10%) + molasses (10%) 9.83±0.37d 18.13±0.20d 11.87±0.08cd 14.17±0.20b wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%) 12.41±0.50bc 19.75±0.44c 12.74±0.16a 15.13±0.13a wilted water hyacinth+ rice straw (10%) + wheat flour (10%) + rice bran (10%) + molasses (10%) 12.59±1.20b 17.33±0.66d 12.39±0.25ab 11.76±0.15cd lsd (p<0.05) 1.49*** 1.60*** 0.51** 0.50*** cv% 7.42 4.40 2.36 2.18 grand mean 11.32 20.41 12.10 12.94 note: cv: coefficient of variation; lsd: least significant difference followed by the same letter in a column are significantly different. ***= treatment means are significantly different at a 0.1% level of significance. in a column, the different letters represents data differs significantly at 5% level of probability. a presented: highest significant mean among treatments. b presented: second highest significant mean among treatments. c presented: significant mean after a and b. d presented: lowest significant means among treatments. agriculture and food sciences research, 2024, 11(2): 36-42 41 © 2024 by the authors; licensee asian online journal publishing group 3.3. palatability of silage the palatability of silage is one of the major criteria when considering its quality. the estimated left-over silage after 3 days of feeding is presented in table 4. the ph value of 3.87 has the highest palatability percentage, i.e. 100%, in wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%). the addition of rice straw primarily decreases the palatability percentage, followed by the addition of wheat flour and rice bran. the preservative content, ph, and cattle acceptance of silage are positively correlated [29]. the ph of water hyacinth silage without additives was 7.33, indicating low quality. however, silage with 15% maize bran or molasses added had a ph of 4.1 and 4.2, respectively, and was well-liked by goats and young steers [30]. table 4. effects of different feed additives on the palatability percentage of silage. treatments left over feed after 3 days of feeding (%) wilted water hyacinth + rice straw (10%) (control) 48.23±6.00a wilted water hyacinth + rice straw (10%) + rice bran (10%) 4.00±2.65c wilted water hyacinth + rice straw (10%) +wheat flour (10%) 8.00±5.29bc wilted water hyacinth + rice straw (10%) + molasses (10%) 15.93±10.50b wilted water hyacinth + rice straw (10%) + wheat flour (10%) + molasses (10%) 3.53±2.60c wilted water hyacinth + rice straw (10%) + wheat flour (10%) + rice bran (10%) 0.00±0.00c wilted water hyacinth+ rice straw (10%) + wheat flour (10%) + rice bran (10%) + molasses (10%) 3.40±3.40c lsd (p<0.05) 11.79*** cv% 55.83 grand mean 11.87 note: cv: coefficient of variation; 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licensee asian online journal publishing group agriculture and food sciences research vol. 9, no. 1, 22-31, 2022 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v9i1.3858 © 2022 by the authors; licensee asian online journal publishing group factors determining where to buy olive oil on the tunisian local market: the importance of quality attributes perception ouertani emna1 dhraief mohamed zied1,2 ( corresponding author) 1higher school of agriculture, mograne, university of carthage, tunis, tunisia. 1email: ouertaniemna2015@gmail.com 1,2rural economics laboratory, national agronomic research institute of tunisia, university of carthage, tunis, tunisia. 2email: dhraief.mz@gmail.com tel: +216 71 230 239 abstract the aim of this paper is to investigate the extent to which factors determining the purchase of olive oil within tunisian consumers also determine their choice of its place of purchase. based on an exploratory survey carried out with tunisian household heads, an exploratory factor analysis was conducted to determine the principal dimensions of quality attributes and indicators. to evaluate factor determining olive oil purchase decision and its place of purchase choice, a binomial and a multinomial logistic regressions are respectively used. consumer sensibility to brand and price have a negative impact on buying decision and on the choice of mills, relatives, and store retail, as suppliers of olive oil. originating from a production area encourage consumers to buy olive oil from friends and relatives. tunisian olive oil industry is therefore able to adopt product and distribution policies based on consumer expectations, to improve local market consumption. keywords: logistic regression, olive oil purchase, place of purchase, quality attributes, indicators. jel classification: e21; m31; q13. citation | ouertani emna; dhraief mohamed zied (2022). factors determining where to buy olive oil on the tunisian local market: the importance of quality attributes perception. agriculture and food sciences research, 9(1): 22-31. history: received: 14 january 2022 revised: 23 february 2022 accepted: 8 march 2022 published: 18 march 2022 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. authors’ contributions: both authors contributed equally to the conception and design of the study. competing interests: the authors declare that they have no conflict of interest. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study followed all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 23 2. literature review ............................................................................................................................................................................ 23 3. methodological approach .............................................................................................................................................................. 24 4. results ................................................................................................................................................................................................ 26 5. discussion .......................................................................................................................................................................................... 30 6. conclusions and implications ........................................................................................................................................................ 30 references .............................................................................................................................................................................................. 31 mailto:ouertaniemna2015@gmail.com mailto:dhraief.mz@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v9i1.3858 https://orcid.org/0000-0002-4234-3293 https://orcid.org/0000-0002-7075-8023 agriculture and food sciences research, 2022, 9(1): 22-31 23 © 2022 by the authors; licensee asian online journal publishing group contribution of this paper to the literature several works explore the effect of olive oil quality attributes on the purchase decision without any focus on their role in the choice of its place of purchase. this paper investigates how these olive oil quality attributes also determine the choice of its favorite sales points. 1. introduction the olive crop is deeply rooted in the traditions of tunisia, both in terms of production and consumption. the consumption of edible oil in tunisia is about 22 kg/capita/year and remains dominated by vegetable oil (soya, colza and corn oils). the domestic consumption of tunisian olive oil is based on family reserves, the direct supply of olive mills or the use of informal circuits. it is difficult to provide a precise estimate of olive oil consumption due to variability in practices and in the channels for sale and purchase. in fact, domestic consumption is not only varying from 20.000 to 60.000 tons/year, but also decreasing. while the olive oil consumption in most of the big consumer countries is improved, it decreases in tunisia from 6 kg/capita/year before 2000 to approximately 3.5-4 kg/capita/year in 2015. this tunisian consumption is very lower than the average in the producing countries (20 kg/capita/year in greece; 12 kg/capita/year in spain and italy) [1, 2]. this low and declining consumption was due to the marginal actions to promote olive oil trade and consumption in the local market. indeed, the promotion is focused on the overseas markets. furthermore, the government set the price of the other vegetable oils (soybean oil), enhance local refining, packaging and trade of vegetable oils such as sunflower and maize oils on the domestic market while leaving the price of the olive oil free to reach five times the price of the other vegetable’s oils. this politics to book the tunisian olive oil to the export and to encourage the consumption of the other subsidized vegetables oils on the local market. so, tunisia was the third olive oil exporter in the world, behind italy and spain. the part of the olive oil domestic consumption in the tunisian olive oil production is about 20%, against 150 % in italy, 70 % in greece and 40 % in spain [1]. furthermore, in the local market, consumers traditionally buy olive oil in bulk and directly from neighbouring olive oil processing units. small quantities are retailed in bottles at supermarkets and hypermarkets located especially in large cities [3]. tunisian producers have realized that to improve market share in a global market, they need also to improve local market consumption and meet consumers’ requirements and perceptions. it is becoming evident for the tunisian olive oil industry to adopt a business strategy based on client preferences, so the objective of this paper is to investigate olive oil consumers’ behaviours and to propose a series of business strategies. therefore, the aims of the present study are: • to explore the behaviour of tunisian olive oil consumers toward olive oil. • to define the way tunisian consumers, perceive the quality attributes and indicators of olive oil, as factor influencing consumer behaviour. • and to discover if factors determining the purchase of olive oil also determine the choice of its place of purchase for tunisian consumers. 2. literature review 2.1. consumers' perception of olive oil quality indicators and their purchase behaviour both endogenous factors (needs and motives, learning, self-concept, personalities, and attitudes) and exogenous factors (culture, reference group, family, and socio-economic situations) affect consumers’ buying behaviour [4, 5]. with reference to olive oil consumption, vlontzos and duquenne [5]; van waterschoot, et al. [6] demonstrate the impact of socio-economic and spatial attributes of consumers households on their olive oil purchase and on the supply modes choices. also, products attributes like, product quality, price, place of sale and country of origin, affect the consumer’s attitude and food purchase decision. more precisely these attributes have "search, experience or credence" properties [7-9]. consumers attach considerable importance to search attributes which are quality indicators that can be tested easily, prior to purchase, by actual inspection of the good. experience quality attributes are those that can only be confirmed after purchase and consumption of the product. credence qualities cannot be verified even after purchase and consumption, they have a positive impact on consumer’s attitude towards a product, and consequently influence consumers buying intentions [10]. for olive oil, different search attributes are proposed for olive oil such as color, smell, taste, transparency, size shape, material and color of the bottle, design of the label and the information released, brand and point of sale [11, 12]. focusing on these search attribute for olive oil, studies on this topic present two groups of factors that allow consumers to evaluate the product: intrinsic attributes and extrinsic attributes. these factors determine perceived quality [12-15]. several studies about olive oil perceived quality revealed that the intrinsic attributes (such as taste, color, flavour, smell) and the extrinsic attributes (packaging, price, place of sale, region of origin) have determine consumers’ purchase decision [16]. however, theses quality attributes are evaluated differently according to the markets. in the greek and dutch markets, quality and color are more important than smell, image-reputation, package, and social influences attributes [3]. whereas krystallis and ness [16]; garcía, et al. [17] point out that greek olive oil consumers grant no importance to the product brand. for the canadian context, studies prove the relative importance of origin, price, and production system compared to color and appearance [3]. studies about uk consumers demonstrate that packaging, size, and price as very determinant factors rodolfo and mónica [18]. rodolfo and mónica [18]; delgado and guinard [19] reveal that the spanish market are divided on two consumers segments: consumers choosing the olive oil because of price and other were guided by the product’s specific attributes, such as organic production. in the usa market, consumers consider the price, available information, and product reputation as determinant factors in the purchase of olive oil [20]. especially for organic olive oil, sandalidou, et al. [20]; sandalidou, et al. [21] attest that greek organic olive oil consumers have a positive perception of health, package, and sensory characteristics such as flavour and taste, but they are dissatisfied about price/quality, promotion, and disposition of this product [21, 22]. while price, geographic origin and certification of extra-virgin olive oil are very important attributes italian consumers of organic olive oil according to cicia, et al. [22]; tsakiridou, et al. [23]. on the other hand, tsakiridou, et al. [23] agriculture and food sciences research, 2022, 9(1): 22-31 24 © 2022 by the authors; licensee asian online journal publishing group verify that socioeconomic consumer characteristics (such as their income, their occupation) deeply affect the demand for organic olive oil [24]. for the tunisian market, mtimet, et al. [2] demonstrate that olive oil consumption is rooted in the tunisian consumers tradition, shoppers are familiar with olive oil attributes, but they are indifferent to the region of origin or any other quality label, and they generally buy olive oil in bulk [3]. oppositely, consumers in northmediterranean olive oil-producing countries are more interested on the region of origin, quality labels and packaging attributes. it should be noted that taste, color and smell can be considered as experience attributes because they are the results of previous consumer experiences. in addition, the choice between different olive oils depends on consumption mode and of food preparations (frying, cooking, for salad dressing…) [25]. besides, cacchiarelli, et al. [24] approved the importance of nutritional and health benefits and positive environmental impacts as credence attributes for olive oil [26]. according to sinha and batra [26] the probability of buying products is lower when the product category is high in experience/credence quality instead of searching through package label information [27]. 2.2. olive oil place of purchase preferences place of purchase is an additional interesting aspect related to purchase habits. based on delgado and guinard [19] and fotopoulos and krystallis [27] studies [14, 20-24, 26-28] reported that most us consumers bought olive oil principally at the supermarket and stores (68%), in contrast with the ways in which mediterranean consumers most frequently buy their olive oil. 41% of cretan consumers buy olive oil at the supermarket, while 38% buy in bulk directly from the producer or farm, and 21% make oil from their own olive orchards [29]. focusing on the italian internal market, giuseppe [3] demonstrate quite differences between the centralnorthern olive oil market and the southern italy [4]. consumers in the central-northern area mainly purchased extra-virgin olive oil at supermarkets or in large-scale retail stores (70%), only 23% purchase olive oil directly from grocery stores (or traditional stores) particularly when they wanted to buy high-quality olive oil. only 5.5% of the sample bought the product at oil mills. in southern italy market, just 27.4% of consumers buy olive oil in largescale retail stores, whereas people who purchase olive oil directly from the producer reached 20.3%. those who bought olive oil in other places of purchase was equally distributed: traditional stores (17.4%), local markets (17.2%) and local oil mills (17.7%). greek olive oil market is dominated by informal markets: more than 45% of household are providing olive oil through relative self-consumption or purchasing olive oil in bulk directly from small producers. factors determining the choice of the place of olive oil purchase are the age of the consumer, the educational level, and the fact of living or not nearby to an olive oil producing area. however, non-significant factors are household's size and family income [6]. moreover, corbeto-fabón, et al. [30] with a focus on the spanish market, demonstrate that the preferences for olive oil attributes depend on the place of purchase [31]. the question arises as to whether the factors determining the purchase of olive oil also determine the choice of its place of purchase. it would be useful to know to what extent those search, experience and credence attributes and those individual socio-economic and demographic characteristics influence the purchase decision of olive oil and its place of purchase selection. a conceptual model to understand factor detraining olive oil purchase and the choice of its place of purchase was proposed in figure 1. figure 1. conceptual framework. 3. methodological approach 3.1. database this study is based on data gathered from a questionnaire survey conducted on a sample of 216 households essentially localized in tunis city centre and its suburbs, the north and the centre of the country, they also originate in the different regions for a better representativeness. in fact, olive oil consumption differs by living environment. the questionnaire was addressed to the household heads. the interviewees were 61% men 68.5% married, with 64% other than 31 years old and 44% with university educational level. the "household size" variable shows that 74% of the families are composed of more than 4 individuals table 1. the income is an important variable in determining the olive oil consumption since it reflects the purchasing power of consumers. it is well distributed among the different income classes selected, with about 45% of the middle classes, whose income is between 1001 and 2000 tnd (euro 3.24tnd). agriculture and food sciences research, 2022, 9(1): 22-31 25 © 2022 by the authors; licensee asian online journal publishing group table 1. socio-demographic profile of the sample (n=216). gender male female 61.1 38.9 age inf 30 31-40 41-50 sup 50 9.3 30.6 33.3 26.9 origin tunis city center and its suburbs north center south 18 41.3 37.1 3.7 marital status married other 68.5 31.5 place of residence north tunis city center and its suburbs center south 32.9 33.3 33.3 0.5 educational level analphabet koranic school primary secondary university 1.9 2.3 20.4 31.5 44 household monthly income ˂500 td 501-1000 1001-1500 1501-2000 2001-2500 2501-3000 ˃3000 5.6 27.3 25.9 18.5 9.3 6.5 6.9 household's size 1 2 3 4 5 ˃6 3.7 9.7 13 26.8 26.8 20 the questionnaire covers all the variables that can affect the consumer behaviour. it is structured into four sections: purchases (place of purchase, quantity, frequency, and budget), consumption preferences, quality attributes and indicators, and socio-demographic and economic variables of the households. for the selected quality attributes and indicators, our assumptions focused on 25 variables that we expect to influence consumer behaviour and measure their perceptions of quality. these variables belong to different quality attributes and indicators defined by branger, et al. [29]; del giudice, et al. [13]; jiménez-guerrero, et al. [14]; ward, et al. [15]; krystallis and ness [16]; garcía, et al. [17]; rodolfo and mónica [18]; delgado and guinard [19]; sandalidou, et al. [20]; sandalidou, et al. [21]; cicia, et al. [22]; tsakiridou, et al. [23]; cacchiarelli, et al. [24]; sinha and batra [26]; fotopoulos and krystallis [27]; corbeto-fabón, et al. [28]; branger, et al. [29]; ding and he [31]. quality indicators are divided into two categories: • extrinsic quality indicators and tangible attributes (6): quality label, packaging, olive oil origin, points of sale, brand name, price. • intrinsic quality indicators (3) such as color, odour and taste. for inviolable attributes, we use 7 credence attributes about health and nutrition and 9 experience attributes (smell, taste, color as experience attributes, mode of uses, and cooking reaction). each of the 25 items, respondents were asked to express, with a score from 1 to 5, the importance of quality attributes and indicators they consider when buying olive oil (1 = "strongly disagree",5 = "strongly agree"). so, the 25 selected quality items are introduced into the database in numerical forms. 3.2. data statistical analysis using spss software (20.0) descriptive statistical analysis was used to determine the socio-demographic profile of the consumers and to describe respondents’ behaviour towards olive oil (perception, purchase, and consumption). an exploratory factor analysis was conducted to determine the principal dimensions among the variables. between the 25 items, 16 were factor-analysed, using principal component analysis (pca) with the varimax rotation method to establish the different dimensions of quality. the varimax rotation was used to maximize the differences between the components extracted and to maintain correlation within the components. the kaiser– meyer–olkin (kmo) measure of sampling adequacy and the bartlett’s test were used to determine the fitness of the data. values of 0.6 or above from the kmo measures indicated that data are adequate for pca [31]. the items factor-analysed sets out details about the different dimensions of quality. to evaluate the impact of socio-demographic factors, search, experience, and credence attributes on olive oil purchase decision (1) and on its place of purchase choice (2), a binomial logistic regression and a multinomial logistic regression are respectively used. for the binomial logistic regression, the outcome variable has only two categories: it is a dichotomous outcome variable describing the purchase or not of olive oil (“1=yes” and “0=no”). for the multinomial logistic regression, the outcome variable has six categories, they describe the six alternatives of olive oil places of purchase: 0= no purchase, 1= olive oil mills, 2=wholesalers, 3= friends and relatives,4= wholesalers, 5= retail stores”. for both binomial and multinomial logistic regression, the predictor variables refer to all the dimensions resulting on the previous factor analysis using principal component analysis. agriculture and food sciences research, 2022, 9(1): 22-31 26 © 2022 by the authors; licensee asian online journal publishing group 4. results 4.1. consuming peculiarities of olive oil in the tunisian market the results of our survey confirm that the purchases of olive oil on the tunisian market are made essentially in bulk. indeed, more than 4/5 of our sample (86.1%) confirm having bought the olive oil and they did it in bulk. the domestic consumption of tunisian olive oil is based on family/friends reserves and the direct supply of olive mills. more than half of the interviewees (53.7%) reported that they bought olive oil primarily from families and friends. 21,3% of interviewed consumers bought it directly from olive oil mills and producers. the market share of wholesalers, retail stores and weekly markets is less important with respective percentages of 4.2 %, 4.6 % and 2.3 % table 2. table 2. the purchasing place choice for olive oil. purchasing place of olive oil (%) no purchase retail stores olive oil producers/mills wholesalers friends and relatives 13.9 6.9 21.3 4.2 53.7 respondents were asked to express, with a score from 1 to 5, what quality attributes and indicators they consider when buying olive oil (table 3). the most important are «taste" (average score 4.83) and "smell" (4.69). in contrast, the less important are attributes about cooking reaction "do not darken" (2.13) and " with high smoking point" (3.3). table 3. quality attributes and indicators perception. quality attributes and indicators minimum maximum mean standard deviation (sd) an oil that do not darken 1 5 2.13 1.057 an oil with high smoking point 1 5 3.3 1.21 brand name 1 5 3.81 1.267 color 1 5 4.57 0.692 cook better 2 5 4.84 0.498 fry better 1 5 1.7 1.068 healthy 4 5 4.97 0.177 multipurpose 1 5 4.27 0.902 nutritional value 2 5 4.94 0.298 origin 1 5 4.37 1.012 packaging 1 5 3.78 1.098 price 1 5 3.89 1.498 purchasing place 1 5 4.19 1.068 quality label 1 5 3.97 1.032 smell 2 5 4.69 0.594 taste 4 5 4.83 0.374 this descriptive analysis summarizes the burden of different quality attributes and indicators on olive oil quality perceptions; it should be deepened by a diagnosis of the determinants of the olive oil purchase on the tunisian market (paragraph 4.3). to this end, an exploratory factor analysis of the olive oil quality attributes and indicators is used to determine the principal dimensions among the variables and to highlight the dimensions of research quality indicators, experience, and beliefs quality attributes. table 4. factor analysis results of the olive oil quality attributes and indicators (varimax rotation). kaiser–meyer–olkin (kmo) measure of sampling adequacy 0.688 bartlett's test for sphericity approx. chi square 803.29 df 120 sig. 0.000 factors 1 2 3 4 5 6 factor 1 extrinsic quality indicators quality label 0.818 0.1 -0.065 -0.084 -0.004 -0.051 packaging 0.803 -0.025 -0.043 0.002 0.06 -0.052 origin 0.778 0.049 0.008 0.29 -0.076 0.233 purchasing place 0.756 0.057 0.002 0.239 -0.14 0.275 factor 2 intrinsic quality indicators smell 0.035 0.846 -0.005 0.074 -0.019 0.016 taste 0.047 0.842 0.219 -0.066 -0.032 0.044 color 0.097 0.614 -0.079 0.303 0.227 -0.018 factor 3 health / nutrition nutritional value -0.053 0.041 0.863 0.011 -0.006 -0.09 healthy -0.042 0.079 0.861 0.118 -0.017 -0.02 factor 4 price/brand price 0.018 0.018 0.172 0.819 0.015 0.024 brand name 0.339 0.191 -0.032 0.654 0.039 -0.028 factor 5 experience attributes mode of use multipurpose -0.129 0.194 -0.167 0.057 0.657 0.162 fry better 0.067 -0.122 0.18 -0.209 0.647 -0.159 cook better -0.046 0.049 -0.033 0.369 0.556 0.027 factor 6 experience attributes cooking reaction an oil with high smoking point 0.041 0.208 -0.102 -0.011 -0.161 0.732 an oil that do not darken -0.179 0.219 0.015 -0.015 -0.275 -0.701 factor statistics eigen values 3.186 2.123 1.623 1.412 1.088 1.023 % of variance 19.912 13.266 10.146 8.827 6.8 6.395 cumulative variance 19.912 33.177 43.323 52.15 58.95 65.346 agriculture and food sciences research, 2022, 9(1): 22-31 27 © 2022 by the authors; licensee asian online journal publishing group 4.2. factor analysis results of the olive oil quality attributes and indicators 16 items were factor-analyzed, using principal component analysis (pca) with the varimax rotation method to establish the different dimensions of quality (table 4). value of 0.688 for the kaiser–meyer–olkin (kmo) measures indicated that data are adequate for pca, and the bartlett’s test was significant. the exploratory factor analysis with varimax rotation of the 16 variables result edit a six-factor solution that explains 65.34% of the total variance. all six factors had eigen values greater than 1 (table 4). the first dimension was labelled as “extrinsic quality indicators”, which explained 19.91% of the total variance; it is determined by attributes related to quality label, packaging, origin and purchasing place. the second dimension “intrinsic quality indicators” includes three indicators (smell, taste and color) and explain 13.26% of the total variance. the third dimension “health / nutrition” focuses on these tow credence attributes and explain 10.14% of the total variance. the fourth dimension, explaining 8.82% of the total variance, is determined by tow attributes “price/brand”. the fifth dimension called “experience attributes about olive oil mode of use” combines the attributes related to the different mode of use of olive oil: a multipurpose oil, an oil that fry better and an oil that cook better. this dimension explicates 6.8% of the total variance. the sixth dimension, which explain6.39% of the total variance, covers two experience attributes about cooking reaction (an oil with high smoking point and an oil that do not darken). knowing that the most common and reliable criterion in extracting factors is the use of eigen values, all factors are with eigen values greater than 1, so they were retained because they were considered significant. in addition, all the items showed factor loading of more than 0.50. these six quality dimensions of olive oil, in addition to socio-economic and demographic consumers’ characteristics, are used as predictor variables to identify factors determining olive oil purchase decision (1) and its place of purchase choice (2). 4.3. factors determining the purchase of olive oil • all six dimensions of the olive oil quality and the socio-demographic and economic consumers’ characteristics (gender, origin, educational level, household's size) were taken into consideration in the binary logistic regression analysis and the dependent variable (categorical variable) was considered “the purchase of olive oil” and could have only two values,1 or 0, as follows: • 1: the consumer purchase olive oil. • 0: the consumer does not purchase olive oil. the values of the regression coefficients and their statistical significance obtained by enter logistical regression method were included in table 4. the logistic regression could use two indicators such as cox and snell r2 and nagelkerke r2that estimates the contribution of predictor variable to the variability of dependent variable. we used the nagelkerke r2 indicator to analyse the contribution of all predictor variables to the variability of the dependent variable. knowing that cox and snell r2 indicator usually underestimates the real value, the test results (table 4) based on the 10 predictor variables could explain in 40,3% the effect of the environment on the artefacts. the results from the classification table showed that the mathematical model predicts 86.6% of cases correctly, so we could conclude that it is a good performing model, and its regression coefficients (β) are shown in “variables in the equation” (table 5). ten predictive variables were analysed by regression analysis, but only three of them had a statistical significance (table 5): educational level, factor 1 referring to extrinsic quality indicators and factor 4 about the consumers’ sensibility to “price and brand”. the remaining predictive variables did not show a statistical significance. by interpretation of the sign of the β coefficients of the independent variables, the educational level, the factor 1 related to consumers’ sensibility to extrinsic quality indicators (quality label, packaging, origin, purchasing place) have a positive effect on olive oil purchase. however, the sensibility of the consumer to “price/brand” quality indicators (factor 4) has a negative effect on olive oil purchase. the educational level of tunisian consumers has a positive and significant effect on the purchase of olive oil: by reference to the odds ratio, it can be argued that when the consumer is with high educational level, the purchasing propensity is 4.7 higher than when he was with primary or secondary educational levels, or also illiterate. in addition, consumers who attach greater importance to extrinsic indicators of quality, particularly the origin of the olive oil, quality labels, packaging, and place of purchase) have a higher probability of purchasing olive oil (odds ratio=2.51). 4.4. factors determining the choice of the purchasing place of olive oil on the tunisian local market a multinomial logistic regression model was used to identify the relationships between dependent variable “choice of the purchasing place of olive oil” and independent variables (the six dimensions resulting on the factor analysis using principal component analysis and four socio-economic and demographic variables). the dependant variable “choice of the purchasing place of olive oil” has m categories(m=5); having no olive oil purchase (and consequently no choice of a place of purchase) was specified as the baseline category (m=1). the probability of membership in other categories is compared to the probability of membership in the reference category. moreover, there will be five predicted log odds, one for each category relative to the reference category. results of the multinomial logistic analysis indicated that the predictor model provides a statistically significant prediction of the choice of place of purchase, -2 log likelihood=391.890, χ2=150.299 and p=0.000. the nagelkerke r2 indicated that the model accounted for approximately 54.6% of the total variance, cox and snell r2=0.501 and mcfadden r2=0.277. prediction success for the cases indicated an overall prediction success rate of 62%. the table 5 presents the regression coefficients, the wald test, adjusted odds ratio [exp(b)], for the odds ratio for each predictor variable used for the four alternative of olive oil place of purchase. the results indicated that some predictors were not statically significant predictors for the four categories of olive oil place of purchase. there was no significant effect such as household's size, factor 2 relative to intrinsic quality indicators perception, agriculture and food sciences research, 2022, 9(1): 22-31 28 © 2022 by the authors; licensee asian online journal publishing group factor3 about health and nutrition as credence quality attributes, factors 5 and 6 concerning consumers' sensibility to experience quality attribute. table 5. the binary logistic regression of the purchase decision model (enter method). model summery step −2 log likelihood cox and snell r2 nagelkerke r2 1 119.501a 0.223 0.403 parameters β s. e.β wald χ2 degrees of freedom p value exp (β) step 1b consumer origin (1 if from production area, 0 if not) 0.630 0.523 1.448 1 0.229 1.877 educational level (1 if with high education. 0 if not) 1.553 0.676 5.286 1 0.021 d 4.727 gender (1 if women, 0 if men) 0.284 0.496 0.328 1 0.567 1.329 household's size(sz) (1 if ≥5 members, 0 if not) -0.332 0.482 0.475 1 0.491 0.717 factor1. extrinsic quality indicators 0.921 0.341 7.303 1 0.007 c 2.512 factor2. intrinsic quality indicators 0.277 0.220 1.589 1 0.208 1.320 factor3. health / nutrition -0.183 0.483 0.144 1 0.704 0.832 factor4. price/brand -1.724 0.468 13.572 1 0.000 c 0.178 factor5. experience attributes mode of use -0.187 0.380 0.242 1 0.623 0.829 factor6. experience attributes cooking reaction -0.077 0.289 0.071 1 0.789 0.926 constant 2.049 0.715 8.219 1 0.004 7.757 note: a. estimation terminated at iteration number 7 because parameter estimates changed by less than 0.001. b. variables entered in step 1: gender, origin, educational level, household's size, factor 1, factor 2, factor3, factor 4, factor5, factor 6. c. statistically significant for p < 1% d. statistically significant for p < 5%. table 6 presents the factors that influence the choice of olive oil points of sales based on the results of a multinomial logit model. for the olive oil producers /mills as first category of olive oil place of purchase, educational level, and the factors one and four describing consumers’ sensibility to extrinsic quality indicators were statically significant. educational level was a significant predictor (p<5%) of mills choice for olive oil purchase with a positive impact. in fact, with a highest educational level, the probability of buying olive oil from mills increase exp (β)=4,8. furthermore, factor 1 describing the importance of extrinsic quality indicators such as quality label, packaging, origin and purchasing place as quality indicators for tunisian consumers, was a significant factor determining the choice of mills as point of purchase (p<1%). it has a positive impact: for consumers finding factor1 very important for quality assessment and for purchase decision, the probability to opt for mills as olive oil supplier increase de 5,02. by buying olive oil from mills, consumers are sure about the origin of the product, the quality label “organic olive oil”, they consider producers/mills as a guarantor of quality. additionally, the importance of brand/price as quality indicators for tunisian consumers (factor 4) is a very significant factor (p<5%), with negative effect on olive oil producer choice: the more the consumer considers the brand and the price as olive oil quality indicators, the less likely it is to buy this oil from mills. indeed, consumers who buy from olive oil from mills, buy olive oil in bulk and in large quantities, at an annual frequency, therefore they pay large sums of money in return. only one predictor is significant for the “wholesalers” as place of purchase of olive oil: the gender is a significant (p<5%) and a main factor determining the choice of wholesalers for the olive oil purchase. in fact, women were 17 times more likely to buy olive oil from wholesalers look for the lowest prices and to save money. when the dependant variable is “friends and relatives”, four predictors are significant: consumer origin, educational level and factors “1 and 4”. the explicative variable “consumers origin” is significant (p<1%) and have a positive impact on buying olive oil from friends and relatives. in fact, the originating from an olive oil producing area are 5,4 times more likely to buy olive oil from friends, family, and relatives. “educational level” is a significant variable (p<5%) with a positive influence on the decision of buying olive oil from relatives; the transition to the high educational level increased the probability of purchasing olive oil from family and friend 4-folds. the first factor describing consumers’ sensibility to extrinsic quality indicators (except brand and price indicators) is statistically significant at 5%. with an exp (β) 2, it has a positive effect on the decision of purchasing olive oil of friends and relatives. especially in the case of consumers belonging to producing countries, when consumers buy olive oil directly from friends and relatives olive oil producers, they are sure about the origin of the product, the quality label “organic olive oil”, they also consider these suppliers as a guarantor of quality. place of purchase is part of the extrinsic indicators determining quality, but also, it is a choice to be made regarding supply areas. as for the olive oil mills, factor 4 "brand/price" as quality indicators for tunisian consumers are significant (p<1%), with negative effect because consumers who buy from friends and relatives, buy olive oil in bulk and in large quantities, at an annual frequency, therefore they pay large sums of money in return. for the retail stores as category of olive oil place of purchase, factor 1 describing the importance of extrinsic quality indicators such as quality label, packaging, origin and purchase area, was a significant factor determining the choice of retail store (p<1%). it has a positive impact: for consumers finding factor1 very important for quality assessment, the probability to opt for retail stores increase de 4,08. agriculture and food sciences research, 2022, 9(1): 22-31 29 © 2022 by the authors; licensee asian online journal publishing group table 6. the results of the multinomial logit model-place of purchase choice. place of purchase a parameter β s. e.β wald degrees of freedom p value exp (β) o li v e o il p ro d u ce rs m il ls constant 0.642 0.825 0.606 1 0.436 factor1. extrinsic quality indicators 1.614 0.433 13.877 1 0.000 5.025 factor2.intrinsic quality indicators 0.341 0.275 1.534 1 0.216 1.406 factor3.health / nutrition 0.098 0.564 0.030 1 0.862 1.103 factor4.price/brand -1.262 0.519 5.900 1 0.015 0.283 factor5.experience attributes mode of use -0.097 0.418 0.054 1 0.816 0.907 factor6.experience attributes cooking reaction 0.285 0.337 0.714 1 0.398 1.329 educational level (1 if with high education. 0 if not) 1.575 0.737 4.563 1 0.033 4.830 gender (1 if women. 0 if men) 0.454 0.577 0.620 1 0.431 1.575 household's size (1 if ≥5 members. 0 if not) 0.001 0.564 0.000 1 0.998 1.001 consumer origin (1 if from production area. 0 if not) -0.097 0.618 0.025 1 0.875 0.908 w h o le sa le rs constant -3.294 1.710 3.709 1 0.054 factor1. extrinsic quality indicators -0.333 0.639 0.271 1 0.603 0.717 factor2. intrinsic quality indicators 0.367 0.427 0.739 1 0.390 1.444 factor3. health / nutrition 0.574 1.765 0.106 1 0.745 1.776 factor 4. price/brand 0.090 0.846 0.011 1 0.916 1.094 factor 5. experience attributes mode of use -0.924 0.702 1.733 1 0.188 0.397 factor 6. experience attributes cooking reaction -0.221 0.539 0.167 1 0.682 0.802 educational level (1 if with high education. 0 if not) 1.803 1.099 2.689 1 0.101 6.065 gender (1 if women. 0 if men) 2.811 1.216 5.348 1 0.021 16.632 household's size (1 if ≥5 members. 0 if not) 0.086 0.913 0.009 1 0.925 1.090 consumer origin (1 if from production area. 0 if not) -1.170 0.955 1.501 1 0.221 0.310 f ri en d s an d r el at iv es constant 0.929 0.797 1.360 1 0.243 factor1. extrinsic quality indicators 0.782 .370 4.459 1 0.035 2.186 factor2. intrinsic quality indicators 0.269 0.241 1.246 1 0.264 1.309 factor3. health / nutrition -0.218 0.523 0.173 1 0.677 0.804 factor4. price/brand -1.928 0.494 15.250 1 0.000 0.145 factor5. experience attributes mode of use -0.093 0.403 0.053 1 0.818 0.911 factor6. experience attributes cooking reaction -0.218 0.309 0.499 1 0.480 0.804 educational level (1 if with high education. 0 if not) 1.572 0.705 4.977 1 0.026 4.818 gender (1 if women. 0 if men) -0.024 0.538 0.002 1 0.964 0.976 household's size (1 if ≥5 members. 0 if not) -0.506 0.519 0.953 1 0.329 0.603 consumer origin (1 if from production area. 0 if not) 1.636 0.605 7.313 1 0.007 5.134 r e ta il s to re s constant 1.348 0.887 2.309 1 0.129 factor1. extrinsic quality indicators 1.406 0.540 6.769 1 0.009 4.078 factor2.intrinsic quality indicators 0.371 0.404 0.844 1 0.358 1.449 factor3.health / nutrition -0.498 0.556 0.803 1 0.370 0.608 factor4.price/brand -1.923 0.587 10.741 1 0.001 0.146 factor5.experience attributes mode of use -0.099 0.491 0.041 1 0.840 0.905 factor6.experience attributes cooking reaction -0.082 0.419 0.038 1 0.845 0.921 educational level (1 if with high education. 0 if not) -0.486 1.053 0.213 1 0.644 0.615 gender (1 if women. 0 if men) -0.939 0.861 1.188 1 0.276 0.391 household's size (1 if ≥5 members. 0 if not) -1.092 0.793 1.897 1 0.168 0.336 consumer origin (1 if from production area. 0 if not) -0.539 0.805 0.449 1 0.503 0.583 note: a. the reference category is: 0 “no purchase”. additionally, the importance of brand/price as quality indicators for tunisian consumers (factor 4) is a very significant factor (p<1%), with negative effect on retail stores choice: the less the consumer considers the brand and the price as olive oil quality indicators, the more likely it is to buy this oil from retail store. in fact, consumers buy olive oil from retail store in small quality but at relatively high prices. these results are in accordance with predictions from previous studies [14, 19]. agriculture and food sciences research, 2022, 9(1): 22-31 30 © 2022 by the authors; licensee asian online journal publishing group 5. discussion olive oil purchase habits of tunisian are based on bulk purchase from family/friends reserves and olive mills. these results confirm those of fotopoulos and krystallis [27] reporting that 38% of cretan consumers buy olive oil in bulk directly from the producer or farm, and 21% make oil from their own olive orchards [28]. this is the same situation in the greek olive oil market where 45% of household are providing olive oil through relatives or directly from small producers van waterschoot, et al. [6]. jiménez-guerrero, et al. [14] establish the same results and state that similar figures can be associated with other mediterranean producing countries [29]. according to theses authors, this habit seems to be a consequence of the consumer experience and a result of their intrinsic cultures. the place of purchase still related to consumers’ culture and purchase habits. the result of the binary logistic regression about factor determining olive oil purchase highlights the positive impact of the educational level and of consumers’ sensibility to extrinsic quality indicators (quality label, packaging, origin, purchasing place), and the negative impact of the consumer's sensibility of to “price/brand”. these findings confirm those of other studies. in fact, jiménez-guerrero, et al. [14] reported that literature review proves that the extrinsic attributes of olive oil are the most important when consumers face the act of purchase [29]. however, intrinsic attributes (e.g., colour or flavour) is on the second place, except for japanese consumers. in the other hand, the probability of buying olive oil is decreasing as consumers' sensitivity to prices and brands increases: the more importance tunisian consumers attach to these extrinsic indicators of quality, the lower the purchasing propensity of olive oil (odds ratio=0.178). differently to the postulates of jiménez-guerrero, et al. [14] the attribute with the highest relative importance is the price for consumers belonging to both producer and non-producer countries [29]. results of the multinomial logistic regression model about the choice of the purchasing place of olive oil underline the non-significant effect of household's size, consumer's sensibility of intrinsic quality indicators and consumers' sensibility to experience and credence quality attributes. indeed, many consumers still do not know olive oil benefits. for this reason, providing information, mainly through advertising, is necessary, to enhance consumers’ awareness of olive oil’s nutritional content and health benefits. this was also recommended by other authors such as sandalidou, et al. [21] and jiménez-guerrero, et al. [14]; cicia, et al. [22]; tsakiridou, et al. [23]; cacchiarelli, et al. [24]; sinha and batra [26]; fotopoulos and krystallis [27]; corbeto-fabón, et al. [28]; branger, et al. [29]. awareness-raising and information regarding olive oil intrinsic quality attribute will also be an asset. for all categories of olive oil purchase except for wholesalers, consumers’ sensibility to extrinsic quality indicators were statically significant, with positive impact for the first factor describing consumers’ sensibility to extrinsic quality indicators other than brand and price and with negative impact for "brand and price" indicators. the gender is a significant and a main factor determining the choice of wholesalers for the olive oil purchase, while educational level is significantly determining the selection of “friends and relatives” and mills for the olive oil procurement. consumer origin is especially significant for supplies carried out from relatives. based on the two models, it can be confirmed that some factors determining the purchase of olive oil also determine the choice of its place of purchase (except for "wholesalers" and "retail store" as purchase places). these factors are "educational level" as socio-demographic factor and factors 1 and 4 relative to all extrinsic quality indicators. indeed, high educational level and consumer sensibility to extrinsic quality indicators such as "quality label, packaging, olive oil origin, points of sale" have a positive effect on olive oil purchase and promote olive oil supply from mills and friends and relatives. on the other hand, consumer sensibility to brand name and price, also extrinsic quality indicators, have a negative impact on purchase decision and on the choice of mills, relatives, and store retail, as suppliers of olive oil. there are also other factor determining the choice of "friends and relatives " as buying place, it's the consumer origin: if consumers come from a production area, the probability of purchase from family and friends increases. in addition, "gender" as socio-demographic factor has a positive and significant impact on olive oil purchase from wholesalers. women, head of household, are more likely to buy from wholesalers. these results confirm the assumptions of corbeto-fabón, et al. [30] about impact of olive oil quality attributes perception on place of purchase choice [31]. in addition, some assumptions of vlontzos and duquenne [5] about the spanish market are confirmed: the educational level and the fact of living or not nearby to an olive oil producing area influence place of purchase choice, while the impact of household's size is not significant [6]. 6. conclusions and implications consumers’ purchasing behaviour is affected to varying degree by different factors: socioeconomic and demographic factors, the relative importance accorded by tunisian consumers to the olive oil intrinsic and extrinsic quality indicators and olive oil experience and credence attributes. the results obtained here prove that having a high educational level and being sensitive to some extrinsic quality indicators such as origin, quality label, packaging and place of purchase have a positive and significant impact on olive oil purchase decision and promote the choice of mills and relatives as olive oil place of purchase. whereas consumers sensibility to brand and price has a negative and significant impact on purchase decision and on the choice of these point of sale of olive oil in the tunisian market. it should be noted that mills and friends/relatives are the main supply areas for tunisian consumers. in addition, being a women head of household increase the probability of olive oil purchase from wholesalers. being originating in an olive oil production area increase the probability of purchasing olive oil from friends and relatives. based on these results, it is recommended that tunisian olive oil institutions work on changing tunisian purchase behaviour from bulk to bottled products, from an annual act of purchase to more frequent purchase of small quantities to meet consumer expectations for quality attributes and to enhance olive oil purchase. this will facilitate the transition from informal (weekly market, friends, and relatives) to formal distribution (supermarket, hypermarket, retail stores, etc.). producers’ marketing strategies must consider the consumer profile (age, gender, origin, and educational level) and behaviour (frequency of purchase, preferred size of the package, awareness about quality attributes) to provide an adequate product, compliant with and met the requirements of the tunisian consumers. this strategy can be based on olive oil’s peculiarities, origin and quality labels. particularly, they have to strengthen the in-store advertising (e.g. tasting sessions as promotion campaigns at supermarkets and hypermarkets) to allow consumers to experience the olive oil properties, so they can make their purchase decisions of bottled and branded olive oil based on sensory factors. in addition, companies can agriculture and food sciences research, 2022, 9(1): 22-31 31 © 2022 by the authors; licensee asian online journal publishing group strengthen consumers’ awareness about olive oil health benefits and mode of use. so, experience and credence quality attributes could become more significant for tunisian consumers. in conclusion, the present study is in effect an exploratory study. sample size and representativeness are its main limitations. it emphasizes the interaction between the olive oil purchase decision and the choice of its place of purchase, based on an investigation quality attributes and indicators perception. references [1] b. karray, j. d. david, and r. m. marsha, "analysis of the tunisian olive sector, fao. retrieved from https://www.fao.org/3/i4104f/i4104f.pdfioc," 2015. 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[31] c. ding and x. he, "k-means clustering via principal component analysis," in proceedings of the twenty-first international conference on machine learning, 2004, p. 29. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.fao.org/3/i4104f/i4104f.pdfioc, http://www.intechopen.com/chapters/27035, http://dx.doi.org/10.1186/s40100-018-0102-8 agriculture and food sciences research issn 2411-6653 vol. 2, no. 2, 32-38, 2015 http://www.asianonlinejournals.com/index.php/aesr * corresponding author 32 climate change and agriculture: modeling the impact of carbon dioxide emission on cereal yield in ghana lawrence amponsah 1* --glory kofi hoggar 2 --samuel yeboah asuamah 3 1 department of general agriculture, sunyani polytechnic, ghana 2 department of general and liberal studies, sunyani polytechnic, ghana 3 department of marketing, sunyani polytechnic, ghana abstract this work is licensed under a creative commons attribution 3.0 license asian online journal publishing group contents 1. introduction ............................................................................................................................................................................... 33 2 research methodology ............................................................................................................................................................... 34 3. empirical results ...................................................................................................................................................................... 34 4. conclusions and policy recommendations ............................................................................................................................. 37 references ...................................................................................................................................................................................... 37 the objective of the paper is to contribute to the body of knowledge in the area of climate change and agriculture by examining the effect of carbon dioxide concentration (co2) on cereal yield using autoregressive distributed lag models (ardl). the research is based on quantitative, descriptive and cross-sectional research using secondary data obtained from world bank data base for the period of 1961-2010. the co-integration test indicates the series are co-integrated. the results on the long run and shorts run elastically co-efficient indicate that there is significant negative link between co2 and cereal yield. there significant positive long run and short run link between cereal yield and income (proxied by real gross domestic product). policy makers and agriculture scientists and environmental scientists should put in place policies to reduce atmospheric temperature increase and pollution to benefit from co2 fertilization in order to ensure food security. the findings indicate that income (proxied by real gross domestic product) positively affect cereal yield. the link between co2 and cereal production should be examine in future studies current study considered cereal yield. keyword: climate change, co2, real gross product, cereal yield. jel classification: q54, q56, q57, q58. http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2015, 2(2): 32-38 33 1. introduction the significant impact of changes in climatic conditions (an increased frequency of climate extreme events; altered precipitation and transportation regimes; higher temperatures; elevated co2 concentration) agricultural production systems now and years to come in developed and developing economies [1]. the role of climate change in agriculture productivity is of much concern to researchers and policy makers [2]. this increased attention results from the fact that crop yield as well as the type of crops to grow is the function of climate change [2-4]. the role of agricultural sector in an economy such as job creation [5] provision of food security; provision of raw material for the industrial sector and supply of export product to generate foreign exchange makes research and policy makers spend a lots of resources in examining the effects of climate change on agriculture. researchers [6] have reported that there is slower growth rate of agricultural sector over the years due to climate change and this becomes more worrisome given the current nature of urbanization and population growth rate. since in the developing economies, such as ghana, agricultural is based largely on the climatic condition [6, 7] there is the need to empirically examine the effect of climate change on agriculture so that the right adaptation measure could be put in place to ensure food security. carbon dioxide (co2) is among the influential climatic factors the influence crop yield and which has empirically attracted attention in the literature. according to researchers [8-12] co2 acts as a fertilizing element (carbon dioxide fertilization) in the growth of crops classified as c3 and c4 crops. co2 is reported to have both direct and indirect effect of plant growth by enhancing photosynthesis processes and changing the climate through greenhouse effect. according to blanc [8] “greater co2 concentration enhances co2 assimilation by crops. this results in faster stomata closures and ultimately lower transpiration rates (i.e. crops loose less water). therefore, co2 concentration increases are most beneficial in sudano-sahel, which has a warm and arid climate”. the implication for these theoretical frameworks is that crop yield is expected to have positive link with carbon dioxide emissions, especially c4 plants, which is a subject of empirical verification such as the current study. the empirical findings on the effects of the link between climate change and agricultural productivity are found in the works of various researchers [6, 8, 12-25]. alaim [6] state that, rising concentration of co2 may cause decline in agricultural productivity and “will act as a fuel to the higher prices of goods and services in the economy”. blanc [8] reported that co2 emissions have significant long run and short run impact on millet yield but not on cassava and maize yields. the results indicate that 1ppm co2 increase in co2 concentration leads to about 4.75% millet yields increase. at the regional level, a1ppm increase co2 concentration leads to about 11.9% increase in millet yields. the results indicated that co2 concentration insignificant impact on sorghum yields. in the study by wang, et al. [26] the results indicate that without the direct effect of co2 the estimated model indicated that all three climate change scenario examined significantly increase biomass production compared to the baseline scenarios whereas the combined effect (climate and co2) increased biomass production much more, increased co2 concentration of 220 and 120 ppm resulted in increased biomass production of 43-45% and 25%, respectively in the study. other researchers such as lobell and field [14] used regression analysis and reported of positive plant responses to responses to co2 fertilization in the 20 largest producing countries of the northern hemisphere. adams [9] established positive plant yields results from co2 fertilization. in a study by de tafur, et al. [27] negative link was reported between co2 and cassava yields. fuhrer [28] states that “ warning accelerate plant development and reduce grain-fill, reduces nutrients use efficiency, and favors c4 weeds over c3 crops… and that warming again reduces the positive co2 effect”. cure and acock [12] used experimental research to report of a positive relationship between maize yields and co2 fertilization. 1.1. the agriculture sector of ghanaian economy according to the state of ghanaian economy [29] the contribution of the agriculture sector to the ghanaian economy in recent past has been low in relation to other sectors. for examples in 2011 the agriculture sector (25.65%) contributed least to the economic growth of the country followed by the industrial sectors (25.9%) and then the service sector (48.5%). the sector is still dominated by the peasant economy without any significant change since independent. statistics indicate that growth rate of the sectors has been falling. for example in 2011 the growth rate was 5.3% but fell to 0.8% in 2011. the sector even experienced negative growth rate in 2007. the contribution to the positive growth rate in 2011 after the rebounce in 2008 and 2009 are the subsectors such as cocoa (14%); livestock (5.1%) and the crop subsector (3.7%). some of the subsectors (forestry and fisheries) have registered negative growth rate. the main cereal crops in ghana are maize; sorghum; millet and rice. the performance of the cereals over the years has not been impressive. in 2011 the total output of the cereal crops did not attain positive growth. in 2011 the output was 186,000 tonnes whereas in 2010 the output was 218,000 tonnes. maize has been the leading subsectors of the cereal crops in terms of yields and cultivated hectares of land. the review of the literature and the agricultural sector indicates that the agricultural sector is important in relation to the economic development of any economy especially in small but open economy such as ghana. examination of factors that are responsible for the growth agricultural sectors in the empirical study is worth doing. the current paper models the long run and short run relationship between cereal yields and co2. the review of the literature indicates mixed findings of the effects of co2 on crop yield. few econometrics studies exist in relation to developing economies such as ghana. the finding on cereal yields and co2 has also been mixed in the literature [8]. the current paper fills the literature gap. policy makers are provided with policy documents in order to adapt to the changes of climate change and to ensure food security. theories of fertilization and climate change are provided with further understanding. agriculture and food sciences research, 2015, 2(2): 32-38 34 the general objective of the study is to contribute to the body of knowledge in the area of climate change and agriculture by modeling the stable long run relationship between carbon dioxide emissions and cereals yields empirically. specifically, the paper examines; (a) the unit root properties of carbon dioxide emissions and cereal yields, (b) the co-integration relationship between carbon emission and cereals yield, (c) the long run and short run parameter estimate of the effect of carbon emission on cereal yield. the study is based on the following research questions; (a) what is the effect of carbon dioxide emissions on cereal yield? (b) what is the nature of long run and short run relationship between carbon dioxide emissions and cereals? the assumptions tested in the current research are; (a) carbon dioxide emissions and cereals yield are cointegrated, (b) carbon emissions are key influential factors explaining cereal yield. data for the study was obtained from world bank database. the estimated model might suffer from errors in variables and data massaging. the study is descriptive in nature and as such, causality issues are not the focus of the current study. the period for the study is between 1961-2010. structural breaks in unit root are not considered in the study. other influential factors such as temperature and precipitation are not considered for non-availability of data. the rest of the study deals with the methods, empirical results conclusions and policy implications. the rest of the paper looks at the methodology, empirical results, discussion, and conclusion 2 research methodology 2.1. design/format/strategy the study modeled the link between co2 and cereal yield in a trivariate, quantitative, descriptive and crosssectional research format. annual time series data for ghana for the period 1961-2011 was used. the period was chosen for availability of data. the data used are co2; cereal yield and economic growth (proxied by real gross domestic product). 2.2. unit root test the examination of unit roots is based on the kwiatkowski, et al. [30] (kpss) and augmented [31] (adf) test models. the adf model is based on the null assumption (h0) that there is a unit root in the levels of the series. the alternative hypothesis (h1) is that the series are stationary in levels. the kpss model is based on the null assumption (h0) that there is no unit root (stationary) in the levels of the series. the alternatives hypothesis (h1) is that the series are unit root (stationary) in levels. 2.3. bound testing approach to cointegration model the cointegration test is based on the autoregressive distributed lag model (ardl) developed by pesaran and shin [32]. series variables in a model are considered to be cointegrated if they are integrated of order one, 1(1) in the presence of non-zero vector which is integrated of order zero, 1(0). the nonstationary series with the same order of integration may be cointegrated if there exist some linear combination of the series that can be tested for stationarity. the cointegration analysis is based on the null hypothesis (ho) that there is no cointegration among the series variables in the models against the alternatives hypothesis (ha) that the variables are cointegrated. h0: b1=b2-b3=… =b k=0. the alternative hypothesis (ha) is not ho. the ardl model estimated is assess for goodness of fit using various diagnostic test such as j-b normality test. breusch-godfred lm test, arch lm test, white heteroskedastically test, ramsey reset. the stability of the model parameters are examined using the cumulative sum of recursive residuals (cusum) and the cumulative sum of squares of recursive residuals (cusumsq). 2.4. the model the conceptual model for the study state that cereal yield (cy) is a function of carbon dioxide emission (co2) with real gross domestic product (rgdp) as the control variable. that is incy= f (lnco2, lnrgdp). the model is estimated in natural log form. 3. empirical results the results are adk; kpss unit roots test results; cointegration test results; long run results; short run results; diagnostic test results and the stability test results. 3.1. time series plots of the series variables the plots of the series variables in levels and in first difference are shown in figures 1to6. the plot of the series in levels indicate the series are not stationary in levels but become stationary in first difference. this calls for formal examination of the stationary properties using the adf test and the kpss test. the results are presented in tables. 3.2. the adf model the unit root results based on the adf test are reported in table 1and table 2. the result of the adf test for unit root in logarithm show that the series are nonstationary in intercept and trend. the null hypothesis of unit root was accepted for the series variables. table-1. adf stationary test results with a constant and time trend variables (logarithm) t-ratio adf p-value results co2 -0.523027 0.9826 not stationary cy -1.74441 0.7315 not stationary rgdp 0.0578461 0.9969 not stationary source: author’s computation, 2014 agriculture and food sciences research, 2015, 2(2): 32-38 35 taking logarithm of the first difference of the series and testing these with intercept and time trend does make the series stationary. that is, the null hypothesis of unit root was rejected. the series become stationary after first difference. these results indicate that the series exhibit unit root processes and are integrated of order one, 1(1) and order zero, 1(0). the results are reported in table 3 table-2. adf stationary test result with constant and time trend variables( 1 st diff. of logarithm) t-ratio adf p-value results δlnco2 -6.01131 1.89e-006 stationary δlncy -4.83595 0.0003711 stationary δlnrgdp -4.7488 0.0005364 stationary source: author computation, 2014 3.3. the kpss model the kpss test results are presented in table 4. the kpss is a reversed test for unit root. it is used in the current study for confirmation of the stationary properties of the series. the series were examined in their logarithm form. the results confirm that of the adf test results. table-3. kpss stationary test results with a constant and time trend variables (logarithm) t-stats results co2 0.28804(<0.01) not stationary cy 0.232247(<0.01) not stationary rgdp 0.306493(<0.01) not stationary source: author computation, 2014 note: 1%; 5% and 10% critical values are 0.213, 0.149 and 0.121 respectively table-4. kpss stationary test results with constant and trend variables( 1 st diff. of logarithm t-stats results δlnco2 0.065819(>0.1) stationary δlncy 0.0478982(>0.1) stationary δlnrgdp 0.100559(>0.1) stationary source: author computation, 2014 note: 1%; 5% and 10% critical values are 0.212, 0.149 and 0.122 respectively in summary, the test results from the adf and the kpss indicate that the series exhibit unit root processes and are integrated of order one, 1(1) and order zero, 1(0). the detection of unit roots in the series indicate that shocks to the series will have permanent effects and not transitory effects. the results indicate that cointegration can be performed. 3.4. cointegration analysis using autoregressive distributed lag (ardl) model/bound approach to cointegration in this section of the paper the results of the relationship between carbon dioxide emissions, infant mortality and real gross domestic product ate presented. the dependent is variable is infant mortality whereas dioxide emission and real gross domestic products are the independent variables. the results on the bound test, long run and short run parameters are presented in table 5, table 6 and table 7. 3.4.1. bound approach to cointegration for infant mortality (fm) the results presented reported in table 5 indicate significant cointegration between carbon dioxide emissions (co2); cereal yield (cy) and real gross domestic product (rgdp) in models 1, 2 and 3. in model 1, the fstatistics value of 11.0045 is greater than the critical value of the upper bounds at the 90%, 95%, and 99% levels of significance, which is an indication of cointegration between carbon dioxide emissions, cereals yields, and real gross domestic product with carbon emissions as the dependent variable. in model2, the f-calculated value of 4.4938 is greater than the critical values of the upper bounds at the 90% and 95% levels of significance, which are indication of cointegration between carbon dioxide emissions, cereals yield, and real gross domestic product with carbon emissions as the dependent variable. in model 3, the f-calculated value of 8.9438 is greater than the critical values of the upper bounds at the 90%, 95%, and 99% levels of significance, which is indication of cointegration between carbon dioxide emissions, cereal yields, and real gross domestic product with real gross domestic product as the dependent variable. the null assumption of no cointegration is rejected in models 1, 2 and 3. models 1 are estimated for long run and short run parameters (elasticity coefficient). table-5. test for cointegration relationship critical bounds of the f-statistic: intercept models 90% level 95% level 99% level 1(0) 1(1) 2.915 3.695 1(0) 1(1) 3.538 4.428 1(0) 1(1) 5.155 6.265 computed f-stats decision 1.flncy(lncy/lnco2, lnrgdp 11.0045[0.001***] cointegrated 2.flnco2(lnco2/lncy, lnrgdp) 4.4938[0.034**] cointegrated 3.flnrgdp(lnrgdp/lnco2, lncy 8.9438[0.003***] cointegrated source: author computation, 2014: note: critical value are obtained from pesaran, et al. [33] and narayan [34]: note*** and** denote significance at 1% and 5% levels of significance. agriculture and food sciences research, 2015, 2(2): 32-38 36 3.4.2. results of longrun elasticities of ardl model the long-run relation between carbon dioxide emissions, cereals yields and real gross domestic product was estimated with cereal yield as the dependent variable. the results are reported in table 6. the results indicated that carbon dioxide concentration is significant (at 10% level) explanatory variable of cereal yield in the long run. the results indicate that increase in carbon dioxide emissions (co2) by 1% will lead to a decrease in cereal yields by about 54.67%. there is positive significant long run relationship between cereal yield (cy) and real gross domestic product (rgdp) in the model estimated. the results indicate that increase in real gross domestic (rgdp) by 1% will lead to an increase in cereal yield by about 76.41%. table-6. estimated long run coefficients. dependent variable is lnfm variable coefficient std.error t-ratio p-value constant 6.2748 1.8826 3.3330 0.002*** trend 0.035723 0.011348 3.1479 0.003*** inco2 -0.54665 0.29027 -1.8833 0.066* inrgdp 0.76406 0.23174 3.29711 0.002*** author’s computation, 2014: ardl (1) selected based on akaike information criterion. note: *** and * denotes statistical significance at the 1% and 10% levels significantly. 3.4.3. result of short-run elasticities of ardl model the results of the short-run dynamic equilibrium relationship coefficients estimated with trend and error correction term (ecm) are reported in table 7. the results on the nature of the short run coefficient are not different from that of the long run parameters. the results indicate that carbon dioxide concentration is statistically significant determinants of cereal crop yields in the short run. the results indicate that increase in carbon dioxide emissions (co2) by 1% will lead to a decrease in cereal yield by about 31.03%. there is significant positive short run relationship between cereal yield (cy) and real gross domestic product (rgdp) by 1%will lead to an increase in cereal yield by about 43.37%. the error correction term is statistically significant and does have theoretical expected sign which is negative. the coefficient of -0.56767 indicate that, after 1 percent deviation or shock to the system, the long run equilibrium relationship of infant mortality is quickly re-established at the rate of about 56.77% per annum. the value does indicate moderate adjustment rate. table-7. short-run representation of ardl model. ardl (1) selected based on akaike information criterion. dependent variable: δlnfm variable coefficient std.error t-ratio p-value trend 0.020279 0.0059051 3.4341 0.001*** δinco2 -0.310331 0.14638 -2.1199 0.040** δinrgdp 0.43373 0.14503 2.9906 0.005*** ecm(-1) -0.56767 0.13033 -4.3557 0.000*** source: author’s computation, 2014. note: *** and* denotes statistical significance at the 1% and 10% levels significantly. 3.4.4. results of diagnostic tests the diagnostic tests of the short-run estimation to examine the reliability of the results of the error correction model are reported in table 8. the null hypothesis of no serial correlation is not rejected using the lagrange multiplier test and f-statistics. the reset test showed evidence of incorrect functional specification of the model. the estimated model did not passed the normality test. the model passed heteroscadasticity test indicating the variances are constant over time. the r 2 (0.81733) and the adjusted r 2 (0.80034) in the table are an indication of very well behave model. the coefficient indicate approximately 81.73% of the variations in cereal yield are attributed to carbon dioxide emissions and real gross domestic product. table-8. short run diagnostic tests of ardl model test statistics lm version f version a: serial correlation chsq(1)=.15519[0.694] f(1, 42) =0.13623[0.714] b: functional form chsq(1)= 5.5570[0.018] f(1, 42)=5.4990[0.024] c: normality chsq(2)=12.1834[0.002] not applicable d: heteroscedasticity chsq(1)=1.4482[0.222] f(1, 46)= 1.4718[0.23] a: language multiplier test of residual serial correlation b: ramsey’s reset test using the square of the fitted value c: based on a test of test skewness and kurtosis of residuals d: based on the regression of squared residual on squared fitted values source: author’s computation, 2014 the stability of the long run estimates was determine by employing the cumulative sum ( cusum) and cumulative sum of squares (cusumg) procedures. this was determined using the residual of the error-correction model indicate by equation (1). the cusum test of stability determines the methodological arrangements of the estimate and its null hypothesis states the coefficients are stable. the null assumption is rejected when the cusum surpasses the given critical boundaries, which demonstrate unstable nature of the estimates. the cusumg determines the stability of the variance. both tests indicate that the estimate and the variance were stable as the residuals and the squared residuals fall within the various 5% critical boundaries. the null assumptions are rejected in both tests. agriculture and food sciences research, 2015, 2(2): 32-38 37 4. conclusions and policy recommendations the objectives of the paper have been achieved. the long run link between co2 and cereal yield has been examined. the series variables in the estimated model are cointegrated. the long run and short run elasticities are statistically significant but do not have the expected signs. the co2 elasticity of cereal yields is negative in the long run and short run. the findings of the current study are in support of the findings of previous studies reported in the literature [6, 8, 27, 35-37]. blanc [8] reported of insignificant effect of co2 concentration on maize yield. fuhrer [28] indicate that the benefits of co2 may be lost in warmer climate. according to j., et al. [37] crop yield (wheat and rice) is decreased as a results of high temperature (above 32) and that higher temperature offset an increase in yield resulting increase atmospheric co2 concentration. amthor [36] indicated benefits of co2 fertilization is largely offsets by pollutants and nutrients limitation. de tafur, et al. [27] reported of negative link between co2 concentration and cassava yields. the findings are in consistent with the findings of researchers [12, 14, 26] who reported of positive significant link between co2 concentration and crop yield. policy makers, agriculture scientists, and environmental scientists should put in place policies to reduce atmospheric temperature increase and pollution to benefits from co2 fertilization in order to ensure food security. agriculture practices should ensure there is no issue of limiting nutrient crop farming. there should by intensive agriculture that will save the potential to adapt to changing climate conditions. farmers should adapt policies such as providing irrigation or increasing its efficiency, maintaining or improving flood control, encouraging agronomic research. the findings indicate that income (proxies by real gross domestic product) positively affect cereal yield. economic policy makers should put in place policies to ensure economic growth so as to be able to attain increase cereal yield. future studies should examine causality issues since the current study is descriptive in nature. the link between co2 and cereal production should be examined in future studies since the current study considered cereal yields. references [1] p. calanca and j. furhrer, swiss agriculture in a changing climate: grassland production and it economic value, in: haurie, a. and viguier, l. 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[37] g. j., b. walker, w. steffen, j. canadell, and j. s. i. ingram, managed production systems. in the terrestrial biosphere and global change: implications for natural and managed systems. cambridge, uk: cambridge university press, 1999. views and opinions expressed in this article are the views and opinions of the authors, agriculture and food sciences research shall not be responsible or answerable for any loss, damage or liability etc. caused in relation to/arising out of the use of the content. 36 © 2021 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 8, no. 2, 36-43, 2021 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2021.82.36.43 © 2021 by the authors; licensee asian online journal publishing group toxicological indices of wistar rats fed formulated chaw of telfairia occidentalis planted on crude oil contaminated and remediated soil reginald c. ohiri1 eugene n. onyeike2 augustine a. uwakwe3 ( corresponding author) 1,2,3department of biochemistry, faculty of science, university of port harcourt, nigeria. 1email: raycohiri@yahoo.com 2email: eugene.onyeike@uniport.edu.ng 3email: augustine.uwakwe@uniport.edu.ng abstract toxicological indices of wistar rats fed formulated chaw of telfairia occidentalis planted on crude oil contaminated and bioremediated soil was studied. farmland, measuring 18 m2 was divided into three lots of 4 m2 and 2 m space was allowed between each lot (a, b and c). two lots were polluted, subjected to 16 weeks of bioremediation and viable seeds of t. occidentalis were planted and grown for 28 days. the harvested vegetable leaves were analyzed. leaves from natural attenuated soil showed high concentration of both aliphatic and polycyclic aromatic hydrocarbons with values of 9.34+0.03 mg/kg and 9.18+0.05 mg/kg for c36 and fluoranthene respectively, while the bioaugmented soil had lead as the highest heavy metal with a value of 0.10+0.02 g/100g. four groups, of 9 rats each (totalling 36 rats) of adult males and females, wealing males and females were subsequently sub-grouped into 3 and allowed to acclimatize for a period of 1 week. these animals were fed formulated rat chaw of 50% dry weight of t. occidentalis, from the three lots (control, bioaugmented and natural attenuated soil) for a period of 28 days. assay of animals’ toxicological index showed elevated concentrations of alkaline phosphatase, alanine and aspartate aminotransferases, urea and creatinine in their sera. a slight increase in serum amylase activity was recorded, while concentrations of their haemoglobin and packed cell volume (pcv) reduced. the obtained results from these biomarkers were confirmed by the histopathological sections of the animal organs. keywords: accumulation, biotransformation, crude oil, excretion, ingestion, tocixity. citation | reginald c. ohiri; eugene n. onyeike; augustine a. uwakwe (2021). toxicological indices of wistar rats fed formulated chaw of telfairia occidentalis planted on crude oil contaminated and remediated soil. agriculture and food sciences research, 8(2): 36-43. history: received: 10 september 2021 revised: 2 december 2021 accepted: 20 december 2021 published: 14 january 2022 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 37 2. experimental materials and methodology ................................................................................................................................. 37 3. results ................................................................................................................................................................................................ 39 4. discussion .......................................................................................................................................................................................... 42 references .............................................................................................................................................................................................. 43 mailto:raycohiri@yahoo.com mailto:eugene.onyeike@uniport.edu.ng mailto:augustine.uwakwe@uniport.edu.ng http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.doi.org/10.20448/journal.512.2021.82.36.43 https://orcid.org/orcid-search/quick-search?searchquery=ajayi%20boboye%20l. agriculture and food sciences research, 2021, 8(2): 36-43 37 © 2021 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by confirming that accumulated hydrocarbons and heavy metals in plants (vegetables) will cause destruction of cellular architecture in animals upon ingestion, thereby causing impairments in cellular activities. 1. introduction one established mechanism by which plants decontaminate oil contaminated soil is by direct absorption of the contaminant hydrocarbons and subsequent accumulation of same in specific plant tissues and organelles. the casparian strip has remain a structural foundation for uptake of substances through the plant roots. this system blocks the possible absorption of soil solution via the plant root to the xylem [1]. substances inside root cells are moved to the aerial parts of the plant. foliar uptake has been the most important pathway for petroleum hydrocarbon uptake [2]. other factors that influence hydrocarbon uptake aside from foliar uptake include aqueous concentration of petroleum hydrocarbon, uptake/exposure period, plants bio-physical and chemical compositions. kipopoulou, et al. [3], reported solubility, octanol-water and octanol-air partition coefficients and vapour pressure as good predictors of petroleum hydrocarbon accumulation in leaf tissues of vegetables. t. occidentalis is a commonly produced crop in the niger delta area of nigeria. this region has recorded exorbitant crude oil pollution since the advent of petroleum exploration in the area and there is a continuous annual elevation of its contaminant load in concord with industrial surge and energy uptake thus necessitating the need for environmental remediation. after remediation of petroleum hydrocarbon pollution within and agricultural area, cultivation and growing of plants are carried out without consideration to hydrocarbon recalcitrance in the post remediated soil. these post remediated hydrocarbons may be absorbed, accumulated and a possible high or low potent toxicant can be generated as an intermediary metabolite and stored in specific plant cells. the photosynthetic activities in leafy vegetable (which t. occidentalis is amongst them) enriches them with protein, minerals and carbohydrates [4]. also referred as fluted pumpkin, t. occidentalis hoof (family: cucurbitaceae) has been a vital commercial crop of cultivated within the humid lowland tropical region situated in the western part of africa [5]. because of its nutritional value, t. occidentalis is prominent in the diet of many animals, including humans [4]. this research was geared towards the evaluation of the toxicological indices of wistar rats fed formulated chaw from t. occidentalis planted on crude oil contaminated and remediated soil. 2. experimental materials and methodology 2.1. source of materials a volume of 30 dm3 of bonny light crude oil was gotten from flow station of shell petroleum development company located at egbema local government area of imo state, nigeria. forty kilograms (40 kg) of poultry manure was obtained from godvine poultry farm located at obio akpor local government area of rivers state, nigeria. viable t. occidentalis seeds were obtained from rumuokoro market, also located in obio akpor local goverment rivers state, nigeria. while a total of 36 healthy wistar rats of 9 rats each of adult males and females, wealing males and females were selected from the research animal house of biochemistry department, university of port harcourt, river state, nigeria. 2.2. source of reagent kits alanine and aspartate aminotransferase, alkaline phosphatase, creatinine and urea kits were sourced from randox laboratories ltd, antrim, united kingdom, while the kit for alpha amylase was from giesse diagnostics, rome, italy. 2.3. description of research site. the research site was geographically located on longitude 70 10″ e and latitude 40 40″ n on the eneka axis of eneka-oyigbo inter-city road in obio akpor, rivers state, nigeria. soils of this research site has an approximate ph of 4.86 + 0.12, it belongs to the ultisols and are made up of humus topsoil with a deep mixture of sandy and clay soil. pipeline vandalization and oil spillage has not been recorded on the research site and its environs. 2.4. pollution and remediation of research site a farmland, measuring 18 m2 was divided into three lots (a, b and c) of 4 m2 and 2 m land space was allowed between each lot. two lots were polluted and remediated as follows: lot b = 4 m2 farmland polluted with 40 dm3 of bonny light crude oil and remediated with 40kg of poultry manure for 16 weeks (bioaugmented soil). lot c = 4 m2 un-remediated farmland polluted with 40 dm3 of bonny light crude oil and allowed for 16 weeks (natural attenuated soil), while lot a = unpolluted 4 m2 farmland (control soil). 2.5. planting and growing and collection of vegetable samples after 16 weeks of remediation, viable t. occidentalis seeds were planted on the 3 lots and 4 weeks of growth were allowed from their germination date. vegetable leaves were harvested at the fourth week with a sterilized razor blade, placed into sterilized cellophane bags and closed with plastic bands. each cellophane bag of harvested samples was marked using a water-resistant marker and subsequently taken to the laboratory within one hour of sample collection for hydrocarbon and heavy metal analysis. 2.6. aliphatic and polycyclic aromatic hydrocarbons (pah) and heavy metals concentration of both aliphatic and polycyclic aromatic hydrocarbons were extracted and determined in t. occidentalis by the method described by the aoac [6], while the method described by jones jr [7], was used to determine heavy metal concentration in the vegetables. agriculture and food sciences research, 2021, 8(2): 36-43 38 © 2021 by the authors; licensee asian online journal publishing group 2.7. formulation of rat chaw harvested leaves of t. occidentalis were dried and weighed using model ab204 mettler toledo weighing balance and a model 4 thomas scientific wiley’s mill was use to grind the vegetable. the ground sample was formed into rat chaw and combined with normal rat feed. 2.8. acclimatization, treatment and sacrificing of research animals four groups of wistar rats made up of 9 rats each (totalling 36 rats) of adult males and females, wealing males and females were subsequently sub-grouped into 3. both the control and test animals were placed at normal room temperature in steel cages and allowed to acclimatize for a period of 1 week while having free access to normal rat feed and portable drinking water. the sub-groups were placed on rat chaw comprising of equal ratio of normal rat feed and dried t. occidentalis from the control, bioaugmented and natural attenuated lots. using chloroform anaesthesia, the animals were sacrificed at the 28th day and blood sample was obtained via cardiac puncture into ethylene diamine tetracetic acid (edta) bottles and taken to the laboratory within one hour for assay of the following toxicological indices: alkaline phosphatase, alanine and aspartate aminotransferases, urea, creatinine, hemoglobin concentration and packed cell volume (pcv). the internal organs (liver, kidney and pancreas) were immediately harvested into a preservative container of 10% physiological saline and subsequently subjected to histological analyses. table-1. concentration of aliphatic hydrocarbon (mgkg-1) in t. occidentalis from crude oil remediated soil. no. of carbon atoms control soil bioaugmented soil natural attenuated soil c8 2.29 + 0.05a 1.30 + 0.03b 5.37 + 0.20c c9 1.60 + 0.03ab 1.50 + 0.03ab 2.78 + 0.04c c10 1.04 + 0.02ac 4.43 + 0.05b 3.02 + 0.03ac c11 1.07 + 0.07a 7.56 + 0.10b 2.11 + 0.04c c12 2.70 + 0.02a 1.23 + 0.01b 1.56 + 0.08c c13 2.05 + 0.02a 2.31 + 0.06b 1.02 + 0.01c c14 1.47 + 0.09ac 8.17 + 0.02b 1.43 + 0.01ac c15 1.66 + 0.09a 1.19 + 0.02b 1.02 + 0.02c c16 1.03 + 0.01a 3.12 + 0.01b 7.90 + 0.18c c17 1.02 + 0.05a 3.02 + 0.02b 5.83 + 0.24c pristane 1.17 + 0.06a 6.65 + 0.21b 5.08 + 0.05c c18 1.32 + 0.03a 2.08 + 0.14b 6.95 + 0.28c phytane 1.61 + 0.01a 1.11 + 0.02b 4.14 + 0.02c c19 1.90 + 0.05a 4.18 + 0.02b 6.83 + 0.06c c20 8.56 + 0.06a 1.65 + 0.02b 6.17 + 0.06c c21 1.53 + 0.08a 9.03 + 0.09b 1.86 + 0.09c c22 2.76 + 0.07a 2.46 + 0.10b 1.19 + 0.04c c23 5.49 + 0.04a 3.07 + 0.14b 1.80 + 0.02c c24 6.83 + 0.08a 2.30 + 0.03b 1.61 + 0.02c c25 3.14 + 0.06a 6.36 + 0.07b 1.04 + 0.00c c26 6.49 + 0.03a 2.02 + 0.02b 7.46 + 0.09c c27 7.80 + 0.09a 9.58 + 0.09b 9.10 + 0.20c c28 1.22 + 0.01a 1.66 + 0.07bc 1.83 + 0.11bc c29 1.59 + 0.05a 6.17 + 0.06b 7.35 + 0.15c c30 1.17 + 0.03a 3.89 + 0.03b 5.11 + 0.01c c31 1.54 + 0.10a 3.87 + 0.05b 5.46 + 0.08c c32 6.09 + 0.03a 9.03 + 0.05b 7.03 + 0.01c c33 5.23 + 0.07a 1.47 + 0.05b 3.11 + 0.01c c34 2.51 + 0.02a 5.08 + 0.03b 6.44 + 0.07c c35 4.18 + 0.10a 3.55 + 0.08b 7.60 + 0.02c c36 2.82 + 0.44a 4.03 + 0.00b 9.34 + 0.03c c37 9.08 + 0.04a 5.97 + 0.21b 2.09 + 0.01c c38 2.26 + 0.08ab 2.19 + 0.04ab 7.18 + 0.02c c39 6.37 + 0.14a 8.79 + 0.04b 5.84 + 0.09c c40 2.55 + 0.08a 6.43 + 0.05b 1.58 + 0.07c note: data presented are mean values + standard deviations of three observations. superscript “a” indicates statistical significance at p < 0.05 when control lot is compared to the other lots. superscript “b” indicates statistical significance at p < 0.05 when bioaugmented lot is compared to the other lots. superscript “c” indicates statistical significance at p < 0.05 when natural attenuated lot is compared to the other lots. superscript “ab” indicates statistical insignificance at p < 0.05 when control and bioaugmented lots are compared to each other, but statistically significant when both are compared to the natural attenuated lot. superscript “ac” indicates statistical insignificance at p < 0.05 when control and natural attenuated lots are compared to each other, but statistically significant when both are compared to the bioaugmented lot.superscript “bc” indicates statistical insignificance at p < 0.05 when bioaugmented and natural attenuated lots are compared to each other, but statistically significant when both are compared to the control lot. 2.9. assay of some toxicological parameters of wistar rats fed with vegetables from bioremediated soil. the diacetyl monoxine method (dam), described by marsh, et al. [8], was used to assay for serum urea concentration while the alkaline picric acid procedure described by slot [9], was used to determine the concentration of creatinine. the 2-chloro-p-nitrophenyl-α-d-maltotrioside (cnpg3) kinetic procedure described by hohenwallner, et al. [10] and the assay procedure of kind and king [11], were used to determine amylase and alkaline phosphatase activities respectively, while the colorimetric end-point procedure of described by reitman and frankel [12] was used to determine both aspartate and alanine aminotransferases. corash [13], procedure was used the haematocrit (pcv) volume and haemoglobin concentration, while histopathological assay of animal agriculture and food sciences research, 2021, 8(2): 36-43 39 © 2021 by the authors; licensee asian online journal publishing group tissues was done using haematoxylin and eosin procedure described by conn [14] and modified by kumar and gill [15]. 2.10. statistical analysis/test of significance the mean + standard deviations and one way analysis of variance were calculated electronically for each group of observations using international business machine (ibm) statistical program for social sciences (spss) 19 software at 95% confidence level, while duncan [16], post hoc multiple range test was used to test for significance. 3. results hydrocarbon and heavy metal content of vegetable samples. the mean values of both aliphatic hydrocarbons, polycyclic aromatic hydrocarbons and heavy metals obtained in t. occidentalis grown on the 3 lots (control, bioaugmented and natural attenuated) are presented in tables 1-3. a significant increase at p<0.05 in aliphatic hydrocarbon content was observed in t. occidentalis harvested from natural attenuated and bioaugmented soils aside c20, c24 and c37 where higher concentrations were recorded in the control vegetables than in the remediated soils polycyclic aromatic hydrocarbon content of t. occidentalis increased in vegetables harvested from both natural attenuated and bioaugmented aside acenaphthene, phenanthrene and indeno(1,2,3-cd)pyrene. higher concentration of acenaphthene was recorded in t. occidentalis from the control lot than in those from both the natural attenuated and bioaugmented soil. phenanthrene concentrations for those grown on the control and natural attenuated soil were equal with a value of 2.67mgkg-1, while phenanthrene concentration in vegetable samples from the bioaugmented soil increased significantly. a significantly lower concentration of indeno(1,2,3-cd)pyrene was observed at p<0.05 in vegetables harvested from the natural attenuated soil than in those from the control soil. a significantly high concentration at p<0.05 of heavy metals were observed in vegetables from the bioaugmented soil as compared to those from both the control and the natural attenuated lots. table-2. polycyclic aromatic hydrocarbon (pah) content (mgkg-1) of t. occidentalis from crude oil bioremediated soil. polycyclic aromatic hydrocarbon control soil bioaugmented soil natural attenuated soil naphthalene 0.55 + 0.07a 3.98 + 0.05b 2.95 + 0.04c acenaphthylene 1.58 + 0.03a 2.87 + 0.01b 3.61 + 0.01c acenaphthene 1.87 + 0.05ab 1.73 + 0.03ab 1.57 + 0.08c fluorene 2.60 + 0.05a 4.44 + 0.04b 5.34 + 0.01c phenanthrene 2.67 + 0.01ac 3.02 + 0.03b 2.67 +0.04ac anthracene 2.33 + 0.01a 3.56 + 0.04b 3.80 + 0.01c fluoranthene 1.57 + 0.01a 7.03 + 0.02b 9.18 + 0.05c pyrene np np np benzo(α)anthracene 1.77 + 0.08a 7.24 + 0.07b 6.09 + 0.04c chrysene 0.07 + 0.01a 2.52 + 0.11b 2.73 + 0.02c benzo(β)fluoranthene 1.69 + 0.06a 1.85 + 0.08b 2.68 + 0.04c benzo(k)fluoranthene np np np benzo(α)pyrene 0.53 + 0.02a 2.98 + 0.10bc 3.02 +0.00bc indeno (1,2,3)pyrene 2.27 + 0.01a 3.28 + 0.03b 4.15 + 0.03c indeno(1,2,3-cd)pyrene 3.47 + 0.07a 6.75 + 0.06b 0.43 + 0.03c dibenz(a,h)anthracene 1.66 + 0.00a 2.58 + 0.04b 1.89 + 0.05c benzo(g,h,i)perylene np np np note: presented are mean values + standard deviations of three observations. np= not present. superscript “a” indicates statistical significance at p < 0.05 when control lot is compared to the other lots. superscript “b” indicates statistical significance at p < 0.05 when bioaugmented lot is compared to the other lots. superscript “c” indicates statistical significance at p < 0.05 when natural attenuated lot is compared to the other lots. superscript “ab” indicates statistical insignificance at p < 0.05 when control and bioaugmented lots are compared to each other, but statistically significant when both are compared to the natural attenuated lot. superscript “ac” indicates statistical insignificance at p < 0.05 when control and natural attenuated lots are compared to each other, but statistically significant when both are compared to the bioaugmented lot. superscript “bc” indicates statistical insignificance at p < 0.05 when bioaugmented and natural attenuated lots are compared to each other, but statistically significant when both are compared to the control lot. table-3. heavy metal concentrations (g/100g) of t. occidentalis from crude oil bioremediated soil. heavy metal control soil bioaugmented soil natural attenuated soil copper 0.01 + 0.0ac 0.03 + 0.00b 0.01 + 0.00ac lead np 0.10 + 0.02b 0.08 + 0.00c chromium np 0.06 + 0.00b np cadmium np 0.03 + 0.00b 0.01 + 0.00c arsenic np np np note: data presented are mean values + standard deviations of three observations. np= not present. superscript “a” indicates statistical significance at p < 0.05 when control lot is compared to the other lots. superscript “b” indicates statistical significance at p < 0.05 when bioaugmented lot is compared to the other lots. superscript “c” indicates statistical significance at p < 0.05 when natural attenuated lot is compared to the other lots. superscript “ac” indicates statistical insignificance at p < 0.05 when control and natural attenuated lots are compared to each other, but statistically significant when both are compared to the bioaugmented lot. agriculture and food sciences research, 2021, 8(2): 36-43 40 © 2021 by the authors; licensee asian online journal publishing group 3.1. indicators of tissue damage and haematological parameters table-4. serum urea and creatinine concentration (mgdl-1) of rats fed t. occidentalis from crude oil bioremediated soil. control soil bioaugmented soil natural attenuated soil urea weanling male 30.19+0.13a 54.35+0.51bc 54.32+0.11bc weanling female 31.40+1.07a 56.80+0.20bc 55.50+0.36bc adult male 33.62+0.18a 52.00+0.63b 58.92+0.47c adult female 32.93+0.19a 58.61+0.30bc 58.66+0.43bc creatinine weanling male 1.19+0.02a 1.73+0.08bc 1.82+0.09bc weanling female 1.11+0.02a 1.79+0.07bc 1.72+0.02bc adult male 0.94+0.03a 1.66+0.03bc 1.71+0.05bc adult female 0.80+0.01a 1.90+0.03bc 1.77+0.08bc note: data presented are mean values + standard deviations of three observations. superscript “a” indicates statistical significance at p < 0.05 when control lot is compared to the other lots. superscript “b” indicates statistical significance at p < 0.05 when bioaugmented lot is compared to the other lots. superscript “c” indicates statistical significance at p < 0.05 when natural attenuated lot is compared to the other lots. superscript “bc” indicates statistical insignificance at p < 0.05 when bioaugmented and natural attenuated lots are compared to each other, but statistically significant when both are compared to the control lot. table-5. serum enzymes concentration (i.ul-1) of rats fed t. occidentalis grown on crude oil bioremediated soil. control soil bioaugmented soil natural attenuated soil alkaline phosphatase weanling male 284.00+5.10a 486.90+10.40b 357.00+3.20c weanling female 268.22+1.49a 479.81+ 0.40b 344.02+5.63c adult male 243.71+1.91a 346.20+ 1.50b 318.07+1.20c adult female 226.53+1.72a 359.70+ 0.84b 304.39+0.75c alanine aminotransferase weanling male 18.00+0.68a 61.28+0.24b 67.42+2.02c weanling female 20.60+0.41a 57.63+0.53b 64.19+1.15c adult male 25.20+0.84a 48.45+1.31b 61.59+0.75c adult female 24.00+0.63a 41.05+0.43b 60.42+0.40c aspartate aminotransferase weanling male 19.00+0.28a 61.00+1.61bc 63.05+0.76bc weanling female 19.22+0.80a 57.37+033bc 58.03+0.07bc adult male 32.81+1.14a 57.60+0.02b 62.40+0.60c adult female 29.70+0.67a 55.00+1.02b 63.81+0.91c amylase weanling male 27.66+0.46a 29.49+0.54bc 29.07+0.23bc weanling female 26.85+0.34a 31.09+0.61bc 31.47+1.03bc adult male 32.83+0.36a 38.18+0.86b 35.17+0.82c adult female 33.51+0.47a 36.89+0.50bc 36.73+0.37bc note: data presented are mean values + standard deviations of three observations. superscript “a” indicates statistical significance at p < 0.05 when control lot is compared to the other lots. superscript “b” indicates statistical significance at p < 0.05 when bioaugmented lot is compared to the other lots. superscript “c” indicates statistical significance at p < 0.05 when natural attenuated lot is compared to the other lots. superscript “bc” indicates statistical insignificance at p < 0.05 when bioaugmented and natural attenuated lots are compared to each other, but statistically significant when both are compared to the control lot. table-6. packed cell volume (pcv) (%) and haemoglobin concentration of rats fed t. occidentalis from crude oil bioremediated soil. control soil bioaugmented soil natural attenuated soil packed cell volume (%) weanling male 31.35+l.33a 24.35+1.53bc 24.00+1.01bc weanling female 31.67+0.58a 25.00+2.04bc 21.68+0.59bc adult male 34.34+2.52a 25.00±3.61bc 25.33+2.52bc adult female 36.33+0.57a 26.36+1.16b 23.67+0.56c haemoglobin (gdl-1) weanling male 10.44 + 0.51a 7.97 + 0.62bc 8.20 + 0.48bc weanling female 10.56 + 0.20a 8.33 + 0.67bc 7.22 + 0.19bc adult male 11.44 + 0.84a 8.34 + 1.21bc 8.44 + 0.84bc adult female 12.11 + 0.19a 8.78 + 0.39b 7.89 + 0.20c note: data presented are mean values + standard deviations of three observations. superscript “a” indicates statistical significance at p < 0.05 when control lot is compared to the other lots. superscript “b” indicates statistical significance at p < 0.05 when bioaugmented lot is compared to the other lots. superscript “c” indicates statistical significance at p < 0.05 when natural attenuated lot is compared to the other lots. superscript “bc” indicates statistical insignificance at p < 0.05 when bioaugmented and natural attenuated lots are compared to each other, but statistically significant when both are compared to the control lot. tables 4 – 6 shows mean concentrations of urea, creatinine, amylase, alkaline phosphatase (alp), aspartate and alanine aminotransferases, packed cell volume and hemoglobin concentrations of rats fed with 50% formulated chaw from the research lots. a significant increase at p<0.05 were observed in all the maker compounds (urea, creatinine, amylase, alkaline phosphatase, alanine and aspartate aminotransferases) of rats fed 50% chaw from the bioaugmented and natural attenuated lots, while a more pronounced decrease in packed cell volume and hemoglobin were recorded in rats fed 50% chaw formulated from vegetables gotten from the agriculture and food sciences research, 2021, 8(2): 36-43 41 © 2021 by the authors; licensee asian online journal publishing group bioaugmented lot than in those fed t. occidentalis chaw formulated from vegetables gotten from the natural attenuated lot. 3.2. sections of rat organs fed with t. occidentalis from crude oil bioremediated lots figure 1 shows histological scores of sections of rat organs fed with t. occidentalis formulated chaw from the crude oil bioremediated lots. the observed lesions in the organ sections were highest in the kidney, liver and pancreas of animals fed t. occidentalis from natural attenuated and bioaugmented lots, while organ sections of those fed t. occidentalis from the control lot had healthier sections with non-significant degrees of lesions. values are means + standard deviations of three determination. figure-1. histological scores of sections of rat organs fed t. occidentalis from crude oil bioremediated soil. sections of rat organs fed t. occidentalis from on crude oil bioremediated lots are presented in figures 2 4. animals fed t. occidentalis from the control lot had healthy kidneys with perfect structures of glomeruli and unaltered tissue architecture figure 2a; while those fed t. occidentalis from both bioaugmented and natural attenuated lots had unhealthy distorted organs with shrunken glomeruli and multicystic spaces figure 2b 2c. figure-2. histology sections of rat kidney fed t. occidentalis from crude oil polluted and remediated soil (a). representative section of the kidney of rat fed t. occidentalis from control lot. [glomeruli perfectly in order (black pointers)]. (b). representative section of the kidney of rat fed with t. occidentalis from bioaugmented lot. [observations: distorted tissue architecture (black pointers), multicystic spaces (blue pointers), glomeruli shrunken (green pointers)]. (c.). representative section of the kidney of rat fed t. occidentalis from natural attenuated lot. [mild oedema (black pointers), tissue stromal proliferation with silt-like channel (blue pointers), glomeruli shrunken (green pointers), distorted tissue architecture (red pointers)]. rats fed t. occidentalis from the control lot had healthy liver sections with well identified hepatocytes and pronounced central vein figure 3a. the presence of slit like channels and proliferation of tissue was noticed around the central veins of the sections of those fed t. occidentalis from the bioaugmented lot figure 3b, while those fed t. occidentalis from the natural attenuated lot had proliferated tissue, enlarged central vein and an altered tissue architecture figure 3c. figure-3. histology sections of rat liver fed t. occidentalis from crude oil polluted and remediated soil. (a). representative section of the liver of rat fed t. occidentalis from control lot. [hepatocytes clearly identified (black pointers), central vein perfectly in order (blue pointers)]. (b). representative section of the liver of rat fed t. agriculture and food sciences research, 2021, 8(2): 36-43 42 © 2021 by the authors; licensee asian online journal publishing group occidentalis from bioaugmented lot. [presence of slit like channels (black pointers), tissue proliferation (blue pointers), central vein blocked (red pointers)]. (c) representative section of the liver of rat fed t. occidentalis from natural attenuated lot. [tissue stromal proliferation (black pointers), distorted tissue architecture (blue pointers), hepatocytes not clearly identified (red pointers), central vein grossly enlarged (green pointers)]. pancreas sections of rats fed t. occidentalis from control lot revealed a healthy, undistorted pancreas figure 4a, while those fed t. occidentalis from both the bioaugmented and natural attenuated lot had scanty stroma, altered tissue architecture and mild oedema figure 4b and 4c. figure-4. histology sections of rat pancreas fed t. occidentalis from crude oil polluted and remediated soil. (a). representative section of pancreas of rat fed t. occidentalis from control lot. [secretory cells perfectly in order (black pointers)]. (b). representative section of pancreas of rat fed t. occidentalis from bioaugmented lot. [scanty stroma (black pointers), stromal abrasions (blue pointers), mild oedema (green pointers)]. (c). representative section of pancreas of rat fed t. occidentalis from natural attenuated lot. [scanty stroma (black pointers), pancreatic acini not clearly seen (blue pointers)]. 4. discussion the accumulation of petroleum hydrocarbons and heavy metals in the leaves t. occidentalis of both the bioaugmented and natural attenuated lots can be linked to partial degradation and recalcitrant hydrocarbons present in the remediated lots (tables 1, 2 and 3). this result corroborates the work of ziółkowska and wyszkowski [17], where contamination of soil with petroleum was reported not to adversely affect the vegetative development of edible plant by can lead to hydrocarbon accumulation in their foliar parts. the increase in concentration of petroleum hydrocarbons observed in the control may be due to trans-boundary contamination caused by horizontal flow of hydrocarbon or due human and animal activities on the research site tables 1 and 2. moreover, ohiri, et al. [18] reported an increase in heavy metal content of crude oil polluted and remediated agricultural soil and they attributed it to heavy metal presence in the petroleum. their report also corroborates that of chicarelli, et al. [19] where heavy metals like nickel and copper are present in petroleum through association with porphyrins. ingestion is the most common route of exposure of bioaccumulated toxic chemicals from edible plants to humans and other animals alike. however, most toxicants diffuse through the cell membrane in the non-ionized form, thus the degree of ionization is important in determining chemical absorptivity [20]. the increased toxicological parameters recorded in this study can be attributed to the ingestion and biotransformation of plant accumulated hydrocarbons and heavy metals by the liver and subsequent excretion via the renal processes of the kidney. (table 4 and 5). the kidney has been responsible for excretion of toxic compounds but in the process of excretion, compounds that are soluble in lipids (e.g. aliphatic and polycyclic aromatic hydrocarbons) are easily reabsorbed through the renal tubule [21], thereby causing accumulation of toxic compounds in the kidney. this accumulation ultimately leads to impairment in renal function, resulting in increased urea and creatinine and destruction of kidney cells as noticed in rats fed t. occidentalis from bioaugmented and natural attenuated lots table 4 and figures 2b and 2c. toxic substances in the liver are biotransformed or secreted into the bile. however, biliary excretion of toxicants does not tantamount to elimination of such toxicants from the body. reabsorption of biliary metabolites results into an enterohepatic cycle, thereby keeping the toxicants in the body and subjecting the livers to more stress conditions. this can lead to destruction of hepatic cells and subsequent elevation hepatic marker enzymes like alkaline phosphatase, aspartate and alanine aminotransferases in the serum. transformation of toxicants by the liver may produce a potential carcinogen [22], which may subsequently lead to cell proliferation as noticed in the livers of rats fed t occidentalis from the bioaugmented and natural attenuated lots figure 3b and 3c. however, the slight increase in serum amylase activity observed in this study indicates minimal destruction of pancreatic cells. table 5 and figures 4a 4c. the pronounced reduction in packed cell volume and hemoglobin concentration recorded this study table 6 may be as a result of destructions of the cells of the kidney and liver. this may cause an impairment in the secretion of erythropoietin thereby resulting to anaemia. erythropoietin in the presence of colony-stimulating factors (csf) regulates the differentiation of stem cell in the bone marrow, which ensures that erythrocyte precursor cells are converted to erythrocytes [21]. aside neurological and developmental anomalies, exposure to elevated lead concentrations may lead to biochemical defects in both humans and animals. the resultant effect of such impairment may be on kidneys, gastrointestinal tract, joints and reproductive tissues, thereby affecting haemoglobin synthesis [23]. this tantamount to the observed decrease in packed cell volume and haemoglobin concentration recorded in rats fed t occidentalis from natural attenuated and bioaugmented lots. conclusively, the agriculture and food sciences research, 2021, 8(2): 36-43 43 © 2021 by the authors; licensee asian online journal publishing group hydrocarbon and heavy metal concentration in crops harvested from crude oil polluted and remediated soil has to be evaluated before crops from such environment can be allowed for both human and animal consumption. references [1] d. w. stevenson, plant uptake of water and mineral nutrients from soil. chicago: encyclopedia britannica, 2009. [2] z. yang and l. zhu, "performance of the partition-limited model on predicting ryegrass uptake of polycyclic aromatic hydrocarbons," chemosphere, vol. 67, pp. 402-409, 2007. available at: https://doi.org/10.1016/j.chemosphere.2006.09.008. [3] a. kipopoulou, e. manoli, and c. samara, "bioconcentration of polycyclic aromatic hydrocarbons in vegetables grown in an industrial area," environmental pollution, vol. 106, pp. 369-380, 1999. available at: https://doi.org/10.1016/s0269-7491(99)001074. [4] leaf, "wikipedia, the free encyclopedia, retrieved from; http://en.wikipedia.org/wiki/leaves.," 2011. [5] fao, "food and agriculture organization minor oil crops," ed rome: fao agricultural services bulletin, 1992, p. 94. [6] aoac, official methods of analysis. virginia, usa: association of official analytical chemists, 2006. 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[20] c. d. klassen, m. o. amdur, and j. doull, the basic science of poison. chicago: encyclopedia britainnica, 2009. [21] j. koolman and k. h. roehm, "colour atlas of biochemistry," 2nd ed new york: thieme stuttgart, 2005, pp. 330 – 331. [22] j. h. ottaway, "the biochemistry of pollution," ed london: edward arnold publication, 1980, pp. 12 20. [23] s. k. karri, r. b. saper, and s. n. kales, "lead encephalopathy due to traditional medicines," current drug safety, vol. 3, pp. 54-59, 2008. available at: https://doi.org/10.2174/157488608783333907. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://en.wikipedia.org/wiki/leaves., 1 © 2021 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 8, no. 1, 1-9, 2021 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2021.81.1.9 © 2021 by the authors; licensee asian online journal publishing group impact of the aerobic mesophilic microorganisms on black sigatoka of bananas according to the cropping systems in the region of kisangani (case of the old secondary forest) gabriel akwakwa hopla1 yang sun2 chunyu sun3 odimba onautshu4 ( corresponding author) 1college of life sciences, jilin agricultural university, changchun, china; faculty of sciences, university of kisangani, drc. 2,3college of life sciences, jilin agricultural university, changchun, china. 4faculty of sciences, university of kisangani, drc. abstract the microorganisms play crucial roles in the cycle of matter and damage the organic substances, sources of electrons, source of carbon, and source of energy for their biosynthesis. in this work, we studied the impact of the aerobic mesophiles microorganisms on the black sigatokaof banana in the old secondary forest. the objective was to count and to identify the microbial diversity of the forest ecosystems, as well as to study their impact on the development of the black sigatoka of banana. the assessment of the microbial populations has been done in an experimental field according to a device in blocks of fischer by the method of successive dilutions of coloration of gram and by the discharge of ascospores. the results showed that the rain season was lower in total microbial biomass (700, 7 colonies) than subdry season (840,3 colonies). the bacillus genera have been more represented more than the coccusgenera. the impact of black sigatoka of banana was raised at the cultivar libangalikale (40%) and low at yangambi 5 km (14%). the distribution of microorganisms in depth in the sub-dry season to the level of surface with vegetation was considerable either 3819, 3. 103 ufc( unit format colony) by gram of soil between 0 and 5 cm against 2754,5.103 ufc by gram of soil between 15 and 20 cm. this suggests that the raised number of microorganisms could have positive impact on soil fertility by decreasing the illnesses in this ecosystem. keywords: aerobic mesophilic, black sigatoka, cropping systems, libanga likale,yangambi 5km, litete, gros michel, mycosphaerellafijiensis morelet, bacillus. citation | gabriel akwakwa hopla; yang sun; chunyu sun; odimba onautshu (2021). impact of the aerobic mesophilic microorganisms on black sigatoka of bananas according to the cropping systems in the region of kisangani (case of the old secondary forest). agriculture and food sciences research, 8(1): 19. history: received: 3 november 2020 revised: 5 february 2021 accepted: 26 february 2021 published: 16 march 2021 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ......................................................................................................................................................................................... 2 2. material and methods ........................................................................................................................................................................ 3 3. results and discussion ...................................................................................................................................................................... 4 4. conclusion and suggestions ............................................................................................................................................................. 6 references ................................................................................................................................................................................................. 6 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2021.81.1.9&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2021.81.1.9&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/2734 https://orcid.org/0000-0002-0343-1143 agriculture and food sciences research, 2021, 8(1): 1-9 2 © 2021 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the objective was to count and to identify the microbial diversity of the forest ecosystems, as well as to study their impact on the development of the black sigatoka of banana. 1. introduction the soil is not just the support in which plants take root and can provide the nutrients essential for their development, but also is a reservoir of microorganisms. these soil microorganisms are important for plant productivity. they are the most abundant living organisms in the soil, and they play an active role in the nutrient and organic matter cycle, soil fertility, plant health and production. over 90% of plants worldwide develop symbiotic associations with at least one type of mycorrhizae [1]. in the soil, there are beneficial bacteria that maintain very intimate relationships with the plant and strengthen its natural defenses against stress of biotic origin. bacteria are of considerable importance in biogeochemical cycles, such as those of carbon or nitrogen with a fundamental role in the fixation of atmospheric nitrogen, a function which has been much studied for several decades [2]. the soil is the outermost layer, marked by living things in the earth's crust. it is the site of an intense exchange of matter and energy between air, water and the global cycles of matter [3].'' it is considered to be one of the most biospheres and is therefore a major reservoir of microbial diversity. it is made up of insoluble mineral debris produced by the fragmentation and alteration of sedimentary rocks (limestone, etc.), organic matter (colloidal humus, etc.), living and dead organic matter, air and water, which allow the proliferation of telluric microorganisms [4]. the physical properties of the soil are determined by the sizes of the particles that compose it. we can classify them by decreasing diameter: sands, silts, clay. the organic part of the soil includes plant debris, animal remains and varying amounts of amorphous organic matter called humus. the gases contained in the soil are mainly oxygen, nitrogen, carbon dioxide (co2) and mineral substances dissolved or dissociated by hydrolysis (mineral ion). oxygen is important for the metabolism of plants and their growth. its presence is also necessary for the development of bacteria, and other organisms. in the soil, there are not only microorganisms that swarm. animals, plants, and viruses are also found [5]. this population of the soil undergoes great fluctuations because of the physicochemical factors of the soil in particular the structure, the organic matters, the textures, the ionic concentration, the temperature, the humidity, the ph, the biotic factors: the types of relations (symbiosis parasitism, predation, and commensalism), the phenomenon of inhibition. to these is added the individual potentiality of each species (reproduction, adaptation, feeding). soil is considered today as a vital, non-renewable source, which should therefore be preserved, although the horticultural uses non-soil growing media. most of the methods developed to ensure soil health apply to horticultural substrates and every effort should be made to limit the use of synthetic chemical inputs and depreciate natural resources such as silt [4]. microorganisms play crucial roles in the cycle of matter and degrade organic matter, sources of electrons, source of carbon, and source of energy for their biosynthesis. by dying, microorganisms in turn help enrich the soil with different carbon compounds [6]. on the other hand, many of them promote the growth of plants, ensure the degradation of pollutants and provide compounds of interest, enzymes, antibiotics or other molecules. microorganisms are found in all types of environment found in nature: they colonize all ecosystems, such as soil, freshwater and marine water, air, but also more hostile environments such as the poles, deserts, geysers, the ocean floor, etc. microorganisms found in extreme environments are called extremophiles [7]. many microorganisms are associated with plants or animals with which they can maintain relationships of symbiosis, commensalism or parasitism. some microorganisms can be pathogenic, that is, cause disease in plants or animals. man very early used the properties of microorganisms (bacteria) to feed, to heal. today the fields of application are very varied. in the food industry, bacteria , such as lactobacillus,lactococcusor streptococcus, yeasts and molds are involved in the production of fermented foods, such as cheese, yogurt, beer [7]. the ability of heterotrophic bacteria to degrade a wide variety of organic compounds is exploited for the treatment of polluted soils in bioremediation strategies or for the treatment of wastewater [8]. bacteria are also used in septic tanks to purify them. in agriculture, some bacteria can be used in place of pesticides in biological control to fight plant parasites [9]. (ex: bacillus thuringiensis), other bacteria will have a beneficial effect on the growth of plants [10]. black sigatoka or black streak disease, caused by the fungus mycosphaerellafijiensismorelet, is considered to be the most devastating disease in the world banana crop [11]. this disease, which has a wide geographic distribution, causes early death of infected leaves and is responsible for more than 50% of the losses in banana production [11]. the banana tree is a giant and perennial herb 1.5 to 9 m tall. it consists of a corm (underground part) with roots and shoots, a pseudo trunk provided with leaves, and a bunch of fruits jones [12]; swennen and vuylsteke [13]. its reproduction is vegetative and ensured by suckers [14]. bananas are monocots without an aerial vegetative stem [15] classified in the order of zingiberal, the musaceaefamily and the genus of musa. the genus musa is characterized by inflorescences with bracts inserted separately from the flowers, and includes 5 sections: australimusa, callimusa, rhodochlamys, eumusa and ingentimusa swennen and vuylsteke [13]. the australimusa include a species cultivated for its fibers, m. textilis and the féhi of the pacific islands bearing an erect inflorescence, fruit to be cooked and a colored sap. the callimusa include a few ornamental species such as m.coccinea. the rhodochlamys also include ornamental species such as m. ornata, m. velutina and m. laterita. edible agriculture and food sciences research, 2021, 8(1): 1-9 3 © 2021 by the authors; licensee asian online journal publishing group bananas belong to the eumusasection and are divided into three categories according to consumption: cooking bananas, dessert bananas and wine bananas. in the democratic republic of congo (drc), it is currently spread in all the provinces [16]. bananas and plantains play an important economic, cultural and nutritional role worldwide, particularly in developing countries in the tropics [17]. they are among the main staple foods in producing countries and constitute a considerable source of employment and income through local and international markets [18]. in the kisangani region, no studies have been done on the contribution of microorganisms to banana fungal diseases. that is why this study was carried out around the masako forest reserve in order to determine the impact of aerobic mesophilic microorganisms associated with black sigatoka of banana. the causative agent of mrn for bananas is m. fijiensismorelet. according to cabi [19], this mushroom is classified as follows: phylum: ascomycota. class: ascomycetes. subclass: dothideomycetidae. order: mycosphaerellales. family: mycosphaerellaceae. genus: mycosphaerella. 2. material and methods 2.1. study area the research was conducted in the kisangani region, around masako forest reserve (0 ° 36'n, 25 ° 13'e, eastern province) [20]. covers an area of 2105 ha of which 1/3 is occupied by the northeast primary forest and at least 2/3 by the secondary forests in the northwest [21]. while the south part of the reserve is occupied by fallow and crops (see figure 1). the masako forest reserve is part of the relatively humid guinean-congolese rain forests. it includes six types of vegetation, including: ruderal vegetation, crop weed, post crop vegetation, musangacercropioid vegetation, gilbertiondendrondewevrei forest, and aquatic and semi aquatic vegetation. the soil of the experimental site is sandy-clay, lumpy, fine to medium, brittle, with many roots, dark brown in color [22]. all the eco-climatic data the position of the city of kisangani are near the equator, and give it an equatorial climate of the af type in the köppen classification [23]. 2.2. experimental set up the experimental plots were established in fallow fields and secondary forest according to the fisher block system [24].with 4 treatments and 5 repetitions per treatment. the blocks, also called repetitions, were made up of a group of plots almost forming a square. five blocks or repeats each containing six plants of the same cultivar was placed in each plot (see figure 2). four locally cultivated varieties (gros michel, libanga likale, litete and yangambi 5km) were used and distributed randomly in the plots of each block. 2.3. biological material the biological material used in this work consisted of soil samples taken under aseptic conditions, using a graduated auger, brand eijkelkamppo box 469872 g, in a field of old secondary forest. the type of soil examined in this work was of the silty-sandy type with a fine structure and a very strong anthropic activity. 2.4. plant material the plant material consisted of 4 cultivars of bananas and plantains, including libangalikale (plantain: musa aab), libangalikale (plantain: musa aab), gros-michel (banana: musa aaa), and yangambi km5 (banana: musa aaa), the most cultivated by farmers in kisangani region. the choice of these cultivars was motivated by their presence in the study environment and this is justified by several reasons mentioned by the inhabitants of the village (figure 3). 2.5. sampling of soil microbial populations we collected the soil under aseptic conditions using an auger. we weighed 10 g of soil under the same conditions, and then fixed the sample in 90 ml of peptone water. a series of successive dilutions was made, but only one dilution for 2 petri dishes due to 1 ml of inoculum per dish was seeded. the liquefied nutrient agar cooled to 45 ° c was poured into the seeded petrie dish and after solidification, a second layer was poured. the incubation was carried out at 30 ° c for 72 hours. the number of cfu (colony-forming unit) was obtained using the following formula: 1).21,01.( dnnvml colonies n + =  where: n: number of cfu / g (ml) of the initial product. ∑ colonies: sum of colonies of interpretable boxes. vml: volume of inoculum = 1 ml. n1: number of boxes considered at d1 selected. n2: number of boxes considered at d2 selected ; d1: factor of the selected d1. 2.6. identification of aerobic mesophilic microorganisms the bacteria were identified using at genus level using by the gram staining method. the bacteria were classified into two groups: gram positive colored purple and gram negative colored pink. gram staining was based on the principle that crystal violet stains of all bacteria purple or dark blue. lugol played the role of an etcher. alcohol acetone crossed the wall of some bacteria and discolored them. however, the wall of other bacteria agriculture and food sciences research, 2021, 8(1): 1-9 4 © 2021 by the authors; licensee asian online journal publishing group remained impassable by alcohol and retained the purple or dark blue color which was well reinforced by lugol. finally, saffron colored pink or red any bacteria which wall was discolored and did not act on bacteria colored purple or dark blue [25]. we took the inoculum and spread it on an object slide to get a smear smearusing a platinum loop. we then dried the smear at laboratory temperature and fixed the preparation by passing the bottom of the slide over the flame. then we covered the entire smear successively with crystal violet, lugol, alcohol and saffron; while leaving a minute of reaction for each dye, except the alcohol which was left for 30 seconds of reaction. before switching from one dye to another, the preparation was washed with tap water. finally, we dried the preparation at laboratory temperature, applied a drop of immersion oil and observed under the microscope at 100x objective. 2.7. mushroom isolates from the samples of collected leaves, the strains of mycosphaerella spp. were isolated by the ascospore discharge technique on agar medium (h2o agar), which were then subcultured on potato dextrose agar medium (pda) [24]. for discharge, the pieces of necrotic leaves were first cut, then soaked in sterile distilled water for 20 minutes to moisten them. the leaf pieces were then placed inside lids of petri dishes which were placed on dishes containing 3% agar. the underside of the leaf was directed upwards, facing the culture medium. the dishes were incubated at 25 ° c overnight. the following day, the ascospores discharged onto the agar overnight were subcultured individually on the pda medium (39 g / l). transplanting was done by observation with an inverted microscope (motic ae31), carefully recovering, one by one, the discharged ascospores using a needle. the cultures were incubated at 25 ° c. and the phenotype of the strains observed after 45 days to evaluate the correspondence with that of m. fijiensis, m. musicola or m. eumusae. the strains obtained were subcultured in petri dishes and in tubes in order to preserve the established collection. 2.8. evolution of the disease data were recorded on only one variable: the incidence of the disease. the incidence of the disease (number of plants infected with mycosphaerella fijiensis expressed as a percentage) was assessed every 2 weeks from 2 months after planting. 2.9. data analysis statistical analysis was performed using r.2.10.0 software. 3. results and discussion the results of our investigations are presented in tables 1 to 4 and illustrated by figures 4 and 5. the total density of the microbial flora according to the depth during the dry season and that of rains is presented in table 2. table-1. number of microbial colonies as a function of soil depth during two seasons. season depth ( cm) number of colonies sub-dry 0-5 1238 5-10 889 10-15 628 15-20 606 average 840,3 rain 0-5 504 5-10 588 10-15 726 15-20 788 average 700,7 by examining table 1 relating to the density of bacteria during the dry season and that of rain, it appears that the number of bacteria varies according to the seasons as well as the depths. however, the rainy season is low in total microbial biomass with an average of 700.7 number of microbial colonies compared to the dry season which showed a high value of the number of colonies is 840.3. furthermore, the number of microorganisms was higher at the surface, 1238 between 0 and 5 cm during the dry season and decreases with depth. a contrary situation was observed during the rainy season when this number increases with depth since we observed 504 colonies on the surface and 788 in depth. this could be due to the unfavorable climatic conditions on the surface during the rainy season allowing the mesophilic bacteria to penetrate in depth and to grow between 20 and 45 ° c. compared to previous work, alexander [26] found 97,510 bacteria per gram of soil, the bacterial titer, while meddah, et al. [27]. found up to 5,4,107. the distribution of bacteria in depth during the dry season and that of rain is illustrated in figure 4. looking at figure 4, it appears that a decrease in the number of microorganisms was observed with depth during the dry season while an increase in this number was observed during the rainy season. in addition, the distribution of microorganisms in depth in the dry season at the level of the planted surface increased from 3819.3.103 cfu to 3184.8.103 cfu per gram of soil. this suggests that the environmental conditions of the environment are recognized as playing a determining role in the dynamics of colonization by the microbial population. in the middle of the forest, the densities are much higher and the highest concentrations are located at the level of the first 10 centimeters and decrease slightly at the last centimeter. this suggests that the high number of microorganisms could have an impact on soil fertility and thereby decrease the incidence of diseases in this ecosystem. furthermore, abiotic factors can also affect the specific composition of microbial communities and their biochemical potential. as a result, we see that there is a drop in microorganisms and we have found that the number of colonies in certain boxes, for example in the last 20 centimeters deep, is higher than on the surface. agriculture and food sciences research, 2021, 8(1): 1-9 5 © 2021 by the authors; licensee asian online journal publishing group the variations recorded are due to physicochemical factors as well as to crops which could exert certain influences on microbial populations in the soil. also, abiotic factors can affect the specific composition of microbial communities and their biochemical potential. the results of the identification of the bacteria by gram staining during the dry season are presented in table 2. table-2. characterization of bacteria after gram staining during the dry season. subsèche. strains form gram staining genus density s1 bacillus gram+ bacillus ++++ s2 coccus gram+ staphylococcus ++ s3 bacillus gram+ bacillus ++++ s4 coccus gram+ staphylococcus ++ s5 bacillus gram+ bacillus ++++ s6 bacillus gram+ bacillus ++++ s7 bacillus gram+ bacillus ++++ s8 bacillus gram+ bacillus ++ s9 bacillus gram+ bacillus + legend: s: strain +: 25% ++ : 50% +++ : 75% ++++ : 100% in light of this table, it appears that during the dry season, the bacillus and shell forms were observed with a large dominance of the first form which also dominates in density. with regard to gram staining, all the strains observed are gram + belonging to the genus bacillus and staphylococcus. the method used being specific to genera, it did not allow us to determine the species. the results of the identification of bacteria by gram staining during the rainy season are presented in table 3. table-3. characterization of bacteria after gram staining in the rainy season. strains form gram staining genus density s1 bacillus gram+ bacillus ++++ s2 bacillus gram+ bacillus ++ s3 bacillus gram+ bacillus +++ s4 bacillus gram+ bacillus + s5 coccus gram+ diplococcus +++ s6 coccus gram+ streptococcus ++ legend: s: strain +: 25% ++ : 50% +++ : 75% ++++ : 100% the observation in table 3 shows that the bacillus form has been more observed (4 strains) than the shell form (2 strains) although all the bacteria are gram +. as for the genus, the bacillus form has been represented by a single genus, bacillus, while the shell form has been represented by the genera diplococcus and streptococcus. figure 5 illustrates the incidence of black sigatoka in bananas and plantains in the old secondary forest. the overall rate for all cultivars was below 50%, and ranged from 14 to 40%. as in previous studies carried out in a hut garden [16]. we find that plantains were more susceptible to this disease and therefore had a high incidence, 40% for libanga likale and 32% for litete than banana trees. in addition gros michel was more sensitive to this disease (34%) while yangambi km 5 (14%) showed tolerance to black sigatoka. table-4. effect of depth on the density (cfu) of bacteria in the rainy season. depth(cm) sample d1 d2 cfu (103) 0-5 b1 260 217 4113,6 b2 271 157 5-10 b1 219 133 2600 b2 105 112 10-15 b1 97 98 1650 b2 89 79 15-20 b1 72 68 1295,4 b2 67 78 légend: b1 et b2 : petridish 1 et 2, cm : centimeter, d : dilution, cfu : colonyfarmat unit. the findings showed that there is a high concentration of the cfu density of bacteria (4113.6) followed by the depth in cfu density of the bacteria of 2600 of 5-10, the depth on the density of ufc of 10-15 with 1650 ufc, and the depth of 15-20cm with 1295.4ufc of bacteria. table 4 revealed that the density of bacteria decreased according to the depth in the rainy season. the density of bacteria was the highest at 0-5cm and lowest at 15-20cm. table 5 showed that the density of bacteria increased according to the depthin the dry season. the bacteria were the highest at 15-20cm (with cfu of 2918.2.103) and lowest at 0-5 cfu of (995.5.103).as shown in tables 4 and 5, the density of bacteria varied differently according to the depth between these two seasons. this result may be explained due to during the dry season,the bacteria moved to moved deep to look for their nutrients. agriculture and food sciences research, 2021, 8(1): 1-9 6 © 2021 by the authors; licensee asian online journal publishing group table-5. effect of depth on the density (cfu) of bacteria in the dry season. depth (cm) sample d1 d2 cfu (103) 0-5 b1 45 47 995,5 b2 69 58 5-10 b1 72 69 1609,1 b2 120 93 10-15 b1 88 105 2145,5 b2 99 180 15-20 b1 111 210 2918,2 b2 103 218 légend: b1 et b2 : petridish 1 et 2, cm : centimeter, d : dilution, cfu : colonyfarmat unit table-6. incidence of black sigatoka in old secondary forest. cultivars libangalikale litete gros michel yangambi km 5 plants n ps pi % n ps pi % n ps pi % n ps pi % bloc 1 6 3 3 50 5 4 1 20 4 2 2 50 5 4 0 0 bloc 2 5 3 2 40 4 3 1 25 5 3 2 40 5 5 0 0 bloc 3 5 4 1 20 4 3 1 25 5 3 2 40 6 4 2 33 bloc 4 4 2 2 50 5 3 2 40 5 4 1 20 5 4 1 20 bloc 5 5 3 2 40 4 2 2 50 5 4 1 20 6 5 1 17 avarage 40 32 34 14 standard deviation 12.2 12.5 13.4 14 legend: n: number of plant, p.s.: plant saint, p.i.: plant infected. table 6 indicated that the highest incidence of black sigatoka in old secondary forest was recorded in cultivar libanga likale (with average of 40%) and the lowest in cultivar yangambi 5km (with average of 14%). we concluded that cultivar yangambi 5km was the most resistant to black sigatoka. 4. conclusion and suggestions the main objective of this work was to count and identify microbial diversity as well as to study the incidence of black sigatoka in old secondary forest near the masako forest reserve in kisangani. the evaluation of microbial populations was carried out in an experimental field using a fischer block device. parameters such as density of microbial population, depth of soil, characterization of microorganisms and incidence of black sigatoka were assessed during the dry and rainy seasons. the results obtained during this work show that: the rainy season is low in total microbial biomass (700.7 colonies) than the season dries up (840.3 colonies). the bacteria rate increases with depth during the rainy season and decreases during the rainy season. the distribution of bacteria in depth in the dry season at the level of the planted area is considerable, at 3,819.3. 103 cfu per gram of soil between 0 and 5 cm compared to 2754.5.103 cfu per gram of soil between 15 and 20 cm. the number of cfus during the rainy season varied from 2,990.9.103 to 3,581.8. 103 cfu per gram of soil between 0-5 cm and 15-20 cm respectively. the incidence of black sigatoka in bananas is high in the cultivar libanga likale (40%) and low in yangambi km 5 (14%). in addition litete and gros michel were presented with intermediate values respectively 32 and 34%. the number of bacteria is relatively high in this ecosystem and would reduce the incidence of black sigatoka in bananas and plantains statistically, the one-factor analysis of variance has shown that there are significant differences between the parameters studied. we suggest that: in-depth studies are carried out to characterize microorganisms at the level of species existing in the soil in order to determine their relationships with soil fertility. that other studies be carried out in more depth on the molecular characteristic of microbial communities in the kisangani region in order to better understand them given their very small size. references [1] a. faugier, "bacterial diversity of soils: access to populations with moniritary numbers “the rare biosphere," unpublished doctoral thesis, ecl, 2010. 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[11] d. h. marin, r. a. romero, m. guzmán, and t. b. sutton, "black sigatoka: an increasing threat to banana cultivation," plant disease, vol. 87, pp. 208-222, 2003. available at: https://doi.org/10.1094/pdis.2003.87.3.208. agriculture and food sciences research, 2021, 8(1): 1-9 7 © 2021 by the authors; licensee asian online journal publishing group [12] d. r. jones, "diseases of banana, abacat and enset, londres," ed uk: cabi publishing, 2000, pp. 4-11, 37-39 et 173-190. [13] r. swennen and d. vuylsteke, "bananier. in: raemaekers, r. h. agriculture in tropical africa," ed brussels: dgci, 2001, pp. 661636. [14] c. w. wardlaw, "banana diseases," ed london: longmans group, 1961, p. 648. [15] a. lassoudiere, "the banana tree and its culture," ed france: edition quae, 2007, p. 384. [16] o. onautshu, "characterization of mycosphaerella fijiensis populations and epidemiology of black sigatoka of banana (musa spp.) in the kisangani region (drc)," unpublished doctoral thesis, ucl, 2013. [17] b. k. dadzie and j. e. orchard, routine post-harvest screening of banana/plantain hybrids: criteria and methods. montpellier, france: inibap, 1997. [18] e. frison and s. sharrock, "the economic, social and nutritional importance of banana in the world. in c. picq, e fouré and e. frison (eds.), bananas and food security," in proceeding of an international symposium held in douala, cameroon, 10 14 november 1998. inibap, montpelleir, france, 1999, pp. 21 – 35. [19] cabi, "crop protection compendium. retrieved from http://www.cabicompendium.org," 2007. [20] a. upoki, "study of the stand of bulbuls (pycnonotidae, passériformes) in the masako forest reserve at kisangani (r.d.congo)," unpublished doctoral thesis, fac. sci., kisangani, 2001. [21] j. lejoly, s. lisowski, and m. njele, "catalog of vascular plants from the kisangani and tshopo sub-regions (haut-zaire)," 3rd ed belgium: labor. botan. syst. phytos. ulb, 1988, p. 122 [22] k. soki, "biology and ecology of termites (isoptera) from the rain forests of northeast zaire (kisangani)," unpublished doctoral thesis, ulb, 1994. [23] m. mate, "growth, phytomass and mineralomass of hedgerows of improving legumes in alley crops in kisangani," unpublished doctoral thesis, 2001. [24] j. carlier, d. de waele, and j. v. escalant, "overall assessment of the resistance of banana trees to fusarium wilt, to leaf diseases caused by mycosphaerella spp. and nematodes," inibap technical guides 7. montpellier, france2003. [25] w. oleko, "course," applied microbiology, vol. 1, p. 92, 2011. [26] m. alexander, introduction to soil microbiology, 2nd ed. united states of america: john wiley & sons, inc, 1977. [27] n. meddah, a. o. touhami, and a. douira, "mycoflora associated with banana (musa accuminata l.), grande naine variety, grown under glass in the gharb region (morocco)," bulletin of the scientific institute, rabat, life sciences section, vol. 32, pp. 1-11, 2010. figure-1. location of the masako forest reserve. source: adaptation of 2010 satellite photo, of lisingi and cft. http://www.cabicompendium.org,/ agriculture and food sciences research, 2021, 8(1): 1-9 8 © 2021 by the authors; licensee asian online journal publishing group figure-2. fisher block device. legend: : gros michel : libanga likale : litete : yangambi km 5 : edge plants figure-3. banana and plantain cultivars used agriculture and food sciences research, 2021, 8(1): 1-9 9 © 2021 by the authors; licensee asian online journal publishing group figure-4. distribution of bacteria by depth over two seasons. figure-5. incidence of black sigatoka in bananas and plantains in old secondary forest in masako in 2014. statistiques analysis result • anova :cfu-saison df sum sq mean sq f value pr(>f) saison 1 990722 990722 1.0264 0.3282 residuals 14 13513548 965253 meansd n rainy 1917.075 755.6602 8 dry 2414.750 1165.9694 8 • anova : cfu-depth df sum sq mean sq f value pr(>f) depth 3 920840 306947 0.2712 0.845 residuals 12 13583431 1131953 meansd n 0-5 cm 2554.55 1800.2359 4 10-15 cm 1897.75 286.0771 4 15-20 cm 2106.80 936.9240 4 5-10 cm 2104.55 572.0964 4 • anova : incidence-cultivar df sum sq mean sq f value pr(>f) cllivars 3 1880.0 626.67 3.6343 0.03581 * residuals 16 2758.9 172.43 -- signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1 meansdn gros michel 34 13.41641 5 libangalikale 40 12.24745 5 litete 32 12.54990 5 yangambi 5km 14 14.22049 5 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 96 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 96-103, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6177 © 2024 by the authors; licensee asian online journal publishing group soils of the baikal region: mapping, use, transformation irina aleksandrovna belozertseva1 daria nikolaevna lopatina2 andrei anatolievich sorokovoi3 ( corresponding author) 1,2,3v.b. sochava institute of geography sb ras, ulan-batorskaya str. 1, irkursk, 664033, russia. 1email: belozia@mail.ru 2email: dariatama@mail.ru 3email: geomer@irigs.irk.ru abstract the article gives a brief overview of the soil cartographic work carried out in the baikal region. the paper contains one of the authors' map of agroecological zoning of soils of the irkutsk region. a fragment of the legend is shown. suitable and unsuitable districts for their use in agriculture have been identified. various types of business activities are recommended. also provided are maps of the use of land of the most developed part of the lands of the baikal region in 1980 and 2023. in the irkutsk-cheremkhov plain with neighboring steppe areas, occupying 11% of the area of the region where the bulk of the farmland is located, relatively good natural and climatic conditions are noted. with the collapse of collective farms and state farms since the 1990s. more than 50% of former agricultural land was abandoned. a positive aspect of the "resetting" of the state system is the emergence of a reserve of agricultural land for use in the form of abandoned land with soils that have restored the level of fertility. the authors proposed a scale of qualitative assessment of the possibility of using land in agriculture, developed an assessment of the state of soil fertility, taking into account their regional characteristics. keywords: conditions, mapping, soil, use. citation | belozertseva, i. a., lopatina, d. n., & sorokovoi, a. a. (2024). soils of the baikal region: mapping, use, transformation. agriculture and food sciences research, 11(2), 96–103. 10.20448/aesr.v11i2.6177 history: received: 21 october 2024 revised: 25 november 2024 accepted: 27 november 2024 published: 2 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the russian science foundation and the ministry of economic development and industry of the irkutsk region (grant number: 23-27-10013 (05-62-824/24)). institutional review board statement: the ethical committee of the v.b. sochava institute of geography, russia has granted approval for this study on 12 march 2023 (ref. no. 78). transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 97 2. results and discussion ................................................................................................................................................................... 97 3. conclusion ....................................................................................................................................................................................... 103 references ............................................................................................................................................................................................ 103 mailto:belozia@mail.ru mailto:dariatama@mail.ru mailto:geomer@irigs.irk.ru https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6177 https://orcid.org/0000-0001-7995-2298 https://orcid.org/0000-0002-2696-0822 https://orcid.org/0000-0001-8987-7068 agriculture and food sciences research, 2024, 11(2): 96-103 97 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature taking into account the new soil classification soil and agroecological zoning of the irkutsk region was carried out. we were complited a land use map of the most developed part in agriculture of the irkutsk region. the authors proposed a scale of qualitative assessment of the possibility of using land. 1. introduction authors group [1-5] carried out ecological zoning of soils in the south of the east european plain, russia and the world. there are few materials on the soils of hard-to-reach mountainous regions [6]. an agroecological assessment of the soils of the irkutsk region and adjacent territories is given in the works of shpedt, et al. [7]; ubugunov [8]; ubugunov, et al. [9]; lopatina and belozertseva [10]; seryshev and somedun [11]; atlas [12] and kalep [13] etc. published maps: "soil zoning. atlas of the baikal region" [14]; "soil and geographical zoning. atlas of the irkutsk region"[15]. any zoning for any purpose reflects the level of knowledge and production of the current time, cannot be unshakable and must be refined and improved as new knowledge of landscape components and conditions for improving production and environmental transformation becomes available. information on the level of fertility, the ecological state of the soil cover of the region is constantly supplemented by new studies. 2. results and discussion as a result of many years of work, together with colleagues from the institute of general and experimental biology of the sb ras, the institute of geography and geoecology of the mongolian academy of sciences, we were completed a series of soil-ecological maps for atlases and two wall maps (soils of lake baikal basin [16]) soils of the baikal natural territory [17]). soils are united by natural zones. the most fertile soils are mollisols (chernozems, umbrisols and other such types of soils) are shown in green and yellow light on the map. this soils are wide spread about 30% of the territory. figure 1. agroecological zoning of soils of the irkutsk region. note: provinces: a east sayan and khamar-daban; b pribaikalye and stanovoy highlands; c irkutsk amphitheater. districts: a1 highland east sayan, a2 middle mountain south-western baikal region, a3 low mountain and foothill; b1 highland baikal and delyun-uransky, b2 mid-mountain district of the stanovoy highlands, b3 patomsky low mountain, b4 foothill and low mountain district of the islands of olkhon and priolkhonya, b5 middle and low mountain primorsky; c1 leno-angarsk medium-altitude plateaus, c2 angarsk and prilensky medium-altitude plateaus, c3 erbogachensky and biryusinsky low plateaus, c4 irkutsk-cheremkhovsky and kansko-rybinsk plain, c5 pre-baikal plain. agriculture and food sciences research, 2024, 11(2): 96-103 98 © 2024 by the authors; licensee asian online journal publishing group created soil maps for areas of research. for example on the slide: osinsky district soil map. heavy metal and essential plant nutrition maps for osinsky district too [18]. soil maps have also been drawn up for other areas of the irkutsk region [19]. various maps of soil pollution in different key areas have been compiled, for example, for such cities as irkutsk and ulan-ude in russia, ulan bator in mongolia [20]. maps of soil degradation and pollution were created: baikal lake basin, baikal natural territory [21].the map shows the natural factors of the potential hazard of soil pollution. areas of actual soil contamination and industrial sources are marked. the symbols show the lands of the mining industry. the degree of soil degradation of agricultural lands is shown. we were created soil and ecological zoning of lake baikal basin [9]. soil groups with relatively similar bioclimatic factors (m. budyko’s dryness index, the sum of biologically active temperatures, type and productivity of vegetation) were combined into soil-ecological provinces, which are dominant in soil forming. soils with the similar lithology and geomorphology features (rocks, relief) were grouped into districts. we created a map of agroecological zoning of soils of the irkutsk region (figure 1). a fragment of the map legend is shown (table 1). the legend of the map shows: soils, soil-forming rocks, vegetation and their productivity, height above sea level, dryness index, the number of "active" temperatures, a period with a negative soil temperature, the duration of the warm period, the spread of permafrost, suitability for agricultural use. suitable and unsuitable districts for their use in agriculture have been identified. various types of business activities are recommended. table 1. map legend fragment "agroecological zoning of soils of the irkutsk region". p ro v in c e s d is tr ic ts № soils breeds vegetation h * к * ∑ tm o re 1 0 0 с * m o n th s* d a y s* p * р v * s u it a b il it y * e as t s ay an a n d k h am ar d ab an h ig hl an d e as t s ay an а 1 regosol, leptosol, gleysols, entic podzols crystalline shales, limestones, dolomites, granitoids, basalts, slag basalts moss-lichen and shrub tundra, thickets of cedar dwarf, fragments of larch shrub woodlands and shrubs, rocky placers. 1 1 7 5 -2 8 7 5 0 ,5 -1 ,0 6 0 0 -1 0 0 0 7 -8 5 0 -6 0 s o li d u p , to 1 0 0 -3 0 0 м 2 0 -4 0 u n su it ab le ir k u ts k a m p h it h ea te r ir ku ts kc he re m kh ov sk y an d k an sk or yb in sk p la in в 4 folic retisols, umbrisols, luvisols, phaeozems, chernozem, histosols siltstones, mudstones, carbonate-free sandstones, redcolored carbonatesilicate deposits, limestones pine and larch-pine forbs, steppe forests, fragments of islands of meadow-steppe vegetation 4 0 0 -7 2 8 1 ,5 -2 ,5 1 4 0 0 -1 8 0 0 5 -6 9 0 -1 0 4 a re ab se n t 4 0 -8 0 m o st ly u sa b le note: *– h height above sea level; к– dryness index as per [22] sum of air temperature more than 100 (∑t) [23]; months period with negative soil temperature; days duration of warm period with air temperature more than 100с; p permafrost distribution; pv – productivity of vegetation by [12]. in most of the vast territory of the region (more than 70%), forestry is recommended under taiga forest vegetation, with the exception of specially protected natural areas, which make up a small area. the most optimal conditions for growing early ripening crops are the irkutsk-cheremkhovsky and kansko-rybinsky lowland dictrict, which occupies 11% of the region's territory, where productive chernozems, umbrisols, fluvisols have formed under meadow-steppe and steppe forest vegetation, covering 33% of the district's area or 3.7% of the irkutsk region. we were complited a land use map of the most developed part in agriculture of the irkutsk region (irkutskcheremkhov plain and adjacent territories) (figure 2), and use maps of certain areas of the irkutsk region have been compiled. agriculture and food sciences research, 2024, 11(2): 96-103 99 © 2024 by the authors; licensee asian online journal publishing group (a) (b) figure 2. agricultural land areas of the most developed part of the irkutsk region in 1988 (a), 2023 (b). note: 1 arable land, 2 deposit, 3 pasture, 4 forest, 5 rivers, 6 – settlement. the area of agricultural land in the region is 4% and, in some areas, up to 48%, of which more than half is not used in agriculture, and in remote areas more than 70%, abandoned since the 1990s. after the "restructuring" of the government system. some of the abandoned land is under pasture. the authors proposed a scheme for assessing the level of soil fertility (table 2) [24]. agriculture and food sciences research, 2024, 11(2): 96-103 100 © 2024 by the authors; licensee asian online journal publishing group table 2. scale for assessing the state of soils by indicators: рнн2о, density, content of physical clay, agronomically valuable aggregates, humus, nitrates, mobile phosphorus and potassium. рнн2о рнkcl nl р2o5, mg/kg к2о, mg/kg no3, mg/kg humus, % s, % c pc, % d g/cm3 sf ∑p <4.5 <4.0 strong <25 <100 <4, >130 <1 <20 verylow >40 >1.5 bad1 <16 4.6-5.5 4.1-4.5 26-50 101-200 4-8 2.1-3.0 20-40 low <10 1.3-1.5 unsatisfactor2 16-20 5.1-5.5 4.6-5.0 average 51-100 201-300 8-15 3.1-5.0 40-60 average 10-20 1.2-1.3 satisfactory3 20-24 5.6-6.0 5.1-5.5 weak 101-150 301-400 15-20 5.1-8.0 60-80 above the average 20-30 1.2-1.1 good4 24-30 >6.1 >5.6 notrequ ired >150 >400 20-130 >8.0 >80 high 30-40 <1,1 excellent5 >30 note: nl need for liming; s content of agronomically valuable structure (%); c content of phosphorus, potassium, nitrates, humus and agronomically valuable aggregates; pc content of physical clay (particles < 0.01 mm); d density was assessed on a scale [25-27]; sf assessment of soil fertility according to the above indicators. p score; ∑ p sum of points. agriculture and food sciences research, 2024, 11(2): 96-103 101 © 2024 by the authors; licensee asian online journal publishing group the assessment of the soil condition was carried out according to the author's methodology in the form of generalizing data and a system of indices of individual indicators (environmental reaction, density, content of the fraction of "physical clay", humus, nitrates, agronomically valuable aggregates, mobile phosphorus and potassium). table fragment «some soil indicators of the main key sites» (table 3). it has been established that the soils of arable land and deposits of remote areas (osinsky, bokhansky, etc.) have the highest values of positive quantitative characteristics. according to agrochemical and agrophysical indicators, they belong to prosperous, the state of their fertility is assessed as "excellent", "good" and "satisfactory". the worst indicators of fertility are the soils of the irkutsk region. the soils of the most developed part of the region (cheremkhovsky, bayandaevsky and ekhirit-bulagatsky districts) are assessed mainly as "good" and "satisfactory" (sometimes "excellent", rarely "non-renewable") according to the main agrophysical and agrochemical indicators. according to such agrophysical indicators as the content of a fraction of physical clay, and sometimes agronomically valuable aggregates and soil density of the most developed areas, they are mainly "unsatisfactory" and "bad" for their use for growing agricultural crops, which is mainly due to natural factors (rocks). the soils of the arable land used near large settlements of almost all regions need the introduction of mineral potash and phosphorus fertilizers, less often nitrogen (mineral or organic). there is a reserve of agricultural land for use in the form of abandoned land, but sometimes they need agrochemical, agrotechnical and other activities. in colder conditions of low plateaus (erbogachensky and biryusinsky okrug, covering about 20% of the region), it is possible to grow early ripening crops of vegetables and greens in greenhouses (closed ground). in the driest and moderately cold conditions (foothill and low-mountain districts of olkhon island and priolkhonya, which make up less than 1% of the area) with rocky soils of low thickness, pasture farming using natural pastures is recommended. about 25% of the land in the highand mid-mountain areas of the region is not recommended for use either in agriculture or in forestry. lands on which endemic and relict vegetation species grow must be included in protected areas. in some cases, the territory of protected areas can be expanded, for example, on the southern coast of lake baikal. agriculture and food sciences research, 2024, 11(2): 96-103 102 © 2024 by the authors; licensee asian online journal publishing group table 3. some soil indicators of the main key sites of the irkutsk, cheremkhovsky, bokhansky, osinsky, bayandaevsky and ekhirit-bulagatsky districts of the irkutsk region, assessment of their fertility level (fragment).. location vegetation, use soils рorizon рн h2o humus, % nh4 mg/kg no3m g/kg р2o5 mg/kg к2о mg/kg pc % d g/cm3 s % ∑p sf irkutskregion near the village of novolisikha arable land agricultural (anthrosol) а 5.8 4 0.6 1 4.75 16.6 4 3 1 22 1 24 4 1.35 2 24.7 2 19 unsatisfactory cheremkhovsky region 1 km north of kasyanovka village background area, grass with sedge steppe meadow chernozems au 6.9 5 12.2 5 30.1 2.5 3 160 5 498 5 43 1 0.61 5 68 2 26 excellent 2.5 km northwest of the village of petrovka land under steam, surepka, weedy vegetation agricultural umbric рu 6.9 5 3.7 3 7.3 11.7 3 35 2 110 2 57 1 1.56 1 60 3 15 unsatisfactory bayandaevsky and ekhirit-bulagatsky region 2 km northwest of ust-orda village arableland, rye agricultural chernozems pu 7.2 5 5.4 4 5.34 1.80 1 39 2 175 2 41 1 1.48 2 24.4 2 14 unsatisfactory 1.2 km northwest of the village of pokrovka land under steam, oats with weedy vegetation agricultural haplic fluvisols pu 8.2 5 19.0 5 12.22 17.20 4 39 2 408 5 55 1 1.21 3 43,0 3 23 good bokhansky, osinsky region chernihiv village arable land, weedy vegetation agricultural chernozems pu 7.9 5 5.4 4 11.3 8.1 3 49 2 200 3 21 4 1.14 2 71 4 27 good zakharovskaya village abandoned land more than 30 years ago, pasture, grass meadow agricultural umbric pu 7.2 5 7.7 4 13.8 8.7 3 325 5 950 5 39 5 0.90 5 71 4 36 excellent note: pc content of physical clay (particles < 0.01 mm); d density was assessed on a scale; s content of agronomically valuable structure (%);∑ p sum of points; sf assessment of soil fertility according to the above indicators. the numerator shows the values of the indicator, the denominator points. agriculture and food sciences research, 2024, 11(2): 96-103 103 © 2024 by the authors; licensee asian online journal publishing group 3. conclusion taking into account the new soil classification [27] soil and agroecological zoning of the irkutsk region was carried out. agroecological zoning of soils took into account the entire set of natural and climatic conditions. in most of the vast territory of the region (more than 50%), forestry is recommended under taiga forest vegetation, with the exception of protected areas, which make up a small area. the most optimal conditions for growing early ripening crops are the irkutsk-cheremkhovsky and kansko-rybinsk lowland district, which occupies 11% of the region's territory, where productive chernozems, umbrisols and fluvisols have formed under meadow-steppe and steppe forest vegetation, covering 33% of the district's area or 3.7% of the irkutsk region. almost all lands suitable for agriculture in soviet times were plowed. however, at this time 45% of the agricultural land of the region is abandoned, and in remote areas more than 70%. according to agrochemical indicators, the soils of most of the abandoned lands are in good and satisfactory condition and can be returned to agricultural circulation. references [1] z. a. antonova, soil and ecological zoning of the ulyanovsk region: abstract of the dissertation of the candidate of biological sciences: 02/03/08, 02/03/13. ulyanovsk: ulyanovsk state university, 2011. 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[24] i. a. belozertseva, d. n. lopatina, n. a. zvereva, a. a. cherkashina, and n. a. skosyrsky, "soils of the most fertile lands of the upper angara region," agrarian russia, no. 8, pp. 24-31, 2024. https://doi.org/10.30906/1999-5636-2024-8-24-31 [25] g. a. vorobyova and n. d. kiseleva, morphoanalytic diagnostics of soils. simulation training on the interpretation of soil research results. irkutsk: igu, 2023. [26] a. y. cheremisinov, a. cheremisinov, and s. plotnikov, "compaction of irrigated soils from the impact of agricultural machines," forestry journal, vol. 4, pp. 156-158, 2013. https://doi.org/10.12737/2198 [27] m. v. butyrin, agrochemical characteristics of agricultural land soils and recommendations for the use of fertilizers in the ust-altan municipal district of the osinsky district of the irkutsk region. irkutsk: fsbi tsas irkutsk, 2009, p. 27. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.19047/0136-1694-2018-92-122-146 https://doi.org/10.24412/0044-3913-2022-1-9-13 https://doi.org/10.34655/bgsha.2020.59.2.005 https://doi.org/10.52575/2712-7443-2023-47-3-392-405 https://elibrary.ru/download/elibrary_24268326_12597939.jpg http://atlas.isc.irk.ru/ https://elibrary.ru/download/elibrary_27561954_79426400.pdf https://doi.org/10.1134/s1064229322010033 https://elibrary.ru/download/elibrary_25970923_45780770.pdf https://archive.org/details/climatelife00mibu https://doi.org/10.30906/1999-5636-2024-8-24-31 https://doi.org/10.12737/2198 105 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 105-112, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.105.112 © 2020 by the authors; licensee asian online journal publishing group redefining global food security: do we really have a global food crisis? percy m. chimwamurombe1 charlie c. luchen2 paidamoyo n. mataranyika3 ( corresponding author) 1,2,3namibia university of science and technology (nust), department of natural and applied sciences, windhoek, namibia. abstract with the climate change effects becoming more and more undoubted in the world populations, the reality of food production trends taking a negative curve is clear. this bring up the questions of whether the farmers will be able to produce food for the sustenance of the world population or not? the rate of developing food shortage coping mechanisms in this regard is slower that the ravaging negative climate change effects of drought and floods on farm performance. this commentary has the aim of requesting a fresh discussion around the fundamentals of what is food, what is a food security and what is nutritional security? it is possible that the humanity has a perception that needs refocusing. this is a perception that some people may choose out of nonfood safety issues not to eat certain foods while other however healthily eat such food. it therefore becomes hard to technically accept that food is in short supply for those the opt not to eat that which is edible. keywords: climate change, food production, food shortage, food security, nutritional security. citation | percy m. chimwamurombe; charlie c. luchen; paidamoyo n. mataranyika (2020). redefining global food security: do we really have a global food crisis? agriculture and food sciences research, 7(1): 105-112. history: received: 27 april 2020 revised: 4 june 2020 accepted: 8 july 2020 published: 22 july 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction .................................................................................................................................................................................... 106 2. what is food security? ................................................................................................................................................................ 106 3. what is food insecurity? ............................................................................................................................................................. 106 4. what is nutritional security? ..................................................................................................................................................... 106 5. what is edible? .............................................................................................................................................................................. 107 6. what is food palatability? .......................................................................................................................................................... 107 7. is it possible to broaden the food base? .................................................................................................................................. 107 8. is it possible to minimize food spoilage? ............................................................................................................................... 108 9. is it possible to avoid food wastage? ..................................................................................................................................... 108 10. can food be recycled? ............................................................................................................................................................... 108 11. is it possible to develop a catalogue of global cuisines? ................................................................................................. 109 12. are some foods superior to others? ...................................................................................................................................... 109 13. what is the role of culture and religion in defining what is food to a society/ to the world population? ....... 109 14. discussion ..................................................................................................................................................................................... 110 15. way forward ................................................................................................................................................................................ 110 references ............................................................................................................................................................................................ 110 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.105.112&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.105.112&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1920 https://orcid.org/0000-0003-1911-1651 https://orcid.org/0000-0002-3093-4635 https://orcid.org/0000-0001-9758-9752 agriculture and food sciences research, 2020, 7(1): 105-112 106 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this paper provides an alternative outlook on the current state of food security across the globe. through review of literature and other published works, this paper brings to light the options to increase food security in the world. 1. introduction climate change effects worldwide are leading to drought in some places and indeed floods in other. some places now have some long cropping seasons while indeed in other places the seasons have become shorter. the overall impact of these changes is the changes in food production in the farms. a lot of farmers are indeed not able to meet the usual food demands of the increasing consumer population. in this commentary, we seek to catalyze a fresh global discussion on what is food shortage, what is food and many definitions concerning food security. our view is that such a discussion can lead to establishing globally accepted food cuisines and a re-evaluation of what people should call food or rather new food. this re-evaluation is expected to redefine food shortage and stimulate innovation around enhancing food security globally, offering new food to other territories instead of relying on pre-set old boundaries of conceptualizing food. 2. what is food security? food security is a concept that has numerous definitions and opinions. the most common definition is the one from the 1996 world food summit that stated: food security, at the individual, household, national, regional and global levels is achieved when all people at all times, have physical and economical access to sufficient, safe and nutritious food to meet their dietary needs and food preferences for an active and healthy life. in europe what is described as food security is known as food safety in the united states [1]. in the european context food security is the regulation and control of food safety chains to monitor food toxicity, hygiene and traceability. kyeyune and turner [2] defined food security as accessibility to food at all times despite production and prices fluctuations. however, the different definitions encompass the various aspects of food from availability, access, production and cost. based on the definitions above, food security was dissected to have 4 major components that are food availability, access, utilization and stability [3]. food availability is best described as the amount of food present at any given time via food production channels. the availability of food is, therefore, usually closely linked to agricultural output. food access on the other hand zones in more on the ability of a household to obtain said food given obstacles like road access and cost [4]. food utilization is the process of preparing and consuming food. this includes the amount of food consumed and the proportions. stability typically refers to circumstances surrounding the accessibility of food which may include violence and price variations [5]. the concept of food security, therefore, should be analyzed on a broader sense as low-income countries have, in addition to growing and purchasing food, resorted to consuming wild fruits, vegetables and crops. plants that are recognized traditionally and accepted within the respective societies [6]. therefore, the notion of largely connecting food to that which only cultivated by some people introduces fundamental errors going forward in the food security discourse. 3. what is food insecurity? the description of food security is incomplete without mentioning food insecurity. food insecurity is described as the insufficient access to or possession of food. food insecurity hangs heavily on the failure to obtain sustainable amounts of nutritious and safe food on a regular basis [6, 7]. the state of food insecurity is often attributed to climate change, however, economic instability and regional conflict in war torn zones also contribute largely. the consequences of which are usually linked to the development of malnutrition and increased mortality rates [8]. high food demand and food production shortfalls results in food scarcity which in turn will give a rise to food insecurity [9]. it should be noted that all the countries flagged by fao as having adverse food crises are also experiencing some form of violent conflict food insecurity [10] and when linked to climate change is usually characterized by low agricultural output due to droughts, diseases, or floods [11]. however, people may still experience food insecurity despite adequate agricultural output. this is because, though food may be available individuals may not have the resources and means to access it, cases that affect both third and first world countries [6]. food insecurity has in some cases been merged with the concept of hunger, making the words food security and hunger interchangeable [1]. the general global increase of food prices and subsequent challenges of accessing food have resulted in some governments in sub-saharan african countries incorporating traditional knowledge systems, strategies and policies in order to improve the situation of food insecurity. many of these attempts target the provision of affordable food and increasing access to food [12]. although food security and insecurity depend on the access to food, one is forced to question whether food access really the point of question. the understanding of what food is within communities is questionable and brings doubt to the idea of food security with respect to access. therefore, can one say food security or insecurity is entirely dependent on the accessibility of food given cultural differences play a significant role in what is socially accepted as food? 4. what is nutritional security? in addition to food security, nutritional security takes into consideration care, health and hygiene practices [1]. therefore, the food and agriculture organization defines nutrition security as “a situation that exits when secure access to an appropriately nutritious diet is coupled with a sanitary environment, adequate health services and care, in order to ensure a healthy and active life for all household members.” although food security and nutritional security are often described as co-dependent and somewhat similar in definition, it should be noted that nutritional security is not always a reflection of food security. nutritional security is highly dependent on food security, however, knowledge of how to implement and utilize available food resources is crucial as nutrition dependent agriculture and food sciences research, 2020, 7(1): 105-112 107 © 2020 by the authors; licensee asian online journal publishing group health complications may arise [13]. nutritional security, like food security encompasses the idea of the availability in sufficient amounts of nutrients that promote positive growth and development [14]. several underutilized foods are known to have good nutritional value but are not mass produced globally and locally neither are they often incorporated into diets such as tylosema esculentum (marama bean) [15]. marama bean, an indigenous plant found in botswana, namibia and south africa is an edible nutritious legume. it is a highly nutritious with high contents of proteins (29-39%) and lipids (32-42%) among other nutrients and antioxidative compounds [16]. nevertheless, despite the high nutritional value of marama bean, it is not consumed much beyond botswana, namibia and south africa. bambara groundnut (vigna subterranea (l.) verdc.), for example is a nutritious pulse grown in several african countries like namibia. despite it not being grown for the global market, it has minimum agricultural input requirements and high nutritional compositions [17]. the functionality of bambara groundnut alludes to the possibility of using underutilized or minor crops and foods in nutritional security. such crops become a solution especially to low income families without regular affordable access to nutritious food. these crops usually require less financial input and in local regions are usually developed to withstand pests and diseases. apart from providing nutritious alternatives and production ease, these minor crops offer other purposes such as fodder, a livestock need not often readily available in the agricultural sector [18, 19]. nutritional security, therefore, requires attentive consideration in itself as more often than not it affects low income families and communities. 5. what is edible? edible is from the latin word that means “to eat”. anything that can be safely consumed is called edible. it is important to define the term edible as there are many cultural and social connotations that may result in alternative definitions. however, that definition is very limiting due to the term “safe to eat”. over the past decades, the number of food allergies cases have increased for both adults and children (by 18%) with the most cases being reported with milk, egg and peanut sensitivities. milk and eggs both being major components of most diets [20, 21]. food allergy is a life-threatening costly condition that reduces the food options of an individual. the term “safe to eat”, therefore, is distorted in that respect. entomophagy, the consumption of insects, is commonly practiced in developing countries and is therefore, nutritious alternatives. some of the most commonly eaten insects like locusts (locusta migratoria), grasshoppers (zonocerus variegatus) and crickets (acheta domesticus) have high protein contents. the protein content of the meal worm for example is 24.6 g/100g [22, 23]. table 1 presents nutrition content of edible insects. table-1. comparative amounts of protein in different foods including insects. food protein (g per 100g) wasps, bees, ants 13-77 true bugs 42-74 beatles 23-66 dragonflies 46-65 crickets 23-65 mopane caterpillar 48-61 termites 32-38 mackerel 16-28 beef 19-24 chicken 23 pork 20 eggs 12 source: (adapted from un report 2013) [24]. 6. what is food palatability? palatable is any food that is acceptable or satisfactory to an individual. it is a perception-based notion. this acceptability is largely determined by our sensory system. our sensory systems are designed to help us detect palatable foods and avoid what they perceive to be non-palatable [25]. therefore, palatability can be said to be subjective as one individual may find a food palatable and the next may not. palatability is also controlled by the absorption, intake, selection and digestion of food [26]. the term palatable refers to the favorable taste a food has that triggers more intake of the food. it may also refer to how one favors the food in question, however, the first definition is usually accepted as an appropriate definition [27]. palatability is also linked to food pleasantness leading to an increased consumption and may influence favor of choice. thereby, possibly prompting intake of new foods [28]. despite this, it is difficult to measure palatability as it is subjective. the influence of it with respect to food consumption may be considered insignificant as a result. 7. is it possible to broaden the food base? yes, the food base can be grown as people’s cultures, traditions and locations strongly dictate what food base is based in that region. for example, in the 1800s, immigrants from italy into the united states introduced foods such as spaghetti and pesto and these got integrated into the us cuisine. however, many different edible plants and animals although nutritious remain underutilized mostly due to ignorance on their existence and cultural beliefs. legumes such as marama bean and bambara groundnut (sometimes referred to as orphan legumes) are consumed in some african countries within certain tribal groups with the benefits of minimal input and great nutrient value, however, these crops remain largely underutilized [15, 17]. it can be argued that growing the food base, although possible, might be a daunting task for example introducing pork to the middle eastern people into their diet is considered impossible as they believe that it is religiously “unholy” [28]. agriculture and food sciences research, 2020, 7(1): 105-112 108 © 2020 by the authors; licensee asian online journal publishing group fish foods also offer a health alternative to nutritional security. different types of seafood are typically rich in vitamin d and other minerals such as calcium, iodine, magnesium, selenium and zinc. in many tropical and coastal countries, they are a readily available and affordable source of food [29]. however, like all food sources now, fish are at risk of becoming scarce as a food source due to climate change and other factors such as overfishing. these factors raise challenges for areas largely dependent on the seafood as a source of food and nutritional security and income. therefore, the effects will impact the economy, migration patterns and population stability [30]. given the potential of many different foods to provide all essential nutrients and be cheaply cultivated or sourced, it is possible to grow the food base if other crops, fruits and animal sources are explored [30]. given the potential of many different foods to provide all essential nutrients and be cheaply cultivated or sourced, it is possible to grow the food base if other crops, fruits and animal sources are explored. 8. is it possible to minimize food spoilage? food spoilage is considered as any sensory change in food that the consumer perceives it to be unacceptable and happens metabolically [31]. spoilage in most food is caused by a high percentage of water and oxygen most especially in the fresh foods [32]. canning is one of the methods in which food spoilage can be avoided when done correctly. it prevents growth of undesirable of microbes, destroys enzymes, and removes oxygen by helping form a high vacuum in the jars [31, 32]. improper storage and handling of food have also been linked to lead to spoilage due to failure to prevent contamination. given the known causes of food spoilage and considering the amount of food discarded each year (estimated to be 40% of all produced food) avoiding food spoilage is of paramount importance and also possible [33, 34]. prevention of spoilage due to microbial activity may be achieved by practicing food preservation techniques like refrigeration, chemical preservatives and drying. improper storage of food may accelerate aging of preservative agents. following storage instructions may, therefore, be a solution to avoiding spoilage [35-37]. it is also vital to explore developments in food technology to prevent food spoilage like the use of cymbopogon citratus oil commonly known as lemongrass oil in english [38]. the conventional food preservation strategies such as the use of refrigerators, heating and the addition of antimicrobial compounds have been reported to contribute to the loss of taste and flavor in the foods. this is replaced using organic agents such as lactic and acetic acid [32]. 9. is it possible to avoid food wastage? food wastage is a very big problem worldwide as it has been reported that about a third of all the food produced globally is wasted [39] as can be seen on the diagram that follows. globally, an estimated 1.3 billion tons of food per year are never consumed with wastage beginning from agricultural production to domestic use in households. on average developed countries have a higher degree of food wasted than developing countries [33]. food wastage unlike food spoilage, is the discarding of food that is still fit for human consumption. as over production and excessive supply result in some produce not being sold in retail markets and subsequently wasted, efforts have been made by some countries to minimize the amount of food wasted. in 2016 france became the first country to ban discarding food considered no longer suitable for sale in favor of donating it to those in need [40, 41]. however, food wastage can be attributed to wasteful habits, therefore, in order to address the problems of and associated with food wastage a couple of actions may be taken. social approaches to reducing wastage include feeding those in need of food and also feeding animals. while introducing fines as punishments for wasting may also be effective. as some food is lost due to spoiling it is essential for consumers to buy food they need only and to also take note of the storing and packaging requirements in order to prolong shelf life [31, 41, 42]. food wastage can be avoided by not over buying, planning ahead, checking the sell-by date as people tend to waste food because they believe it has gone bad. retailers can help the consumers buy the right amounts of food and help them with proper storage conditions [43]. food wastage can also be avoided and reduced by raising the awareness on the impact of food wastage on food security. less food wastage would lead to more efficient land use and better water resource management with positive impacts on climate change and livelihoods (figure 1). figure-1. the figure presents the distribution of food lost in the world post-harvest. the figure presents overall percentages of fruits and vegetables, different meats, dairy and cereals. source: adopted from food and agriculture organization food loss index [44]. 10. can food be recycled? the idea of recycling is to maximize the use of any given item with overall intention to save the environment, minimize costs and improve resource utilization. with respect to food production, most food is recycled as 10 25.5 12 22 8 0 5 10 15 20 25 30 other roots,tubers and oil-bearing crops meat and animal products fruits and vegetables cereals and pulses percentage food loss food loss from post harvest to distribution2016 agriculture and food sciences research, 2020, 7(1): 105-112 109 © 2020 by the authors; licensee asian online journal publishing group production stage with the intent (in most cases) to use it as fodder. recycled food may also be used as fertilizer though it can be difficult to sort when in large quantities risking inclusion of food packaging [33, 42]. food waste may also be used to clean water by biodegradation and bioabsorption as in the case of dye removal in water. it is, therefore, possible and advisable to recycle food [45]. there have been reports of food being recycled from manufacturing and processing industries and this food being used as feed [46]. recycling of left-over food helps in the management of food waste, as it can be utilized in aerobic and anaerobic biotechnological processes [46]. in most cases the major fraction of food waste, however, may not be qualitatively good enough in terms of hygienic issues to reuse it as food. therefore, the hydrolysis of organic materials and recycling of them as secondary raw materials in processes for materials and chemicals productions seems the only resource efficient way to deal with it. 11. is it possible to develop a catalogue of global cuisines? yes, it is possible, with the integration and sharing of different food catalogues around the globe. in the last century there has been an increase in interrelationships between cultures and countries thereby allowing for exchange of cultural systems including food and food habits [47]. tourists eager to try different cuisines are attributed to the introduction to different cuisines. an increased number of tourists are travelling seeking to try out different foods in what is now known as culinary tourism. diversity, palatability and unique presentation encourage information exchange of ideas and recipes and all variations available while visiting [48]. restaurants that serve foreign cuisines also encourage cultural flow of global foods [47]. therefore, given these facts, it is possible to develop a global cuisine with the incorporation of different cultures, lesser known or underutilized foods and improved food preparation techniques that prevent or limit food spoilage and wastage. although with such a catalogue a debate would arise on which foods to include as one that would include all the foods consumed globally will be impossible to construct. in addition, some foods might not be considered a good fit for the catalogue as they would have not passed the food safety agents tests for them to be considered non risky for consumption. for example, in europe you have the european food safety authority that provides scientific advice on foods that they say helps protect the consumer animals and the environment from food-related risks. 12. are some foods superior to others? superiority is being dominant or better than the next object or individual. therefore, to say some foods are better than others is a question of preferences hence an answer to this question will not have any objectivity tied to it. some nutritionists have said some foods are superior compared to others due to them having a higher percentage of essential minerals as compared to other foods [49]. soybeans for example are legumes considered superior based on their high protein content which ranges between 34.96% – 36.07% (or higher) [50]. the term superfoods may also be coined to describe nutritious foods with other health benefits like disease prevention. other foods may have phenolic compounds and antioxidants that improve general health [51]. in western europe, hungary in particular, they had been reports of “inferior” foods being shipped into the country. one such case is when the hungarian authorities found a certain brand of pizza having less cheese than the same pizza from the same brand being sold in germany and austria. inferior food can be synonymous with inferior good. with an inferior good being a good whose demand goes down when the income of a population goes down hence foods such as noodles and canned foods are said to be inferior foods as they are normally consumed by people from low income households. the superiority of food can be determined by nutritional value and health benefits. foods with high nutritional value are considered superior based on their ability to provide essential nutrients in sufficient amounts. the idea of superiority of foods can also be considered subjective as it can be judged according to flavor and taste resulting in favor of particular foods. this will lead to some foods being eaten more than others depending on an individuals’ preference [52]. 13. what is the role of culture and religion in defining what is food to a society/ to the world population? there is a saying that goes, “you are what you eat” we can see how culture largely influences what we consume as when we are consume some certain kind of traditional food we are grouped to belong to a certain culture. food then plays an important role in defining the cultural identity of a group of people [53]. religion and food are also connected which adds meaning and significance to our lives. attempts to address and rectify malnutrition challenges based on food security have been, in some cases, noted to be hindered by culture and beliefs related to edible foods [3]. the un documented cases in which nutritious and edible foods were rejected based on cultural aspects not considered [24]. over the turn of the 21st century it has been found that american fast demand has declined while asian demand has increased due to its diversity and uniqueness [54]. for example, muslims fast during ramadan and during this fasting month they eat and drink before dawn and after sunset. also, the jews follow a strict diet rule which they refer to as a kosher diet [53]. many followers of buddhism, hinduism, and jainism are vegetarian this is in part due to them following a doctrine that teaches noninjury or nonviolence. therefore, abstinence from eating meat in these traditions stems from the desire to avoid harming other living creatures. in this regard we can understand that food conveys religious sentiments of the people, making them unique and having their own identity. the following table 2 shows this interrelatedness. culture and religion play a huge role with respect to food intake. culturally or religiously acceptable foods contribute to the discord relating to the definition of edible. within the hindu religion, it is thought disreputable or to some extent punishable by religious law to consume beef as cattle are a symbol of a deity. then, within the confines of the term “safe to eat”, the definition is distorted given that beef is consumed by many other cultural groups all over the world. pork is another such example widely shunned by individuals who practice the islamic religion and some christian groups. it is important then to try and conjure an accommodating definition of what is edible with regards to and the contributions to food security. agriculture and food sciences research, 2020, 7(1): 105-112 110 © 2020 by the authors; licensee asian online journal publishing group table-2. various religious sects and their food preferences. type or religion practice or restriction rationale buddhism refrain from meat while vegetarian diet is preferred. moderation in all foods. fasting required of monks. natural foods of earth are considered most pure. monks avoid all solid food after noon. eastern orthodox christianity restrictions on meat and fish. fasting selectively observance of holy days includes fasting and restrictions to increase spiritual progress hinduism beef prohibited. all other meat and fish restricted or avoided. alcohol avoided. numerous fasting days. cow is sacred and cannot be eaten, but products of the "sacred" cow are pure and desirable. fasting promotes spiritual growth. islam pork and certain birds prohibited. alcohol prohibited. coffee/tea/stimulants avoided. fasting from all food and drink during specific periods. eating is for good health. failure to eat correctly minimizes spiritual awareness. fasting has a cleansing effect of evil elements. judaism pork and shellfish prohibited. meat and dairy at same meal prohibited. leavened food restricted. fasting practiced land animals that do not have cloven hooves and that do not chew their cud are considered as unclean (e.g. hare, pig, camel). mormonism alcohol and beverages containing caffeine prohibited. moderation in all foods. fasting practiced. caffeine is addictive and leads to poor physical and emotional health. fasting is the discipline of self-control and honoring god. protestants few restrictions to food or fasting observations. moderation in eating, drinking and exercise is promoted. god made all animal and natural products for humans' enjoyment. gluttony and drunkenness are sins to be controlled. rastafarian meat and fish restricted. vegetarian diets only, with salts, preservatives and condiments prohibited. herbal drinks permitted; alcohol, coffee and soft drinks prohibited. marijuana used extensively for religious and medicinal purposes pigs and shellfish are scavengers and are unclean. foods grown with chemicals are unnatural and prohibited. biblical texts support use of herbs (marijuana and other herbs) roman catholicism meat restricted on certain days. fasting practiced restrictions are consistent with specified days of the church year. seventh-day adventist pork prohibited and meat and fish avoided. vegetarian diet is encouraged. alcohol, coffee and tea prohibited diet satisfies practice to "honor and glorify god" source: adapted from faqs religion and dietary practices [55]. 14. discussion when talking of a world food crisis it is important to redefine terms such as edible and palatable only then can we confidently talk of having a food crisis. food is termed edible if it is safe to eat while some foods may not be edible in some regions of the word, such as most grasses that are eaten in most parts of africa are not known as food or consumed in western countries. another example is the marama bean which was mentioned earlier in the report. therefore, growing the food base in these areas that have reported a large population of its people not having sufficient food would be one way to tackle the food crisis. on the other hand, this may not be accepted by some people because most of the foods that are eaten by the local population in africa have not undergone any scientific tests therefore some individuals may not deem them edible or safe to eat. food wastage which has been extensively defined can be avoided by taking critic steps along the production to consumption pipeline. wastage should be minimized during the production, farmers have their produce wasted due to it not meeting consumer standards on a yearly basis and most of the food is lost in this way. this could be avoided by farmers only breeding/growing the consumer appealing or marketable foods. another way to curb the food wastage problem would be to give the excess food in households and supermarkets to the less privileged before the food spoils and make use of preservation techniques such as refrigeration. although reducing food waste by giving the surplus food to the underprivileged looks like a simple solution some sections of the population feel this is not the way to go. their arguments lie in the belief that this is a morally distressing and problematic solution to food wastage, as it provides and illusion of solving hunger while in the actual fact it is not addressing the underlying problem of poverty. 15. way forward despite all the information presented above, the questions raised surrounding food security do not present a clear picture of the state of food resources in many communities. food security though largely dependent on climatic conditions and consequentially the economic status of the country, does not put much consideration into underutilized plants and animals as well as unknown edible plants and crops. the basis of food security, therefore, ignorantly lies on recognized available foods. in conclusion, it is important to acknowledge the differences in definitions of the terms presented in this paper, however, the original definitions though insightful are largely limiting. many questions are raised therefore by unifying the limitations of the definitions. there is need, thus, to re-evaluate the definition of food security and given the many possibilities present on the different types of foods globally available. references [1] a. d. jones, f. m. ngure, g. pelto, and s. l. young, "what are we assessing when we measure food security? a compendium and review of current metrics," advances in nutrition, vol. 4, pp. 481-505, 2013. available at: https://doi.org/10.3945/an.113.004119. 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[accessed september 28, 2019]," 2020. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.fao.org/food-loss-and-food-waste/flw-data http://www.faqs.org/nutrition/pre-sma/religion-and-dietary-practices.html http://www.faqs.org/nutrition/pre-sma/religion-and-dietary-practices.html 8 © 2023 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 10, no. 2, 8-13, 2023 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v10i2.5186 © 2023 by the authors; licensee asian online journal publishing group invitro analysis of antifungal effects of botanicals on sclerotinia sclerotiorum causing white mold disease prashant gyanwali1 renuka khanal2 netra prasad pokharel3 bhuwan tharu4 rajan koirala5 sandesh paudel6 rajan paudel7 ( corresponding author) 1,2,3,4,5,6,7institute of agriculture and animal science, paklihawa campus, tribhuwan university, nepal. 1email: prashantgyawali7@gmail.com 2email: renukhanal57@gmail.com 3email: netrapokharel73@gmail.com 4email: tharubhuwan44@gmail.com 5email: rajankoirala567@gmail.com 6email: paudels761@gmail.com 7email: rajan@iaas.edu.np abstract white mold, sclerotinia sclerotiorum, is a devastating fungal plant pathogen that has affected many crop species worldwide. using chemicals to control the disease has been practiced over the years, whose prolonged application has negatively impacted the environment, thus finding an organic solution is crucial. the analysis quantifies the effect of 5 different local plants that have been proven to have fungicidal properties; artemisia vulgaris l., azadirachta indica l., zingiber officinale roscoe, allium sativum l., and lantana camara l. poisoned food technique was used to study the inhibition effect, carried out by inoculating and growing the fungus on pda media infused with botanical extracts. the data of mycelial mat diameter was recorded till the control plates were fully occupied. the growth-inhibiting capacity was found as 100% by allium sativum, 28% & 43% by azadirachta indica, 22.44% & 44% by zingiber officinale, and 16.33% & 8.55% by lantana camara at 10% and 20% concentrations respectively. only a slight difference between the overall inhibition effect of the two concentrations was found with 37.22% inhibition by 20% concentration and 33.38% inhibition by 10% concentration. no inhibition effect was observed from artemisia vulgaris which could be due to heat neutralization of the active constituent during sterilization. further research needs to be conducted using the botanical with different sterilization techniques. this in-vitro study identified garlic as a critical antifungal alternative to conventional fungicides. field experiments need to be done to prove its effectiveness. keywords: fungal-growth inhibition, invitro analysis, phytochemicals, phytopathology, plant extracts, plant protection, white mold. citation | gyanwali, p., khanal, r., pokharel, n. p., tharu, b., koirala, r., paudel, s., & paudel, r. (2023). invitro analysis of antifungal effects of botanicals on sclerotinia sclerotiorum causing white mold disease. agriculture and food sciences research, 10(2), 8– 13. 10.20448/aesr.v10i2.5186. history: received: 1 september 2023 revised: 27 october 2023 accepted: 13 november 2023 published: 30 november 2023 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: the ethical committee of the institute of agriculture and animal science, paklihawa, tribhuvan university, nepal has granted approval for this study on 24 october 2022. transparency: the authors state that the manuscript is honest, truthful, and transparent, that no key aspects of the investigation have been omitted, and that any differences from the study as planned have been clarified. this study followed all writing ethics. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ......................................................................................................................................................................................... 9 2. materials and methodology .............................................................................................................................................................. 9 3. result .................................................................................................................................................................................................. 11 4. discussion .......................................................................................................................................................................................... 11 5. conclusion ......................................................................................................................................................................................... 12 references .............................................................................................................................................................................................. 12 mailto:prashantgyawali7@gmail.com mailto:renukhanal57@gmail.com mailto:netrapokharel73@gmail.com mailto:tharubhuwan44@gmail.com mailto:rajankoirala567@gmail.com mailto:paudels761@gmail.com mailto:rajan@iaas.edu.np https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v10i2.5186 https://orcid.org/0000-0001-6428-491x https://orcid.org/0009-0001-6929-4284 https://orcid.org/0009-0001-9237-3992 https://orcid.org/0009-0006-0783-4245 https://orcid.org/0009-0004-0663-2380 https://orcid.org/0009-0006-9293-2762 https://orcid.org/0000-0001-5108-6113 agriculture and food sciences research, 2023, 10(2): 8-13 9 © 2023 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the research provides information that differs in botanicals used and their preparation from other similar research and is a unique study against white mold. the paper adds the knowledge of how different botanicals fare against white mold and guides further research regarding its organic control. 1. introduction sclerotinia sclerotiorum (lib.) de bary (helotiales: sclerotiniaceae) is known by multiple names but its most common name recognized worldwide is ‘white mold’ as it is characterized by cotton-like fluffy white mycelial growth seen on infected plants’ stems. the pathogen exhibits its symptoms by creating water-soaked lesions on leaves that move downwards to the stem where it develops into necrotic tissues before developing its characteristic cottony mycelium [1]. sclerotinia is a polyphagous, necrotrophic, homothallic pathogen that is rampant worldwide, especially in temperate regions [2]. it is a devastating plant pathogen that has infected over 75 families, 278 genera, 408 species, and 42 subspecies of plants. a more pronounced effect of sclerotinia is seen in dicot plants like peas, soybean, sunflower, rapeseed, and occasionally in monocot crops and grasses [3]. s. sclerotiorum can cause varying degrees of yield losses to different crops, ranging from 20 to 50% loss in canola production [4] and 10 to 50% loss in lettuce production [5]. plant pests have been interfering with and preventing crops from reaching their full potential yield since the beginning of agriculture. their damage to plant parts, systems, physiological processes, and ultimately yield has been one of the contributing factors to food scarcity in the world [6, 7]. almost 40% loss of crop production and 20% loss in post-harvest is incurred every year due to insect pests, weeds, and diseases in the world [8]. to combat the losses, many chemical pesticides have been developed over the years which on one hand have helped reduce the pests’ effect while on the other hand have developed a host of problems like resistant pests, persistent pollutants, biological life hazards, and loss of biodiversity due to death of non-target organisms [9, 10]. so, there have been many attempts at developing alternatives to chemical pesticides like biopesticides, botanical extracts, and microbial pesticides. these alternatives have the benefit of being biodegradable, extremely efficient and accurate, and less harmful to the health of humans and the environment [11]. many studies have been done on controlling sclerotinia using fungicides [12, 13] from which numerous negative effects arise due to their prolonged use [14]. therefore many studies are being conducted to measure the efficacy of different biopesticides in the control of fungi [15]. so, farmers are in dire need of appropriate and sustainable approaches to the management of such pests and pathogens [16]. the purpose of this research is to evaluate the efficacy of some locally available plants of nepal with fungicidal effects against white mold in-vitro, which could be later incorporated into the farmers’ fields with further processing and development. 2. materials and methodology 2.1. plant extract preparation the plant extracts were produced through a simple crude aqueous extraction similar to the process used by timila and manandhar [17]. first, the fresh botanicals were washed to clean any dirt or external chemicals present in them. then after shade drying, 100 g each of fresh plant parts were taken and ground into paste separately using a mixer. each resultant paste was mixed with 100 ml of distilled water. extra care was taken to create a constant weight-to-volume ratio of botanicals to water so that their concentration would be the same throughout each extract. the mixture was then filtered through a muslin cloth and we obtained the botanical extracts in the form of filtrate as shown in figure 1. the aqueous extracts kept in separate flasks were autoclaved at 121 °c @ 15 psi for 20 minutes to eliminate any microorganisms present within. figure 1. aqueous botanical extracts of (from left) a. vulgaris, a. sativum, a. indica, z. officinale and l. camara. 2.2. media preparation the media of choice for the analysis was pda (potato dextrose agar) prepared by mixing ready-made pda powder with distilled water. the analysis was carried out using the “poisoned food technique” as described by balouiri, et al. [18] where two concentrations (10% and 20%) of each plant extract were infused with pda, creating a total of 10 unique growth media. to create the two concentrations of 10% and 20%, 10 ml and 20 ml botanical extracts were mixed with 90 ml and 80 ml molten pda respectively. unaltered pda media was kept separately as control and four replications for each unique media and control were made by pouring molten pda 20 ml each in petri dishes. agriculture and food sciences research, 2023, 10(2): 8-13 10 © 2023 by the authors; licensee asian online journal publishing group 2.3. fungal culture preparation sclerotinia for the analysis was isolated from the garden pea (pisum sativum) pod shown in figure 2. the pda media was inoculated with threads of mycelia initially and it was allowed to grow at 24 °c for a week in an incubator. the inoculating processes were repeated two more times from the obtained growth to ensure the exclusion of any other contaminants and obtain a pure culture. the final pure culture of 7 days was then used to inoculate all the different media and control with the help of a cork borer sterilized in an open flame. mycelium discs of 5 mm diameter were punched out from the pure culture and placed on the media at the center of each petri dish. the discs were placed with the mycelial side facing down to assist the pathogen in establishing on the new media. the whole inoculation process was done under aseptic conditions inside the laminar air-flow cabinet, and the inoculated dishes were incubated at 24 °c in dark. figure 2. infection of s. sclerotiorum on garden pea. 2.4. experimental design and antifungal activity assay the experiments were conducted in completely randomized design (crd) with 5 treatments of 2 concentrations viz: 10% and 20%. four replications each of treatments and control were kept resulting in a total of 44 petri plates. the fungicidal effects of botanicals were recorded by daily (24 hours) observation and measurement of the radial growth of mycelium, till the control plates were completely covered by the mycelial mat. for the antifungal activity assay, the percentage of mycelium growth inhibition was calculated by using the formula mentioned by al-samarrai, et al. [19]; bekker, et al. [20] and javed, et al. [21]. percentage inhibition = (𝐶−𝑇) 𝐶 × 100% where, c = colony diameter (cm) of the control. t = colony diameter (cm) of the test plate. the measurement of mycelial diameter was taken in cm and an average of 4 replications was taken for each treatment. the inhibition percentage was calculated for both concentrations of all the botanical extracts for 3 days until the control plate was fully occupied by the mycelial mat and the results are presented in table 1. 2.5. statistical analysis the data were recorded in ms excel 2016, along with calculation of mean and standard error of mean. the analysis of variation (anova) was done through r-stat version 4.2.2. fisher-lsd test was used to compare means at 0.05 significance level. table 1. growth inhibition of s. sclerotiorum by the use of botanicals. treatments 10% concentration 20% concentration mean growth inhibition % av. diameter (cm)* inhibition %* av. diameter(cm)* inhibition %* a. indica 6.48 28 5.13 43 35.5 a. vulgaris 9 0 8.88 1.33 0.67 a. sativum 0 100 0 100 100 z. officinale 6.98 22.44 6.03 33 27.72 l. camara 7.53 16.33 8.23 8.55 12.44 control 9 --------9 --------------- lsd (0.05) 9.80 3.34 sem 1.45 0.494975 cv % 19.47 5.95 grand mean 33.38 37.22 note: * significant difference at 0.05 probability level; lsd: least significant difference; sem: standard error of mean; cv: coefficient of variation. agriculture and food sciences research, 2023, 10(2): 8-13 11 © 2023 by the authors; licensee asian online journal publishing group 3. result after 72 hours of inoculation, the control petri plate was fully covered by the mycelia while both concentrations of a. sativum did not have any growth, keeping their inhibition at 100%. a. indica and z. officinale inhibited the growth by 28% & 22.44% in 10% concentration and 43% & 33% in 20% concentration respectively. l. camara also showed its antifungal property by inhibiting the growth by 16.33% & 8.55% in 10% & 20 % concentrations respectively. but a. vulgaris only suppressed the growth by 1.33% in the 20% concentration while no effect was seen in the 10% concentration. there was little difference between the mean inhibition effect of the two concentrations. 20% concentration was only slightly better with 37.22% mean inhibition compared to 33.38% of 10% concentration. overall, allium sativum performed the best among all the botanicals, completely inhibiting the growth of sclerotinia sclerotiorum, with a. indica, z. officinale, and l. camara showing mild to low levels of inhibition against the pathogen with 35.5%, 27.72%, and 12.44% mean inhibition across the two concentrations respectively. the inhibition levels are graphically shown in figure 3 and the petri plates after 72 hours of inoculation can be seen in figure 4. figure 3. percentage of inhibition of s. sclerotiorum by the use of botanicals (72 hours). figure 4. mycelial growth after 72 hours in (a): a. indica, (b): a. vulgaris, (c): a. sativum, (d): z. officinale, i: l. camara, (f): control, where (i): 10% concentration and (ii): 20% concentration. 4. discussion the world’s increasing population demands greater food production every year. this arises a challenge to protect the crops and reduce food loss, especially from pests and pathogens. globally 30% of crop products are lost due to fungal plant pathogens, and the toxins produced by the fungal growth and changes in color, odor, and taste, degrades food on a large scale [22]. fungicide use can decrease such losses of food, but in the wake of organic food agriculture and food sciences research, 2023, 10(2): 8-13 12 © 2023 by the authors; licensee asian online journal publishing group in recent years, their usage has been limited [23]. therefore, many attempts have been made in finding organic solutions to the fungal problem, one of which is the use of plant botanicals. the botanicals selected for the analysis have potential antifungal properties which have been shown by various studies throughout the years; zingiber officinale [24] azadirachta indica [25, 26] artemisia vulgaris [27] allium sativum [28] lantana camara [29]. these plants have been used for our plant pathological needs and purposes through multiple generations spanning thousands of years [30]. in that course of time, their foundation, and the key constituents responsible for their beneficial uses have been documented. the compounds responsible for the antifungal nature of the botanicals are presented in table 2. the concentration, amount, and quality of these phytochemicals depend vastly on the genotype, environmental condition, soil condition, plant parts harvested, harvesting season, and the stage at which they are harvested [31]. the plant parts which have been used for this analysis have all been collected in the locality of rupandehi, nepal, and so these parameters are likely to change along with the efficacy against sclerotinia in comparison to native plants of any other region. table 2. chemical constituents of the botanicals used for the analysis. plant family key constituent references artemisia vulgaris (mugwort leaf) asteraceae α-thujone, ascaridole β-thujone 1,8-cineole satyal, et al. [32]; blagojević, et al. [33] and cetin, et al. [34] azadirachta indica (neem leaves) meliaceae quercetin azadirachtin sithisarn and gritsanapan [35] and mahmoud, et al. [36] zingiber officinale (ginger rhizome) zingiberaceae gingerols terpenes park, et al. [37] and yeh, et al. [38] allium sativum (garlic cloves) amaryllidaceae allicin ajoenes leontiev, et al. [39] and el-saber, et al. [40] lantana camara (leaves) verbenaceae germacrene-d, e-caryophyllene bicyclogermacrene, isocaryophyllene passos, et al. [41] and sousa, et al. [42] the most effective among all the botanicals used was found to be allium sativum, followed by azadirachta indica, zingiber officinale, and lantana camara. similar results have been obtained in many experiments done worldwide [15, 17, 43]. a. vulgaris did not show any significant effect and failed to inhibit fungal growth. the antifungal property of artemisia species, just like the other botanicals, is governed by its secondary metabolites. one of the secondary metabolites present in artemisia is a compound ascaridole [32, 44]. it is a thermolabile compound [45] so it decomposes when subject to extreme heat. this could be the reason for the negative inhibition shown by a. vulgaris as the compound could have been destroyed when it was being autoclaved for sterilization. 5. conclusion the world is battling with food insecurity and the use of chemicals. while the short-term immediate benefits are favorable, continued chemical usage can be devastating for our food supply in the future and the use of biorational compounds is imperative for sustainable agriculture. so, it is important in identifying the best organic alternative to some of the major diseases of crops. these identified botanicals can be then used as a source to isolate the specific chemical constituent and develop a much more effective remedy for agricultural diseases. such analysis can help identify hidden control measures of our locality and further aid in developing new medicines for diseases of plants. the obtained result of the antifungal nature of the botanicals used here, needs to be further confirmed in the field condition. many other local botanicals can be examined for their efficacy against sclerotinia sclerotiorum and efforts can be made to isolate the chemical compounds. further experimentations with different extraction and application methods need to be conducted before ruling out any potential plant. references [1] m. d. bolton, b. p. thomma, and b. d. nelson, "sclerotinia sclerotiorum (lib.) de bary: biology and molecular traits of a cosmopolitan pathogen," molecular plant pathology, vol. 7, no. 1, pp. 1-16, 2006. https://doi.org/10.1111/j.13643703.2005.00316.x [2] g. s. saharan and n. mehta, sclerotinia diseases of crop plants: biology, ecology and disease management. the netherlands: springer science & business media, 2008. 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licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 1-6, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.1.6 © 2020 by the authors; licensee asian online journal publishing group efficiency of agriculture and water sector and the reality of food security in arab countries (2010-2017) omer allagabo omer mustafa expert of economics and banking, general union of arabs experts (morocco), assistant professor of economics, banking and finance, deputy secretary of academic affairs-sudan academy for banking and financial sciences (sabfs-sudan), khartoum, sudan. abstract the main aim of the paper is to presents the reality of food security in arab countries (that means literally the arab peninsula and the north african arab states); in the light of the efficiency of agriculture and water sector during the period (2010-2017). the descriptive analytical approach was used. secondary data were collected and used to describe challenges that faced arab food security during that period. the results found that agricultural production is insufficient to meets the demand for food in arab countries because it highly depends on rain-fed irrigation which causes low productivity; in connection with poor strategic planning. further, it is found that, poor exploitation and miss-management of water resources, conversion of farmland to urban uses, application of old technology, population growth, rising demand for food and degradation of natural resources have contributed to widening of food gap. the study recommends that, if arab countries want to achieve the food security and self-sufficiency they have to consider the following: use of modern technology to improve the productivity, cooperation in agricultural research, good utilization of water resources and effective agricultural and financing policies. keywords: efficiency, agriculture, water sector, reality, food security, arab countries. citation | omer allagabo omer mustafa (2020). efficiency of agriculture and water sector and the reality of food security in arab countries (2010-2017). agriculture and food sciences research, 7(1): 1-6. history: received: 4 november 2019 revised: 6 december 2019 accepted: 9 january 2020 published: 18 february 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ......................................................................................................................................................................................... 2 2. environmental and climatic characteristics of the arab region ............................................................................................ 3 3. food gap and self-sufficiency in arab countries ....................................................................................................................... 4 4. conclusions .......................................................................................................................................................................................... 5 5. recommendations ............................................................................................................................................................................... 5 references ................................................................................................................................................................................................. 5 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.1.6&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.1.6&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1327 https://orcid.org/0000-0002-1697-8877 agriculture and food sciences research, 2020, 7(1): 1-6 2 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by presenting the reality of food security in arab countries (that means literally the arab peninsula and the north african arab states); in the light of the efficiency of agriculture and water sector during the period (2010-2017). 1. introduction arab world still suffers from a chronic food gap. agriculture sector is heavily dependent on rainwater where irrigated area is approximately 21.5% of the cultivable area .also, demand for food has long exceeded domestic agricultural production [1]. the per capita of water availability in arab region is annually estimated at 790m3, which is less than the water poverty line defined globally by 1000m3.moreover, estimates of 60% of irrigation water is wasted and about 1.3 billion tons of materials used for human consumption are lost. 1.1. the problem statement in recent years, importations of food commodities by arab countries issue of food security have been increased. this situation raises questions about the reality of arab food security and the role of agricultural and water sector in reducing of the arab food gap? 1.2. the research significance the study focuses on the agriculture and water sector as important factor in achieving food self-sufficiency for arab countries. it also concentrates on the reality of food security and challenges that are facing the bridqe of food gap. 1.3. the research objectives • to recognize the reality of food security in arab countries. • to explain the relation between food security and efficiency of agriculture and water sector. • to highlight challenges that face arab food security. • to track and analyze the food gap in arab countries. 1.4. the research questions • what is the reality of food security in arab countries? • what are the challenges facing arab food security? • are agriculture and water sector effective in reducing the food gap in arab countries? 1.5. the research methodology to answer the questions and achieve the objectives, the study draws on the descriptive-analytical approach in explaining the reality of food security in arab countries and its relation to efficiency of agriculture and water sector. data were collected from secondary sources. 1.6. literature reviews fao [2] defined the concept of food security as a situation that exists when all people, at all times, have physical, social and economic access to sufficient, safe and nutritious food that meets their dietary needs and food preferences for an active and healthy life. mohammad [3] stated that there is a need to differentiate between the relative food security and absolute food security when defining food security. relative food security is the ability of a state to provide the regular needs of its people from food commodities fully or partially. talukder [4] used time series data to examine the relationship between food security and self-sufficiency status of bangladesh 1999-2000. the analysis showed that in relation to standard nutritional norm of food intake, bangladesh virtually remained a surplus producer of food grains from the year 1999-2000. awatif, et al. [5] assessed the challenges confronting the expansion of agricultural production in sudan. the results stressed the critical role of agricultural bank of sudan in enhancing of agricultural production. najib [6] discussed food security, efficiency and shifting of dietary habits in arab countries. the results pointed out that arab countries, in their quest to enhance food self-sufficiency, face serious challenges emanating from a backdrop of constraining factors, including aridity, limited cultivable land, scarce water resources and serious implications of climate change . al-fawwaz and ahmed [7] investigated the reality of food security in the arab world. the study concludes that the reality of food security is unstable and fluctuated; there is no self-sufficiency to cover the needs of the arab states. in addition, food security is connected to the power of water security, as most of water resources come from non-arab states. water shortage may lead to the food gap for arab countries. lee, et al. [8] suggested that the water-energy nexus might be a serious issue in future sustainable planning and policies, given the impacts of climate change in the world in general and in arab world particularly. hameed, et al. [9] reviewed the 21st century challenges that face food, energy and water (few) security in the middle east. the study found that, most of the studied countries are facing few resource insecurities. also the study suggested that climatic and socioeconomic factors have contributed to the subsequent stress on few resources, particularly the water sector. despite, a number of research bodies have discussed the dilemma of food security in the world, this paper seeks to highlight efficiency of both agriculture and water sector to identify the reality of food security in arab countries during the period (2010-2017). agriculture and food sciences research, 2020, 7(1): 1-6 3 © 2020 by the authors; licensee asian online journal publishing group 2. environmental and climatic characteristics of the arab region arab region is characterized by environmental diversity caused by several factors, of which are: geographical location, wide area and the diversity of its topography. these factors led to the diversity of the environment in terms of temperature and the amounts of rain. arab region is divided into three main climatic regions, the largest of which is the arid desert region, which covers about 75% of the total area. it is characterized by a hot continental climate in summer, cold in winter with scarcity of rains (less than 100mm per year). followed by the semi-wet / semi-arid tropical region, which covers the southern parts of the arab world with rainfall in average btween100mm-600mm annually. the mediterranean region includes the coastal strip of arab countries bordering the mediterranean sea characterized by mild climate and annual rainfall of 1000mm. it also includes land south of the coastline, with area of one million km2 and annual rainfall ranged between 600mm to 1000mm [10]. 2.1. agricultural land in arab countries total arable area in arab world is about 197 million hectares, representing 14.7% of the total area of 1344 million hectares. this area is well below the global average of 37%. the cultivated area is estimated at 75 million hectares in 2016-2017. it represents about 38.1% of the arable area and 5.6% of total area of arab region [11]. table-1. agricultural and wastelands in arab countries (2010-2017). (million hectares) year cultivated area rainfed agricultural lands irrigated agricultural lands uncultivated lands seasonal rainfed sustainable rainfed rainfed as % of cultivated seaso nal irriga ted sustain able irrigate d irrigated as % of cultivated wastela nd uncultivate d as % of cultivated 2010 68.48 35.47 5.93 60.5 10.44 3.05 19.7 13.572 19.8 2011 68.75 34.20 6.01 58.5 10.61 3.09 19.9 14.804 21.6 2012 68.97 32.90 6.09 56.5 10.79 3.13 20.2 16.037 23.3 2013 69.69 36.70 5.70 60.8 11.03 3.70 21.1 12.553 18.0 2014 69.92 33.58 5.74 56.2 10.15 3.77 19.9 16.664 23.8 2015 77.21 39.06 5.79 58.1 9.920 3.86 17.8 18.520 23.9 2016 75.08 31.25 5.84 49.4 9.770 3.84 18.1 24.380 32.5 2017 76.14 35.15 5.81 53.8 9.840 3.85 17.9 21.450 28.2 source: amf [12]. table 1 shows that: • arab countries depend on rain-fed agriculture, where the percentage of rain-fed land to total arable area ranged 60.5% in 2010 to 49.4% in 2016. • during the period 2010-2017, the percentage of irrigated land did not exceed 22% of the total agricultural land, this, might led to expansion of food gap. • the percentage of uncultivated land ranged between 18% and 24%, indicating inefficiency uses of cultivated. 2.2. performance of agricultural sector in arab countries (2010-2017) usually the performance of the agricultural sector is measured by its contribution to gdp and by the average per capita income of agricultural output. table-2. agricultural product in arab countries (2010-2017). million dollars growth rate of agricultural product growth rate of arab gdp per capita income of agriculture product($) agriculture product as % of arab gdp agriculture production arab gdp year 9.4 17.0 395 6.2 124419 2004730 2010 6.7 17.9 386 5.6 132802 2365373 2011 -2.0 5.08 372 4.9 130144 2485542 2012 8.5 11.7 394 6.4 141280 2775394 2013 3.7 1.15 399 5.3 146553 2807440 2014 -3.2 -12.4 392 5.9 141876 2457553 2015 0.35 -3.10 383 6.1 142373 2381302 2016 2.83.78 366 5.6 138320 2471390 2017 source: aoad [13]. figure-1. growth rate of arab gdp (2010-2017). source: based on table 2. agriculture and food sciences research, 2020, 7(1): 1-6 4 © 2020 by the authors; licensee asian online journal publishing group table 2 and figure 1 shows that: • agricultural sector has low contribution to arab gdp and it ranged between 4% and 7%. • during 2010-2017, arab gdp achieved highest growth rate of 17.9% in 2011, while it achieved highest negative growth rate of 12.4% in 2015. • during 2010 – 2017, per capita of agricultural gdp not exceed $ 400. • during 2010-2017, arab agricultural product achieved highest growth rate of 9.4% in 2010, while it achieved highest negative growth rate of 3.2 % in 2015. 2.3. water resources in arab countries most of arab region is located in arid and semi-arid climatic zones and it considered one of the poorest in the world in terms of water resources. annual rate of water per capita is about 700m3 which is less than the water poverty line which is 1000 m3. this rate expected to decrease to 500 m3 in 2025 in light of population growth.water resources are distributed between renewable surface water and groundwater reserves. all these resources are estimated at 350 billion m3 per year in addition to non-conventional resources, including desalination and purification water, which is estimated at 10 billion m3per year [14]. 2.3.1. surface water resources surface water resources in arab countries are estimated at 296 billion m3per year of total water resources, of which about 50% is used. agricultural sector, houses and industrial sector utilize about 88%, 7% and 5% respectively. surface irrigation contributes to wasting quantities of water, increasing soil salinity, depleting nutrients and reducing land productivity [15]. 2.3.2. groundwater resources due to increases of demand for groundwater in arab countries, groundwater reserves estimated at 7734 billionm3. also,based on the potential increase of population, volume of water resources allocated to agricultural is expected to rise to 378 billion m3in 2025.this indicates that water crisis in arab countries is putting its weight on the issue of achieving arab food security, which requires development and rationalization of the means of using water resources to intensify agricultural production. as well as work to developing and training of human resource and raising awareness of efficiency use of water among farmers [16]. 3. food gap and self-sufficiency in arab countries food deficit in arab countries is related to the difference between domestic production and import (net imports) of different food commodities. the continued disparity between growth rate of agricultural production and the demand for food commodities led to a food gap in 2016 of $ 32981 million. average food gap during the period (2010-2016) arrived$ 33959 million and the gap in flour group constitutes 71.2% of the total value of arab food gap in 2016. table-3. food gap and self-sufficiency of major commodity groups (2010-2016). million dollars 2016 2015 2014 2013 2012 2011 2010 statement food gap selfsuff% food gap selfsuff% food gap selfsuff% food gap selfsuff% food gap selfsuff% food gap selfsuff % food gap selfsuff% 32981 34932 38086 35600 36700 36000 35209 total 18271 37.8 20783 45.1 25604 45.2 22296 51.2 24106 54.7 23003 52.1 17479 44.6 flour -58 92.4 -78 103.2 -234 107.9 -100 105.1 -108 109.1 -105 106.4 -22 101.2 potato 2960 47.8 3227 33.8 3419 37.6 2430 32.7 2823 34.7 2623 33.7 2989 33.4 sugar (refined) 860 50.9 911 56.2 575 63.2 730 65.5 815 72.5 750 69.5 507 55.5 legumes 4379 32.2 3699 37.4 4471 29.1 4587 35.4 5780 38.4 5140 36.8 3987 36.8 oils and greases -1509 101.4 -2891 107.0 -3663 111.1 -2643 108.9 -2819 111.3 -2791 109.1 -2007 102.7 vegetables -1085 101.1 -1120 104.7 -2398 109.8 -950 107.3 -973 109.9 -911 107.9 -1136 97.5 fruits 6957 74.8 7997 73.3 6865 78.4 7370 95.7 8760 97.1 8459 96.0 6018 95.5 meat 3453 78.1 3125 74.9 4252 74.2 1129 83.0 1340 86.0 1270 81.0 2088 77.7 dairy products 417 89.1 180 95.4 114 96.6 96 97.1 98 98.5 97.0 97.3 50.0 95.6 eggs -1666 116.5 -912 101.5 -918 99.2 -558 101.5 -685 108.5 -613 105.3 -499 100.7 fish source: aoad [13] ,(-) means surplus, food gap for the years 2011 and 2012 is estimated. agriculture and food sciences research, 2020, 7(1): 1-6 5 © 2020 by the authors; licensee asian online journal publishing group figure-2. food gap in arab countries (2010-2016). source: based on table 3. table 3 and figure 2 shows that: • food gap in the consumer goods group reached highest value in 2014 of $ 38086 million and it declined to $ 32981 million in 2016.this reflects some efforts to improve the performance of agricultural sector. • food gap continued in commodity groups include flour, sugar, legumes, oils and fats, meat, dairy products and eggs. • food gap in flour group constituted the highest during the period 2010-2017, so arab countries failed to achieve self-sufficiency in this group. • arab countries in 2016 achieved self-sufficiency of 116%, 101.4%, 101.1% and 92.4% with surplus of $ 1666 million, $1509 million, $1085million and $58million in production of fish, vegetable, fruits and potatoes respectively. 4. conclusions • despite the contribution of agricultural sector in provision of food in arab countries, but it continued to record a weak performance indicator, as evidenced by low self-sufficiency rates and the persistence of food gap in basic commodity groups. • low performance of agricultural sector can be attributed to high dependence on rain irrigation, which covers about 55% of the total arable areas. • inefficient use of water resources contributed to wasting amounts of water that could have been directed to agricultural sector to help in reducing the food gap. • there is a growing in the food gap, which is worth paying attention, if arab countries want to achieve food security and self-sufficiency. 5. recommendations in order to integrate agriculture and water sector to achieve arab food security and reduce food gap, it is necessary to consider the following: • work to use modern technology because traditional agriculture leads to poor productivity. • agricultural policies should be more effective in developing agricultural sector. • cooperation in the establishment and development of a scientific centers specialized in the field of agricultural research at the level of arab countries. • it is important to pay attention to effective management of water resources to be supportive and integrated with agricultural policies. • financing policies should be formulated to accommodate the development of agricultural and water sector in arab countries. references [1] fao, "arab horizon 2030: prospects for enhancing food security in the arab region, technical summary. food and agriculture organization of the united nations (fao). retrieved from https://www.unescwa.org/publications/arab-horizon-2030-prospectsenhancing-food-security-arab-region. [accessed 25th nov, 2019]," 2017. [2] fao, the state of food insecurity in the world 2001. rome: food and agriculture organization of the united nations (fao), 2002. [3] w. a. mohammad, "concepts related to food security. special files. retrieved from: https://www.aljazeera.net/specialfiles/pages/3422ae4c-5f01-4a63-a108-a2349e1dd104. [accessed 25th nov, 2019]," 2003. [4] r. talukder, "food security, self-sufficiency and nutrition gap in bangladesh," bangladesh development studies, vol. 31, pp. 35-62, 2005. [5] m. awatif, i. saefalnasr, and a. m. omer, "policies to enhance agricultural production in sudan. retrieved from http://www.reading.ac.uk/web/files/apd/apdsrs_sabf.pdf. [accessed 28th nov, 2019]," 2011. [6] s. najib, "food security in arab countries: efficiency, productivity, and shifting dietary habits, international centre for advanced mediterranean agronomic studies, france," pp. 14. retrieved from: https://www.iamm.ciheam.org/publications/183/wl32.pdf [accessed 19th nov, 2019], 2015. [7] a. al-fawwaz and a. ahmed, "the reality of food security in the arab world," international journal of asian social science, vol. 6, pp. 251-261, 2016.available at: https://doi.org/10.18488/journal.1/2016.6.4/1.4.251.261. http://www.unescwa.org/publications/arab-horizon-2030-prospects-enhancing-food-security-arab-region http://www.unescwa.org/publications/arab-horizon-2030-prospects-enhancing-food-security-arab-region http://www.aljazeera.net/specialfiles/pages/3422ae4c-5f01-4a63-a108-a2349e1dd104 http://www.reading.ac.uk/web/files/apd/apdsrs_sabf.pdf http://www.iamm.ciheam.org/publications/183/wl32.pdf agriculture and food sciences research, 2020, 7(1): 1-6 6 © 2020 by the authors; licensee asian online journal publishing group [8] m. lee, a. a. keller, p. c. chiang, w. den, h. wang, c. h. hou, and j. yan, "water-energy nexus for urban water systems: a comparative review on energy intensity and environmental impacts in relation to global water risks," applied energy, vol. 205, pp. 589-601, 2017.available at: https://doi.org/10.1016/j.apenergy.2017.08.002. [9] m. hameed, h. moradkhani, a. ahmadalipour, h. moftakhari, p. abbaszadeh, and a. alipour, "a review of the 21st century challenges in the food-energy-water security in the middle east," water, vol. 11, pp. 1-20, 2019.available at: https://doi.org/10.3390/w11040682. [10] amf, "the joint arab economic report. arab monetary fund(amf),united arab emirate," pp. 5. retrieved from: https://www.amf.org.ae/en/content/joint-arab-economic-report-2018-brief-english-version. [accessed 14th nov, 2019], 2018. [11] ibid, "the joint arab economic report," p. 49, 2018. [12] amf, "the joint arab economic report for the years 2010-2017.arab monetary fund (amf),united arab emirate. retrieved from: https://www.amf.org.ae/ar/jointrep," 2010-2017. [13] aoad, "report on the state of agriculture in arab countries, arab organization for agricultural development (aoad). retrieved from http://www.aoad.org/eng/enews24nov2019-11.htm," 2018. [14] amf, "the joint arab economic report for the year. arab monetary fund (amf), united arab emirate," p. 54. retrieved from: https://www.amf.org.ae/ar/jointrep, 2016. [15] ibid, "the joint arab economic report," p. 51, 2016. [16] ibid, "the joint arab economic report," p. 55, 2016. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.amf.org.ae/en/content/joint-arab-economic-report-2018-brief-english-version http://www.amf.org.ae/ar/jointrep, http://www.aoad.org/eng/enews24nov2019-11.htm, http://www.amf.org.ae/ar/jointrep 38 © 2020 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 7, no. 1, 38-45, 2020 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2020.71.38.45 © 2020 by the authors; licensee asian online journal publishing group developing calibration model for prediction of malt barley genotypes quality traits using fourier transform near infrared spectroscopy yadesa abeshu1 ashagrie zewdu2 ( corresponding author) 1food science and nutrition research program, holeta agricultural research center, ethiopian institute of agricultural research; addis ababa, ethiopia. 2center for food science and nutrition, college of natural and computational sciences, addis ababa university; addis ababa, ethiopia. abstract ethiopia is the second largest barley producer in africa. but the quality traits are always influenced by the cultivar itself and growing environment. thus, the study was targeted on developing calibration model for predicting malt barley quality traits of genotypes grown at different locations using near infra-red spectroscopy for selection purposes in barley breeding program. for this purpose, 60 barley samples were collected from holeta, debre-birhan and bekoji. samples were chemically analyzed in duplicate for 5 barley traits. the calibration model was developed based on 120 samples spectral data and 60 chemistry data results using the calibration software of the ft-near infrared spectroscopy. the barley protein calibration model having (r²c= 0.97; rpd=5.7 and r2 c=0.94; rpd=4.16) respectively, can be regarded as broadly applicable; extract and friability (r²c= 0.96; rpd=4.54 and r2 c=0.95; rpd=4.36) respectively were accepted as useable with good prediction capability; whereas β-glucan calibration model (r²c= 0.90; rpd=3.18) allowed only for screening purpose in some applications. barley grain dry matter with model parameters result (r2 c=0.86; rpd=2.69)shown usable with caution only for rough screening purposes. hence near infrared spectroscopy is fast and cost-efficient, the breeding program can increase the intensity of variety selection using calibration models reflected good predicting performances except models for dry matter and β-glucan. keywords: barley, genotypes, malt, trait, near infrared spectroscopy, calibration, validation. citation | yadesa abeshu; ashagrie zewdu (2020). developing calibration model for prediction of malt barley genotypes quality traits using fourier transform near infrared spectroscopy. agriculture and food sciences research, 7(1): 38-45. history: received: 7 january 2020 revised: 10 february 2020 accepted: 17 march 2020 published: 13 april 2020 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: authors would like to express their gratitude to rebeka g/tsadik (phd) giz contact person and coordinator in ethiopia and berhanelakew (phd) senior barley breeder at holeta research center for their kind supports to the successful attainment for the study. their special thanks also go to wilde peer (phd) kws breeding company senior breeding advisor and christian utschig (phd) for coordinating laboratory facility at vlb, berlin and assisted them in providing nirs statistical materials. besides they would like to thank addis ababa university, center for food science and nutrition as well as holetta agricultural research center barley breeding team members and food science and nutrition research program for their support in sample collection and laboratory analysis. funding: the authors would like to thank the giz-ssap project and kws breeding company of germany which supported them in financial and technical assistance. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 39 2. materials and methods ................................................................................................................................................................... 39 3. results and discussion ................................................................................................................................................................... 41 4. conclusion ......................................................................................................................................................................................... 44 5. recommendation.............................................................................................................................................................................. 45 references .............................................................................................................................................................................................. 45 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.38.45&domain=pdf&date_stamp=2017-01-14 http://crossmark.crossref.org/dialog/?doi=10.20448/journal.512.2020.71.38.45&domain=pdf&date_stamp=2017-01-14 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ https://www.asianonlinejournals.com/index.php/aesr/article/view/1503 http://orcid.org/0000-0002-4648-2370 agriculture and food sciences research, 2020, 7(1): 38-45 39 © 2020 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study was targeted on developing calibration model for predicting malt barley quality traits of genotypes grown at different locations using near infra-red spectroscopy for selection purposes in barley breeding program. 1. introduction barley (hordeum vulgare l.) is the fourth most important cereal crop worldwide after wheat, corn and rice [1]. but it is the fifth most important grain crop in ethiopia and has diverse ecologies being grown from1800 to 3400m altitude in different seasons and production systems which makes ethiopia being the second largest producer in africa, next to morocco. also recognized as one of the country’s most ancient food crop, which is believed to have first domesticated about 10,000 years ago from its wild relatives in the fertile crescent of the near east and center of diversity in ethiopia [2]. it is most widely used for the production of malt worldwide [3]. barley malt also mainly used as a source of fermentable sugars for alcoholic fermentation like production of beer [4]. barley isa complex mixture of many organic components that include protein, starch, oil, polysaccharides and sugars. the amount of each of the constituents will vary due to both the genetic background and the environmental conditions during grain development. the malting process of barley in particular, modifies the grain components during the controlled steeping, germination and drying processes. malt is an essential ingredient in beer production where soluble components of the malt are extracted into a liquid broth called wort [5]. the ability to predict grain quality for different purposes in early generations would be of great benefit to breeders and industries, allowing for selection of suitable lines to deliver product of the highest quality. at later stages in the barley breeding programs, micro-malting can be carried which requires large barley sample sizes, destructive and requires experienced personnel. for this purpose, near infrared spectroscopy is an ideal technique as it is fast, reliable and nondestructive which does not require large sample sizes [6]. near infrared spectroscopy is a type of vibration spectroscopy that employs photon energy in the range of 7.96 x 10-20 to 2.65 x 10-19 j. the range is higher than necessary to promote molecules to their lowest excited vibrational states and lower than typical values necessary for electron excitation in molecules [7]. hence, food nir spectra comprised of broad bands arising from overlapping absorptions corresponding mainly to overtones and combinations of vibration modes involving c–h, o–h, n–h and sh chemical bonds [8]. thus, sophisticated mathematical techniques, termed chemometrics, are heavily employed to allow calibration for reliable extraction of relevant information encoded in the nir spectral data [9]. the aim of this study is to determine constituents of particular interest for the brewing of barley grain criterion important traits by using nirs. this was performed using near infra-red reflectance (nir) technology which is effective in simultaneously predicting of multiple constituents in agricultural products. 2. materials and methods 2.1. materials the samples used in this study were from barley breeding program trials of ethiopian institute of agricultural research centers having the mandate of barley growing and breeding potential around central highland areas of ethiopia. thus, barley samples were collected representing a range of breeding generations fully-fledged at different environments throughout barley growing highland areas, specifically from holeta, debre birhan and bekoji locations. about 60 samples from 2018 year of barley growing season were collected from the pre-specified growing areas for the study. particularly, from the plot of the breeding program-controlled trials depending on genotypes, location, type (food or malt barley) and quality variability. 2.2. sample preparation barley samples which were collected from different locations and different genotypes of breeding trials were selected purposively from different plots depending on agronomical data and source of genotypes from where they originated as well as the history of their quality data. for barley reference and spectral data analysis 150g per sample was taken after manually cleaned and graded. then the samples were packed into plastic bag. prior to reference samples chemical analysis, malt(after malted)were ground using a laboratory sample mill3100 (perten instruments, hagersten, sweden) to pass through 0.5mm sieve for calibration reference data. but for malt friability determination the malt sample was not grounded, because the friabilimeter machine itself grounds the sample for the ratio of friability measurement.before malt quality traits analysis, the malt barley samples were malted according to phoenix automated micro malting system (phoenix bios stems, adelaide, australia) designed to process 300g of 24 barley samples per batch [10]. after kilning the rootlets were removed from the malted samples by using mechanical malt cleaner that had been reconfigured to simultaneously process eight 250 g samples (fraser fabrications pty ltd, malaga western australia). 2.3. wet chemistry analysis malt quality traits of malt barley were chemically analyzed for reference data at holeta food science and nutrition laboratory, eiar (ethiopian institute of agricultural research) in collaboration with vlb institute in berlin (germany) for the traits mentioned below. but simple quality traits were analyzed at holetta, eiar cereal quality laboratory. blank and known concentration sample were analyzed with the samples to control the analysis biases. the samples also duplicated to reduce the reproduced errors in each sample chemical analysis. 2.3.1. maltextract content malt extract content was determined according to a small-scale version of the european brewery convention [11] methods manual, section 4.9.1. fine grind malt was extracted using a hot water mashing bath (siemens mashing machine, germany).for extraction, 50 g of finely ground malt was mixed with 200 ml of distilled water and mash at 45°c with continuous stirring. after 30 minutes of mashing, the temperature was increased by 1°c/min until 70°c. as temperature reaches 70°c, there was added of 100 ml distilled water. after 1 hour, the mash was cooled to 30°c and adjusted to a volume of 515 ml or a weight of 450g. the extract was filtered using agriculture and food sciences research, 2020, 7(1): 38-45 40 © 2020 by the authors; licensee asian online journal publishing group whatman 12cm filter paper into 500ml cylinder and specific gravity was measured at 20oc using a dma5000 density meter (anton paar gmbh, graz, austria). therefore the following formula was used to put the end result.e = p(800+m)/(100-p); where, e= extract content, p= wort density (oplato), m= malt moisture content. 2.3.2. malt total protein the malt protein content was determined using kjeldhal method (digester sbs 2000, distillation unit 5000dl, foodalytgimbh, germany) according to aoac [12]. for analysis one gram ground sample of malt barley was measured and transferred into completely dry kjeldhal flask. ten gram of kjeldhal tablet was added to the sample inside the flask. twenty milliliter of 98% concentrated sulphuric acid was mixed with the sample. the sample digestion was started by connecting the kjeldhal flasks with the digestion rock. the digestion was completed when the brown color of the sample completely disappeared. after the digested sample was cooled, 100 ml of distilled water and 80 ml of sodium hydroxide (32%) were added and distilled into 25 ml of excess boric acid containing 0.5 ml of screened indicator. the distillate was titrated with 0.1n hydrochloric acid to the methyl red end point. the protein was calculated by using this formula: cp%= [(t-b)*14*6.25)]/[w(100-mc)]; where cp=crude protein, t= volume of hcl used in titration, b= blank used as control and w= weight of sample taken for analysis. 2.3.3. malt friability malt grain samples were analyzed using a friability measuring machine (pfeufferfriabilimeter gmbh, germany) which is used as pressure roller to grind the sample against a rotating screen. low, medium and high friability malts were tested according to ebc method 4.15 [11]. 50g malt sample was run in the friabilimeter for 8 min and the non-friable fraction was weighed to get the final result. 2.3.4. malt-glucan the malt -glucan content was determined using the megazyme kit method (megazyme, bray, ireland)according to european brewery convention [11] method 4.16.1.for the analysis 100 mg sample was suspended and hydrated in a buffer solution of ph 6.5 and incubated with purified lichenase enzyme and filtered. an aliquot of the filtrate was then hydrolyzed to complete with purified β-glucosidase. the d-glucose produced was assayed using a glucoseoxidase/peroxidase reagent. the final prepared aliquot was measured by spectrophotometer at absorbance 510 nm against reagent blank within one hour. finally the beta-glucan was calculated using the formula;β-glucan (%w/w) = ∆a * (f/w) * 27;where, ∆a=absorbance afterβ-glucosidase treatment (reaction) minus reaction blank absorbance, f=factor for the conversion of absorbance values to μg of glucose, w= the calculated dry weigth of the sample analyzed in mg. 2.3.5. dry matter content barley grain dry matter content was determined according to aoac [12] international standard method from grain flour prepared using the above sample preparation method. 5g of barley flour was taken using a sensitive analytical balance and oven dried at 105oc temperature for 3 hours. after the dried sample was cooled in a desiccator, the final measurement was taken using the same analytical balance to get the result using the following known formula for moisture content. mc% = (wi-wf)/wi*100; where wi is initial weight, wf is final weight; dm=(100-mc)%. 2.3.6. samples spectral data acquisition before scanning the samples, the spectroscopic performance of the device was checked using gold standard (1.038) and light trap standard (0.00065) provided by bruker company of germany. then 60 barley samples were scanned twice for spectral data using near infrared spectrometer (tango2017, bruker optics gmbh, germany) which uses scan and rotating mode within 16 seconds, sample presentation with rotating accessory having 600 ml sample cuvette. it used32 amounts of scans between 867 –2535 nm wave length ranges with 16 cm-1 wave number band resolution. the nir sensor used in this experiment is capable of recording spectra by diffusive reflectance measurements at the mentioned spectral wave length range. spectra data figure-1 shown below was acquired from untreated barley grains before malting using a bruker tango software. it is the raw spectral data which needs to be treated statistically to correct spectral biases. the device recorded spectra in diffusive reflectance by using an integrating sphere. measurements were done in duplicate for each sample, leading to a total number of 120 spectral data. figure-1. barley samples component matrix raw spectral data collected by nir. agriculture and food sciences research, 2020, 7(1): 38-45 41 © 2020 by the authors; licensee asian online journal publishing group 2.4. spectral data pre-processing there were spectral biases and overlaps of wave length bands due to matrix effect, different particle size and spectroscopic condition. for this matter spectral data obtained according to figure-1 above must be pre-processed using the mathematical manipulation of the nir machine as follows. the spectral data shown at figure-2 below was pre-processed using chemometrics system of ft-ir’s opus software version 7.5.1. therefore, first derivative plus standard normal variate (1st d/ve+snv) transformations, with 17 smoothing points were found to be the best method for the above spectral data treatment/pre-processing. the above figure 1 raw spectral data was changed to the spectral data shown below in figure 2. the shape and the resolution of the spectral data was completely changed as reflected below after preprocessed. so, the main objective of spectral data pre-processing was to remove biases, unwanted outliers and increase the resolution of overlapped wave length bands that could result errors in development of calibration model. figure-2. samples pre-processed spectral data. 2.5. developing calibration and validation the laboratory reference data was correlated with spectral data using the software opus version7.5.1 of tango (bruker, optics gmbh, germany). in total, 120 spectral data were used for calibration and validation. from 120 spectral data, with approximately two thirds of the samples were used in the calibration set and one third in the validation set. validation method used to check the performance of the calibration model was test set validation method. the calibration set was checked to cover the whole variation in terms of spectral data, traits and locations. the following numbers of factors were used for the traits of interest: protein content 8, extract content 8, friability 9, ß-glucan 10 and dry mater 10.the calibration model result was evaluated by using opus software statistical systems (chemometrics) [13]. 2.6. calibration model statistical evaluation statistical tools were used to evaluate the efficiency of nir calibrations and various terms are important in understanding the performance of a calibration model as described by williams [14]. this includes statistics of calibration as well as statistics of validation. for developing calibration model a partial least square (pls) regression was used which was performed using opus software. the coefficient of determination (r2), root mean square error estimation (rmsee), root mean square error of prediction (rmsep),ratio of performance to deviation (rpd), standard deviation (sd) and standard error of prediction) good parameters to evaluate the model performance depending on the reference data and spectra. the calibration was automatically tested by test set validation method. the final calibration was determined from an optimization routine of opus after the removal of the outliers. during the optimization step, various frequency regions and also spectral pre-treatments was systemically tested to determine the optimal calibration that was directly stored in the device memory for future quantitative analysis as used by krapf, et al. [13]. 3. results and discussion calibration model development highly depends on selecting a set of good calibration samples data obtained by wet chemistry analysis method. the set of calibration samples used in this study also contained the range of chemical and physical variations for which calibration model was applied. accordingly, the calibration experiment was established using a mathematical relationship between the nir spectrum and physical/chemical properties determined by reference methods. similarly, comparing the chemical reference data variability and model accuracy is vital point. 3.1. calibration model nirs calibration models were developed for five barley traits determining quality for malt and food use. the model performance was assessed by the following parameters: coefficient of determination of the calibration (r²c) and validation set (r²v), standard errors of prediction (sep) as well as root mean square error of prediction (rmsep). the rpdv value indicates the suitability of the calibration for the prediction. with a higher rpdv value the calibration will more likely be able to predict the right sample values. according to the generally used classification in agriculture based on rpdv-values and r2 c-values, the calibration for protein can be regarded as successful calibration [15]. agriculture and food sciences research, 2020, 7(1): 38-45 42 © 2020 by the authors; licensee asian online journal publishing group the calibration and validation statistical description was very important to see the feasibility of each traits data to build calibration model. table 1 implies the general features of descriptive statistics of traits to detect the right data variability for developing strong calibration model. good data variability was observed in all traits except for dm as shown in table 1 below. therefore, expected calibration model is good for all traits if could not be strong model for dm only i.e. not implying the good fitness between reference data and spectral data, it is all about to capture the range of traits value obtained from different barley samples. table-1. descriptive statistics for calibration and validation. calibration data validation data traits units mean range sd nc mean range sd nv extract % 78.1 73-83.5 2.93 120 78.1 69.9-84 3.12 60 protein % 10.4 7-14.1 1.88 120 10.5 6.8-13.3 2.08 60 friability % 61.0 29-97 18.96 120 59 20-98 22.13 60 β-glucan mg/l 685.0 158-1000 279.5 120 699 50-1000 358.6 60 dm % 91.34 90.6-92.3 0.4 120 91.28 90.8-91.9 0.26 60 note: dm=dry matter; sd=standard deviation; nc=no of spectral data used for calibration; nv=no of spectral data used for validation. the following barley traits analyzed chemically using wet chemistry methods were assigned for calibration model to fit the true value with predicted spectral value using pls statistical regression. spectral data was pretreated using appropriate statistical tool to remove outliers and to check the status of the chemistry and spectral data for developing calibration model as planned. 3.2. extract content model extract content prediction delivered good calibrations as in table 2 for whole grain samples (r2 c= 0.956; rpdc= 4.54 with variable samples). the model major parameters r2 and rpd reflect acceptable for most applications and screening purposes. results from this study was comparable and much better than that of previous researchers who developed promising calibrations for predicting the extract of whole grain (r2 c =0.78 0.85) [16] and ground barley (r2 c = 0.77 -0.96) [17]. because this property is highly influenced by the malting process since enzyme activity during malting influences the malt extract which limits the accuracy of any nir prediction based on not malted barley. this is why different calibration model performance is reported by different scientists, even if the accuracy of reference sample analysis data is very important. additionally, the following figure 3 expressed the calibration curve obtained from chemistry/reference data and nir predicted spectrum data of extract content. figure-3. barley extract content calibration curve. 3.3. protein model similarly calibration model results were obtained for whole barley grain samples protein content with (r2 c= 0.97; rpd= 5.7 of variable samples)as in table 2. the model is more acceptable than the models for other traits which could be usable in most applications, quality assurances and quality control. this prediction of nitrogen content from whole grain barley is well established in the literature and the results from this study compared well with those of previous reports for whole grain barley with r2 c = 0.94 [18] and r2 c = 0.95 [19]. because nir prediction is more effective in predicting biochemical properties than physical properties according to my observation from different literatures report. in addition to model parameter information, the following figure 4 expressed the calibration curve obtained from reference data and nir predicted spectrum data of protein content. figure-4. barley protein content calibration curve. agriculture and food sciences research, 2020, 7(1): 38-45 43 © 2020 by the authors; licensee asian online journal publishing group 3.3.1. friability model friability prediction performance model shown in table 2 of this study was (r2 c=0.95; rpd=4.36), which was excellent like the protein model which could be used in most applications including in quality assurances, but in our case it was trustful model to identify excellent barley friability for malt factories at breeding final stage. almost similar performed model was also reported in the literature with parameters value (r2 c=0.91; rpd=3.33). therefore, the friability calibration model was not as such challenging like in dry matter and beta-glucan as observed from similar model reported by different authors [15]. additionally, figure 5 expressed the calibration curve obtained from chemistry/reference data and nir predicted spectrum data of friability content. this curve was built after exaggerated outliers were removed from the spectrum data manually as well as by the opus system. figure-5. barley friability calibration curve. 3.3.2. β-glucan model beta-glucan prediction model was successful in this study for whole grain barley samples having (r2 c=0.90; rpd=3.18) as in result table 2. according to williams [14] this type of model performance is usable with caution specially for screening purposes such like in early stage breeding lines. but a similar study have been reported in the literature with much lower (r2 c=0.25)as compared to this study by black and panozzo [16]. on the other hand [20] reported (r2 c=0.61) and he referred that the poor distribution of reference values in the sample range may be the reason for the poor results in beta-glucan model of barley grain that were obtained in his study. but in this study the good modifying malting process, excellent sample variability selection and moderate accurate chemistry analysis made the model better as compared to the reported literature. additionally, the figure 6 expressed the calibration curve obtained from chemistry/reference data and nir predicted spectrum data of β-glucan content. figure-6. barley beta-glucan calibration curve. 3.3.3. dry matter model moisture predictions model from the same whole barley grain samples were good as in table 2, but was only acceptable for some screening purposes, because (r2 c = 0.86; rpdc=2.69) was less as compared to the recommended range by williams [14]. results of moisture content from this study are not comparable well to literature reports, this is due to the smaller sample moisture content ranges (90.6-92.3 % dm) utilized compared to those used by previous researchers. similar problem was also observed in literatures with the small sample range (78.4 83.4% dm) with in reference values obtained as studied by roux [20]. the range of samples needs to be expanded in order to obtain acceptable calibrations model. additionally, the figure 7 expressed the calibration curve obtained from chemistry/reference data and nir predicted spectrum data of dry matter content. agriculture and food sciences research, 2020, 7(1): 38-45 44 © 2020 by the authors; licensee asian online journal publishing group figure-7. moisture content calibration curve. generally, calibration model for the three traits extract content, protein content and friability were promising model for value predicting in some applications while beta-glucan and dm was delivered not trusted model as observed in the result table 1 below. but calibration model alone does not enough to judge traits model to be applicable for targeted purpose. it is very important to see the validation parameters result side by side with the calibration parameters result as clearly shown in table 2 for all selected traits in this study. table-2. calibration and validation statistical parameters results. calibration statistical parameters validation statistical parameters traits r2 sec rmsee rpd slope r2 v sep rmsep rpd bias extract 95.62 0.65 0.67 4.54 0.95 0.81 1.37 1.34 2.28 0.062 protein 96.93 0.33 0.34 5.7 0.97 0.93 0.55 0.54 3.78 -0.03 friability 94.75 4.35 4.5 4.36 0.95 0.59 14.10 13.8 1.57 0.32 β-glucan 90.11 87.90 91 3.18 0.90 0.49 254.33 249 1.41 -22.6 dm 86.23 0.15 0.153 2.69 0.86 0.51 0.18 0.29 1.43 -0.001 note: sd=standard deviation of reference data; r2 =coefficient of determination; sec=standard error of calibration; standard error of prediction; rmsee=root mean square error of estimation; rpd=ratio of performance to deviation. 3.4. traits validation model model evaluation was performed by test set validation; because test set validation is a more independent validation method than cross-sectional validation as williams [14]. in this method, the set of calibration samples is divided into a calibration set and validation set. according to this study from 60 samples,20 samples spectral data and chemistry data was set for validation. using this process, the models were validated and checked for their prediction capacities. the principle was predicting quantitative value using the model from the validation set spectral data and comparing the predicted value against the chemistry data set for validation. validation parameters for each barley traits were acceptable as in table 2 for extract content, protein content and friability having(r2 v= 0.81& rpd=2.28), (r2 v=0.93 &rpdv=3.78) and (r2 v=0.59& rpdv=1.57) values respectively. these models could be applied for quality control and screening purposes in breeding programs or any other applications. but as previously observed in calibration model above validation result for beta-glucan (r2 v= 0.49 with rpdv = 1.41) and dm (r2 v= 0.51 with rpdv = 1.43) was not reflected good model even though could be used in some rough screening purposes. this kind of model was not widely usable for prediction as black and panozzo [16] reported unless for simple rough screening purpose. other parameters like sep\rmsep and bias with lower value reflected that the model was well performed. these validation parameters showed very convincing value that the user could believe the model to use for prediction of protein, extract and friability in general. similarly, a study by [18] delivered excellent prediction models for these traits that could be used in most applications. this should be due to the nature of the sample and less modifying malting process. 4. conclusion barley is the fourth most important cereal crop worldwide. also, it is a crop of ancient origin in ethiopia and is considered as a center of diversity for barley, because of the presence of great diversity in ecology. most of the ethiopian barley is consumed as food at home. at the same time using barley for malt production establishes new value-added chains from which ethiopian small holders can benefit significantly. the main objective of this study was enabling the breeding programs to select the appropriate genotypes easily by developing calibration model using near infrared spectroscopy. depending on the objective samples wet chemistry for barley (extract, protein, friability, beta-glucan and moisture content) were analyzed using the international official methods described in materials and method part. likewise, calibration models were developed for all above mentioned traits using nir opus software statistical parameters. these wet chemistry and calibration model results demonstrated a realistic approach to predict quality traits of ethiopian barley protein content, extract content, friability, ß-glucan and dry matter by applying nirs. therefore, this prediction model will enable the selection of appropriate food and malt barley genotypes for future end goals. since nirs is fast and cost-efficient the barley breeding programs can increase the intensity of selecting superior candidate lines for variety verification. also with regard to broadening calibration performance ethiopian barley landraces were included in the samples to capture the whole ethiopian barley variability. agriculture and food sciences research, 2020, 7(1): 38-45 45 © 2020 by the authors; licensee asian online journal publishing group 5. recommendation malt quality traits that could have direct or indirect correlation with the identified traits should be included for model development to reduce matrix effect. for some of the traits like dry matter and beta-glucan further study is very important to distinguish between sample variability range and wet chemistry analysis inaccuracy which could contribute to results less trusted model observed in most studies. also, to develop most excellent prediction method for barley, excellent sample variability, accurate wet chemistry data and having good instrument condition is very important. references [1] s. marwat, m. hashimi, and k. khan, "barley (hordeum vulgare l.) a prophetic food mentioned in ahadith and its ethnobotanical importance," american-eurasian journal of agricultural & environmental sciences, vol. 12, pp. 835-41, 2012. [2] fao, "food balance sheets. faostat. rome. retrieved from: http://faostat3.fao.org/download/fb/fbs/e," 2014. [3] s. e. ullrich, genetics and breeding of barley quality attributes. in: barley science: recentadvances from molecular biology to agronomy of yield and quality (edited by g.a. slafer, j.l. molina-cano, r. savin, j.l. araus& i. romagosa). binghamton, new york: the haworth press, 2002. [4] s. kreisz, malting. in: handbook of brewing: processes, technology, markets (edited by h.m. eßlinger). weinheim: wiley-vch verlaggmbh& co. kgaa, 2009. [5] d. e. briggs, hough. js, stevens, r. & young, tw,‘. malting and brewing science'. london: chapman and hall, 1981. [6] t. woodcock, g. downey, and c. p. o'donnell, "better quality food and beverages: the role of near infrared spectroscopy," journal of near infrared spectroscopy, vol. 16, pp. 1-29, 2008. available at: https://doi.org/10.1255/jnirs.758. [7] c. pasquini, "near infrared spectroscopy: fundamentals, practical aspects and analytical applications," journal of the brazilian chemical society, vol. 14, pp. 198-219, 2003. available at: https://doi.org/10.1590/s0103-50532003000200006. [8] h. huang, h. yu, h. xu, and y. ying, "near infrared spectroscopy for on/in-line monitoring of quality in foods and beverages: a review," journal of food engineering, vol. 87, pp. 303-313, 2008. available at: https://doi.org/10.1016/j.jfoodeng.2007.12.022. [9] w. wang and j. paliwal, "near-infrared spectroscopy and imaging in food quality and safety," sensing and instrumentation for food quality and safety, vol. 1, pp. 193-207, 2007. available at: https://doi.org/10.1007/s11694-007-9022-0. [10] m. nilsen and j. panozzo, in: leaping ahead with near infrared spectroscopy. ed by batten, g.d., flinn, p.c., welsh, l.a. and blakeney, a.b. australia: nir spectroscopy group, 1995. [11] european brewery convention, analytica ebc, 5th ed. nurnberg, germany: verlag hans carl, 1998. [12] aoac, official methods of analysis, 17th ed. usa: association of official analytical, 2000. [13] l. c. krapf, a. gronauer, u. schmidhalter, and h. heuwinkel, "near infrared spectroscopy calibrations for the estimation of process parameters of anaerobic digestion of energy crops and livestock residues," journal of near infrared spectroscopy, vol. 19, pp. 479-493, 2011. available at: https://doi.org/10.1255/jnirs.960. [14] p. c. williams, implementation of near-infrared technology. in: near-infrared technology inthe agricultural and food industries (edited by p.c. williams & k.h. norris). usa: st. paul, minnesota, 2001. [15] g. fox, m. g. borgognone, p. flinn, and d. poulsen, "genetic and environmental analysis of nir feed quality predictions on genotypes of barley (hordeum vulgare l.)," field crops research, vol. 120, pp. 380-386, 2011. available at: https://doi.org/10.1016/j.fcr.2010.11.007. [16] c. black and j. f. panozzo, "utilising near infrared spectroscopy for predicting malting quality in whole grain barley and whole grain malt," in proceedings of the 10th australian barleytechnical symposium, canberra, act, australia, 2001. [17] s. tragoonrung, p. hayes, and s. broich, "near-infrared reflectance estimates of grain protein and malt extract in hill and row plot evaluations of spring malting barley," canadian journal of plant science, vol. 70, pp. 71-78, 1990. available at: https://doi.org/10.4141/cjps90-008. [18] m. edney, j. morgan, p. williams, and l. campbell, "analysis of feed barley by near infrared reflectance technology," journal of near infrared spectroscopy, vol. 2, pp. 33-41, 1994. available at: https://doi.org/10.1255/jnirs.29. [19] m. sohn, d. s. himmelsbach, f. e. barton, c. a. griffey, w. brooks, and k. b. hicks, "near-infrared analysis of whole kernel barley: comparison of three spectrometers," applied spectroscopy, vol. 62, pp. 427-432, 2008. available at: https://doi.org/10.1366/000370208784046768. [20] e. roux, "near infrared (nir) spectroscopy for selection of malting barley in south african breeding programmes," doctoral dissertation, stellenbosch: university of stellenbosch, 2011. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://faostat3.fao.org/download/fb/fbs/e, 15 © 2021 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 8, no. 2, 15-19 2021 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/journal.512.2021.82.15.19 © 2021 by the authors; licensee asian online journal publishing group postharvest losses of tomato (solanum lycopersicum) in the open markets in ibadan metropolis ojimi, k. o1 oyediran, w. o2 salawu, m. b3 abiodun, y4 alaka f. a5 otufale, g. a6 ojo, o. m7 ( corresponding author) 1,3,4department of agricultural extension and management, federal college of animal health and production technology, moor plantation, ibadan, nigeria. 1email: ojimikafilatolaide@gmail.com 2department of agricultural extension and rural development, federal university of agriculture, abeokuta, nigeria. 5department of agricultural education, school of secondary education (voc. & tech. programmes), fces, oyo, oyo state, nigeria. 6department of agricultural education, tai solarin college of education, omu-ijebu, ogun state, nigeria. 7department of agricultural extension and rural development, ladoke akintola university of technology, ogbomosho, oyo state, nigeria. abstract postharvest losses of tomato pose serious threat on the economic viability of actors in the tomato production chain in nigeria. therefore, postharvest losses of tomato among marketers in ibadan metropolis of oyo state nigeria were investigated by the researchers. one hundred and twenty tomato marketers were chosen from the five food markets in ibadan metropolis through a simple random sampling technique. the study adopted linear regression and descriptive statistics to analyze data. the outcomes of the investigation show that females were 92.5% and 88.3% were married. capital is primarily raised through cooperative societies (57.5%). postharvest losses of tomato accounted for 18.9% of purchase which represents ₦124,800/day. the major causes of phls are poor packaging (100%) and poor storage (100%) by the tomato marketers. meanwhile, all the respondents (100%) adopted sorting to reduce phls of tomato. the regression analysis showed that education has a significant effect to reduce phls of tomato. the study concluded that huge postharvest and monetary losses of tomato are experienced by the tomato marketers. the study recommends provision of quality postharvest handling education to tomato marketers as a way of reducing phls of tomato in ibadan. keywords: tomato, postharvest losses, education, food, marketers, metropolis, packaging, sorting, storage, ibadan. citation | ojimi, k. o; oyediran, w. o; salawu, m. b; abiodun, y; alaka f. a; otufale, g. a; ojo, o. m (2021). postharvest losses of tomato (solanum lycopersicum) in the open markets in ibadan metropolis. agriculture and food sciences research, 8(2): 15-19. history: received: 12 august 2021 revised: 15 october 2021 accepted: 10 november 2021 published: 8 december 2021 licensed: this work is licensed under a creative commons attribution 3.0 license publisher: asian online journal publishing group acknowledgement: all authors contributed to the conception and design of the study. funding: this study received no specific financial support. competing interests: the authors declare that they have no conflict of interests. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study was reported; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study follows all ethical practices during writing. contents 1. background of the study ................................................................................................................................................................ 16 2. research methods ............................................................................................................................................................................ 16 3. conclusion ......................................................................................................................................................................................... 19 references .............................................................................................................................................................................................. 19 https://www.doi.org/10.20448/journal.512.2021.82.15.19 https://orcid.org/0000-0003-4490-5736 mailto:ojimikafilatolaide@gmail.com http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ agriculture and food sciences research, 2021, 8(2): 15-19 16 © 2021 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this paper contributes to the knowledge on postharvest losses because it looks at the dimension of tomato marketers in the city, unlike most previous studies that focus on rural areas. the study identified infrastructural deficit as a major setback to the proper handling of tomato hence, tomato phls are increasing while the income and profits of tomato marketers are seriously affected in the city. 1. background of the study agricultural losses are one of the greatest problems facing agricultural production in nigeria. according to oyediran and omoare [1] phls regularly happen along the food supply chain, from the ranch door through till it gets on the table of the final consumer. losses are experienced along the chain in the dealing with, capacity, transportation and preparing, in this way bringing about a decrease in the amount, quality and market worth of horticultural products [2]. the fundamental driver of phls incorporate mechanical harm, physiological decay and organic, that is, postharvest diseases and insect pests [3]. rodents and birds additionally cause postharvest misfortunes in tomatoes [4]. losses are because of unpleasant dealing with, helpless bundling and quality misfortunes brought about by temperature stress [5]. additionally, the degree of misfortunes is essentially impacted by pre-collect conditions and field activities, for example, cultivar and soil types, crop the executives rehearses, helpless climate conditions, creepy crawly bug control projects and reaping just as bundling and dealing with rehearses [6]. postharvest losses are predominant in fruit and vegetables among which is tomato crop. postharvest losses of this crop ranges between 20 40% [7]. tomato (solanum lycopersicum) is highly perishable crops with attendant high postharvest losses. the bulkiness of tomato adds to transportation inconveniences and storage difficulty. however, it is perhaps the most mainstream and generally developed natural product on the planet including africa [8]. it is local to south america [9]. the harvest was brought into west africa by portuguese brokers and liberated slaves from west indies [10]. tomato is developed and eaten from one side of the planet to the other. it is utilized differently, remembering crude for plates of mixed greens, and prepared into tomato soup. it is the second most significant vegetable around the world, as far as the measure of nutrients and minerals it adds to the eating routine [11]. tomato is plentiful in nutrients, minerals and lycopene [12]. it is an amazing cell reinforcement that assists with lessening the danger of prostate and bosom malignant growth [13]. in nigerian markets, tomato is sold fresh to the consumers. many times, marketers particularly in the northern nigeria preserved this crop by sun drying and sold in bags and baskets. in the southern nigeria, the crop is usually grinded, boiled and put inside refrigerators or in jars. however, the marketing of tomato is influenced by various issues notwithstanding certain components of cultivating [14]. these incorporate the irregularity of creation, which subjects a country's creation to changes dependent on deficient water system offices. losses likewise happen on the way because of significant distance to business sectors, poor and insufficient foundations, and the strategy for transportation of tomato [15]. it is against this foundation that this examination evaluated postharvest misfortunes of tomato among marketers in ibadan metropolis, oyo state. 1.1. objective of the examination the broad objective of this examination was to evaluate the postharvest losses of tomato among marketers in the investigation region. 1.2. explicit goals the particular goals of the investigation were to; i. describe the financial attributes of the respondents in the investigation region. ii. estimate the worth of misfortune experienced by the tomato advertisers in the investigation region. iii. identify the significant reasons for postharvest misfortune in tomato advertising in the investigation region. iv. determine the financial variables impacting postharvest misfortunes of tomato in the examination region. v. assess the procedures took on by tomato dealers towards lessening postharvest misfortunes in the investigation region. 2. research methods 2.1. study region the examination was done in ibadan, a city in south western piece of nigeria, capital of oyo state, situated around 110 km (around 70 mi) upper east of lagos. ibadan is a significant travel point between the coast and regions toward the north. the number of inhabitants in ibadan city region expanded at a development pace of 3.9% per annum from 1952 to 1963 when the populace rose to 1,258,625, then, at that point to 1,829,300 of every 1999 at a development pace of 1.65% from 1963 [16]. the populace development is said to have moved step by step to the lesser city with a development pace of 4.7% per annum somewhere in the range of 1991 and 2006. it exists in the scope 7oc 19' 08" and 7o 29' 25" of the equator and longitude 3o 47' 50" and 4o 0' 22". 2.2. sampling procedure and sample size purposive sampling technique was used for this study. ibadan metropolis was purposively selected because of predominance of tomato markets in the city. out of 9 major tomato markets in ibadan, five (5) were randomly selected which are, shasha, bodija, orita merin, oja oba and oje. twenty-four were arbitrarily chosen from each market, making the complete number of 120 respondents for this investigation. 2.3. measurement of variables and data analysis the estimates of tomato losses on daily basis was calculated by the differences between the total quantity (kg) of tomato bought and quantity of tomato sold by the respondents, and multiplying it by the price per kg which is ₦400.00. the information was broke down utilizing both elucidating insights and direct relapse. agriculture and food sciences research, 2021, 8(2): 15-19 17 © 2021 by the authors; licensee asian online journal publishing group 2.4. respondents’ qualities age distribution in table 1 shows that 45% of the tomato marketers were 31 to 40 years, followed by 25.83% that were in age category 41 to 50 years, and just 9.17% were between 51 – 60 years. the mean age was 38.44 years which implies the tomato sellers are in their productive age. majority of the respondents are females, with 92.5%, while the remaining (7.5%) were male tomato sellers. most (88.3%) of the respondents were married while others were single (7.5%), divorced (3.3%) and widowed (0.8%). the average household size of the respondents was 4 6 members for 58.33%, 1 – 3 members for 30.83% and more than 6 members for 10.83%. secondary occupation distribution reveals that 91.67% of the respondents did not engage in secondary occupation other than selling of tomato. table-1. distribution of respondents by socio-characteristics (n = 120). variables category frequency percentages mean age < 30 24 20.00 38.44 31-40 54 45.00 41-50 31 25.83 51-60 11 9.17 sex male 9 7.50 female 111 92.50 marital status single 9 7.50 married 106 88.30 divorce 4 3.30 widow 1 0.80 religion islam 70 58.30 christianity 49 40.80 traditional 1 0.80 household size 1-3 37 30.83 4.00 4-6 70 58.33 > 6 13 10.83 secondary occupation yes 10 8.3 no 110 91.67 source: field survey, 2018. 2.5. sources of credit and accessibility by the tomato marketers as shown in table 2, credit is marginally accessible by 45.8% of the respondents while 54.2% did not have access to credit. this reflects in the source of funds distribution which reveals that 57.5% of the respondents get credit from cooperative societies, and 30% used money accrued from their personal savings. access to credit by almost sixty percent of the respondents is due to the fact that majority (73.3%) of the respondents were members of cooperative society. average earning by the respondents was ₦55,575 in which 45% of the respondents earned between ₦30,000 to ₦40,000. fifty percent of the respondents had spent between 6 – 10 years in the marketing of tomato while the average years of experience was 6.4 years. table-2. distribution according to sources of credit and accessibility by the tomato marketers (n = 120). credit sources category frequency percent mean credit accessibility yes 55 45.80 no 65 54.20 source of fund cooperative society 69 57.50 bank 13 10.80 personal savings 36 30.00 membership of cooperative society yes 88 73.30 no 28 23.30 mode of earning income daily 108 90.00 weekly 7 5.80 monthly 3 2.50 monthly earning (₦) < 30,000 14 11.67 55,575 30,001 – 50,000 54 45.00 50,00170,000 27 22.50 70,001 – 90,000 13 10.83 90,001 > 12 10.00 marketing experience (yrs.) 6.39 < 5 55 45.83 6-10 60 50.00 6.4 11 -15 5 4.17 source: field survey, 2018. 2.6. estimate of the value of loss experienced by tomato marketers the results in table 3 revealed that the estimated value of tomato purchased was ₦657,200.00 while the quantity sold was ₦532,400.00. the difference which is ₦124,800.00 represented the tomato postharvest losses in the study area; these losses constitute 18.9% of total quantity tomato bought. agriculture and food sciences research, 2021, 8(2): 15-19 18 © 2021 by the authors; licensee asian online journal publishing group table-3. average estimates of values of tomato losses daily basis by the marketers. tomato quantity loss (kg) value (₦) % tomato bought 1643 657,200.00 tomato sold 1331 532,400.00 tomato loss 312 124,800.00 18.99 note: price per kg = ₦400.00. 2.7. significant reasons for postharvest misfortunes in tomato marketing the significant reasons for misfortunes in tomato promoting are introduced in table 4. the outcomes showed that poor packaging (100%) and poor storage (100%) of tomatoes are the two major causes of postharvest losses among tomato marketers. this implies that marketers need to be educated on how to best package and preserve tomato properly, as this may reduce tomato postharvest losses in the study area. it was also found that poor sanitation causes of postharvest losses by 99.2%. this is followed by the problem of over-ripeness (98.3%), poor handling (97.5%), and tomato bruises (97.5%). results in table 4 further revealed that 95% postharvest losses were caused by pathogen infestation while 91% of postharvest losses were due to bad roads during transportation of tomatoes from farm to market. however, 41.7% and 34.2% indicated that riot and limited alternative use of produce respectively were mild causes of postharvest losses. table-4. causes of postharvest losses in tomato marketing (n = 120). major causes frequency (%) rank yes no poor packaging 120 (100) 0 (%) 1 poor storage 120 (100) 0 (%) 1 poor sanitation 119 (99.2) 1 (0.8) 3 lack of processing unit 118 (98.3) 1 (0.8) 4 over ripeness 118 (98.3) 2 (1.7) 4 poor handling 177 (97.5) 3 (2.5) 6 bruises 177 (97.5) 3 (2.5) 6 pathogen infestation` 114 (95.0) 6 (5.0) 8 bad roads 110 (91.7) 10 (8.3) 9 poor pre-harvest 99 (82.5) 21 (17.5) 10 bad state arrival at market 90 (75.0) 30 (25.0) 11 malformed fruit 70 (58.3) 50 (41.7) 12 non removal of field heat 59 (49.2) 61 ( 50.8) 13 drought 55 (45.8) 64 (53.3) 14 riot 50 (41.7) 68 (56.7) 15 limited alternative use of produce 41 (34.2) 79 (65.8) 16 source: field survey, 2018. 2.8. techniques adopted by tomato marketers towards reducing postharvest losses the results indicated that proper sorting and grading (100%) is the most adopted technique by tomato marketers to reduce amount of tomato postharvest losses (table 5). this suggests that proper sorting and grading has been an effective technique adopted by the marketers to checkmate postharvest losses in the study area. also, 95%, 94.2% and 92.5% reported that proper packaging, proper handling, and transporting the tomatoes on a good road respectively reduce tomato postharvest losses. however, refrigerating (57.5%) and covering of tomato with paper (47.5%) were reported to be ineffective technique towards reducing tomato postharvest losses, and these are the least adopted techniques by tomato marketers. table-5. techniques adopted by tomato marketers to reducing losses. techniques frequency (%) rank yes no proper sorting & grading 120 (100) 0 (0.00) 1 proper packaging 114 (95.0) 6 (5.0) 2 proper handling 113 (94.2) 7 (5.8) 3 good transportation 111 (92.5) 9 (7.5) 4 purchasing unripe tomatoes 75 (62.5) 44 (36.7) 5 refrigerator storage 69 (57.5) 51 (42.5) 6 covering tomato with paper 57 (47.5) 63 (52.5) 7 source: field survey, 2018. 2.9. determination of the factors influencing tomato phls linear regression estimates of factors influencing phls of tomato are presented in table 6. the r is 79.8%, which shows that there is high degree of correlation between the variables. the results further showed that only two variables in the model has a significant effect on value loss of tomato, and these are educational level and secondary occupation of the respondents. though there is an inverse relationship between education level and value of tomato loss, that is, a unit increase in education level will reduce value of loss of tomato by 89.97%. this shows that education can promote best ways to package and preserve tomato and it is a very vital factor that can reduce the value of tomato postharvest losses in the study area. results in table 6 also revealed engaging in a secondary occupation is significantly positive to value of tomato loss at 1%. this indicates that there is a direct relationship between the two variables. this infers that, a unit increase in having secondary occupation will increase value of tomato losses by 55.86. this suggests that the more tomato marketers have a secondary occupation, the value losses of tomato experienced increase. other variables, however, are found not significant to the tomato postharvest losses. agriculture and food sciences research, 2021, 8(2): 15-19 19 © 2021 by the authors; licensee asian online journal publishing group table-6. factors affecting tomato misfortunes from the relapse gauges. socio-economic factors coefficient std. error t sig. constant 997.425 279.226 3.572 0.001 sex 59.977 102.274 0.586 0.559 age 4.971 4.119 1.207 0.230 educational status -89.974*** 35.27 -2.551 0.035 household size -47.465 39.619 -1.198 0.233 secondary occupation 55.86** 27.249 2.05 0.004 marketing experience -17.036 13.464 -1.265 0.208 r = 79.8%; note: *** 1 percent sig., ** 5 percent sig. 3. conclusion this examination discovered that respondents are in their dynamic age and literates. but, they experienced high postharvest losses with attendant huge monetary losses on daily basis which is attributed to poor packaging and storage. regression results showed that educational level of the respondents has inverse relationship but significant to the phls. the study therefore recommends that quality education on postharvest handling should be offered to the tomato marketers in order to reduce phls of tomato in the study area; electricity supply should be stable to encourage adoption of refrigeration in the tomato marketing; and government should construct good roads so as to ease the movement of the marketers and reduce tomato damages. references [1] w. o. oyediran and a. m. omoare, "potentials and challenges of agricultural education in reducing postharvest losses and food insecurity in ogun state, nigeria," american research journal of agricultural, pp. 1 – 10, 2016.available at: https://doi.org/10.21694/2378-9018.16003. [2] l. h. aramyan and j. b. gouh, "reducing postharvest food losses in developing economies by using a network of excellence as an intervention tool," presented at the 2014 ifama and cca food and agribusiness world forum people feed the world’ in cape town, south africa, june15–19, 2014. [3] i. b. adeoye, o. m. o. odeleye, s. o. babalola, and s. o. afolayan, "economic analysis of tomato losses in ibadan metropolis, oyo state, nigeria," african journal of basic & applied sciences, vol. 1, pp. 87–92, 2009. [4] helvetas, "consequences of post-harvest losses to vegetable farmers in tanzania: a case of singida region," helvetas swiss inter cooperation survey report 2014, 2014. [5] world food logistics organization, "identification of appropriate postharvest technologies for improving market access and incomes for small horticultural farmers in sub-saharan africa and south asia," wflo grant final report, university of california, uc, davis, usa. 32010. [6] y. kahsay, f. abay, and d. belew, "intra row spacing effect on shelf life of onion varieties (allium cepa l.) at aksum, northern ethiopia," world journal of agricultural research food safety, vol. 1, pp. 34–42, 2013. [7] a. kader, "effects of postharvest handling procedures on tomato quality," presented at the symposium on tomato production on arid land, 2003. [8] g. n. agrios, plant pathology. new york, usa: academic press, 2003. [9] o. osemwegie, a. oghenekaro, and l. owolo, "effects of pulverized ganoderma spp., on sclerotium rolfsii sacc and post-harvest tomato (lycopersicon esculentum mill.) fruits preservation," journal of applied sciences research, vol. 6, pp. 1794-1800, 2010. [10] e. okwori and h. c. dikko, "tomato processing and utilization, extension bulletin. 241," 2010. [11] e. c. bombelli and e. r. wright, "tomato fruit quality conservation during post-harvest by application of potassium bicarbonate and its effect on botrytis cinerea," agricultural science and research, vol. 33, pp. 167-172, 2006.available at: https://doi.org/10.7764/rcia.v33i3.346. [12] d. john, r. t. suthin, r. s. usha, and r. udhayakumar, "role of defense enzymes activity in tomato as induced by trichoderma virens against fusarum wilt caused by fusarium oxysporum f f. sp lycopersici," journal of biopesticide, vol. 3, pp. 158-162, 2010. [13] e. giovannucci, "tomatoes, tomato-based products lycopene, and cancer: review of the epidemiologic literature," journal of the national cancer institute, vol. 91, pp. 317-331, 2000.available at: https://doi.org/10.1093/jnci/91.4.317. [14] v. o. ebong, agribusiness management in developing countries: the nigeria perspective. uyo: dorand publishers, 2000. [15] d. mukaminega, "hybrid dryer (solar and biomass furnace) to address the problem of post-harvest losses of tomatoes in rwanda," research project submitted to larenstein university of applied sciences, wageningen, the netherlands, 2008. [16] national population census (npc), "national bureau of statistics official gazette. (fgp 71/52007/2 500 (ol.24) abuja. retrieved from: http//www.nigerianstat.gov.ng," 2007. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.nigerianstat.gov.ng,/ 222 © 2024 by the author; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 222-227, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6254 © 2024 by the author; licensee asian online journal publishing group pre-grafting treatments of plant fatty acid extract, seaweed (ascophylum nodosum l.) and micronized calcite increase the grafting success in grapevine propagation ali sabir selcuk university faculty of agriculture department of horticulture, konya, türkiye. email: asabir@selcuk.edu.tr abstract globally increasing interest in environmentally sound viticulture in precision agriculture promoted the viticulturists to find eco-friendly treatments in both vineyards and nurseries. the present study was conducted to evaluate the possible effects of plant oil extract (as plant activator), seaweed (ascophylum nodosum l.) extract, calcium oxide and iron chelate on callusing degree on graft union point and final take of ‘trakya ilkeren’ table grape cultivar grafted on 110 richter (berlandieri x rupestris) rootstock. bud break and shoot emergence commenced earlier in grafts subjected to pregrafting immersion into iron chelate and plant oil extract during graft union room duration than those of nontreated control grafts. the greatest degree in callusing of graft union point was obtained from plant oil extract (3.8), which was closely followed by seaweed (3.7), while, on the other hand, control and immersing the graft materials into iron chelate had the lowest effect with the same value (2.2). the highest percentage of graft final take was obtained from plant oil extract (75%), followed by seaweed treatment (55%), while control and iron chelate grafts were as low as 45 and 44%, respectively. overall findings indicated that immersing the scion canes and rootstock cuttings into plant seed oil before grafting could be recommended to increase the nursery grafting success. keywords: cambium callogenesis, final take, grafting compatibility, grapevine propagation, nursey practices, omega grafting, precision viticulture. citation | sabir, a. (2024). pre-grafting treatments of plant fatty acid extract, seaweed (ascophylum nodosum l.) and micronized calcite increase the grafting success in grapevine propagation. agriculture and food sciences research, 11(2), 222–227. 10.20448/aesr.v11i2.6254 history: received: 20 november 2024 revised: 19 december 2024 accepted: 24 december 2024 published: 26 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. contents 1. introduction .................................................................................................................................................................................... 223 2. materials and methods ................................................................................................................................................................. 223 3. results and discussion ................................................................................................................................................................. 224 4. conclusion ....................................................................................................................................................................................... 226 references ............................................................................................................................................................................................ 226 mailto:asabir@selcuk.edu.tr https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6254 https://orcid.org/0000-0003-1596-9327 agriculture and food sciences research, 2024, 11(2): 222-227 223 © 2024 by the author; licensee asian online journal publishing group contribution of this paper to the literature vticulture has been carried out in vineyards established using grape saplings grafted on different rootstocks to prevent the adverse effects of soil-borne pests such as phylloxera and nematodes. the rootstock currently used in viticulture have been bred from various north american native species. therefore, the rootstock genotypes show a great variability in grafting affinity with cultivars. some of them frequently failure to form adequate unity at graft point. for a sustainable viticulture, a good affinity between cultivar and rootstock has been desired to obtain healthy and abundant number of saplings. the present study revealed that plant extract effectively improved the graft success in grapevines. therefore, the results could be useful for increasing the grapevine propagation potential with an environmentally friendly practical approach. 1. introduction grape cultivars (vitis vinifera l.) are important plants highly consumed for fresh table purpose as well as for the production of other foods such as raisin (dried berries), juice and wines. various grape cultivars have long been cultivated throughout the subtropical and temperate regions around the world, especially in warm sunny climates with mild winters and dry summer periods during berry ripening. however, recently ever-increasing stress factors exacerbated by climate change, have been restricting the sustainability of grape production over the world [1]. for a precision viticulture under stress factors, grapevine cultivar and rootstock breeding studies have been promoted by different councils. general purposes in breeding of grapevine rootstocks are to provide resistance to soil born insect phylloxera (daktulosphaira vitifoliae fitch), improper soil conditions, diseases or other environmental problems [2]. north american vitis species can effectively tolerate the phylloxera pest and can provide varying degrees of prevention of such soil-borne problems [3]. therefore, the breeding and unitization of environment-adopted rootstock to cope with problematic areas should be based on many criteria such as soil features of vineyard, genetically rooting ability of the rootstock, genetic potential in vine vegetative development, usefulness in grafting and scion/rootstock graft compatibility [4-6]. grafting practice combines two separate plant parts, a scion material (the grape cultivar selected for crop production) and a rootstock (part of the grapevine serves as its root system). grafting the a grape cultivar onto disease-resistant rootstocks now extends to many horticultural plants [7]. however, mistakes in any process of grafting operation can inevitably cause failure in grafts success as well as serious subsequent problems difficult to compensate. many factors play pivotal roles on the success of grafting such as exogenous synthetic hormonal applications [8] grafting technique [9] cold treatment of the cuttings before grafting, time of grafting [10] cutting shape of grafting parts [11] and environmental conditions [12]. in order to increase the graft success, synthetic chemical-based hormones such as iba (indole butyric acid) have widely been used to provide greater callusing at graft point and faster vegetative growth [13]. however, excessive use of chemicals causes problems not only in terms of financial production cost but also in terms of the environment and public health. therefore, the development and application of ecofriendly sustainable approaches have been gained a particular attention recently. in this context, organic plant activators such as seaweed (ascophylum nodosum l. extract), fatty acid-based extracts and micronized calcium have been reported to induce vegetative development. therefore, this study was performed to investigate pre-grafting treatments of various organic treatments on grafting success in grapevine nursery propagation. 2. materials and methods the present experiment was established at the department of horticulture, faculty of agriculture, university of selcuk in konya türkiye, using a glasshouse equipped with central heating unit and roof-wall ventilating systems. in dormant season hardwood canes of grape rootstock, 110 richter with about five nodes were collected from the labelled vines as experimental rootstock material. ‘trakya ilkeren’, an early ripening hybrid hybrids of ‘alphonse lavallee’ x ‘perlette’, was used as the scion cultivar. preparation of rootstock cuttings of 110 r and ‘trakya ilkeren’ cultivar was performed from the grapevine germplasm repository of selcuk university. cuttings for grafting were taken from vigorous, healthy, well nourished, and mature canes generated from the previous summer season’s development. the rootstock cuttings were prepared from middle portion of cane to contain 4-5 nodes. the average length and diameter cuttings prepared for grafting was 18-24 and 0.8 to 1.2cm, respectively and the cut at the base (lower end) of the cutting was performed just below the basal node while the top was cut about 3 – 4 cm above a node. the one-year-old scion (‘trakya ilkeren’) canes were taken from healthy dormant vines from vineyard. the scion cuttings of matching thickness were selected for the grafting experiments. all the rootstock and scion cuttings were treated with fungicidal solution containing carbendazim 2 g per liter by dipping them for 10 minutes and then shade dried for about 20 minutes. the scion cuttings with singly winter bud (node) were taken for grafting. following treatments were performed just before grafting; immersing the scion and rootstock cuttings into 1) calcium oxide at 2% concentration for 2 h, 2) seaweed (ascophylum nodosum l.) extract solution at 2% concentration for 2 h, 3) plant oil extract (plant activator) at 2% concentration for 2 h, 4) iron chelate solution at 2% concentration for 2 h and 5) nontreated cuttings were used as control. after treatments, omega grafting techniques was used by using graft machine. the scion and the rootstock pieces for grafting were of similar diameter to guarantee a complete contact between the cambium of both graft pieces. grafting operation was performed in nursery facilities, following the commercial protocol established in the nursery and described by sabır and ağaoğlu [8]. briefly, one-bud (approx. 4-5 cm) hardwood cuttings of ‘trakya ilkeren’ were grafted onto approximately 25 cm de-budded canes of 110 r rootstock. the graft points were wrapped in parafilm wax applied as a dip covering the scion and graft union to prevent eternal disease inoculation and to fix graft parts tightly. melted wax was maintained at a constant temperature of 60 to 70 °c. waxed scion cuttings were just dipped into the cold tap water to cool. grafting combinations were replicated 3 times, comprising a total of thirty grafted plants. then, grafted plants were placed in a callogenesis chamber for 25 days. after callogenesis stage, the grafted plantlets were transplanted into the pots following a randomized complete block design after their uprooting for evaluation. in order to evaluate the effects of grafting methods on grafting success, the following main parameters were examined as illustrated by celik [14] and sabır and ağaoğlu [8]. agriculture and food sciences research, 2024, 11(2): 222-227 224 © 2024 by the author; licensee asian online journal publishing group • callus formation (0-4 scale) and callusing rates at graft union point after callogenesis stage: a scale ranging from 0 to 4 was used (0=no callus, 1=25%, 2=50%, 3=75% and 4=100% callus formation on graft union surface) to determine the callus formation level at graft union point. • rates of the grafted cuttings with different callusing levels were groped as partial callusing rate (the percentages of the grafts having the grade 1 and 2 at graft union point) and proper callusing rate (the percentages of the grafts having the grade 3 and 4 at graft union point). the percentages of the grafts having the grade 3 and 4 at graft union point was recorded as proper callusing rate (%). • final take rate (%) at nursery stage: percentages of the survived young grafted vines that have an adequate vegetative development until the end of the vegetation period. • shoot length (cm) after nursery development: the length of the scion shoot was measured by a meter with a sensitivity of 1 mm when the vegetative development of the plant was near to cease. • regression analysis (p<0.01) was carried out to assess the correlations between the investigated parameters. 2.1. statistical analysis analysis of data variance was performed with spss 13.0 software program. the least significant difference (lsd) was calculated at the level of p<0.05. 3. results and discussion mean value of callusing degree at graft point, rates of partial or proper callusing levels obtained after the callogenesis chamber before transplanting were presented in figure 1. the highest callusing degree was obtained from the grafts treated with plant activator (3.8), which was followed closely by seaweed (3.7) with the same significance level. micronized calcite provided significant improvement in callusing of graft point. on the other hand, the lowest callusing degrees were determined in control (2.2) and iron chelate (2.2) treatments. these findings are in accordance with the previous reports on different cultivar7rootstock combinations [8, 15]. anatomical union between the rootstock and the scion parts of graft is aided by a so-called tissue “callus”. wound-repair xylem is anatomically the first differentiated tissue for a proper unity of rootstock and cultivar tissues. aside from whatever role the auxin (iaa) plays in promoting cell division at graft union point (callus formation) [16] iaa also plays an important role in xylem formation across the callus bridge between stock and scion. during this union stage, callus is formed in varying degrees under the effects of various factors such as grafting type and callusing capacities of scion and rootstock genotypes. present findings implied that callusing capacity of the genotypes should be improved by exogenous supply of plant activators such as plant essential oil extract that contains bioactive compounds and phytohormones. the callus formation grade at graft union point is an essential factor determining the graft compatibility level between scion cultivar and rootstock [8, 14]. in addition to plant fatty acid extract, seaweed (ascophylum nodosum l.) remarkably improved the callusing of grafts during callogenesis chamber. figure 1. changes in callusing degree (1-4 scale) at graft union point grafts in response to pre-grafting cane immersion treatments. note: each column represents the mean of three replicates with ten grafts per replicate. error bar stands for the standard deviation of that mean. the proper callusing rate (%) of grafts was significantly improved by plant activator, seaweed and micronized calcite treatments with the percentages 83%, 80% and 73%, respectively (figure 2). iron chelated had no significant effect on proper callusing rate with the similar percent with control (43% and 42%, respectively). seaweed extract has been proven to induce plant development and this promoting effect has been attributed to its betaine content [17]. agriculture and food sciences research, 2024, 11(2): 222-227 225 © 2024 by the author; licensee asian online journal publishing group figure 2. changes in proper callusing rate (%) at graft union point grafts in response to pre-grafting cane immersion treatments. note: each column represents the mean of three replicates with ten grafts per replicate. error bar stands for the standard deviation of that mean. the final take, ultimate indication of graft success, significantly differed in response to pre-grafting treatments (figure 3). the highest final take percentage was obtained from the grafts subjected to pre-grafting soaking of plant oil extract activator (75%). this percentage is higher than those of singh and kaur [18] whose general rates was around 48% and lower than those of ghojage, et al. [19] recorded around 80-81%. such differences could be due to the genotypic variation as well as grafting condition. cambial continuity after callusing between graft partners has also an essential role in graft survival rate [20]. seaweed and micronized calcite treatments had also significant contributions to improvement of final take with the respective values of 55% and 51%. the lowest final take rates were determined in control (45%) and iron chelate (44%) treatments. from a practical perspective, plant activator yielded promising results as an environmentally safe and sustainable approach. figure 3. changes in final take percentage (%) of grafts in response to pre-grafting cane immersion treatments. note: each column represents the mean of three replicates with ten grafts per replicate. error bar stands for the standard deviation of that mean. effects of various pre-grafting treatments on scion shoot length have been illustrated in figure 4. shoot development pattern of the scions of grafts displayed a similar pattern to that of final take. the highest shoot length was obtained from plant activator which was closely followed by seaweed. on the other hand, iron chelate and micronized calcium immersions had no significant effects on shoot development with similar values. a strong correlation between shoot growths, stock to scion ratio, and callus development indicates that growth of grafted grapevine could be different in respect to grades of callus development [14] and rootstock genotype [21]. properly callusing at the graft union had direct influence on subsequent shoot development, an indicating of good graft compatibility [22]. adequate shoot growth and proper lignification lengthwise the rootstock and scion trunk would determine the cold hardiness level of the saplings during the first years in vineyards for especially continental climate conditions where winter chilling injury frequently occurs. agriculture and food sciences research, 2024, 11(2): 222-227 226 © 2024 by the author; licensee asian online journal publishing group figure 4. changes in scion shoot length (cm) in response to pre-grafting cane immersion treatments. note: each column represents the mean of three replicates with ten grafts per replicate. error bar stands for the standard deviation of that mean. according to regression analysis (p<0.01), there are significant correlations between the investigated parameters, with the greatest correlation between callusing degree (1-4 scale) and percentage of proper callusing rate (%) at graft union point (table 1). final take, as main indication of grafting success, showed remarkable correlations with callusing degree and percent of proper callusing rate. a proper callusing at grafts union point, a genetically controlled mechanism [23] under the effects of environmental conditions such as exogenous treatments is one of the preconditions of successful grafting [24]. there was a strong positive correlation between final take and shoot length. in arid and semiarid regions, a strong and well-lignified shoot development is desired for coping with the harsh environmental conditions [25]. plant oil extract, serving as an activator, promoted the callusing at graft wound probably due to its phenolic acid and flavanol contents as indicated by assunção, et al. [22]. table 1. correlation between the investigated parameters. parameters callusing degree proper callusing final take shoot length callusing degree 0.969 0.686 0.846 3.-4. degree 0.617 0.704 final take 0.768 4. conclusion overall investigations in the present study indicated that plant activator (plant oil extract) was the most effective pregrafting treatment with the greatest effects on graft callusing, scion shoot growth and final take (nursery survival rate) probably due to its bioactive compound such as phenolic acids and flavanols as indicated in literature. therefore, its application could be recommended as an environment-friendly sustainable approach in grapevine grafting. further studies on different genotypes and various doses would yield extensive information about the use of environment-friendly substances in commercial plant propagation. in particular, graft success in the rootstocks hard to graft, such as certain berlandieri hybrids (41 b), would be improved by proper use of plant activators. references [1] y. gayretli, s. abdulhadi, o. demirkeser, i. turkoglu, b. aslan, and a. sabir, "physiological responses of different grapevine genotypes (vitis spp.) to variable temperatures artificially established as climate change scenery," agrolife scientific journal, vol. 13, no. 1, pp. 98-105, 2024. [2] p. c. s. leao, j. h. b. nascımento, d. s. moraes, and e. r. souza, "yield components of the new seedless table grape ‘brs isis’ as affected by the rootstock under semi-arid tropical conditions," scientia horticulturae, vol. 263, p. 109114, 2020. [3] a. j. winkler, j. a. cook, w. m. kliewer, and l. a. lider, general viticulture. berkele: university of california press, 1974. [4] a. sabir, "improvement of grafting efficiency in hard grafting grape berlandieri hybrid rootstocks by plant growth-promoting rhizobacteria (pgpr)," scientia horticulturae, vol. 164, pp. 24-29, 2013. https://doi.org/10.1016/j.scienta.2013.08.035 [5] s. tedesco, a. pina, p. fevereiro, and f. kragler, "a phenotypic search on graft compatibility in grapevine," agronomy, vol. 10, no. 5, p. 706, 2020. https://doi.org/10.3390/agronomy10050706 [6] p. spiegel-roy and s. lavee, "performance of table grape cultivars on different rootstocks on arid climate," vitis, vol. 10, no. 3, pp. 191-200, 1971. [7] g. loupit, l. brocard, n. ollat, and s. j. cookson, "grafting in plants: recent discoveries and new applications," journal of experimental botany, vol. 74, no. 8, pp. 2433-2447, 2023. https://doi.org/10.1093/jxb/erad061 [8] a. sabır and y. ağaoğlu, "the effects of different iba and naa applications on grafting success of some cultivar/rootstock combinations in potted grape sapling production," alatarım, vol. 8, no. 2, pp. 22-27, 2009. [9] s. verma, s. singh, h. krishna, and v. patel, "comparative performance of different grafting techniques in grape cv. pusa urvashi," indian journal of horticulture, vol. 69, no. 1, pp. 13-19, 2012. [10] h. çelik and f. odabaş, "the effects of the grafting time and types on the success of the grafted grapevine production by grafting under nursery conditions," turkish journal of agriculture and forestry, vol. 22, no. 3, pp. 281-290, 1998. [11] c. alley and a. koyama, "grapevine propagation. xvi. chip-budding and t-budding at high level," american journal of enology and viticulture, vol. 31, no. 1, pp. 60-64, 1980. https://doi.org/10.5344/ajev.1980.31.1.60 [12] n. baydar and m. ece, "comparison of different variety/rootstock combinations in grafted grapevine sapling production in isparta conditions.," süleyman demirel üniversitesi fen bilimleri enstitüsü dergisi, vol. 9, no. 3, pp. 49-53, 2005. [13] z. p. zhang peiyu, x. d. xiang dianfang, l. j. lu junting, g. m. guo mingjun, w. x. wu xueren, and q. y. qi yongshun, "effects of plant growth regulators on the cutting rooting of fenghuang 51 grape variety," china-fruits, vol. 1, no. 2, pp. 28-29, 1997. https://doi.org/10.1016/j.scienta.2013.08.035 https://doi.org/10.3390/agronomy10050706 https://doi.org/10.1093/jxb/erad061 https://doi.org/10.5344/ajev.1980.31.1.60 agriculture and food sciences research, 2024, 11(2): 222-227 227 © 2024 by the author; licensee asian online journal publishing group [14] h. celik, "the effects of different grafting methods applied by manual grafting units on grafting success in grapevines," turkish journal of agriculture and forestry, vol. 24, no. 4, pp. 499-504, 2000. [15] a. sabir and z. kara, "nursery evaluation of different grafting technigues for a sustainable viticulture using 99r and 5bb rootstock," presented at the 2nd international symposium on sustainable development, 2010. [16] h. yin et al., "graft-union development: a delicate process that involves cell–cell communication between scion and stock for local auxin accumulation," journal of experimental botany, vol. 63, no. 11, pp. 4219-4232, 2012. https://doi.org/10.1093/jxb/ers109 [17] g. blunden, t. jenkins, and y.-w. liu, "enhanced leaf chlorophyll levels in plants treated with seaweed extract," journal of applied phycology, vol. 8, pp. 535-543, 1996. https://doi.org/10.1007/bf02186333 [18] n. singh and g. kaur, "study on time and method of grafting on the graft success in grape," j krishi vigyan, vol. 6, no. 2, pp. 264271, 2018. [19] a. ghojage et al., "effect of season on softwood grafting in jamun (syzygium cumini skeels.)," acta horticulturae, vol. 890, pp. 123127, 2011. https://doi.org/10.17660/actahortic.2011.890.15 [20] a. sabir, "comparison of green grafting techniques for success and vegetative development of grafted grape cultivars (vitis spp.)," international journal of agriculture and biology, vol. 13, no. 4, pp. 628–630, 2011. [21] c. j. soar, p. r. dry, and b. loveys, "scion photosynthesis and leaf gas exchange in vitis vinifera l. cv. shiraz: mediation of rootstock effects via xylem sap aba," australian journal of grape and wine research, vol. 12, no. 2, pp. 82-96, 2006. https://doi.org/10.1111/j.1755-0238.2006.tb00047.x [22] m. assunção, s. canas, s. cruz, j. brazão, g. c. zanol, and j. eiras-dias, "graft compatibility of vitis spp: the role of phenolic acids and flavanols," scientia horticulturae, vol. 207, pp. 140-145, 2016. https://doi.org/10.1016/j.scienta.2016.05.020 [23] m. feng, f. augstein, a. kareem, and c. w. melnyk, "plant grafting: molecular mechanisms and applications," molecular plant, vol. 17, no. 1, pp. 75-91, 2023. https://doi.org/10.1016/j.molp.2023.12.006 [24] h. schaefer, "physiological investigations on grafting affinity and callus formation of vines ii," analysen des kallus. wein wissenschaft, vol. 37, pp. 84-89, 1982. [25] f. ferlito, g. distefano, a. gentile, m. allegra, a. lakso, and e. nicolosi, "scion–rootstock interactions influence the growth and behaviour of the grapevine root system in a heavy clay soil," australian journal of grape and wine research, vol. 26, no. 1, pp. 68-78, 2020. https://doi.org/10.1111/ajgw.12415 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1093/jxb/ers109 https://doi.org/10.1007/bf02186333 https://doi.org/10.17660/actahortic.2011.890.15 https://doi.org/10.1111/j.1755-0238.2006.tb00047.x https://doi.org/10.1016/j.scienta.2016.05.020 https://doi.org/10.1016/j.molp.2023.12.006 https://doi.org/10.1111/ajgw.12415 32 © 2022 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 9, no. 1, 32-38, 2022 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v9i1.3862 © 2022 by the authors; licensee asian online journal publishing group we are how much we eat: nutrient-specific versus calorie-based adult-equivalent scales mohammad ali1 kira m. villa2 ( corresponding author) 1centenary college of louisiana, united states. email: mali@centenary.edu tel: (+1)505-720-9017 2university of new mexico, united states. email: kmvilla@unm.edu tel: (+1)505-277-5304 abstract the accurate conversion of household-level food availability into individual-level nutrient availability depends on which adult-equivalent scale is employed. the purpose of this paper is to come up with a set of adult-equivalent scales based on the specific daily intake requirement for macroand micronutrients. using t-tests on household-level data from nepal, we also attempt to find whether on average there are differences between the individual-level nutrient availability estimates when they are calculated through nutrient-specific versus calorie-based, per capita, or organization for economic cooperation and development (oecd) adult-equivalent scales. the results suggest that on average there are significant differences between the individual-level nutrient availability estimates depending on which adult-equivalent scale is used. finally, we find that nutrient-income elasticities calculated through different adult-equivalent scales are statistically different from each other. thus, the nutrient-specific adult-equivalent scales derived in this paper have the potential to reduce measurement error in future studies. keywords: nutrient availability, adult-equivalent scales, nepal, micronutrients, nutrient-income elasticities, food expenditures. citation | mohammad ali; kira m. villa (2022). we are how much we eat: nutrient-specific versus calorie-based adult-equivalent scales. agriculture and food sciences research, 9(1): 32-38. history: received: 10 february 2022 revised: 15 march 2022 accepted: 30 march 2022 published: 19 april 2022 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. authors’ contributions: both authors contributed equally to the conception and design of the study. competing interests: the authors declare that they have no conflict of interest. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study followed all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 33 2. methods ............................................................................................................................................................................................. 33 3. results ................................................................................................................................................................................................ 35 4. conclusion ......................................................................................................................................................................................... 37 references .............................................................................................................................................................................................. 37 mailto:mali@centenary.edu mailto:kmvilla@unm.edu https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v9i1.3862 https://orcid.org/0000-0002-7477-2674 https://orcid.org/0000-0002-1540-2892 agriculture and food sciences research, 2022, 9(1): 32-38 33 © 2022 by the authors; licensee asian online journal publishing group contribution of this paper to the literature our contribution in this research is to calculate nutrient-specific adult-equivalent scales that will enable researchers to more accurately translate food availability data from household surveys to macroand micronutrient availabilities per adult male equivalent. using these nutrient-specific adult-equivalent scales will potentially reduce measurement error in future studies. 1. introduction household-level surveys often contain food modules that collect information on household food availability (i.e., the total amount of food available to a household at a given time) in the form of food expenditures [1]. although not as precise as food frequency questionnaires (ffqs), household-level food expenditure modules are commonly employed because they are cheaper and less timely to collect than individual food consumption data [2, 3]. food expenditure can then be used to estimate individual food/nutrient availability, where individual-level availability is calculated as the total amount of food/nutrients available to the household divided by the number of household members [4-6]. however, per capita food/nutrient availability is not comparable across households with different demographic compositions as the cost of an optimal consumption basket for an individual household member depends on that member’s age and sex. consequently, many use some version of a cost-based adult equivalency scale [7]. commonly used cost-based equivalence scales, such as the organization for economic co-operation and development (oecd) scale [8] are generally developed for overall household consumption or expenditure (not just household nutrient availability). they also typically assume the existence of economies of scale (e.g., buying in bulk can be cheaper per unit than buying in small amounts). for example, the oecd scale assumes that additional adults are equivalent to 0.5 of the first adult. while this is a reasonable assumption for cost-based measures and analysis, economies of scale do not exist for nutrient availability. moving from a one adult to a two adult household (each with the same nutrient requirements) requires that the household’s nutrient availability must double (not increase by 0.5). for these reasons, cost-based adult equivalency scales are generally not appropriate tools for measuring nutrient availability in a household. while cost-based adult equivalency scales are the most common, some instead employ nutrition-based equivalency scales. however, the only nutrition-based adult-equivalent scales, that we are aware of e.g. claro, et al. [9] are based on the calorie requirements of different age groups and sex. calorie-based adult-equivalent scales are an improvement from per capita measures and cost-based equivalency scales and are perfect for calculating individual-level calorie availability however, they still can lead to unreliable estimates of the availability for other nutrients such as macro(fats, carbohydrates, and proteins) and micronutrients (vitamins and minerals). this gap is notable as micronutrient deficiency is currently a bigger problem than calorie deficiency in most developing countries [10]. therefore, employing a measure using only caloric conversion rates may miss an important dimension of nutrient availability. any adult-equivalent scale based on caloric requirements by sex and age will not correctly calculate the individual-level micronutrient availability per adult equivalent within a household. therefore, having accurate conversion factors for other vital nutrients is important to better understand disparities in nutrient availability and food security in developing countries and is necessary for comparing nutrient availability across households with different compositions and size [11, 12]. in this paper, we propose nutrient-specific adult-equivalent scale using the daily nutrient intake guidelines provided by the institute of medicine [13]. this scale provides a more accurate representation of individual-level nutrient availability within a household relative to the nutrient needs of that household. we also calculate the magnitude of the difference in estimated daily nutrient availabilities when using our nutrient-specific scale compared to calorie-based, per capita, and oecd adult equivalent scales with data from the third round of the nepal living standards survey (nlss iii). thus, our contribution is two-fold. first, the calculation of nutrient-specific adultequivalent scales will enable researchers to accurately translate food availability data from household surveys to macroand micronutrient availabilities per adult male equivalent. second, this paper also provides an estimate of the difference in the calculation of daily individual-level nutrient availabilities if the commonly used calorie-based, per capita, and oecd adult equivalent scales are used instead of those that are nutrient-specific. we find that on average there are significant differences in the individual-level daily nutrient availability estimates when they are calculated using the nutrient-specific adult-equivalent scale proposed in this paper compared to the other commonly used (calorie-based, per capita, and oecd) adult equivalent scales. importantly, we find that the average difference is also not just in one direction, as in the use of calorie-based, per capita, and oecd adult equivalent scales overor under-calculates the mean nutrient availability depending on each specific nutrient. thus, our study provides a much more accurate benchmark for future studies using household survey data to calculate individual-level nutrient availability estimates. 2. methods 2.1. data to empirically explore variation in measured nutrient availability based on different adult equivalency scales, we use data from nlss iii collected in 2010-2011. nlss iii consists of a total of 5,988 household-level observations but missing observations reduces our sample to 4,425 households. nlss iii includes a food module containing information on weekly household food availability based on foods obtained through home production, market purchases, and in-kind receipts. we use this food availability data to calculate daily household-level nutrient availability using the reference tables provided by the united states department of agriculture (usda) national nutrient database for standard reference [14]. the nutrients for which we calculate the daily household nutrient availability are proteins, carbohydrates, vitamins a, c, b1 (thiamin), b2 (riboflavin), b3 (niacin), b6, b9 (folic acid), b12, calcium, iron, and zinc. we select these macroand micronutrients, as they are important protective and growth nutrients needed to maintain healthy bodily functions such as bone growth and brain activity [15]. after calculating the daily household-level nutrient availability, we convert it into four measures of average individual-level availability for each household using a calorie-based scale (proposed by claro, et al. [9]) a per capita agriculture and food sciences research, 2022, 9(1): 32-38 34 © 2022 by the authors; licensee asian online journal publishing group approach, the oecd scale, and the nutrient-specific scale we propose in this paper. using these, we construct three variables equal to the difference between individual-level daily nutrient availability constructed with our nutrientspecific adult-equivalent scale and that based on the calorie-based, per capita, and oecd adult-equivalent scales respectively. further explanation of how these difference variables are used is provided in the next sections of this paper. 2.2. nutrient-specific adult-equivalent scales we construct nutrient-specific male adult equivalent scales using the daily nutrient intake guidelines provided by the institute of medicine [13]1. these guidelines provide the daily intake recommendations for all the macro and micronutrients analyzed in this paper, accounting for age and sex differences. based on the age-specific guidelines proposed by institute of medicine [13] we construct our scale for the following age groups: 1-3, 4-8, 9-13, 14-18, 19-30, 31-50, and over 50 years of age. for each age group, other than the 1-3 year-olds, the daily intake recommendations also differ by sex. to estimate the nutrient-specific adult-equivalent scales, we use the daily nutrient recommendation for males in the 19-30 age group for each nutrient as the reference value to which the daily recommendation for all other age and sex groups is compared—making our scale a measure of nutrient availability per adult male equivalent in the household. accordingly, the adult-equivalent conversions for each nutrient were computed by dividing the daily recommendation for each age and sex group with the reference value for that specific nutrient. since, according to the institute of medicine’s guidelines, the vitamin d and carbohydrates recommendations do not change with age or sex, we do not include these nutrients in table 1 as their conversion scale are equal to one for all categories. table 1 details the nutrient-specific adult equivalent scales calculated using the above methodology. a close look at table 1 suggests that the male adult equivalent conversion scales for every sex and age group vary considerably across the different nutrients. this implies that even the use of a calorie-based adult-equivalent scale can overor underestimate nutrient availability per male adult equivalent depending on the nutrient. table 2 provides the caloriebased and oecd adult-equivalent scales for different ages. because a per-capita measure simply divides householdlevel nutrient availability by household size, the conversion scale under this approach is simply one for all demographic categories. it is important to keep in mind that these scales do not differentiate between different macro and micronutrients. table 1. nutrient-specific adult-equivalent scales. ages (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) proteins a c b1 b2 b3 b6 b9 b12 calcium iron zinc child (1-3) 0.23 0.33 0.17 0.42 0.38 0.375 0.38 0.375 0.375 0.7 0.875 0.27 female (4-8) 0.34 0.44 0.28 0.5 0.46 0.5 0.46 0.5 0.5 1 1.25 0.45 male (4-8) 0.34 0.44 0.28 0.5 0.46 0.5 0.46 0.5 0.5 1 1.25 0.45 female (9-13) 0.61 0.67 0.5 0.75 0.69 0.75 0.77 0.75 0.75 1.3 1 0.73 male (9-13) 0.61 0.67 0.5 0.75 0.69 0.75 0.77 0.75 0.75 1.3 1 0.73 female (14-18) 0.82 0.78 0.72 0.83 0.77 0.875 0.92 1 1 1.3 1.875 0.82 male (14-18) 0.93 1 0.83 1 1 1 1 1 1 1.3 1.375 1 female (19-30) 0.82 0.78 0.78 0.92 0.85 0.875 1 1 1 1 2.25 0.73 male (19-30) 1 1 1 1 1 1 1 1 1 1 1 1 female (31-50) 0.82 0.78 0.78 0.92 0.85 0.875 1 1 1 1 2.25 0.73 male (31-50) 1 1 1 1 1 1 1 1 1 1 1 1 female (>50) 0.82 0.78 0.78 0.92 0.85 0.875 1.15 1 1 1.2 1 0.73 male (>50) 1 1 1 1 1 1 1.31 1 1 1 1 1 table 2. calorie-based and oecd adult-equivalent scales. ages (1) ages (2) calorie-based oecd infants (0-1) 0.29 household head 1 child (1-3) 0.51 child (0-3) 0.3 child (4-6) 0.71 child (4-6) 0.3 child (7-10) 0.78 child (7-10) 0.3 female (11-14) 0.86 female (11-14) 0.3 male (11-14) 0.98 male (11-14) 0.3 female (15-18) 0.86 female (15-17) 0.3 male (15-18) 1.18 male (15-17) 0.3 female (19-24) 0.86 female (18-24) 0.5 male (19-24) 1.14 male (18-24) 0.5 female (25-50) 0.86 female (25-50) 0.5 male (25-50) 1.14 male (25-50) 0.5 female (>50) 0.75 female (>50) 0.5 male (>50) 0.9 male (>50) 0.5 2.3. differences between cost-based, calorie-based, and nutrient-specific scales we compare average household nutrient availability per adult male equivalent based on three different equivalency scales. specifically, we examine the difference between estimated average individual daily nutrient availability based on our nutrient-specific equivalent scale and that based on calorie-based (via claro, et al. [9]) per capita, and the oecd adult equivalent scales respectively. we do this for each nutrient and test whether the means of these differences are equal to zero. thus, our hypotheses can be defined as: 1 we do not calculate a nutrient-specific adult-equivalent scale for fats in this paper as the institute of medicine does not provide fat intake recommendations across demographic categories. agriculture and food sciences research, 2022, 9(1): 32-38 35 © 2022 by the authors; licensee asian online journal publishing group 𝐻0: 𝐷𝑖𝑓𝑓_𝑁𝑢𝑡𝑟𝑖𝑒𝑛𝑡ℎ𝑖 = 0 (1) where 𝐷𝑖𝑓𝑓_𝑁𝑢𝑡𝑟𝑖𝑒𝑛𝑡 is the difference obtained by subtracting measures of daily individual-level nutrient availability calculated with the calorie-based, per capita, and oecd adult equivalent scales from the daily individuallevel nutrient availability calculated with the nutrient-specific scale for each nutrient 𝑖 in household ℎ. the t-statistics obtained from the testing of these hypotheses will reveal whether the difference between the individual-level nutrient availabilities calculated through the nutrient-specific adult-equivalent scale are on average significantly different from those calculated through other (calorie-based, per capita, or oecd) adult-equivalent scales. thus, if the null hypotheses are rejected, it would imply that nutrient availability estimates obtained from the above-mentioned adultequivalent scales are on average different from each other. 3. results table 32 reports average daily nutrient availability per household male equivalent based on each of the three adult equivalency scales—specifically, the nutrient-specific (column 2), calorie-based (column 3), per capita (column 4), and oecd (column 5) adult-equivalent scales. column 1 of table 3 reports the daily recommendation of each nutrient for an adult male, for reference. because the average nutrient availability units reported in columns 3-5 are in adult male equivalents, estimated availabilities can be compared to the daily recommendations reported in column 1. thus, if a household’s nutrient availability calculated with the nutrient-specific scale is lower than the daily recommendation reported in column 1, then the household is deficient in that nutrient (i.e., the household does not have enough of that nutrient available to meet the daily requirement of all of its members). strikingly, we see that depending on the nutrient some equivalence scales indicate that the household has a surplus (shortage) of the nutrient when the household is actually deficient (sufficient) in it. for example, the oecd scale indicates average vitamin c availability is sufficient when it is actually below the daily recommendation when using the more accurate nutrientspecific scale. conversely, the per capita scale indicates that households are, on average, deficient in protein when there is actually a small surplus of protein, on average. table 3. summary statistics for nutrient availability from different scales. nutrients daily requirement nutrient-specific calorie based per capita oecd (1) (2) (3) (4) (5) carbohydrates (g) 130 337.0 373.6 337.0 596.7 (160.1) (171.9) (160.1) (280.3) proteins (g) 56 62.17 53.32 48.14 84.82 (30.34) (24.97) (23.38) (39.34) vitamin a (mcg) 900 133.5 117.7 106.7 182.9 (138.9) (120.9) (114.8) (178.1) vitamin c (mg) 80 65.47 53.30 48.62 83.53 (71.59) (57.40) (54.29) (87.58) vitamin d (mcg) 15 0.416 0.456 0.416 0.721 (0.559) (0.580) (0.559) (0.909) vitamin b1 (mg) 1.2 2.229 2.113 1.909 3.369 (1.045) (0.962) (0.907) (1.547) vitamin b2 (mg) 1.3 0.848 0.772 0.697 1.220 (0.529) (0.486) (0.447) (0.729) vitamin b3 (mg) 16 23.45 21.83 19.70 34.90 (11.51) (10.32) 9.609 (16.91) vitamin b6 (mg) 1.3 2.187 2.232 2.013 3.565 (1.148) (1.136) (1.057) (1.847) vitamin b9 (mcg) 400 599.2 589.2 538.5 927.6 (626.2) (611.4) (573.9) (948.4) vitamin b12 (mcg) 2.4 1.005 0.994 0.898 1.556 (0.998) (0.993) (0.913) (1.488) calcium (mg) 1000 281.5 328.9 296.1 517.6 (229.2) (261.1) (236.9) (387.9) iron (mg) 8 10.04 14.59 13.20 23.22 (5.859) (7.474) (7.049) (11.80) zinc (mg) 11 12.58 10.85 9.763 17.33 (6.597) (5.434) (4.967) (8.832) specifically, when we compare average individual-level nutrient availability constructed via our nutrient-specific scale with that via a calorie-based scale (columns 2 and 3 of table 3, respectively), we find that, on average, estimated availability is greater for proteins, vitamins a, c, b1, b2, b3, b9, b12, and zinc when calculated using the nutrientspecific scale. this indicates that on average a calorie-based scale will underestimate the availability of these nutrients within a household. for the rest of the nutrients, the average availability is greater when using the calorie-based scale, indicating this scale will underestimate their availability, on average. comparing the nutrient-specific scale with a per capita scale, the per capita scale underestimates nutrient availability for most nutrients with the exception of calcium and iron. finally, average nutrient availability estimated via the oecd equivalency scale substantially overestimates individual-level nutrient-availability for all nutrients examined. table 43 reports the average difference between nutrient availability based on the nutrient-specific scales and the other three scales and tests whether or not those differences are statistically different from zero. all reported average 2 the figures in parenthesis are the standard deviations of the daily nutrient availability per household male equivalent based on each of the three adult equivalency scales. 3 table 4 shows missing means for the difference between availabilities calculated through per capita and nutrient-specific adult-equivalent scales for carbohydrates, and vitamin d. this is because according to the institute of medicine’s guidelines, the requirements for these nutrients do not change with age and sex. the missing values in table 5 and 6 are also because of the same reason. agriculture and food sciences research, 2022, 9(1): 32-38 36 © 2022 by the authors; licensee asian online journal publishing group differences are statistically significant at one percent level of significance. the only exception is that there is no difference between the nutrient-specific and the per capita scales when estimating household nutrient-availability in carbohydrates or vitamin d. this is because daily recommendations for these two nutrients are the same across all the demographic cohorts. some of the average differences reported in table 4 are quite substantial as a proportion of the daily recommendation. for example, average differences in estimated protein availability range from approximately 14% to 40% of the daily recommendation. average estimated differences in vitamin b9 range from 2% to 82% of the daily recommendation. table 4. difference in means between nutrient-specific and other nutrient availabilities. variables calorie-based per capita oecd carbohydrates -36.64*** -259.8*** proteins 8.853*** 14.03*** -22.65*** vitamin a 15.74*** 26.80*** -49.45*** vitamin c 12.16*** 16.85*** -18.06*** vitamin d -0.0403*** -0.305*** vitamin b1 0.117*** 0.320*** -1.139*** vitamin b2 0.0757*** 0.151*** -0.371*** vitamin b3 1.619*** 3.753*** -11.45*** vitamin b6 -0.0446*** 0.174*** -1.378*** vitamin b9 9.947*** 60.66*** -328.5*** vitamin b12 0.0106*** 0.107*** -0.552*** calcium -47.39*** -14.62*** -236.1*** iron -4.550*** -3.156*** -13.18*** zinc 1.736*** 2.819*** -4.746*** note: *** p<0.01. to further illustrate this point, we take an example representative household from the nlss iii dataset, comprising of two males, aged 48 and 15 years old, and four females aged 67, 43, 12, and 4 years old. these ages represent the average age of husbands, wives, sons, daughters, and elderly individuals in the nlss iii dataset. furthermore, we add a 4-year-old female to our representative household to see the effect of additional children on the calculated individual-level nutrient availability. we also calculate the average nutrient availability per household for every nutrient in the dataset while assuming that if these amounts of every nutrient were available to the representative household described above, how the use of calorie-based, per capita, and oecd scales will compare with the use of nutrient-specific scale. table 5 shows these results where a negative number means that the scale in question overestimates the individual-level availability for that specific nutrient while a positive number suggests underestimation of the same. for example, the calorie-based scale underestimates the individual-level availability of proteins while overestimating the individual-level availability of calcium for the representative household in comparison with the nutrient-specific scale. the results in table 5 clearly show that the use of a calorie-based, per capita, or oecd adult-equivalence scale for all macroand micronutrients will either underestimate or overestimate the true nutrient availabilities for the representative household in the sample. therefore, the nutrient-specific adult-equivalent scales developed in this paper provide a better way of converting the food purchases or consumption data from household surveys into reliable measures of individual-level nutrient availabilities, potentially reducing the measurement error in future studies. table 5. overand underestimation of individual-level nutrient availability from different adult-equivalent scales for a representative household. variables calorie-based per capita oecd carbohydrates (grams) -30.4 -357.41 proteins (grams) 11.08 15.34 -34.76 vitamin a (micrograms) 18.56 27.27 -75.07 vitamin c (milligrams) 15.35 19.37 -27.81 vitamin d (micrograms) -0.04 -0.41 vitamin b1 (milligrams) 0.16 0.33 -1.66 vitamin b2 (milligrams) 0.09 0.15 -0.56 vitamin b3 (milligrams) 2.13 3.9 -16.94 vitamin b6 (milligrams) 0.05 0.23 -1.91 vitamin b9 (micrograms) 25.53 70.21 -455.2 vitamin b12 (micrograms) 0.04 0.12 -0.77 calcium milligrams) -58.48 -32.99 -332.73 iron (milligrams) -4.21 -3.05 -16.75 zinc (milligrams) 1.96 2.84 -7.53 3.1. nutrient-income elasticities from different scales as a robustness check for the finding that individual-level nutrient availabilities obtained from different adultequivalence scales are significantly different from each other, we calculated the nutrient-income elasticities for all macroand micronutrients using ordinary least squares (ols). in our econometric model comprising of separate regression equations, the dependent variables are the natural log of the individual-level nutrient availabilities for each macroand micronutrient calculated using the nutrient-specific, calorie-based, per capita, and oecd adultequivalence scales. the main variable of interest is the natural log of per capita household income, while the control variables include educational dummy variables for the household head, the number of male and female children (less than 17 years old) in the household, the number of male and female adults in the household, and dummy variables indicating whether the household is brahmin caste (upper caste), lives in an urban center, and whether the household head is male. community controls consist of spatial features including distance to market and regional dummies. we agriculture and food sciences research, 2022, 9(1): 32-38 37 © 2022 by the authors; licensee asian online journal publishing group also control for community fixed effects. for further discussion on the variables used, refer to ali, et al. [6]. table 6 shows these results. table 6. nutrient-income elasticities for macroand micronutrients from different scales. nutrient-income elasticities (1) (2) (3) (4) (5) nutrient-specific calorie-based per capita oecd chi-squared carbohydrates 0.204*** 0.198*** 0.121*** 47.80*** (0.0148) (0.0147) (0.0157) proteins 0.318*** 0.350*** 0.356*** 0.274*** 226.46*** (0.0142) (0.0135) (0.0134) (0.0147) vitamin a 1.061*** 1.082*** 1.085*** 1.015*** 330.76*** (0.0387) (0.0382) (0.0378) (0.0399) vitamin c 0.617*** 0.655*** 0.659*** 0.585*** 209.21*** (0.0296) (0.0288) (0.0286) (0.0301) vitamin d 0.558*** 0.579*** 0.626*** 423.49*** (0.0210) (0.0218) (0.0260) vitamin b1 0.229*** 0.247*** 0.251*** 0.180*** 128.91*** (0.0121) (0.0117) (0.0117) (0.0131) vitamin b2 0.425*** 0.438*** 0.435*** 0.401*** 552.88*** (0.0128) (0.0122) (0.0119) (0.0140) vitamin b3 0.216*** 0.236*** 0.242*** 0.160*** 86.70*** (0.0140) (0.0136) (0.0136) (0.0147) vitamin b6 0.141*** 0.180*** 0.185*** 0.110*** 69.14*** (0.0135) (0.0129) (0.0130) (0.0141) vitamin b9 0.618*** 0.638*** 0.644*** 0.561*** 238.45*** (0.0250) (0.0247) (0.0246) (0.0257) vitamin b12 0.789*** 0.800*** 0.781*** 0.829*** 609.62*** (0.0241) (0.0238) (0.0230) (0.0275) calcium 0.544*** 0.532*** 0.538*** 0.455*** 242.45*** (0.0215) (0.0215) (0.0214) (0.0224) iron 0.347*** 0.318*** 0.323*** 0.243*** 110.67*** (0.0147) (0.0145) (0.0145) (0.0156) zinc 0.214*** 0.235*** 0.241*** 0.161*** 99.59*** (0.0142) (0.0136) (0.0136) (0.0147) observations 4,425 4,425 4,425 4,425 4,425 note: robust standard errors in parentheses. *** p<0.01. columns (1) to (4) of table 6 show that using a nutrient-specific in comparison to calorie-based, per-capita, or oecd adult-equivalence scale results in changes in estimated nutrient-income elasticities. column (5) of table 6 also shows that the null hypothesis of equality of the coefficients in each row was rejected at the 1% level of significance. therefore, we can conclude that the choice of adult-equivalence scale in the calculation of individuallevel nutrient availability might matter when it comes to the estimation of nutrient-income elasticities. further analysis is needed to see if the choice of adult-equivalence scale in the calculation of nutrient availabilities makes a difference in other kinds of studies involving their use. 4. conclusion household surveys usually contain data on food purchases or food consumption that have been used widely used to construct of calorie and nutrient availability in a household. while this approach provides a good measure of household-level nutrient availability, it is problematic to translate this measure into individual-level nutrient availability when nutrient requirements differ depending on demographic cohorts and households differ in their demographic make-up. to address this problem, adult equivalency scales are often employed in which household nutrient availability is divided by the number of adult equivalents in the household. different equivalency scales, such as per capita, the oecd, or calorie-based scales, result in different estimated numbers of adult equivalents. while each of these scales are appropriate for certain situations they cannot be universally applied to every macroand micronutrient. for example, when individual-level calorie availability is calculated using a calorie-based scale, none of them provide accurate estimates of household nutrient availability per adult male equivalent for other important macroand micronutrients. in this paper, we propose a set of adult-equivalent scales for macro(carbohydrates and proteins) and micronutrients (vitamins and minerals) based on the specific daily intake requirement for each nutrient across different demographic cohorts. these nutrient-specific scales can be used in future research relying on household food expenditure/availability surveys to arrive at individual-level nutrient availability measures. using this more accurate scale for nutrient-specific adult equivalents will reduce measurement error in these studies. we find that other commonly used adult-equivalent scales can result in drastically different estimates of nutrientavailability per adult equivalent. moreover, they can misrepresent whether a household has enough of a nutrient available to meet its needs or if it is actually deficient in that nutrient. therefore, it is necessary to use equivalency scales that are appropriate for the nutrient of interest and the demographic make-up of the household. references [1] a. naska, v. vasdekis, and a. trichopoulou, "a preliminary assessment of the use of household budget survey data for the prediction of individual food consumption," public health nutrition, vol. 4, pp. 1159-1165, 2001. [2] a. trichopoulou and a. naska, "european food availability databank based on household budget surveys: the data food networking initiative," european journal of public health, vol. 13, pp. 24-28, 2003. available at: https://doi.org/10.1093/eurpub/13.suppl_1.24. agriculture and food sciences research, 2022, 9(1): 32-38 38 © 2022 by the authors; licensee asian online journal publishing group [3] r. engle-stone and k. h. brown, "comparison of a household consumption and expenditures survey with nationally representative food frequency questionnaire and 24-hour dietary recall data for assessing consumption of fortifiable foods by women and young children in cameroon," food and nutrition bulletin, vol. 36, pp. 211-230, 2015. available at: https://doi.org/10.1177/0379572115587272. 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[8] oecd, "what are equivalence scales. retrieved from https://www.oecd.org/els/soc/oecd-note-equivalencescales.pdf," 2021. [9] r. m. claro, r. b. levy, d. h. bandoni, and l. mondini, "per capita versus adult-equivalent estimates of calorie availability in household budget surveys," public health notebooks, vol. 26, pp. 2188-2195, 2010. available at: https://doi.org/10.1590/s0102311x2010001100020. [10] j. hoddinott, m. rosegrant, and m. torero, investments to reduce hunger and undernutrition. copenhagen: copenhagen consensus, 2012. [11] j. r. tedford, o. capps jr, and j. havlicek jr, "adult equivalent scales once more—a developmental approach," american journal of agricultural economics, vol. 68, pp. 322-333, 1986. available at: https://doi.org/10.2307/1241434. [12] a. s. deaton and j. muellbauer, "on measuring child costs: with applications to poor countries," journal of political economy, vol. 94, pp. 720-744, 1986. available at: https://doi.org/10.1086/261405. [13] institute of medicine, dietary reference intakes: the essential guide to nutrient requirements. washington, d.c: the national academies press, 2006. [14] usda, "national nutrient database for standard reference. usda. retrieved from http://ndb.nal.usda.gov/ndb/," 2018. [15] usfda, "guidance for industry: a food labeling guide. u.s. food and drug administration. retrieved from http://www.fda.gov/food/guidanceregulation/guidancedocumentsregulatoryinformation/," 2018. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.oecd.org/els/soc/oecd-note-equivalencescales.pdf, http://ndb.nal.usda.gov/ndb/, http://www.fda.gov/food/guidanceregulation/guidancedocumentsregulatoryinformation/, 84 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 2, 84-92, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i2.6960 © 2025 by the authors; licensee asian online journal publishing group comprehensive analytical report on crop performance using ratio-based metrics mahalakshmi kovvuri1 bharat khushalani2 ( corresponding author) 1,2shri vishnu engineering college for women, bhimavaram, india. 1email: bharat@svecw.edu.in 2email: 23b01a4266@svecw.edu.in abstract this comprehensive study aims to provide a detailed comparative analysis of crop performance by investigating the behavior of crops classified as exhibiting either high or low kilograms per hectare (kgh) productivity through the lens of three fundamental ratio-based indicators: production-to-area, yield-to-area, and yield-to-production. these ratios are critical metrics that help quantify land-use efficiency, productivity levels, and output stability, thereby offering a multi-dimensional perspective on agricultural performance. such ratios indicate how crops behave under varying agronomic and environmental contexts. this study delves into crop performance by distinguishing them into two primary categories based on variance characteristics. high kgh crops are identified as those more susceptible to significant fluctuations due to factors such as climatic variability, market shifts, and biological vulnerabilities. conversely, low kgh crops display relatively stable and predictable patterns of output, making them more reliable under standard farming conditions. the research utilizes both line and bar charts to effectively visualize the inter-crop differences and temporal trends in these ratios, highlighting patterns of consistency and volatility that characterize various crops. the findings aim to support enhanced data-driven decision-making in crop planning, agricultural land management, and policy formulation, particularly in regions facing significant variability in agricultural outputs due to climatic, environmental, or socio-economic factors. by carefully analyzing these ratios, the study not only sheds light on crops with superior resilience and efficiency but also identifies those prone to instability, offering valuable insights for stakeholders in the agricultural sector. keywords: agricultural productivity, bar chart analysis, crop performance, crop planning and policy, crop yield stability. citation | kovvuri, m., & khushalani, b. (2025). comprehensive analytical report on crop performance using ratio-based metrics. agriculture and food sciences research, 12(2), 84–92. 10.20448/aesr.v12i2.6960 history: received: 19 june 2025 revised: 21 july 2025 accepted: 25 july 2025 published: 4 august 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: both authors contributed equally to the conception and design of the study. both authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 85 2. methodology ..................................................................................................................................................................................... 85 3. ratio metrics and analysis ............................................................................................................................................................ 86 4. key findings ..................................................................................................................................................................................... 91 5. policy implications and recommendations................................................................................................................................ 91 6. conclusion ......................................................................................................................................................................................... 92 references .............................................................................................................................................................................................. 92 https://www.doi.org/10.20448/aesr.v12i2.6960 mailto:bharat@svecw.edu.in mailto:23b01a4266@svecw.edu.in https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ agriculture and food sciences research, 2025, 12(2): 84-92 85 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature by analyzing the defined ratios, our study not only sheds light on crops with superior resilience and efficiency but also identifies those prone to instability, offering valuable insights for stakeholders in the agricultural sector. the terminology itself is new and not available in previous studies. 1. introduction agricultural productivity remains a cornerstone of food security, economic resilience, and rural development. conducting a quantitative evaluation through ratio-based metrics provides a nuanced perspective on how crops behave under varying agronomic and environmental contexts. this study delves into crop performance by distinguishing them into two primary categories based on variance characteristics. high kgh crops are identified as those more susceptible to significant fluctuations due to factors such as climatic variability, market shifts, and biological vulnerabilities. on the other hand, low kgh crops display relatively stable and predictable patterns of output, making them more reliable under standard farming conditions. this classification supports targeted agricultural decision-making, particularly when assessing risk, input optimization, or crop selection for specific agro-ecological zones. the dataset employed in this study originates from india’s agricultural statistics and encompasses state-wise data for a diverse range of principal crops. the temporal coverage spans financial years 2001–02 through 2015–16, allowing a longitudinal analysis of trends and variability in crop performance. this dataset includes key variables such as the area under cultivation (hectares), total production (metric tonnes), and yield per hectare for each crop across multiple states. the data was compiled from government records and agricultural statistical publications (source to be specified), providing a comprehensive basis for calculating ratio-based metrics that reflect productivity and efficiency. this rich dataset enables an in-depth assessment of how crop outputs evolve over time across different agro-climatic zones and socio-economic conditions, making it instrumental for data-driven agricultural policy and planning. a global dataset of crops is considered in iizumi and sakai [1] and the authors perform an analysis over a time period of around 35 years. they take wheat, rice, soybean, and maize and obtain the yield time series of these crops. senapati and goyari [2] have divided their study period into three distinct phases: pre-green, early phase, and post-green revolution period, and compared the growth rates during these periods. the development and regulatory challenges of genetically modified crops in india are discussed in shukla et al. [3]. their work highlights both scientific progress and policy barriers. this study extends the discussion by visually analyzing crop performance across indian states. reddy et al. [4] improved crop simulation in clm5 using long-term site-level data for indian crops. their work enhanced model accuracy for wheat and rice growth patterns. building on this, our study visually analyzes crop performance across indian states for better planning. long-term trends in millet cultivation across india are examined in yamuna et al. [5]. they reported declining area and production but stable or improving yields. this study builds on their insights through visual analysis of crop performance across states. the extremes of crop yields in india using extreme value theory are analyzed in lakshmi kumar et al. [6]. their study highlights regional differences in yield variability and stresses the need to consider extreme events in agricultural planning to improve food security. state-wise yields of major food crops in india for 2014–15 using statistical methods are analyzed in talukdar et al. [7]. the study revealed significant regional differences in productivity, emphasizing the need for targeted strategies to improve crop yields. the growth and sustainability of major crops in haryana are studied in bagaria and jatav [8], highlighting positive trends due to water-efficient practices. their findings stress the need for sustainable agriculture to ensure long-term food security. changing crop patterns and diversification in koch bihar, west bengal, focusing on six key crops, are considered in islam and das [9]. their analysis revealed significant shifts influenced by environmental and socio-economic factors. an analysis of four decades of data to assess trends in the area, production, and yield of key crops in bangladesh is done in akhter et al. [10]. their findings reveal significant growth in rice and wheat yields, while jute cultivation experienced a decline, highlighting the need for adaptive agricultural strategies. a survey of agricultural datasets to enhance crop yield prediction using machine learning algorithms is conducted in tripathi et al. [11]. their study emphasizes the importance of accurate yield forecasting for precision agriculture and food security. the growth patterns of oilseed crops in india are considered in reddy and immanuelraj [12], revealing significant spatial and temporal disparities. their study highlights the need for targeted strategies to enhance oilseed productivity and reduce reliance on imports. a comprehensive global analysis of crop yield trends and variability using 8,088 country-crop yield series from fao data is conducted in arata et al. [13]. employing robust statistical methods, they identified a slowdown in yield growth and increased variability in certain regions, highlighting implications for global food security and the need for adaptive agricultural policies. a systematic review on the application of data analytics in crop management is conducted in chergui and kechadi [14]. their study emphasizes how digital agriculture leverages big data and advanced technologies to enhance productivity, optimize resource use, and promote sustainable farming practices. global crop yield variability from 1981 to 2010 is analyzed in iizumi and ramankutty [15], attributing significant changes to climate change. their study highlights the increasing impact of climate factors on yield stability, emphasizing the need for adaptive agricultural strategies. long-term trends in foodgrain yield, area, and production in india are examined in shaharshad et al. [16]. their analysis highlights how both short-term and long-term factors influence agricultural output dynamics. growth trends and structural changes in india’s commercial crop production from 1980 to 2020 are studied in kutty [17], highlighting the impact of policy reforms on agricultural productivity. a dynamic-statistical biomass model to predict crop yields in kazakhstan, utilizing 21 years of regional data (2000–2021), is developed in sadenovaa et al. [18]. the model demonstrated strong correlations with official statistics, indicating its robustness against meteorological variability. 2. methodology the analytical framework employed in this study is centered on a comprehensive evaluation of three critical productivity ratios production-to-area, yield-to-area, and yield-to-production across the two major crop categories distinguished by their kgh behavior. each ratio serves a unique purpose in unraveling different aspects agriculture and food sciences research, 2025, 12(2): 84-92 86 © 2025 by the authors; licensee asian online journal publishing group of crop performance and land-use efficiency. the methodology involves systematically processing the dataset to compute these ratios for every crop and every state-year combination within the study period. these calculations form the backbone of subsequent visual analyses. both line and bar plots are employed as key visualization tools to analyze the temporal and cross-sectional trends of these ratios. bar plots provide an intuitive understanding of inter-crop performance by clearly displaying the magnitude of ratios in selected years, thereby facilitating easy comparison of productivity and efficiency among crops. on the other hand, line plots reveal the temporal evolution of these ratios, capturing fluctuations, peaks, and troughs that characterize the highand low-performing crops over time. this dual approach helps to uncover both steady trends and volatility, which are essential for understanding the underlying factors driving crop performance. special attention is given to evaluating performance thresholds, such as the 5 kgh benchmark, which helps differentiate between highly productive and moderately performing crops. this threshold also assists in policy prioritization by flagging crops that exceed or fall below critical productivity levels. by integrating these ratio analyses with visual tools, the study provides policymakers, researchers, and farmers with actionable insights, enabling more informed decisions related to crop selection, resource allocation, and resilience building in agriculture. 3. ratio metrics and analysis 3.1. ratio of production to area the ratio of production to area serves as a fundamental indicator of output efficiency, measuring how much crop yield (typically in tons) is produced per unit of land (e.g., hectare). this ratio is essential in assessing the agricultural productivity of land, especially in contexts where cultivable land is scarce or subject to degradation. by comparing output relative to the area under cultivation, this metric helps evaluate the intensity and effectiveness of land use, offering valuable insights into which crops optimize production under prevailing environmental and agronomic conditions. in the case of high kgh crops, the data visualizations, particularly line plots, reveal substantial instability and fluctuation, as shown in figure 1. the peaks and troughs observed highlight the unpredictable nature of production outcomes, which can be attributed to erratic rainfall, pest infestations, soil fertility variations, and inconsistent input application. these fluctuations, visualized further in figure 2, make high kgh crops less dependable from both an economic and policy standpoint. figure 1. production/area – high kgh crops. figure 2. production/area – high kgh crops (bar). agriculture and food sciences research, 2025, 12(2): 84-92 87 © 2025 by the authors; licensee asian online journal publishing group in contrast, low kgh crops exhibit a much more predictable trend in their production-to-area ratio. bar charts, such as in figure 4, demonstrate uniform and steady performance, while line plots in figure 3 display smooth, consistent curves, indicating resilience and adaptability. these characteristics imply that low kgh crops benefit from stable environmental conditions, optimized agronomic practices, and improved seed genetics, making them ideal for land productivity enhancement. figure 3. production/area – low kgh crops. figure 4. production/area – low kgh crops (bar). mathematical formula: this ratio measures how many kilograms of crop are produced per hectare, reflecting land productivity. statistical classification: the production-to-area ratio is used to classify crops into high kgh and normal kgh groups based on a threshold value of 5 kg/ha. crops with p/a values exceeding 5 kg/ha are designated as high kgh, indicating exceptionally high land productivity but often accompanied by higher variance (greater year-to-year fluctuations). examples include sugarcane, cotton, and banana, which frequently surpass this threshold, with sugarcane reaching peaks over 70 kg/ha. crops consistently below the 5 kg/ha threshold fall into the normal kgh category. these crops, such as rice, maize, and wheat, generally show lower but steadier production per hectare. statistically, high kgh crops exhibit higher variance (standard deviation) in p/a values, reflecting sensitivity to environmental factors and management practices, while normal kgh crops show tighter clustering around their mean p/a, indicating stability. the production-to-area ratio plot clearly demonstrates which crops utilize land most efficiently in terms of raw output. crops like sugarcane and banana dominate this visualization, with sugarcane frequently exceeding 70 kgh and peaking beyond 75 kgh in certain years, showing extremely high land productivity. cotton follows with notable consistency, often surpassing the 5 kgh threshold, classifying it within the high-efficiency crop group. this visual behavior confirms sugarcane’s and cotton’s suitability for regions prioritizing land maximization. however, the statistical distribution of these values reveals significant year-to-year fluctuations, as evidenced by the irregular bar heights across the timeline, indicating high variance. such variability underlines environmental dependency and sensitivity to factors like rainfall, fertilizer input, and agronomic management. on the other hand, crops like maize, wheat, and paddy remain steadily clustered in the range of 2–4 kgh, with minimal deviation across years, reflecting stable and predictable land performance. this contrast, visually and agriculture and food sciences research, 2025, 12(2): 84-92 88 © 2025 by the authors; licensee asian online journal publishing group statistically apparent in the plot, serves as a direct basis for classifying crops into high and normal kgh categories and is critical when prioritizing land allocation strategies in fluctuating agricultural zones. 3.2. ratio of yield to area the ratio of yield to area is another crucial performance metric, reflecting the efficiency of land use in producing a normalized crop output. unlike the production ratio, this metric often accounts for factors such as weight, quality, or moisture content. it is particularly useful in modern agriculture, where maximizing both quantity and quality from available land is critical. for high kgh crops, this ratio shows substantial inconsistency. line plots, such as in figure 5, display sharp spikes followed by abrupt declines, indicating sensitivity to environmental and market changes. figure 6 is a bar chart representation that highlights this volatility even further. peaks may reflect favorable weather or robust input use, while dips often suggest stressors like droughts, pest attacks, or supply shortages. these erratic trends make planning around such crops a risky endeavor. figure 5. yield/area – high kgh crops. figure 6. yield/area – high kgh crops (bar). conversely, low kgh crops display minimal variation in yield-to-area ratios. this is clearly depicted in figure 7 and figure 8, where both line and bar plots reflect a smooth and consistent performance trend. this stability arises from controlled agronomic conditions, including dependable irrigation, the use of hybrid seeds, and reliable fertilizer access. such uniformity enables better input planning, more predictable income for farmers, and improved policy effectiveness. agriculture and food sciences research, 2025, 12(2): 84-92 89 © 2025 by the authors; licensee asian online journal publishing group figure 7. yield/area – low kgh crops. figure 8. yield/area – low kgh crops (bar). mathematical formula: this ratio accounts for yield adjusted by quality factors, normalized per hectare. statistical classification: in the statistical context, the yield-to-area ratio is analyzed to distinguish crop groups based on their average ratio and variance. crops with y/a values notably above 0.1 (or similarly high thresholds relevant to the data range) and high inter-annual variability are classified as high kgh crops. for instance, ginger shows y/a spikes around 0.42 with sharp drops, indicating high variance and unstable yield performance. normal kgh crops tend to have y/a ratios clustered within a narrower band (e.g., 0.02 to 0.035) and exhibit low variance, signifying consistent yield quality per hectare. statistically, the difference between high and normal kgh is evaluated using measures like standard deviation and coefficient of variation, identifying crops with stable versus volatile yield efficiency. the yield-to-area ratio plot emphasizes quality-adjusted production relative to land use, visually distinguishing high-value crops from more stable staples. crops such as ginger, black pepper, and turmeric stand out with bar heights reaching 0.42 and sometimes even higher, particularly in years like 2003–04 and 2005–06 for ginger. these values are significantly above those of other crops, where the majority such as rice, maize, and wheat cluster consistently between 0.02 and 0.035. the wide spread and sharp fluctuations among the high y/a crops suggest strong input responsiveness but also highlight volatility. this variability is evident from the irregular and steeply rising or falling bars across years, indicating sensitivity to climatic or economic stressors. in contrast, the flat and even distribution of y/a ratios in staples demonstrates agronomic stability, achieved through regulated irrigation, hybrid seed use, and consistent cultivation practices. these statistical trends in the bar plot underscore how this ratio can be used not only to assess land performance but also to inform precision agriculture decisions, guiding investments into crops that either demonstrate stable returns or require controlled environments to mitigate risks. 3.3. ratio of yield to production the ratio of yield to production offers insights into how yield performance aligns with total output. while less commonly used, it reveals whether gains in yield translate proportionately into production or if inefficiencies such as post-harvest losses are present. this metric is especially useful for identifying gaps in crop conversion efficiency. agriculture and food sciences research, 2025, 12(2): 84-92 90 © 2025 by the authors; licensee asian online journal publishing group high kgh crops tend to display wide variability in this ratio. line and bar charts, as presented in figure 9 and figure 10, show erratic values. these issues could stem from increased area not leading to proportional yield gains or losses in the supply chain. the discrepancies suggest that yield improvements in these crops do not always result in scaled production, limiting their reliability. figure 9. yield/production– high kgh crops. figure 10. yield/production– high kgh crops (bar). in contrast, low kgh crops typically exhibit stable and higher yield-to-production ratios, as seen in figure 11 and figure 12. this alignment indicates that improvements in yield translate directly into overall production, which points to greater operational efficiency. such crops are more suited for scale-up and offer a better return on investment. figure 11. yield/production– low kgh crops. agriculture and food sciences research, 2025, 12(2): 84-92 91 © 2025 by the authors; licensee asian online journal publishing group figure 12. yield/production– low kgh crops (bar). mathematical formula: this ratio reflects the consistency or proportionality between yield and production. statistical classification: statistically, the yield-to-production ratio is used to assess operational efficiency beyond raw yield or production numbers. crops with y/p ratios fluctuating widely ranging, for example, from 0.02 to 0.12 are classified as high kgh due to irregular efficiency patterns. black pepper and tobacco demonstrate such variable y/p ratios, indicating inconsistencies such as post-harvest losses or processing inefficiencies. in contrast, crops exhibiting a near-linear relationship in y/p, with ratios consistently clustered around lower stable values (e.g., 0.015 to 0.035), fall under normal kgh. this stability suggests effective conversion of yield gains into production increments, marking operational efficiency. variance metrics, trend analysis, and standard deviation are used statistically to differentiate these categories, highlighting crops with consistent versus inconsistent yield-to-production relationships. the yield-to-production ratio plot provides a clear visualization of the internal relationship between yield and total output. high values in this plot such as those for black pepper and mesta, often reaching 0.1 to 0.12 indicate that the yield (possibly quality-adjusted) constitutes a significant portion of total production, highlighting potential inefficiencies in scaling up or post-harvest processing. these elevated bars contrast with the more consistent lower ratios for crops like maize, wheat, and rice, where values range between 0.015 and 0.035. this suggests that in these staple crops, increases in yield proportionally translate into overall production, reflecting operational efficiency. from a statistical perspective, the high variability observed in the bar heights for crops like turmeric and ginger points to inconsistency in yield-to-output translation, which may result from unpredictable losses or input/output mismatches. visually, the irregular bar heights for such crops differ markedly from the even, linear patterns seen in staples, reinforcing their classification as high-variance crops and highlighting their suitability for more controlled, investment-intensive cultivation systems. this ratio, as depicted in the plot, is essential for diagnosing systemic inefficiencies and guiding research or infrastructure investments. 4. key findings the comparative analysis of high kgh and low kgh crops through the lens of the three-ratio metrics reveals clear patterns and distinctions. high kgh crops exhibit significant volatility across all ratios, indicating inconsistent performance. while they may demonstrate high yields or production levels in certain years, these instances are not reliable and often arise under specific, favorable conditions. this unpredictability limits their potential for stable food supply planning, commercial investment, and policy support. the inherent risk associated with high kgh crops calls for careful consideration before large-scale promotion. in contrast, low kgh crops consistently show a strong alignment between area, yield, and production. this makes them more dependable for agricultural stakeholders, from farmers and supply chain actors to policymakers. their performance across all three ratios particularly in yield-to-area and yield-to-production suggests high efficiency, resilience, and scalability. these attributes make them excellent candidates for subsidy allocation, research funding, and national food security strategies. among the three metrics, the yield-to-area ratio emerged as the most consistent and reliable across both crop categories, offering a clear indicator of land-use efficiency. meanwhile, the yield-to-production ratio proved particularly useful in identifying internal inconsistencies and assessing readiness for large-scale farming operations. combined, these ratios create a comprehensive framework for evaluating crop sustainability and productivity. 5. policy implications and recommendations based on the analysis of ratio-based crop performance metrics, several policy and strategic recommendations emerge. first, government subsidies and agricultural support programs should prioritize crops that demonstrate consistent and high performance across both yield and production dimensions. these crops not only ensure stable returns for farmers but also contribute to food security and economic resilience at the national level. high kgh crops, despite their risks, should not be dismissed entirely. instead, they should be the focus of targeted innovation efforts, including the development of pest-resistant or drought-tolerant varieties, the agriculture and food sciences research, 2025, 12(2): 84-92 92 © 2025 by the authors; licensee asian online journal publishing group promotion of adaptive farming techniques, and the establishment of insurance schemes that cushion farmers against environmental shocks. research funding and public-private partnerships can play a key role in making these crops more reliable performers. the three ratios analyzed in this report, production-to-area, yield-to-area, and yield-to-production, should be integrated into broader agricultural management systems. for example, they can serve as the foundation for crop insurance models, where premiums are based on volatility, or in credit scoring systems that assess the risk of lending to specific agribusinesses based on crop consistency. additionally, coupling these metrics with climate, soil, and satellite datasets will enable the development of advanced predictive models, supporting precision agriculture and smarter resource allocation. 6. conclusion this multi-ratio analytical approach offers a robust and comprehensive decision-support framework for evaluating crop performance under varying agronomic and environmental conditions. by systematically examining the interrelationships among production, yield, and area through three distinct ratios production-to-area, yield-toarea, and yield-to-production this methodology provides granular insights into the efficiency, consistency, and potential scalability of agricultural outputs. through this lens, the contrast between high kgh and low kgh crops becomes particularly evident. while high kgh crops may demonstrate occasional surges in productivity, their unpredictable nature limits their reliability for long-term planning and investment. in contrast, low kgh crops consistently deliver stable performance across all key metrics, making them more suitable for scaling, strategic support, and policy prioritization. the integration of visual tools such as bar and line plots not only enhances interpretability but also enables temporal and cross-sectional analysis. this visual representation aids researchers, agronomists, and policymakers in identifying both risk-prone crops that require targeted interventions and resilient varieties that merit broader promotion. when combined with complementary datasets such as climate patterns, soil conditions, and technological inputs, this framework can be further strengthened to guide evidence-based agricultural planning. ultimately, this approach promotes a more sustainable, scalable, and stable agricultural ecosystem. it equips stakeholders with the clarity needed to make informed decisions, allocate resources efficiently, and drive innovation in crop management and development. references [1] t. iizumi and t. sakai, "the global dataset of historical yields for major crops 1981–2016," scientific data, vol. 7, no. 1, p. 97, 2020. https://doi.org/10.1038/s41597-020-0433-7 [2] a. k. senapati and p. goyari, "agricultural growth and production variability of principal crops in india: an empirical investigation," advances in plants & agriculture research, vol. 8, no. 1, pp. 45-51, 2018. [3] m. shukla, k. t. al-busaidi, m. trivedi, and r. k. tiwari, "status of research, regulations and challenges for genetically modified crops in india," gm crops & food, vol. 9, no. 4, pp. 173-188, 2018/10/02 2018. https://doi.org/10.1080/21645698.2018.1529518 [4] k. n. reddy et al., "improving the representation of major indian crops in the community land model version 5.0 (clm5) using site-scale crop data," geoscientific model development, vol. 18, no. 3, pp. 763-785, 2025. https://doi.org/10.5194/gmd-18-7632025 [5] n. yamuna, d. kolekar, and n. rakesh, "trend and instability analysis of area, production, productivity of millets in india," journal of scientific and reports, vol. 30, no. 8, pp. 686-999, 2024. https://doi.org/10.9734/jsrr/2024/v30i82290 [6] t. lakshmi kumar, h. barbosa, s. madhu, and k. koteswara rao, "studies on crop yields and their extreme value analysis over india," sustainability, vol. 11, no. 17, p. 4657, 2019. https://doi.org/10.3390/su11174657 [7] b. talukdar, p. k. chakravartty, and b. sharma, "analysis of state wise yield of major food crops in india for the year 2014-15," vol. 44, no. 2, 2023. https://doi.org/10.53555/jaz.v44i2.4543 [8] n. bagaria and s. s. jatav, "growth dynamics and sustainability of major crops in haryana, india," indian journal of extension education, vol. 61, no. 2, pp. 80-84, 2025. https://doi.org/10.48165/ijee.2025.61215 [9] k. islam and t. das, "changing patterns of crop combination, crop concentration and crop diversification regions: a case study based on six principal crops of koch bihar district, west bengal, india," egyptian journal of agricultural research, vol. 103, no. 2, pp. 188-204, 2025. https://doi.org/10.21608/ejar.2024.282439.1536 [10] s. akhter, j. r. sarker, and k. r. das, "growth and trend in area, production and yield of major crops of bangladesh," international journal of economics, finance and management sciences, vol. 4, no. 1, pp. 20-25, 2016. https://doi.org/10.11648/j.ijefm.20160401.13 [11] a. tripathi, b. s. rathore, and d. singh, "survey paper on agricultural dataset for improving crop yield prediction using machine learning algorithms," international journal of computer applications, vol. 184, no. 46, pp. 28-34, 2023. https://doi.org/10.5120/ijca2023922571 [12] v. k. reddy and k. t. immanuelraj, "area, production, yield trends and pattern of oilseeds growth in india," economic affairs, vol. 62, no. 2, pp. 327-334, 2017. https://doi.org/10.5958/0976-4666.2017.00016.x [13] l. arata, e. fabrizi, and p. sckokai, "a worldwide analysis of trend in crop yields and yield variability: evidence from fao data," economic modelling, vol. 90, pp. 190-208, 2020. https://doi.org/10.1016/j.econmod.2020.05.006 [14] n. chergui and m. t. kechadi, "data analytics for crop management: a big data view," journal of big data, vol. 9, no. 1, p. 123, 2022/12/23 2022. https://doi.org/10.1186/s40537-022-00668-2 [15] t. iizumi and n. ramankutty, "changes in yield variability of major crops for 1981–2010 explained by climate change," environmental research letters, vol. 11, no. 3, p. 034003, 2016. https://doi.org/10.1088/1748-9326/11/3/034003 [16] c. shaharshad, k. manikandan, m. priyadharshini, and m. rishad, "trends in yield, area, and production of foodgrains in india," international journal for multidisciplinary research, vol. 7, no. 3, 2025. https://doi.org/10.36948/ijfmr.2025.v07i03.44350 [17] k. kutty, "growth trends of commercial crops production, area, and yield in india: an appraisement of the structural stability regression model," estudios de economía aplicada, vol. 41, no. 1, p. 27, 2023. https://doi.org/10.25115/sae.v41i1.8625 [18] m. a. sadenovaa, n. a. beisekenova, m. rakhymberdinaa, p. s. varbanovb, and j. j. klemešb, "mathematical modelling in crop production to predict crop yields," chemical engineering, vol. 88, pp. 1225-1230, 2021. https://doi.org/10.3303/cet218820 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1038/s41597-020-0433-7 https://doi.org/10.1080/21645698.2018.1529518 https://doi.org/10.5194/gmd-18-763-2025 https://doi.org/10.5194/gmd-18-763-2025 https://doi.org/10.9734/jsrr/2024/v30i82290 https://doi.org/10.3390/su11174657 https://doi.org/10.53555/jaz.v44i2.4543 https://doi.org/10.48165/ijee.2025.61215 https://doi.org/10.21608/ejar.2024.282439.1536 https://doi.org/10.11648/j.ijefm.20160401.13 https://doi.org/10.5120/ijca2023922571 https://doi.org/10.5958/0976-4666.2017.00016.x https://doi.org/10.1016/j.econmod.2020.05.006 https://doi.org/10.1186/s40537-022-00668-2 https://doi.org/10.1088/1748-9326/11/3/034003 https://doi.org/10.36948/ijfmr.2025.v07i03.44350 https://doi.org/10.25115/sae.v41i1.8625 https://doi.org/10.3303/cet218820 49 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 49-53, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6818 © 2025 by the authors; licensee asian online journal publishing group the potential of specific profilin based fingerprinting differs in legume species adam kovacik1 lenka kucerova2 alzbeta jauschova3 jana ziarovska4 ( corresponding author) 1,2,3,4institute of plant and environmental sciences, faculty of agrobiology and food resources, slovak university of agriculture in nitra, slovakia. 1email: adam.kovacik@uniag.sk 2email: xkucerova@uniag.sk 3email: xjauschova@uniag.sk 4email: jana.ziarovska@uniag.sk abstract one requirement for using germplasm in agricultural development initiatives is its characterization, which necessitates knowledge of the genetic polymorphism and relationships among the individual varieties. up to now, different dna markers were utilized for this purposed, one of the newest are those for coding regions. here, we aimed to investigate polymorphism and genetic relationships among 24 varieties of cicer arietinum l. and 23 varieties of pisum sativum l. using profilin based fingerprinting. pcr approach was used to generate homologue amplicons of plant profilins and upgma grouping for visualization of obtained fingerprint similarity. amplification results showed different results for analysed legume species, where the higher polymorphism at the level of 96% was obtained within the accessions of pea varieties, as for chickpea fingerprints a very similar profiles were generated with only a limited amplicons of a total of three different length of 150 bp, 182 bp and 348 bp. this information could be useful in breeding strategies for the improvement of chickpea and pea accessions. keywords: cicer arietinum l., length polymorphism, pisum sativum l., profilin, dna markers, fingerprinting. citation | kovacik, a., kucerova, l., jauschova, a., & ziarovska, j. (2025). the potential of specific profilin based fingerprinting differs in legume species. agriculture and food sciences research, 12(1), 49– 53. 10.20448/aesr.v12i1.6818 history: received: 15 november 2024 revised: 28 may 2025 accepted: 13 june 2025 published: 25 june 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the grant agency of faculty of agrobiology and food resources, slovak university of agriculture in nitra, slovakia (grant number: ga fapz 12/2024). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 50 2. materials and methods ................................................................................................................................................................... 50 3. results and discussion ................................................................................................................................................................... 51 references .............................................................................................................................................................................................. 52 mailto:adam.kovacik@uniag.sk mailto:xkucerova@uniag.sk mailto:xjauschova@uniag.sk mailto:jana.ziarovska@uniag.sk https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6818 https://orcid.org/0009-0004-7474-6056 https://orcid.org/0009-0001-9997-8214 https://orcid.org/0009-0008-2929-6403 https://orcid.org/0000-0002-0005-9729 agriculture and food sciences research, 2025, 12(1): 49-53 50 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature allergen based dna marker techniques provide a valuable insight to polymorphism of plant coding regions. here, profilin based polymorphism was firstly applied to analyse the variability among pea and chickpea germplasm. 1. introduction genetic diversity of plants and its description is still of great importance in the research. plants are inevitable parts of our daily lives, but many of their characteristics are still unknown. the group of legumes provide a basic part of human nutrition in many areas worldwide [1] that is why the interest in their germplasm genetic variability is actual and different studies were published previously [2-5]. especially molecular and dna markerbased characterization of legume germplasm has the great potential in determining of genetic diversity and identification of unique variants that can be used in breeding programmes. in this study, selected varieties of cicer arietinum l. and pisum sativum l. were analysed. both of this species are well characterized in their germplasm variability using different dna based markers. investigation of chickpea gerplasm by rapd (randomly amplified polymorphic dna) provided a low degree of polymorphism with high coefficients of genetic similarity [6]. intersimple sequence repeat (issr) markers were found to be dependent on the characteristics of their 3´anchores sequences in the ability to generate polymorphism in chickpea accessions and a combination of the principal component anaysis (pca) together with cluster analysis in data analysis is important for informativeness of generated data and extracting the valuable informations [7, 8]. in this technique, more primers need to be used to analyse germplasm thoroughly [9]. retrotransposon based dna marker techniques such as ipbs (inter primer binding sites polymorphism) were reported to be able to generate up to the 100% polymorphism in chickpea variability analysis [10]. start codon targeted (scot) polymorphism is efficient in genetic diversity analysis of chickpea depending on the primer combinations and the genetic similarity can range from 84 to 98%, but many of primer combination can be monomorphic in its results [11]. investigation of pea germplasm by rapd were performed previously with good results in obtained polymorphism where clear pattern of clustering according to the source of germplasm were showed in different studies [12-14]. microsatellite based markers were reported to differentiate successfully among parent and hybrid genotypes of pea [15] and provided a high level of polymorphism among pea genotypes [16]. scot are more informative marker compared to issr and rapd markers for discrimination and identification of p. sativum subspecies [17]. retrotransposon based marker techniques were proved to assess distinctness of pea germplasm, especially in cases where the time frame plays an important role. rbip technique (retrotransposon based information polymorphism) was reported to be robust and easy to score method, while multilocus irap (inter retrotransposon amplified polymorphism) produced informative fingerprint already in a single analysis [18]. dna markers are actually developed based on in silico prediction and the of potential of these markers was reported to be efficient [19]. in silico approach in allergen coding-based dna markers is based on the identification of conserved parts in the amino acid and genomic sequences of individual homologs of plant allergens. finding the similarity in alignments, specific as well as degenerate primers are designed [20]. plant allergens and their protein and nucleotide sequences share a high degree of homology [21, 22] what allow to predict dna markers for them, too. previously, bet v 1 based amplified polymorphism was conformed to be informative in various of plant species, as the genomic sequences of this pollen allergen of birch has the high level of conservation in its epitops [23, 24]. here, an abundant plant allergen profilin wase used based on the designation of primer pair to its specific conservative part. when using a nonspecific profilin based primer pairs, the profilin based amplified polymorphism method (pbap) was effective generating of specific fingerprints in all of the analysed genotypes of soybean and groundnut [25, 26] here, specific one was used. the aims of the present study were (1) analyzing of the potential of specific legume profilin based marker technique to reveal the polymorphism among legume species and genotypes, (2) characterizing and comparing the polymorphism based on the specific profilin based fingerprints generated for cicer arietinum l. and pisum sativum, l. 2. materials and methods 2.1. biological material seeds of legume species cicer arietinum l. and pisum sativum l. were obtained from genebank of slovak republic, piešťany. different numbers of individual varieties were randomly selected to be able analyse the natural intraspecie variability as old land, ancient as well as modern varieties were represent in a final number of 24 for chickpea and 23 for pea. young plants were obtained in vitro from sterilized seeds in basal murashige and skoog medium [27] with day-length 15h and 20°c. 2.2. dna extraction total genomic dna was extracted by genejet™plant genomic dna purification mini kit (thermo scientific, waltham, ma, usa) following the manufacturer instruction. quantity and quality of extracted dna was analysed spectrophotometrically by nanophotometer™ (implen) and functionality in pcr was checked by its primers [28]. 2.3. specific profilin based polymorphism analysis primers for specific profilin based polymorphism fingerprints were designed using the in silico analysis of their conserved sequences [29]. in pcrs, dreamtaq™ dna polymerase together with 600 nmol of each primer was utilized. pcr conditions of time and thermal profile was: 95 ◦c for 5 minutes (95 ◦c for 45 seconds; 55 ◦c for 45 seconds; 72 ◦c for 35 seconds) 40x plus final 72 ◦c for 10 minutes. obtained amplicons were separated in 2% agarose gel stained by gelred®. amplified fingerprints were transformed into binary matrices and based on this, agriculture and food sciences research, 2025, 12(1): 49-53 51 © 2025 by the authors; licensee asian online journal publishing group distance matrices and dendrograms of genetic dissimilarity were prepared using upgma analysed based on jaccard index [30]. dendrograms of profilins were created by dendroupgma software [31]. 3. results and discussion in the case of obtained fingerprints for cicer arietinum l., a very similar profiles were generated with only a limited amplicons of a total of three different length of 150 bp, 182 bp and 348 bp. a total of six groups were obtained in the constructed dendrogram, that are part of three branches with the sparation of varieties calia, sefiros and farihame with the same fingerprint profile (figure 1). the first group of genotypes were characterized with only one amplified fargment with the length of 150 bp from which genotypes yialousa and kalika were separeted having one more amplicon with the length of 348 bp. second branch of the dendrogram comprises from groups with amplicon of the 182 bp length and those that have this one plus amplicon with the lenght of 348 bp. figure 1. dendrogram of chickpea accession using the profilin specific fingerprinting. for pisum sativum l., very different specific profilin fingerprints were obtained. for individual varieties, from 7 to 12 amplicons were generated with the length within the range from 95 bp up to the 1350 bp. the polymorphism in the analysed set of pea varieties was of 96%. different groups were obtained in the constructed dendrogram, with a separation of varieties lancet, libochovicky urodny, frostar and parade (figure 2). the most of analysed accessions were grouped with the average jaccard coefficient of genetic similarity of 0.47 and two of analysed pea varieties (cicero and skagid) have the same profile of generated amplicons. an amplicon of the length 95 bp was amplified in all of the analysed pea varieties. profilins belong to actin binding molecules and are defined as plant panallergens, what provide them as markers universal in their use [32]. for legume species soybean and groundnut genotypes, profilins were previously utilized by using degenerated primer pair in pbap (profilin based amplified polymorphism) [25, 26]. for both of this species, polymorphic profiles were generated. in the set of groundnut accessions, all were distinguishable but profiling based marker technique, but in soybean, a couple of varieties (sciaming and krajina) produced the same fingerprint profiles. pbap fingerprints of arachis hypogaea l. were distributed in the length ranging from 78 bp up to the 1642 bp and for soybean from 118 bp up to 1000 bp. this correspond to the finding of this study, that profilin based fingerprints are specie specific for legumes with different levels of polymorphism generated. such differences in the ability of generating polymorphism and distinguishing of analyzed genotypes are in concordance with natural variability of profiling homologs in plants. multiple proteins belonging to profilins were found to have their sequences in higher plants. they are divided into two classes that differ in their distinct expression patterns, mainly in vegetative and reproductive plant tissues [33]. actuallly, more than four hundred plant profilin proteins sequences are stored at ncbi (national center for biotechnology information) gene database [34]. it was reported previously that even small changes in the sequence of amino acid can alter the agriculture and food sciences research, 2025, 12(1): 49-53 52 © 2025 by the authors; licensee asian online journal publishing group biochemical properties of profilin substantially and a results in sequence diversity among profiling genes with their evolutionary origin in the polyphyletic mode [35]. figure 2. dendrogram of pea accession using the profilin specific fingerprinting. the results obtained in this study provide data that profilin based generated polymorphism characteristics provide a good source of germplasm variability in the analysis of legumes. references [1] 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[35] d. k. pandey and b. chaudhary, "evolution of functional diversity among actin-binding profilin genes in land plants," frontiers in cell and developmental biology, vol. 8, p. 588689, 2020. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 42 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 42-48, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6570 © 2025 by the authors; licensee asian online journal publishing group understanding cowpea yield: a comprehensive analysis of physiological traits' contribution through path analysis ankrumah emmanuel1 ogbonna peter ejimofor2 onyia vincent nduka3 omoigui lucky4 kamara alpha yahya5 ndifon elias m’jaika6 ( corresponding author) 1,6department of crop science, alex ekwueme federal university ndufu-alike, ebonyi state, nigeria. 1email: ankrumahemmanuel@gmail.com 6email: emndi4nn@yahoo.com 2,3department of crop science, university of nigeria nsukka, enugu state, nigeria. 2email: ogbonna@unn.edu.ng 3email: onyia@unn.edu.ng 4,5project-seed system, international institute of tropical agriculture, kano station, sabo bakin zuwo road kano, kano state, nigeria. 4email: l.omoigui@cgiar.org 5email: a.kamara@cgiar.org abstract this study investigates the physiological processes affecting the grain yield of cowpea (vigna unguiculata), a key protein, vitamin, and mineral source in human diets. gaining an understanding of these mechanisms can be crucial for developing high-yielding cowpea varieties in breeding programs. a field experiment was conducted with 30 treatments, including three sowing dates (early august, late august, early september) and ten cowpea genotypes (uam091051-1, uam09-1046-6-1, uam14-126-l33, it99k-573-1-1, it89kd-288, uam14-126-l6, uam14-122-17-7, uam14-123-18-3, uam14-127-20-1-1, and uam14-130-20-4). these treatments were arranged in a split-plot design within a randomized complete block design, replicated three times. key physiological traits like leaf area index (lai), intercepted photosynthetically active radiation (ipar), stomatal conductance, photosynthetic rate, transpiration rate, and chlorophyll content were measured. data collected were analyzed using correlation and path coefficient methods; the results showed significant positive correlations between grain yield and traits like lai, stomatal conductance, and photosynthetic rate. in contrast, the transpiration rate negatively correlated with yield. path analysis revealed that the net photosynthetic rate had the most direct impact on grain yield, highlighting its role in photosynthesis and grain filling. the study suggests that cowpea breeding efforts should focus on improving photosynthetic efficiency and optimizing traits like lai and stomatal conductance to boost cowpea grain yields. keywords: breeding program, correlation coefficient, cowpea, grain yield, path analysis, physiological traits, planting date, yield partitioning. citation | emmanuel, a., ejimofor, o. p., nduka, o. v., lucky, o., yahya, k. a., & m’jaika, n. e. (2025). understanding cowpea yield: a comprehensive analysis of physiological traits’ contribution through path analysis. agriculture and food sciences research, 12(1), 42–48. 10.20448/aesr.v12i1.6570 history: received: 22 november 2024 revised: 26 march 2025 accepted: 7 april 2025 published: 9 april 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 43 2. materials and methods ................................................................................................................................................................... 43 3. results and discussion ................................................................................................................................................................... 44 4. conclusion ......................................................................................................................................................................................... 47 references .............................................................................................................................................................................................. 47 mailto:ankrumahemmanuel@gmail.com mailto:emndi4nn@yahoo.com mailto:ogbonna@unn.edu.ng mailto:onyia@unn.edu.ng mailto:l.omoigui@cgiar.org mailto:a.kamara@cgiar.org https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6570 https://orcid.org/0000-0001-9983-2453 https://orcid.org/0000-0003-4434-7922 https://orcid.org/0009-0004-9446-251x https://orcid.org/0000-0001-8473-2775 https://orcid.org/0000-0002-1844-2574 https://orcid.org/0000-0001-6027-4714 agriculture and food sciences research, 2025, 12(1): 42-48 43 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study is original in its use of path coefficient analysis to quantify the direct, indirect, and total contributions of physiological traits, such as lai, ipar, chlorophyll content, and gas exchange parameters to cowpea grain yield. it provides new insights into trait interactions, supporting targeted breeding for higher, stress-resilient yields. 1. introduction cowpea (vigna unguiculata l. walp) is widely cultivated in the tropical and subtropical regions of africa, primarily for its edible seeds, green pods, and nutritious leaves, which are consumed as vegetables. it serves as a crucial source of calories and protein for millions of people in developing countries [1, 2]. additionally, cowpea is an affordable and important source of protein, vitamins, and minerals in the human diet, and it also serves as healthy fodder for livestock feed [3, 4]. cowpea is grown on 14.13 million hectares globally, yielding 4.51 million metric tons with an average productivity of 387.45 kg/ha [5]. africa, particularly west africa, accounts for over 90% of the world’s production, with nigeria being the leading producer and consumer in the region, generating 3.6 million tons in 2021 [6]. cowpea is an important grain legume that is highly adapted to various agro-ecological environments [7]. its resilience under marginal environmental conditions and its capacity for symbiotic nitrogen fixation make it a critical food security crop in regions prone to environmental stress [8]. despite these attributes, improving cowpea yield remains a significant challenge, especially due to climate variability and soil nutrient limitations [3]. cowpea yield is an intricate trait that is affected by multiple yield-contributing traits controlled by different genes and environmental influences [7]. a correlation study was conducted to examine the relationships between different traits and their respective contributions to yield [9]. several researchers have explored the connection between these traits to identify key factors for selecting high-yielding varieties [10]. the yield of grain legumes is a complex quantitative trait influenced by the interaction of various growth and physiological processes [11]. therefore, cowpea yield improvement requires an understanding of the physiological traits that influence yield performance. furthermore, it was noted that physiological traits, including the leaf area index (lai), the intercepted photosynthetically active radiation (ipar), the concentration of chlorophyll, and the gas exchange parameters (stomatal conductance, photosynthetic rate, and transpiration rate), have a major impact on the accumulation of biomass and the potential yield of grain legumes [12, 13]. however, there is inadequate information on the complex relative contributions of these traits, whether direct or indirect, to grain yield. the path coefficient analysis is essential for use as a criterion for enhancing yield prediction, management, and selection in the improvement of grain yield in cowpea. this could aid in the strategic selection of desirable traits for breeding. this study could fill this gap and gather information on the contributions of various physiological parameters associated with grain yield and yield-related traits. therefore, this study on path analysis was carried out to dichotomize the direct, indirect, and total contributions of key physiological traits on cowpea grain yield, which is an essential approach to identify the critical traits driving yield improvement and to provide an understanding of their interconnectedness. 2. materials and methods 2.1. experimental site field studies were conducted during the 2021 and 2022 main cropping seasons at the teaching and research farm of the department of crop breeding and genetics, joseph sarwun tarka university (jst-uam) in makurdi (7°44ˈn, 8°30ˈe, 686 m asl) in the northern guinea savanna. prior to the establishment of the field trial, soil samples were taken from the location and characterized according to the analytical procedures of the soil laboratory of the department of soil science, university of nigeria, nsukka. weather information was collected from the accuweather stations installed at the trial site. 2.2. cowpea varieties, sowing date and experimental design ten (10) cowpea varieties and three sowing dates were evaluated. the experimental design was a randomized complete block in a split-plot arrangement with three replications. the main plot consisted of the sowing dates of early august, late august, and early september. the cowpea varieties were assigned to the subplot. four mediummaturing and semi-determinate varieties (it99k-573-1-1, uam14-126-l33, uam14-126-l6, and uam09-10511, which mature within 75–80 days), and six indeterminate varieties (it89kd-288, uam14-122-17-7, uam14123-18-3, uam14-127-20-1-1, uam14-130-20-4, uam09-1046-6-1, which mature within 85–90 days) were used. eight of these varieties were developed by the jst-uam, while the other two were developed by the iita. the subplots were 3 × 4 m and consisted of four ridges with 75 cm spacing between the ridges and 20 cm between plant stands on each ridge. the field was disc-harrowed and ridged before planting. the trial was planted as per the scheduled sowing dates: on the 5th of august, 19th of august, and 2nd of september, 2021, respectively. in 2022, it was done on the 4th of august, the 18th of august, and the 5th of september, respectively. seeds of the cowpea varieties were planted at a depth of 3 cm. four seeds were planted and later thinned to two stands per hill. thinning was performed 14 days after sowing. at sowing, the recommended fertilizer rate for legumes in the nigerian savannas of 50 kg of p2o5 in the form of ssp was applied. a mixture of pendimethalin (500 g l−1) and gramoxone (1:1-dimethyl-4,4bipyridinium dichloride), at a rate of 1 l ha−1 each, was applied immediately after sowing using a knapsack sprayer. this was followed by hoe weeding 4 weeks after sowing. agriculture and food sciences research, 2025, 12(1): 42-48 44 © 2025 by the authors; licensee asian online journal publishing group 2.3. data collection and evaluation the two middle ridges were used for data collection. at the vegetative stage, leaf area index (lai) and intercepted photosynthetically active radiation (ipar) were measured simultaneously at the full bloom stage using the accupar model lp-80 par/lai ceptometer, following the procedure reported by kamara et al. [7]. the net photosynthetic rate, stomatal conductance, and transpiration rate were measured simultaneously at the full flowering and podding stages of cowpea using a portable photosynthesis system, model li-6400xt (li-cor biosciences, usa), after calibration according to the manufacturer’s instructions. relative leaf chlorophyll content was measured at the full bloom and podding stages using the konica minolta spad 502 chlorophyll meters, where a minimum of five different measurements was conducted on different plants, and average readings were recorded. at pod maturity, the two middle ridges were used for measuring yield components. pods from the two middle ridges (net rows) were harvested, threshed, and seed weight per plot was obtained. the moisture content of grain samples from each plot was determined using a v-tech 3.5 to 40% grain moisture tester, model sf00000885. grain yield was extrapolated from the total seed weight in grams per plot and adjusted based on moisture content (12%) to obtain grain yield in kilograms per hectare (kgha-1) using the relation below. grain yield (kg haˉ1) = 𝑆𝑒𝑒𝑑𝑠 𝑤𝑒𝑖𝑔ℎ𝑡 𝑝𝑒𝑟 𝑝𝑙𝑜𝑡 (𝑔) 1000 ∗ 100 − moisture (%) 88% ∗ 10000m² 4.5𝑚² simple correlation coefficients between the grain yield (y) and physiological characters (x) and within the physiological traits themselves were worked out using the following equation after [14]. 𝑟𝑥ʏ = 𝑛(∑xy) − (∑x)((∑y) √[𝑛∑х2 − (∑х)2][𝑛∑ү2 − (∑ү)2] where: rxʏ = correlation coefficient between variables x and y, n = number of observations, σxy = sum of the product of x and y values, σx, σy = sums of x and y values, respectively, σx2, σy2 = sums of squared values for x and y. the calculated coefficients were further used to develop the following simultaneous equations, from the general path equation using the poolman equation, in order to partition the correlations into cause and effect by working out the path coefficients (pi). for the dependent variable y (grain yield), the path equation is: y = pxi x1 + px2x2 + px3x3 + ⋯ + pxnxn + residual. where: pxi is the path coefficient (direct effect) of variable xi on y. xi is the independent variables, and residual (pr) is the unaccountable variance by the model. 𝑟𝑌1 = 𝑃𝑌1 + 𝑃𝑌2 × 𝑟12 + 𝑃𝑌3 × 𝑟13 + 𝑃𝑌4 × 𝑟14 + 𝑃𝑌5 × 𝑟15 + 𝑃𝑌6 × 𝑟16 (1) 𝑟𝑌2 = 𝑃𝑌1 × 𝑟21 + 𝑃𝑌2 + 𝑃𝑌3 × 𝑟23 + 𝑃𝑌4 × 𝑟24 + 𝑃𝑌5 × 𝑟25 + 𝑃𝑌6 × 𝑟26 (2) 𝑟𝑌3 = 𝑃𝑌1 × 𝑟31 + 𝑃𝑌2 × 𝑟32 + 𝑃𝑌3 + 𝑃𝑌4 × 𝑟34 + 𝑃𝑌5 × 𝑟35 + 𝑃𝑌6 × 𝑟36 (3) 𝑟𝑌4 = 𝑃𝑌1 × 𝑟41 + 𝑃𝑌2 × 𝑟42 + 𝑃𝑌3 × 𝑟43 + 𝑃𝑌4 + 𝑃𝑌5 × 𝑟45 + 𝑃𝑌6 × 𝑟46 (4) 𝑟𝑌5 = 𝑃𝑌1 × 𝑟51 + 𝑃𝑌2 × 𝑟52 + 𝑃𝑌3 × 𝑟53 + 𝑃𝑌4 × 𝑟54 + 𝑃𝑌5 + 𝑃𝑌6 × 𝑟55 (5) 𝑟𝑌6 = 𝑃𝑌1 × 𝑟61 + 𝑃𝑌2 × 𝑟62 + 𝑃𝑌3 × 𝑟63 + 𝑃𝑌4 × 𝑟64 + 𝑃𝑌5 × 𝑟65 + 𝑃𝑌6 (6) from the above equations py1, py2, py3, py4, py5, and py6 are the path coefficients (direct effect) representing lai, ipar, chlorophyll content, stomatal conductance sc, transpiration rate (tr), and photosynthetic rate (pr), while py2r12, py3r13, py4r14, py5r15, py6r16, ……. py4r64, py5r65 are the indirect effects, while r12 …… r65 are the correlation coefficients. the individual percentage contributions are computed using. 𝑃𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝐷𝑖𝑟𝑒𝑐𝑡 𝐶𝑜𝑛𝑡𝑟𝑖𝑏𝑢𝑡𝑖𝑜𝑛 = ( pi 𝑅2 ) 2 𝑋 100 where, pi = direct path coefficient of the ith independent variable. 𝑅2 = coefficient of determination (sum of the direct and indirect contributions of all variables). while the percentage combined contribution (%) of any two characters were also computed using the following relation. 𝐶𝑜𝑚𝑏𝑖𝑛𝑒𝑑 𝐶𝑜𝑛𝑡𝑟𝑖𝑏𝑢𝑡𝑖𝑜𝑛 = ( (𝑃ᵢ + 𝐼ᵢ)² 𝑅² ) 𝑋 100 dewey and lu [15] where: iᵢ is the sum of the indirect effects of the ith variable on the dependent variable through other variables. pi + iᵢ is the total effect (direct + indirect) of the independent variable. the residual effect (re) was calculated using the relation: 𝑅ₑ = √1 − 𝑅² r2 is the coefficient of determination, which measures the proportion of the total variance in the dependent variable that is explained by the independent variables. 3. results and discussion 3.1. correlation coefficients (r) between grain yield and physiological traits in cowpea the results of the correlation analysis revealed some highly significant (p < 0.01) positive associations between the physiological components assessed and the grain yield of cowpea (table 1). lai showed a strong positive correlation with stomatal conductance (r = 0.746) and photosynthetic rate (r = 0.720), which indicates that increased leaf area contributes to better gas exchange and photosynthetic efficiency. this is in agreement with the studies by oluwasemire and odugbenro [16], who demonstrated that higher lai enhances the plant’s ability to intercept light and perform photosynthesis, leading to improved growth and development. however, the negative correlation with transpiration rate (r = -0.561) suggests that as the leaf area increases, water loss through transpiration decreases, likely due to a more efficient water use system or stomatal regulation. this aligns with the findings of bouranis et al. [17], who discussed the trade-offs between leaf area expansion and water use efficiency in crops. the positive correlations between chlorophyll content and both stomatal conductance (r = 0.495) and photosynthetic rate (r = 0.353) are expected, as higher chlorophyll content is indicative of better light-harvesting capabilities and enhanced photosynthetic activity [18]. the weak negative correlation with transpiration rate (r = -0.329) might suggest that plants with higher chlorophyll content manage water more efficiently, similar to findings in studies on drought-tolerant varieties of crops [19]. the strong positive association recorded between stomatal conductance and photosynthetic rate (r = 0.838) undoubtedly aligns with the physiological role of stomata in regulating co₂ uptake for photosynthesis. as stomatal conductance increases, more co₂ is available for agriculture and food sciences research, 2025, 12(1): 42-48 45 © 2025 by the authors; licensee asian online journal publishing group photosynthetic reactions, which has been well-documented in studies on plant gas exchange [13]. the significant negative relationship between stomatal conductance and transpiration rate (r = -0.684) reflects the role of stomata in balancing water loss and carbon uptake, a common feature in plants adapted to water-limited environments [12]. a strong negative correlation between transpiration rate and photosynthetic rate (r = -0.642) revealed the potential for cowpea varieties with lower transpiration rates to maintain higher photosynthetic efficiency. this finding is supported by research into the water-use strategies of leguminous crops, where reduced water loss through transpiration helps sustain photosynthesis under water stress [12, 16]. there were significant correlations between lai (r = 0.759), chlorophyll content (r = 0.490), stomatal conductance (r = 0.684), photosynthetic rate (r = 0.645), and grain yield, suggesting that traits improving carbon assimilation and water use efficiency directly contribute to yield formation. this agrees with several studies emphasizing the importance of these traits in yield improvement under various stress conditions [16, 19-21]. conversely, the negative correlation between transpiration rate and grain yield (r = -0.572) further reinforces the idea that efficient water use is critical for optimizing yield in cowpea, a finding supported by studies on the physiological responses of cowpea to drought [21]. however, the lack of a significant relationship between ipar and grain yield in this study contrasts with some literature that highlights the importance of ipar in determining crop yield potential [18, 22, 23]. nevertheless, the weak negative association with chlorophyll content suggests that under certain conditions, higher radiation interception does not always translate to better photosynthetic performance, especially if other limiting factors, such as nutrient or water availability, constrain growth. table 1. matrix of correlation coefficients (r) showing the association between grain yield and some physiological parameters of cowpea. parameters lai ipar chlorophyll content stomatal conductance transpiration rate photosynthetic rate grain yield lai 1 ipar -0.087 1 chlorophyll 0.487** -0.271** 1 stomatal conductance 0.746** -0.057 0.495** 1 transpiration rate -0.561** 0.001 -0.329** -0.675** 1 photosynthetic rate 0.720** -0.038 0.353** 0.838** -0.642** 1 grain yield 0.759** -0.060 0.490** 0.684** -0.572** 0.645** 1 note: ** correlation is significant at the 0.01 level. ipar: intercepted photosynthetic rate. lai: leaf area index 3.2. direct, indirect and total contributions of some physiological characters to grain yield in cowpea the results of the path coefficient revealed that leaf area index (lai) and intercepted photosynthetically active radiation (ipar) recorded weak to moderate negative direct effects (-0.187 and -0.330, respectively), but strong positive total contributions to grain yield (0.759 and 0.760, respectively) (table 2). the negative direct effects suggest that dense canopy architecture and suboptimal light interception can cause yield reduction as a result of mutual shading and resource competition [20]. however, their high indirect contributions through photosynthetic rate emphasize the importance of canopy architecture in driving efficient utilization of light intercepted [20]. even though the negative direct effect (-0.217) of chlorophyll content on grain yield exists, its strong positive total association (0.890) suggests that its contribution is highly effective (0.822) via indirect paths, mainly through improvement in photosynthetic rate. this finding is in line with the role of chlorophyll as the primary pigment for light absorption during photosynthesis [24]. this means that increasing chlorophyll content alone may not directly increase the grain yield of cowpea; however, maintaining optimal chlorophyll levels is vital for optimizing light interception and carbon assimilation [19]. the study also revealed that stomatal conductance displayed a strong positive direct effect (0.572) on cowpea grain yield (table 2), indicating its critical role in co₂ and water vapor exchange regulation between the leaves and the atmosphere [25]. the current study aligns closely with the findings of chikov and akhtyamova [19], who reported that efficient stomatal conductance can improve photosynthetic rates by ensuring adequate co₂ uptake while minimizing water loss, which is crucial in maintaining photosynthesis in varying environmental conditions. the highly strong indirect effect through the net photosynthetic rate (0.915) confirmed the independent report by boukar et al. [8], who stated that the reinforcement in stomatal conductance indirectly impacts crop yield performance by optimizing the photosynthetic machinery. however, under extreme drought conditions, high stomatal conductance may lead to high transpiration rates and a potential negative impact on grain yield, as reported by bouranis et al. [17]. the transpiration rate had a moderate positive (0.339) direct effect; however, it had a strong negative (-0.572) total contribution to grain yield (table 2). this conflicting effect highlights the dual responsibility of transpiration in plant physiology. while transpiration is necessary for cooling and nutrient uptake, according to bouranis et al. [17], a higher transpiration rate can deplete water resources, especially under drought conditions. the highly strong negative (-0.701) indirect contribution through the photosynthetic rate emphasizes that high transpiration may affect stomatal regulatory function by closure, leading to a decline in the influx and uptake of co₂, thus resulting in a decrease in photosynthesis. the path analysis showed that the photosynthetic rate has the highest (1.092) direct effect on grain yield (table 2). this finding aligns with the central role of photosynthesis in biomass accumulation and grain filling, as it is the fundamental process converting light energy into chemical energy [26]. previous studies have shown that enhancing photosynthetic efficiency can significantly increase yield, especially in c3 plants like cowpea, which often suffer from photorespiration losses [10, 22]. additionally, the strong indirect effects of the photosynthetic rate through physiological traits like lai (0.786) and stomatal conductance (0.915) indicate the importance of strengthening photosynthetic capacity while optimizing canopy architecture and efficiency in gaseous exchange to achieve higher cowpea grain yields. agriculture and food sciences research, 2025, 12(1): 42-48 46 © 2025 by the authors; licensee asian online journal publishing group table 2. path analysis for direct (diagonal), indirect (off diagonal) and total contributions effect of some physiological traits to grain yield of cowpea planted during 2021 to 2022 rainfed seasons. parameters lai ipar chlorophyll content stomatal conductance transpiration rate photosynthetic rate total corr. lai -0.187 0.029 -0.105 0.426 -0.190 0.786 0.759** ipar 0.016 -0.330 0.059 0.318 0.000 0.697 0.760** chlorophyll content -0.091 0.090 -0.217 0.397 -0.112 0.822 0.890** stomatal conductance -0.139 -0.184 -0.150 0.572 -0.229 0.915 0.784** transpiration rate 0.105 0.000 0.071 -0.386 0.339 -0.701 -0.572 photosynthetic rate -0.134 -0.211 -0.163 0.479 -0.218 1.092 0.845** note: lai = leaf area index. ipra = intercepted photosynthetic active radiation. ** significant probability at 0.01 level. 3.3. direct and combined contributions (%) of yield components to grain yield in cowpea the highest percentage (119.2%) direct contribution of photoassimilate to grain yield of cowpea was exhibited by the net photosynthetic rate (figure 1). this is expected because the capacity to produce high amounts of photoassimilates and efficient partitioning of carbon compounds towards harvestable organs has shown a major impact on crop yield [18, 20]. an increase in the net photosynthetic rate translates into improved dry matter accumulation, translocation, and efficient partitioning of photoassimilates for optimal grain filling [22, 27]. however, its negative percentage indirect contribution (-24.7%) may indicate competitive allocation of assimilates to vegetative growth and possible potential imbalances in the source-sink dynamics [22]. the percentage contribution of stomatal conductance was significant, with a 32.7% direct impact on grain yield (figure 1). this explains the efficient regulatory role of stomatal conductance in optimizing the rate of co₂ entry into the leaf mesophyll membrane and the exit of water vapor, thus influencing photosynthetic efficiency [18, 28]. high stomatal conductance is associated with better gas exchange and higher transpiration rates, which often improve yield in sufficient moisture ecology [22]. the moderate indirect contributions of about 21.2% suggest the interconnectedness of stomatal conductance with other physiological processes in cowpea for the maintenance of metabolic gaseous exchange equilibrium and efficient nutrient uptake. the contributions of about 11.5% of the transpiration rate (figure 1) directly into cowpea grain are physiologically linked to its role in relatively maintaining leaf temperature and driving nutrient flow through the plant [10]. however, about -91.1% negative indirect contribution indicates that under extreme transpiration, certain conditions, like drought, especially at critical cowpea developmental phases, can lead to excessive loss of water through the stomata, resulting in poor grain filling and a subsequent negative impact on grain yield. even though intercepted photosynthetic active radiation contributed relatively 10.9% modestly directly to grain yield, in contrast to about 109% high indirect contribution. canopy architectural characteristics have been reported to primarily affect light interception efficiency of plants [16, 18, 20]. the huge indirect contributions recorded imply the magnificent role displayed by ipar through the synergistic interactions with other physiological traits like leaf area and chlorophyll content, which translate to canopy light-use efficiency [16]. the percentage direct contribution (4.7%) of chlorophyll was significantly low (figure 1); however, the very high indirect contribution of about 110.7% to the grain yield highlights that chlorophyll concentration alone is not a direct driver of grain yield in cowpea, but rather enhances the photosynthetic capacity jointly with other physiological traits like lai and ipar [16, 18, 20]. a high chlorophyll content has been reported to correspond to better quality light absorption by the chlorophyll pigments, which facilitates more efficient carbon assimilation [17]. the result of the current study showed that lai contributed the lowest percentage (3.5%) of direct contribution to grain yield, but had a strong indirect effect (94.5%) on cowpea grain yield. lai determines the total leaf area available for light capture and gaseous exchange, thus playing a fundamental role in determining crop growth potential [10]. its strong indirect effect could be attributed to its interaction with ipar and photosynthetic traits, which together define the crop’s radiation-use efficiency [29]. figure 1. direct and indirect contributions (%) of physiological traits on cowpea grain yield. figure 2 presents a summary path diagram illustrating the associations between various physiological traits and their effects on cowpeas' grain yield. agriculture and food sciences research, 2025, 12(1): 42-48 47 © 2025 by the authors; licensee asian online journal publishing group figure 2. summary path diagram showing association of some effects of physiological traits to grain yield of cowpea. 4. conclusion the simple correlation analysis revealed a highly significant and positive association between and among most of the physiological components assessed and grain yield. the path coefficient analysis explained the contribution of physiological traits to cowpea grain yield through direct and indirect pathways. photosynthetic rate and stomatal conductance had highly significant direct effects, making them primary targets for breeding programs aimed at enhancing yield. on the other hand, traits like lai, ipar, and chlorophyll content contribute mainly through indirect interactions, suggesting that these traits are critical for improving the overall efficiency of the photosynthetic system and canopy structure. references [1] b. singh, h. a. ajeigbe, s. a. tarawali, s. fernandez-rivera, and m. abubakar, "improving the production and utilization of cowpea as food and fodder," field crops research, vol. 84, no. 1-2, pp. 169-177, 2003. https://doi.org/10.1016/s03784290(03)00148-5 [2] a. y. kamara et al., "integrating planting date with insecticide spraying regimes to manage insect pests of cowpea in north-eastern nigeria," international journal of pest management, vol. 56, no. 3, pp. 243-253, 2010. https://doi.org/10.1080/09670870903556351 [3] c. musvosvi, "morphological characterisation and interrelationships among descriptors in some cowpea genotypes," j. afri. crop sci., vol. 9, pp. 501-507, 2009. 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[28] m. m. barbour, s. bachmann, u. bansal, h. bariana, and p. sharp, "genetic control of mesophyll conductance in common wheat," new phytologist, vol. 209, no. 2, pp. 461-463, 2016a. https://doi.org/10.1111/nph.13628 [29] m. m. barbour, j. r. evans, k. a. simonin, and s. von caemmerer, "online co 2 and h2o oxygen isotope fractionation allows estimation of mesophyll conductance in c4 plants, and reveals that mesophyll conductance decreases as leaves age in both c4 and c3 plants," new phytologist, vol. 210, no. 3, pp. 875-889, 2016b. https://doi.org/10.1111/nph.13830 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.4236/ajps.2014.55080 https://doi.org/10.1111/j.1365-3040.2005.01493.x http://dx.doi.org/10.1371/journal.pone.0073921 https://doi.org/10.2135/cropsci2007.07.0397 http://dx.doi.org/10.4236/as.2014.54028 https://doi.org/10.1111/j.1365-3040.2009.02016.x https://doi.org/10.1078/0176-1617-00887 https://doi.org/10.1111/nph.14171 https://doi.org/10.1111/nph.15923 https://doi.org/10.1111/nph.13628 https://doi.org/10.1111/nph.13830 104 © 2024 by the author; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 104-109, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6178 © 2024 by the author; licensee asian online journal publishing group evaluation of physico-chemical properties of unripe plantain peels as affected by different drying temperature regimes enobong okon umoh department of agricultural engineering, akwa ibom state university, ikot akpaden, uyo, nigeria. email: enobongumoh@aksu.edu.ng abstract this study is aimed at evaluating the physico-chemical properties of unripe plantain peel, as affected by different drying temperature regimes. the plantain peels were subjected to different drying temperatures of 60, 75, 90 and 105 oc for 12 hours, using a laboratory oven to produce flour samples. standard analytical methods were used in evaluating the flour sample parameters. results of the physico-chemical analysis of the flour samples showed that samples dried at 60, 75, 90 and 105 oc had moisture contents of 2.67, 2.00, 1.38 and 0.66%, respectively; ash contents of 15.83, 15.35, 15.08 and 14.97%, respectively; fibre contents of 12.84, 13.34, 13.68 and 14.10%, respectively; protein contents of 9.80, 8.40, 7.35 and 7.00%, respectively; lipid contents of 77.43, 7.55, 7.80 and 7.94%, respectively; carbohydrate contents of 54.09, 55.02, 56.32 and 56.43%, respectively; caloric values of 321.47, 322.63, 324.32 and 325.74 kcal, respectively. there was a significant difference (p < 0.05) in moisture, ash, fibre, protein, lipid and carbohydrate contents, as well as the caloric value of the unripe plantain peel flour samples. the different drying temperature regimes had significant effect on the physico-chemical properties of the unripe plantain peel. moisture, ash, fibre and protein contents decreased with corresponding increase in drying temperature, while lipid content, carbohydrate content and caloric value increased with increase in drying temperature. keywords: evaluation, physico-chemical properties, plantain peel, temperature regimes, unripe plantain, flour. citation | umoh, e. o. (2024). evaluation of physico-chemical properties of unripe plantain peels as affected by different drying temperature regimes. agriculture and food sciences research, 11(2), 104–109. 10.20448/aesr.v11i2.6178 history: received: 17 october 2024 revised: 14 november 2024 accepted: 20 november 2024 published: 2 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. contents 1. introduction .................................................................................................................................................................................... 105 2. materials and methods ................................................................................................................................................................. 105 3. results and discussions ................................................................................................................................................................ 106 4. conclusions ..................................................................................................................................................................................... 108 reference.............................................................................................................................................................................................. 108 mailto:enobongumoh@aksu.edu.ng https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6178 https://orcid.org/0000-0003-0970-3493 agriculture and food sciences research, 2024, 11(2): 104-109 105 © 2024 by the author; licensee asian online journal publishing group contribution of this paper to the literature this research paper contributes to the literature in the area of food processing technology, food value addition, and converting agricultural waste materials to useful food product, which ultimately contributes in food security. 1. introduction plantain (musa paradisiaca) with a world production index of 20.5 million tons per year is a very important food of the tropics and subtropics, where both ripe and unripe foods are consumed [1]. plantain is a versatile and very nutritious food staple that is widely consumed, either ripe or unripe. they are a great source of carbohydrates, having a nutritional value that is similar to that of yam or potato. in nigeria, they are eaten as snacks in the form of chips, as plantain amala, or dodo ikire [2]. unripe and mature plantain could be ground into flour. typically, in nigeria, flour is turned into amala, a low-glycemic gruel that is frequently advised for diabetics. plantain is rich in healthy components such as phenols, carotenoids and dietary fiber [3-5]. the peel of foods contains a higher proportion of bioactive compounds compared to the pulp [6]. moreover, the antioxidant activity of the peel is greater than that of the food itself [7]. research on some nutritional properties of thirteen plantain cultivars after proximate analysis revealed that moisture content in cultivars (peels) ranged from 78.74% to 87.33%; ash content varied between 0.87% and 2.38%; protein content ranged from 1.67% to 4.2%; lipid content varied from 0.84% to 2.24%; fibre values oscillated between 2.38% and 3.72%; dry matter content fluctuated between 12.67% and 21.26%; and carbohydrate content ranged from 88.84% to 92.91% [8]. the chemical makeup of banana and plantain peels, which is influenced by the ripeness of the fruit, determines their ultimate purpose. on a dry basis, unripe fruit peel contains 33–43% total dietary fiber, 6–10% protein, 6–12% ash, 2–6% lipids, and 11–39% starch [9]. additionally, dopamine, a potent antioxidant, is abundant in plantain peels. the antioxidant in plantains is thought to be mostly responsible for catecholamine [6]. plantain peel has been used for generations as a traditional treatment in various countries, according to imam and akter [10]. plantain peel extract has the potential to be utilized as a functional component in the food sector, according to pereira and maraschin [11]. to enhance the functionality of bioactive substances, like phenolic compounds found in plantain peel, they must be extracted from the peel and then encapsulated to prevent deterioration and increase storage effectiveness. for the production of functional plantain peel powder, optimal extraction and effective encapsulation are consequently crucial. plantain peels are important raw materials for several products yet termed useless and unproductive which makes the cost of the raw material reliant on the food excluding the peels in the market because consumers do not recognize the value of its peels, thus they are thrown away. an environmental issue is the careless way in which this peel trash is being disposed of. peels from a wide range of fruits and meals are now being promoted as a natural source of nutritional fiber and antioxidants. for these reasons, plantain peel has gained popularity as new studies indicate that it is a highly beneficial source of antioxidants and dietary fiber. the aim of this study is to evaluate the physico-chemical properties of unripe plantain peels, as affected by different drying temperature regimes. 2. materials and methods 2.1. sample collection and preparation fresh unripe plantain was bought from the local market in ikot akpaden, mkpatenin l.g.a., akwa ibom state, nigeria. the unripe plantain was cleaned, peeled and the peels made into chips, of uniform sizes. the fresh chips oven-dried for 12 hours at 60, 75, 90 and 105 oc, respectively, andthen milled using food grade model sk30-ss disc attrition mill, manufactured by munson machinery co. inc., new york, sieved with laboratory sieve of 600 µm aperture size, and the sample stored for laboratory analyses [2]. 2.2. determination of physico-chemical properties 2.2.1. moisture content determination moisture content was determined according to the standard method of analysis of the aoac [12] as highlighted by umoh and iwe [13]. a beaker that had been cleaned and dried in the oven was weighed after it had cooled in a desiccator (a). the beaker was weighed after adding two grams (2 g) of the flour mix sample, and the weight of the beaker plus sample was recorded (b). after being dried for four hours at 105 °c in the oven, the beaker and its contents were swiftly placed into a desiccator to cool before being weighed again. this process was carried out repeatedly until a consistent weight, (c) was achieved. equation 1 was used to compute the moisture content. calculation: moisture content (%) = loss in weight due to drying weight of sample taken = b−c b−a × 100(1) where: a = weight of empty beaker (g) b = weight of beaker + sample (g) c = weight of beaker + sample after drying (g) 2.2.2. ash content determination a muffle furnace (model skl-1200) was used to ignite a crucible with a lid at 105 °c for one hour. after that, it was weighed and placed in a desiccator to cool (a). the pre-weighed crucible was filled with two grams (2g) of the flour sample, and the weight of the crucible and its content taken (b). in a fume cupboard, bunsen flame was used to burn the crucible and its contents until the smoking stopped. the crucible and content were moved into a muffle furnace and heated to 550 °c for two hours, or until a white ash formed. the crucible was then cooled, covered, and put in a desiccator, before it was weighed (c). equation 2 was used to determine the amount of ash content in the sample [12]. ash (%) = b−c b−a × 100(2) agriculture and food sciences research, 2024, 11(2): 104-109 106 © 2024 by the author; licensee asian online journal publishing group where: a = weight of crucible (g), b = weight of crucible + sample (g), c = weight of crucible + ash (g). 2.2.3. crude fibre determination petroleum ether was used to defatten two grams (2 g) of the sample for two hours. 200 milliliters of sulfuric acid (h2so4) solution were used to boil it for 30 minutes. it was then filtered through linen on a fluted funnel and rinsed with hot water until the washings were no longer acidic. after being moved to a beaker, the residue was heated for a further thirty minutes with 200 milliliters of naoh solution. it was then filtered, and boiling water was used to wash the residue multiple times until it was clear of base sodium hydroxide (naoh). ultimately, the residue underwent two methanol washes, was quantitatively moved into a crucible that had been weighed beforehand, and was oven dried at 105 °c (io). it was weighed (ia), chilled in a desiccator, and burned at 550 °c. after burning, the weight loss was also recorded. using equation 3, the crude fiber content was determined [12]. crude fibre (%)= ia−io weight of original sample taken x 100 (3) 2.2.4. crude protein determination crude protein was determined using the kjeldahl method, as explained by umoh and iwe [13]. a standard 250 ml kjeldahl flask was filled with precisely one gram (1g) of the wheat sample, 1.5 g of catalyst (cuso4), 1.5 g of na2so4, and 5 ml of concentrated h2so4. to prevent foaming, the digesting (kjeldahl) flask was placed on a heating mantle and heated gradually over a number of hours to create a clear, blue solution. the digested solution was quantitatively moved to a 100 ml standard flask and filled with distilled water to the proper level after being allowed to cool. an equivalent volume of 40% naoh solution was added to a 20 milliliter (20 ml) portion of the digest after being pipette into a semi-micro kjeldahl distillation unit. a 100 ml conical flask containing a 10 ml saturated boric acid solution and two drops of tashirus indicator (double indicator) were added to the ammonia after it had undergone a steam distillation procedure. the distillation process was continued until around two thirds of the original volume was recovered after the condenser tip was immersed in the boric acid-double indicator solution. after titrating the distillate with 0.1m hydrochloric acid (hcl) solution until a purple-pink end-point was observed, a few milliliters of distilled water were used to cleanse the condenser's tip. with the exception of excluding the sample, a blank determination was also carried out in the same manner as described above. equation 4 was used to determine the crude protein content [12]. (sample titre−blank titre)0.1×0.014 weight of sample × 100 20 × 100 1 × 6.25 (4) 2.2.5. crude fat determination the extraction thimble was filled with two grams (2 g) of the flour samples after it had been cleaned, dried in the oven, and slightly blocked with cotton wool. a round bottom flask with a capacity of 500 ml was filled with 150 ml of petroleum ether, which has a boiling point ranging from 35 to 60 ℃. the round-bottom flask was placed on a heating mantle and the soxhlet extractor was inserted. after being put together, the soxhlet device was left to reflux for almost four hours. a dried, pre-weighed beaker (w1) was filled with the extract, and the thimble was then cleaned with some ether before being returned to the beaker. after being heated in a steam bath to remove extra solvent, the beaker was cooled in a desiccator and weighed (w2). the crude fat content was calculated using equation 5 [12]. crude fat (%) = weight gain in flask weight of sample × 100 1 = w2 − w1 weight of sample × 100 1 (5) 2.2.6. carbohydrate content determination determination of carbohydrate content of the sample was carried out by subtracting the total estimations (in %) made of nitrogen (protein), fat, ash and crude fibre contents from 100%. the resultant value gave the percentage carbohydrate content by difference, using equation 6 [12]. carbohydrate (%) = 100 − (a + b + c + d) (6) where, a = percentage protein content. b = percentage lipid content. c = percentage ash content. d = percentage fibre content. 2.2.7. caloric value determination this was obtained using the atwater factor method. the values of the crude protein, fat, and carbohydrate obtained were multiplied by 4, 9, 4 respectively, and the sum of the products was taken as the caloric or energy value, (in kcal) of the sample, using equation 7 [12]. energy value (kcal) = (x × 4) + (y × 9) + (z × 4) (7) where, x = protein content. y = lipid content. z = carbohydrate content. 2.3. statistical analysis the triple-checked data were analyzed using the statistical analysis of variance (anova), with a 5% threshold of significance applied to the mean separation using the duncan's multiple range test. 3. results and discussions the results of the physico-chemical properties of unripe plantain peel flour are presented in table 1. agriculture and food sciences research, 2024, 11(2): 104-109 107 © 2024 by the author; licensee asian online journal publishing group table 1. physico-chemical properties of unripe plantain peels flour. sample temp (oc) moisture content (%) ash (%) fibre (%) protein (%) lipid (%) carbohydrate (%) caloric value (kcal) a 60 2.67±0.003a 15.83±0.010a 12.84±0.022d 9.80±0.003a 7.43±0.004d 54.09±0.011d 321.47±0.002c b 75 2.00±0.011b 15.35±0.022b 13.34±0.001b 8.40±0.022b 7.55±0.006c 55.02±0.002c 322.63±0.026d c 90 1.38±0.001c 15.08±0.002c 13.68±0.006c 7.35±0.020c 7.80±0.032b 56.32±0.022a 324.32±0.041a d 105 0.66±0.100d 14.97±0.004d 14.10±0.021a 7.00±0.201d 7.94±0.024a 56.43±0.006b 325.74±0.021b note: values are mean ± standard deviation of triple determination. mean values with different superscripts, a, b, c, d, are significantly different (p< 0.05) from each other. agriculture and food sciences research, 2024, 11(2): 104-109 108 © 2024 by the author; licensee asian online journal publishing group 3.1. moisture content results of the physico-chemical analysis of the unripe plantain peel flour samples indicated that sample dried at 60, 75, 90 and 105 oc had moisture contents of 2.67, 2.00, 1.38 and 0.66%, respectively. sample dried at the lowest temperature (sample a) recorded the highest moisture content, while the highest drying temperature produced the sample with the lowest moisture content (table 1). there was a significant difference (p < 0.05) in the moisture contents of the samples. the moisture content of the unripe plantain peel decreased with increase in the drying temperature. therefore, drying temperature significantly affected the moisture content of the unripe plantain peel. moisture content of samples is presumed as one of the most important determination of shelf stability. the moisture content values for the flour samples ensure adequate storage in packages [14]. 3.2. ash content the unripe plantain peel recorded ash contents of 15.83, 15.35, 15.08 and 14.97% for samples dried at 60, 75, 90 and 105 oc, respectively (table 1). these recorded values are higher than 3.94 to 4.28%, earlier reported for green plantain flour [2]. the high ash content is an indication that unripe plantain peel is a rich potential source of mineral elements. similarly, there was a significant difference (p < 0.05) in the ash contents of the unripe plantain flour samples. increase in drying temperature resulted in decrease in ash content of the sample. thus, drying temperature had a significant effect on the ash content of the unripe plantain peel. 3.3. fiber content fibre contents of the unripe plantain peel were 12.84, 13.34, 13.68 and 14.10% for samples dried at 60, 75, 90 and 105oc, respectively (table 1). these values are higher than the range of values, 1.00 to 2.76%, earlier reported for aerial yam-soybean flour [14]. there was a significant difference (p < 0.05) in the fibre contents of the samples. fibre content of the sample increased with a corresponding increase in drying temperature. the drying temperature significantly affected the fibre content of the unripe plantain peel. 3.4. protein content the unripe plantain peel flour samples recorded protein contents of 9.80, 8.40, 7.35 and 7.00% for samples dried at 60, 75, 90 and 105 oc, respectively (table 1). these recorded values are higher than 3.85 to 4.68%, earlier reported for green plantain flour [2]. there was a significant difference (p < 0.05) in the protein contents of the unripe plantain peel. however, the protein content decreased with increase in drying temperature. this may be attributed to the process of protein denaturation. therefore, the drying temperature significantly affected the protein content of the unripe plantain peel. 3.5. lipid content results of the physico-chemical evaluation of the unripe plantain peel flour samples showed that lipid contents were 7.43, 7.55, 7.80 and 7.94% for samples dried at 60, 75, 90 and 105 oc, respectively (table 1). there was a significant difference (p < 0.05) in the lipid contents of the unripe plantain peel. increase in drying temperature resulted in increased lipid content of the sample. the drying temperature had a significant effect on the lipid content of the unripe plantain peel. 3.6. carbohydrate content the unripe plantain peel recorded carbohydrate contents of 54.09, 55.02, 56.32 and 56.43% for samples dried at 60, 75, 90 and 105 oc, respectively (table 1). these recorded values are lower than 85.66 to 86.81%, earlier reported for green plantain flour [2]. the low carbohydrates contents imply that unripe plantain peel flour could be recommended as a diet for the diabetics. there was a significant difference (p < 0.05) in the carbohydrate content of the samples. the carbohydrate content increased with increase in the drying temperature. the drying temperature significantly affected the carbohydrate content of the unripe plantain peel. 3.7. caloric value the caloric value of the unripe plantain peel ranged from 321.47 to 325.74 kcal. this range of values is a bit lower than 421.75 to 429.34 kcal for aerial yam-soybean flour, earlier reported by umoh [14] and 380.29 to 385.23 kcal for green plantain flour [2]. samples dried at 60, 75, 90 and 105 oc had caloric values of 321.47, 322.63, 324.32 and 325.74 kcal, respectively (table 1). there was a significant difference (p < 0.05) in the caloric values of the unripe plantain peel flour samples. the caloric value increased with a corresponding increase in the drying temperature. consequently, the drying temperature had a significant effect on the caloric value of the unripe plantain peel. 4. conclusions this study has shown that different drying temperature regimes has significant effect on the physico-chemical properties (moisture, ash, fibre, protein, lipid, carbohydrate and caloric value) of unripe plantain peel. moisture, ash, fibre and protein contents decrease with corresponding increase in drying temperature, while lipid content, carbohydrate content and caloric value increase with increase in drying temperature. reference [1] i. macdonald and j. low, fruit and vegetables. nairobi: evans brothers ltd, 1990. [2] e. o. umoh, i. uko, i. a. akpan, and s. p. edem, "effect of different drying methods on the proximate composition and energy value of green plantain (musa paradisiaca) flour," aksu journal of agriculture and food sciences, vol. 8, no. 2, pp. 154-162, 2024. https://doi.org/10.61090/aksuja.2024.026 [3] r. baskar, s. shrisakthi, b. sathyapriya, r. shyampriya, r. nithya, and p. poongodi, "antioxidant potential of peel extracts of banana varieties (musa sapientum)," food and nutrition sciences, vol. 2, no. 10, pp. 1128–1133, 2011. 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[9] e. agama-acevedo, j. a. sañudo-barajas, r. vélez de la rocha, g. a. gonzález-aguilar, and l. a. bello-peréz, "potential of plantain peels flour (musa paradisiaca l.) as a source of dietary fiber and antioxidant compound," cyta journal of food, vol. 14, no. 1, pp. 117-123, 2016. https://doi.org/10.1080/19476337.2015.1055306 [10] m. z. imam and s. akter, "musa paradisiaca l. and musa sapientum l.: a phytochemical and pharmacological review," journal of applied pharmaceutical science, vol. 1, no. 5, pp. 14-20, 2011. [11] a. pereira and m. maraschin, "banana (musa spp) from peel to pulp: ethnopharmacology, source of bioactive compounds and its relevance for human health," journal of ethnopharmacology, vol. 160, pp. 149-163, 2015. https://doi.org/10.1016/j.jep.2014.11.008 [12] aoac, official methods of analysis of the association of official analytical chemists, 15th ed. arlington, virginia, usa: aoac, 2010. [13] e. o. umoh and m. o. iwe, "effects of extrusion processing on the proximate compositions of aerial yam (dioscorea bulbifera)soybean (glycine max) flour blends using response surface methodology," journal of food research, vol. 11, no. 1, pp. 38-52, 2022. https://doi.org/10.5539/jfr.v11n1p38 [14] e. o. umoh, "evaluation of proximate composition, functional properties and antinutritional factors of aerial yam-soybean flour," international journal of food science and nutrition, vol. 5, no. 1, pp. 70-74, 2020. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.jff.2017.11.006 https://doi.org/10.1016/s0308-8146(02)00186-3 https://doi.org/10.1080/19476337.2015.1055306 https://doi.org/10.1016/j.jep.2014.11.008 https://doi.org/10.5539/jfr.v11n1p38 19 © 2025 by the author; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 19-24, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6555 © 2025 by the author; licensee asian online journal publishing group effect of extrusion process parameters on the overall acceptability of snacks produced from aerial yam and soybean flour mixture umoh, enobong okon department of agricultural engineering, akwa ibom state university, ikot akpaden, p.m.b., 1167, uyo, nigeria. email: enoumoh40@gmail.com abstract this study was conducted to evaluate the effect of extrusion cooking parameters on the overall acceptability of the snacks produced from aerial yam and soybean flour mixture. design expert (version 11.0.1) was used in the experimental design, with a three-factor experimental setup at five levels each. the aerial yam and soybean flour mixture was formulated in the ratio of 1:3 and extruded using a laboratory-scale single-screw extruder. response surface methodology was adopted in analyzing the effect of the independent variables on the dependent variable. results showed that the overall acceptability scores ranged from 4.20 to 7.10. response surface analysis revealed that an increase in barrel temperature, screw speed, and feed moisture resulted in an increase in overall acceptability. the overall acceptability of the snacks was significantly (p < 0.05) affected by barrel temperature and feed moisture, while the screw speed had no significant (p > 0.05) effect on overall acceptability. the interaction of the extrusion cooking parameters had a significant (p < 0.05) effect on the overall acceptability of the snacks. the high range of scores recorded for the snacks indicates that the aerial yam and soybean composite flour extruded snacks were generally well accepted by the panelists. keywords: aerial yam flour, extrusion cooking, overall acceptability, response surface, single-screw extruder, soybean flour, snacks. citation | okon, u. e. (2025). effect of extrusion process parameters on the overall acceptability of snacks produced from aerial yam and soybean flour mixture. agriculture and food sciences research, 12(1), 19–24. 10.20448/aesr.v12i1.6555 history: received: 21 february 2025 revised: 24 march 2025 accepted: 28 march 2025 published: 4 april 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. contents 1. introduction ...................................................................................................................................................................................... 20 2. materials and methods ................................................................................................................................................................... 20 3. results and discussions .................................................................................................................................................................. 21 4. conclusion ......................................................................................................................................................................................... 23 references .............................................................................................................................................................................................. 23 mailto:enoumoh40@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6555 https://orcid.org/0000-0003-0970-3493 agriculture and food sciences research, 2025, 12(1): 19-24 20 © 2025 by the author; licensee asian online journal publishing group contribution of this paper to the literature this paper presents an empirical study on the effect of extrusion process parameters (barrel temperature, screw speed, feed moisture) on the acceptability of snacks derived from aerial yam and soybean flour mixtures. the study is relevant to food science and agricultural processing, particularly in formulating/developing new extruded food products. 1. introduction mixing, shearing, heating, pumping, shaping, and sizing are just a few of the unit operations that are uniquely combined in the efficient, continuous extrusion process. a high-temperature-short-time (htst) process called extrusion cooking technology is being used more and more in the food industry to create new cereal-based snacks, such as breakfast cereals, baby foods, high-dietary-fiber foods, and modified starch from cereals [1-5]. extrusion cooking, like other methods of heating food, can affect nutritional content in both positive and negative ways. benefits include minimizing lipid oxidation and contaminating microbes, increasing soluble dietary fibers, gelatinizing starch, destroying anti-nutritional elements, and preserving the food's natural colors and flavors. notwithstanding these benefits, the process's drawbacks include the loss of heat-labile vitamins and the occurrence of the maillard reaction, which lowers the protein's nutritional value. for an extruded product to be nutritionally balanced, production factors must be controlled [5, 6]. establishing a trustworthy dataset that can tell processors, vendors, and customers about their preferences depends heavily on the sensory qualities of food [7]. according to umoh et al. [8], sensory qualities that are perceptible to the human senses, such as appearance, flavor, texture, scent, taste, and general acceptability of foods, are commonly used to evaluate the quality of food. kothakota et al. [9] investigated how the extrusion procedure affected the extrudates' overall acceptability. the extruded product, comprising broken rice flour, dehydrated pineapple waste pulp powder, and red grain powder, has an overall acceptance rating of 5.2 to 7.5, according to the investigations. the total acceptability value of the extrudates made from rice flour, red grain powder, and pineapple waste pulp powder was also found to rise quadratically as screw speed and barrel temperature increased, before falling linearly as these parameters decreased. as the feed moisture content rose, so did the total acceptance value, according to kothakota et al. [9]. upadhyay et al. [10] also observed similar findings. the barrel temperature and their interactions, screw speed, and feed moisture have been reported to have a significant effect on the overall acceptability of the extrudates [11]. a perennial, semi-wild food crop, the aerial yam (dioscorea bulbifera) is a member of the dioscoreaceae family of yams and grows on vines that ascend to poles and trees. after cooking, the hard back of the bulb is peeled off and consumed. among its many common names are bitter yam, aerial yam, potato yam, air yam, and air potato. originating in west africa, dioscorea bulbifera is a hardy climbing plant that is grown for its bulbils, which are eaten after being cooked like potatoes in oily water or roasted with a local sauce made of palm oil and additional spices [12]. aerial tubers or bulbils are harvested by manual plucking from the vine. they are included in the roots and tubers, which are widely distributed throughout the tropics, with only a few in the temperate regions of the world [13]. there are roughly 50–60 species of yams (dioscorea spp.) in nigeria, but only 5–6 of them are significant food crops. regrettably, some of these food crops, like dioscorea bulbifera, have been underutilized for their nutritional value [12, 14, 15]. soybeans are cultivated mainly for their seeds, which are used to produce oil and as food for humans and animals. in the food industry, soy food sources are probably the ones that are developing the fastest. products range from traditional soy food assortments to protein trimmings, dairy and meat substitutes, and various western and traditional food assortments that have advanced with soybean flour and its components [16, 17] soybeans are nutrient-dense and have a distinct chemical makeup on an average dry matter basis; they have the highest protein concentration of any legume, with roughly 40% protein and 20% oil [16]. soybeans have high levels of both quantity and quality in their protein and oil components. because soy oil has a high percentage of unsaturated fatty acids, including linolenic and linoleic acids, it is considered a healthy oil. furthermore, soybeans contain a variety of trace chemicals called phytochemicals that have been demonstrated to have special health advantages. the greatest amount of protein obtained comes from soy meals because of their high protein content and high protein utilization [15, 16]. additionally, the use of aerial yam (dioscorea bulbifera) in conjunction with known food processing techniques, such as extrusion cooking, would introduce new food processing technology and food products, providing consumers with variety [15]. the potential application of aerial yam and soybean blending and/or extrusion cooking of the blend in food product formulation has not been sufficiently studied [18]. this study aims to evaluate the impact of extrusion cooking on the overall acceptability of snacks produced from aerial yam and soybean composite flour, using response surface analysis. 2. materials and methods 2.1. procurement of soybean seeds and aerial yam bulbs the aerial yam bulbs and soybean seeds were acquired from the uyo urban market in the uyo local government area of akwa ibom state, nigeria. 2.2. sample preparation in this study, flour samples were made at the department of agricultural engineering's crop processing laboratory at akwa ibom state university, ikot akpaden, akwa ibom state, nigeria. agriculture and food sciences research, 2025, 12(1): 19-24 21 © 2025 by the author; licensee asian online journal publishing group 2.3. preparation of aerial yam flour in accordance with the procedure outlined by umoh et al. [12], aerial yam flour was made by cleaning and sorting the bulbs to remove any unwanted materials, then peeling them with a knife, washing them with clean water, and cutting them into chips that were 10 mm thick. the chips were then dried in a laboratory oven set at 60°c for 12 hours, after which they were milled using an italian-made mf120 hammer mill and sieved through a laboratory sieve with an aperture size of 600 µm. the flour that was produced was then sealed in a polyethylene bag for later use. 2.4. preparation of soybean flour the process outlined by umoh and ekanem [5] was used to prepare the soybean flour. after screening the seeds to remove splits, foreign materials, and damaged beans, they were cleaned and roll-boiled at 100°c for 30 minutes, oven-dried at 70°c for 12 hours, and then ground in a disc attrition mill. the milled full-fat soybean was then sieved using a 100-mesh standard sieve, and the flour was sealed in an airtight polyethylene bag at room temperature for later use. 2.5. preparation of flour mixture sample aerial yam and soybean flour mixture was made in a 1:3 ratio, which consists of 25% aerial yam flour and 75% soybean flour. 2.6. preparation of the extruded snacks a laboratory-scale single-screw extruder was used for this purpose. two hundred grams (200 g) of the composite flour sample were precisely measured, preconditioned to the appropriate moisture levels, and left for approximately two minutes (2 min) to ensure uniform hydration of the raw material. the extruder was turned on, and the barrel temperatures and screw speeds were adjusted accordingly. the raw material was fed into the extruder through the hopper, and the extruded snacks were collected as they came out of the die, oven-dried, and then placed in airtight zip-lock polyethylene bags for further laboratory analysis [5]. 2.7. determination of overall acceptability the overall acceptability (sensory quality attribute) of the aerial yam and soybean composite flour extruded snacks was determined using a 9-point hedonic scale ranging from 1 (extremely dislike), 5 (neither like nor dislike), to 9 (extremely liked) [15]. a ten-member semi-trained panel evaluated the samples and accordingly scored the overall acceptability of the snacks. 2.8. experimental design/statistical analysis the experimental design was created using design expert (version 11.0.1). barrel temperature (bt), screw speed (ss), and feed moisture levels (fm) were chosen as the independent factors in a three-factor experiment that had five levels each. the impact of the independent variables or factors on the dependent variable (the response) was examined using response surface methodology (rsm). 3. results and discussions the results of the overall acceptability of snacks produced from aerial yam and soybean flour mixture are presented in table 1. table 1. overall acceptability of aerial yam and soybean composite flour snacks. s/n bt (oc) ss (rpm) fm (%) overall acceptability 1 105 115 31 5.51 ± 0.011 2 105 115 35 6.43 ± 0.031 3 105 115 35 6.39 ± 0.031 4 105 85 35 5.76 ± 0.111 5 100 130 33 6.42 ± 0.008 6 110 100 37 7.10 ± 0.006 7 100 100 37 6.00 ± 0.021 8 110 130 33 5.45 ± 0.008 9 105 115 35 6.45 ± 0.031 10 115 115 35 5.60 ± 0.001 11 95 115 35 4.89 ± 0.021 12 105 115 39 6.17 ± 0.043 13 105 115 35 6.47 ± 0.031 14 100 100 33 4.20 ± 0.002 15 110 130 37 5.13 ± 0.002 16 110 100 33 5.98 ± 0.021 17 105 145 35 4.99 ± 0.006 18 100 130 37 6.11 ± 0.011 19 105 115 35 6.41 ± 0.031 20 105 115 35 6.44 ± 0.031 note: values are mean ± standard deviation of triplicate determination. bt = barrel temperature, ss = screw speed, fm = feed moisture. table 1 depicts the overall acceptability results of the snacks made with a composite flour of aerial yam and soybean. according to the recorded ratings, the snacks' overall acceptability ranged from 4.20 to 7.10. this range of values is lower than 7.22 to 8.33 for pulse-based snacks [11] but higher than 4.00 to 6.44 previously reported for sorghum-based extrudates supplemented with defatted soy meal flour by tadesse et al. [19]. agriculture and food sciences research, 2025, 12(1): 19-24 22 © 2025 by the author; licensee asian online journal publishing group however, the range of values for overall acceptability for the extruded snacks made from aerial yam and soybean composite flour falls between 5.2 and 7.5 for rice flour-pineapple waste pulp powder-red grain powder extrudates those previously reported by kothakota et al. [9] and those reported to be 5.94 to 7.02 for selected aerial yam cultivars and african breadfruit extruded snacks by olatoye and arueya [20]. the snacks produced at 100°c barrel temperature, 100 rpm screw speed, and 33% feed moisture had the lowest overall acceptability score of 4.20, while the snacks made at 110°c barrel temperature, 100 rpm screw speed, and 37% feed moisture yielded the highest score (7.10). the wide range of scores recorded for the snacks indicates that the extruded aerial yam and soybean composite flour was generally well accepted by the panelists. 3.1. effect of extrusion process on overall acceptability the response surface plot displayed in figures 1 through 3 illustrates how the extrusion process factors (barrel temperature, screw speed, and feed moisture) affect the overall acceptability of the aerial yam and soybean composite flour extruded snacks. figure 1. response surface plot showing the impact of barrel temperature and screw speed on overall acceptability. figure 2. response surface plot showing the impact of barrel temperature and feed moisture on overall acceptability. agriculture and food sciences research, 2025, 12(1): 19-24 23 © 2025 by the author; licensee asian online journal publishing group figure 3. response surface plot showing the impact of screw speed and feed moisture on overall acceptability. the overall acceptance score of the snacks increased when both the barrel temperature and the extruder's screw speed increased, according to the response surface plot for the effects of both variables on the extrudates' overall acceptability in figure1. the findings of kothakota et al. [9] are comparable to this discovery. the response surface plot for the impact of feed moisture and barrel temperature on the snacks' overall acceptability score is displayed in figure 2. the overall acceptability increased when the extruder's barrel temperature was raised. in the meantime, the overall acceptability of the snacks increased in tandem with an increase in feed moisture. this claim supports the earlier report of kothakota et al. [9]. consistent with the findings of kothakota et al. [9] the response surface plot on the effect of screw speed and feed moisture on overall acceptability in figure 3 showed that an increase in both factors led to a corresponding increase in the overall acceptability of the snacks. table 2. analysis of variance for overall acceptability at a 5% significance level. source sum of squares df mean square f-value p-value barrel temp 2.472 3 0.824 1.609 0.0255 screw speed 1.975 3 0.658 1.285 0.338 feed moisture 1.398 3 0.466 0.910 0.0474 error 4.611 9 0.512 total 704.359 20 corrected total 9.339 19 barrel temperature, screw speed, and feed moisture all had p-values of 0.0255, 0.338, and 0.0474, respectively, according to an analysis of variance (anova) at the 5% significance level (table 2). this suggests that screw speed had an insignificant (p > 0.05) impact on the snacks' overall acceptability, whereas barrel temperature and feed moisture had a significant (p < 0.05) impact. the overall acceptability of the snacks was shown to be significantly (p < 0.05) impacted by the extrusion process parameters and their interactions, according to the "test of betweensubjects effects" of the parameters. 4. conclusion this study has demonstrated that the extrusion process parameters—barrel temperature, screw speed, and feed moisture—as well as how they interact, affect the snacks' quality features in both positive and negative ways. the overall acceptability of the aerial yam and soybean composite flour extruded snacks increased as a result of increases in barrel temperature, screw speed, and feed moisture. while screw speed had no discernible impact on overall acceptance, barrel temperature and feed moisture had a considerable impact on the snacks' overall appeal. the overall acceptance of the snacks was significantly impacted by the interactions of the extrusion parameters. references [1] l. sebio and y. chang, "effects of selected process parameters in extrusion of yam flour (dioscorea rotundata) on physicochemical properties of the extrudates," food/nahrung, vol. 44, no. 2, pp. 96-101, 2000. https://doi.org/10.1002/(sici)15213803(20000301)44:2<96::aid-food96>3.0.co;2-9 [2] i. pardeshi and p. chattopadhyay, "whirling bed hot air puffing kinetics of rice-soy ready-to-eat (rte) snacks," journal of ready to eat food, vol. 1, no. 1, pp. 01-10, 2014. 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[11] h. alemayehu, s. a. emire, and c. henry, "effects of extrusion process parameters on the quality properties of ready-to-eat pulsebased snacks," cogent food & agriculture, vol. 5, no. 1, p. 1641903, 2019. https://doi.org/10.1080/23311932.2019.1641903 [12] e. o. umoh, m. o. iwe, and p. ojimelukwe, "optimization of extrusion process parameters for the anti-nutritional composition of aerial yam (dioscorea bulbifera)-soybean (glycine max) flour blends using response surface methodology," international journal of food science and nutrition, vol. 6, no. 6, pp. 62-69, 2021. [13] e. umoh, "evaluation of proximate composition, functional properties and antinutritional factors of aerial yam-soybean flour," evaluation, vol. 5, no. 1, pp. 70-74, 2020. [14] m. ogbuagu, "nutritive and antinutritive composition of the wild (in-edible) species of dioscorea bulbifera (potato yam) and dioscorea dumentorum (bitter yam)," journal of food technology, vol. 6, no. 5, pp. 224-226, 2008. [15] e. o. umoh and k. s. abasiekong, "production and quality evaluation of pasta from aerial yam and soybean flours blend using singlescrew extruder," research journal of food science and quality control, vol. 2756, p. 5483, 2024. https://doi.org/10.56201/rjfsqc.v10.no4.2024.pg24.41 [16] m. o. iwe, the science and technology of soybean: chemistry, nutrition, processing, utilization. enugu, nigeria: rojoint communication services ltd, 2003. [17] e. o. umoh and m. o. iwe, "influence of extrusion process conditions on bulk density, water absorption capacity and oil absorption capacity of extruded aerial yam-soybean flour mixture," african journal of food science, vol. 17, no. 6, pp. 111-121, 2023. [18] e. o. umoh and m. o. iwe, "effects of extrusion processing on the proximate composition of aerial yam (dioscorea bulbifera)soybean (glycine max) flour blends using response surface methodology," journal of food research, vol. 1, no. 1, pp. 38-52, 2022. https://doi.org/10.5539/jfr.v11n1p38 [19] s. a. tadesse, g. bultosa, and s. abera, "functional and physical properties of sorghum-based extruded product supplemented with soy meal flour," cogent food & agriculture, vol. 5, no. 1, p. 1707608, 2019. https://doi.org/10.1080/23311932.2019.1653617 [20] k. olatoye and g. arueya, "chemical and sensory characteristics of extruded snack from selected aerial yam (d ioscorea bulbifera) cultivar and african breadfruit (treculia africana) seed," journal of culinary science & technology, vol. 21, no. 3, pp. 449-465, 2023. https://doi.org/10.1080/15428052.2021.1955795 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.62154/ajasfr.2024.016.010409 https://doi.org/10.22392/actaquatr.1560683 https://doi.org/10.70112/ajsat-2024.13.2.4246 https://doi.org/10.5897/ajfs2013.1065 https://doi.org/10.1080/23311932.2019.1641903 https://doi.org/10.56201/rjfsqc.v10.no4.2024.pg24.41 https://doi.org/10.5539/jfr.v11n1p38 https://doi.org/10.1080/23311932.2019.1653617 https://doi.org/10.1080/15428052.2021.1955795 59 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 59-64, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6076 © 2024 by the authors; licensee asian online journal publishing group agri-food fraud in africa vis-à-vis global health wilfred angie abia1,2 taty brandy perkwang1,2 kaoke matheiu djoussou3 ( corresponding author) 1department of biochemistry, faculty of science, university of yaounde 1, bp 812, yaounde, cameroon. email: abiawilfred@gmail.com 2agri-food safety and one health authority, yaounde, cameroon. email: tatybrandy13@gmail.com 3university of maroua, bp 814 maroua, cameroon. email: kaomath3@gmail.com abstract this paper reports on the extent of agri-food fraud in africa, its occurrence across the africa-eu trade borders, and associated implications for global health. it revealed that agri-food fraud in africa and across africa-eu trade borders may be a concern. meanwhile, a few suspected cases have been reported, there may be more unchecked/unnoticed agri-food fraud practices ongoing in africa requiring food trade border checks and rapid decisions making processes to reduce exposures. exposure to adulterants can have significant implications for global health, as consumers unknowingly pay for, and consume, products that may be adulterated with substances which constitute a risk factor to one or more unchecked hazards against consumers' health protection. as global trade in agri-food products expands, the supply chain vulnerability to fraudulent activities becomes increasingly evident. this may be worse partly due to the inadequate food safety laws in african countries or poor enforcement. there is a need for food safety regulations in africa that aim to ensure the safety and authenticity of food products, protect public health, and maintain consumer confidence. finally, there is a need for rapid detection tools to make quick decisions on the field and agri-food traceability systems. keywords: africa-eu trade border, agri-food fraud, authenticity, consumers’ health, traceability, global health. citation | abia, w. a., perkwang, t. b., & djoussou, k. m. (2024). agri-food fraud in africa vis-à-vis global health. agriculture and food sciences research, 11(2), 59–64. 10.20448/aesr.v11i2.6076 history: received: 2 september 2024 revised: 14 october 2024 accepted: 22 october 2024 published: 6 november 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: brought the idea and coined and designed the study and proofread, w.a.a.; carried out the literature review and performed data analysis, w.a.a. and t.b.p.; prepared the draft manuscript, t.b.p. and k.m.d.; read and approved the final manuscript, t.b.p., k.m.d. and w.a.a. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 60 2. methodology ..................................................................................................................................................................................... 60 3. results and discussion ................................................................................................................................................................... 60 4. conclusion ......................................................................................................................................................................................... 63 references .............................................................................................................................................................................................. 63 mailto:abiawilfred@gmail.com mailto:tatybrandy13@gmail.com mailto:kaomath3@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6076 https://orcid.org/0000-0003-3796-8616 https://orcid.org/0009-0001-2167-988x https://orcid.org/0009-0005-6001-6658 agriculture and food sciences research, 2024, 11(2): 59-64 60 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the agri-food fraud situation in africa and incidence across trade boarders – a scenario which may be largely unchecked requiring authenticity assessment and fraud detection studies in africa. the potential bearing of agri-food fraud in africa on consumers health globally as per the food supply chains. 1. introduction as the world population grows exponentially, nowadays, consumers can hardly ascertain the origin or authenticity of the several scarce and long-supply chain agricultural commodities or foods we pay for. the rate of agri-food fraud is alarming as many sellers, retailers, and individuals nationally and internationally have indulged in this deceiving practice for economic gain. agri-food fraud poses a significant challenge to the integrity of the global food supply chain, with potentially severe consequences for human health. according to the european commission [1] agri-food fraud is “a non-compliance concerning any suspected intentional action by some fraudsters to deceive purchasers and gain an undue advantage thereof, in violation of the rules referred to in article 1(2) of regulation (eu) 2017/625” [2]. the repercussions of agri-food fraud extend far beyond economic losses to profound concerns for consumers’ health [3]. agri-food fraud, encompassing a range of deceptive practices within the agricultural and food sectors, jeopardizes the safety and authenticity of food products [4]. the consequences of consuming adulterated or misrepresented food items can be severe, leading to adverse health effects and compromised nutritional value [5]. however, there is inadequate information on suspicious agricultural commodities or food products crossing the african trade borders. 2. methodology this scoping review paper has reviewed and summarized evidence from existing literature on agri-food fraud in africa and across the africa-european union borders. the quality of each existing study of interest was not rigorously assessed. the search was based on agri-food fraud speculations involving any agri-food commodities, and cases of suspicions whether in africa or abroad as long as the origin of the commodities could be confirmed to have originated from africa. additionally, this review probed into potential associated health implications that may arise should consumers consume such manipulated agri-food commodities. keywords such as agri-food fraud, africa, exposure, and health were used to search for relevant datasets on various search engines such as google. the review was conducted using data obtained from online publications such as google scholar, web of science, and scopus. in addition, a manual search for relevant literature was carried out using reference lists to include relevant studies, published in the english language, and published between 2000 and 2024. when necessary, tables and figures were added to the narrative presentation of the results. 3. results and discussion 3.1. agri-food fraud suspicious incidence across the africa-eu trade borders of agri-food fraud in africa have revealed the magnitude of deceptive practices that span from the farm (production) to the market (processing and trade) making food susceptible to fraud at any stage’ [6]. for example, at the primary production stage, instances such as apiculture demonstrate how adulteration can occur. for example, a finished product of apiculture, honey, which is a highly valued product consumed worldwide as a sweetener and medicine has had a decreased rate of interest and trust because of its vulnerability to adulteration. honey is easily manipulated by illegally mixing it with syrups or sugars to increase volume and profitability [7, 8]. this not only diminishes the quality and nutritional value of the honey but also poses health risks to consumers who may unknowingly ingest substandard products. as products move from the farm to the market, fraud extends to finished goods, thus the longer the food supply chain, the higher the risk of fraud occurring. an example is the mislabeling of olive oil, wherein lower-grade oil is labeled as premium "extra virgin" olive oil [9]. this misrepresentation not only deceives consumers but also undermines the reputation of genuine producers and affects trade relations [8]. similarly, herbal medicine and spices are susceptible to substitution, with inferior or cheaper herbal medicine and spices being presented as premium variants [10, 11]. moreover, counterfeiting of dairy products where fake versions of branded items are sold [12] not only jeopardizes consumer health but tarnishes the reputation of business brands. as the world population grows exponentially, the pressure to meet food demand may serve as a motivation for perpetrators of food fraud practices. furthermore, mislabeling, a prevalent form of food fraud, has increased from 33% in 2015 to 47% in 2019 globally’ [13]. as predicted by the consumers brand association, food fraud may cost between 10 15 billion usd loss to the global food industry, hence affecting 10% of all world food products that are being commercially sold [14]. there is little evidence of agri-food fraud across the african and european trade borders. this fraudulent activity encompasses a range of practices, including mislabeling, adulteration, counterfeiting, and misrepresentation of food products (table 1 and 2). some food categories particularly vulnerable to fraud in cross-border trades were identified (table 1 and 2). notably, dairy products, seafood, spices and herbs, olive oil, honey, and fresh produce emerged as high-risk categories, with documented fraud incidents. generally, several common agricultural commodities or foods are vulnerable to fraud, each with a spectrum of potentially fraudulent practices (table 1). additionally, some instances of manipulated agri-food products have been detected in some african markets/countries (table 2), some of which might have been destined to cross the trade borders between africa and europe. these incidents involve mislabeling seafood species, adulteration of olive oil with lower-grade oils, and the presence of contaminants in spices and herbs. agriculture and food sciences research, 2024, 11(2): 59-64 61 © 2024 by the authors; licensee asian online journal publishing group table 1. incidence of agri-food fraud in farm and market settings in africa. stage fraudulent practices total percentage (%) examples citations farm adulteration in apiculture 111 2.2 mixing honey with syrups or sugars fakhlaei, et al. [7]; handford, et al. [12] and marvin, et al. [15] production site adulteration of beer and alcoholic beverages nd nd addition of illegal substances like strychnine shears [16] market adulteration of milk and milk product 526 10.5 addition of urea, formalin, detergents, ammonium sulphate, boric acid, caustic soda, benzoic acid, salicylic acid, hydrogen peroxide, sugars and melamine in milk marvin, et al. [15] and azad and ahmed [17] mislabeling of olive oil labeling lower-grade oil as extra virgin olive oil girelli, et al. [9] substitution of spices and herbal medicine 99 2.0 substituting cheaper spices and herbs for premium ones counterfeit dairy products (poultry meat and poultry product) 384 7.6 selling fake versions of branded dairy products osman, et al. [11]; kamboj [10]; handford, et al. [12] and marvin, et al. [15] red palm oil addition of sudan dye iv (a carcinogenic compound / azo dye) eggs and egg products 63 1.6 contains harmful substances such as benzoic acid, sodium alginate handford, et al. [12] and marvin, et al. [15] table 2. reported agri-food fraud incidents in some african countries and speculated health implications. country, sub-african region food type agri-food fraud practice potential health implications citations cameroon, central africa red palm oil -use of azo coloring dye (sudan dye iv) sudan dye iv is well known as a carcinogen toukap and abia [18] nigeria, west africa fat and oil, alcoholic and nonalcoholic drinks, honey, sugar, mılk honey being diluted with syrup and milk counterfeited with no animal protein loss of consumers' trust in the product. opia [19] and nagel [20] ghana, west africa red palm oil sudan dye iv is being mixed in palm oil sudan dye iv is a carcinogen. it has the potential to cause cancer kenya, east africa vegetable oil vegetable oil made from recycled oil which is harmful to humans deceive consumers and give less confidence to the brand. nagel [20] egypt, north africa wheat illegal addition of wrong or bad amount of wheat deceive consumers of the choices they want mabrouk, et al. [21] morocco olive oil olive oil counterfeiting by adulteration with cheaper oils ellyat [22] nigeria and ethiopia honey substandard honey diluted with sugar syrup orina [23] african nations (such as ghana, morocco, senegal, the gambia, egypt, kenya, togo, madagascar, uganda) shrimps, sole (fish product), vegetables (e.g. spinach leaves) and fruits (e.g. oranges), etc shrimps to which sulphites (e 220 – e 228) were added as additives, and noncompliance in cases of vegetables and fruits e.g. being tainted with pesticide residues european commission (ec) [24] agriculture and food sciences research, 2024, 11(2): 59-64 62 © 2024 by the authors; licensee asian online journal publishing group recently, according to the european commission’s first (january 2024) monthly agri-food fraud suspicions report on the eu agri-food fraud suspicions [24] majority of suspicions were at border control followed by market control. several suspicious commodities were from africa. for example, shrimps from senegal were suspected to be tampered with using sulphites (e 220 – e 228) (471 mg/kg) as notified by spain and classified as additives not compliant with eu maximum limits (mls). likewise, sole (fish product) from gambia was manipulated using cynoglossus senegalensis instead of synaptura spp as notified by italy and classified as adulteration (ingredient substitution) [24]. furthermore, a series of non-compliances (implicit claim violations) classified as residues of pesticides not compliant with eu maximum residue limits (mrls) mainly involving fruits and vegetables originating from africa specifically, egypt (vine leaves, carrots, mandarins, oranges, and strawberries), kenya (mangetout peas and peppers), ghana (pineapples), madagascar (cowpeas), togo (spinach leaves), and or uganda (peppers) were suspected in europe as notified by belgium, netherlands, slovenia, romania, italy or germany [24]. meanwhile, there may be more unchecked and unnoticed food fraud situations, these further emphasize the need for food trade border checks for rapid decisions toward reducing exposures and protecting” consumers’ health [25]. therefore, “one fraud anywhere may equate to several consumers in one world – the one fraud to consumers in one world concept – abia 2024”. this further complements the relevance of the potential agri-food fraud across africa-eu trade borders and calls for action. the trade relationship between africa and the european union (eu) is a pivotal nexus in the global food supply chain, facilitating the exchange of agricultural and food products. this trade corridor does not only support economic growth but also presents opportunities for cross-cultural cooperation and development. agriculture commodities or foods are traded across the africa-eu trade borders from african countries to the european markets or from the european countries to the african nations, thus constituting an import-export traffic situation. african countries export a diverse range of products to the eu, including fruits, vegetables, coffee, cocoa, nuts, and various processed foods while european countries also export products to africa such as machinery, pharmaceuticals, and industrial goods [26, 27]. however, within this trade relationship, vulnerabilities to agri-food fraud emerge, as products traverse complex supply chains, diverse regulatory frameworks, and varying economic conditions. agri-food fraud, a multifaceted issue characterized by deceptive practices in the food supply chain, has emerged as a critical challenge in the global trade of agricultural commodities and food products [28]. the rate of agri-food fraud has become an increasing problem globally in the food industry and for consumers with sellers seeking to capitalize on financial gains [29]. in this context, the trade relationships between africa and the european union (africa-eu) have garnered attention as a hotspot for agri-food fraud. these trade borders form a critical juncture in the global food trade, where products navigate complex supply chains, regulatory variations, and socio-economic disparities. this dynamic trade relationship presents opportunities and challenges, making it an important focal point for understanding the intricacies of agri-food fraud and its implications. given that agricultural commodities and food products are traded commonly across the africa-eu trade borders, certain food categories are particularly susceptible to fraudulent activities in this trade relationship. this may partly be due to factors such as high demand, diverse supply sources, and challenges in traceability [13]. for instance, in the export of fruits and vegetables, mislabeling or substitution can occur, leading to the misrepresentation of origin or variety. commodities like coffee and cocoa, where quality and origin play a crucial role, may also face adulteration or false labeling, affecting the authenticity and value of the products [30]. additionally, processed foods, such as olive oil and spices, may be susceptible to mislabeling, substitution, or dilution, impacting both consumer health and the reputation of producers. 3.2. agri-food fraud and global health agri-food fraud is a potential threat to consumers' health, wherever globally a consumer consumes manipulated agri-food, jeopardizing global health [25]. global health mainly focuses on worldwide health improvement (including mental health), reduction of disparities, and protection against global threats that disregard national borders, including the most common causes of human death and years of life lost from a worldwide perspective [31]. exposure to agri-food fraud can have significant implications for global health, as consumers unknowingly pay for, and consume products that may be illegally adulterated with substances (or that are being mislabeled) which constitute a risk factor to one or more unchecked hazards against consumers' health protection [8, 25]. these fraudulent practices compromise the integrity of the food supply chain and can lead to adverse health effects. for instance, adulterating foods with inferior ingredients or contaminants exposes consumers to potential allergens, toxins, or harmful chemicals that can trigger allergic reactions, foodborne illnesses, or long-term health issues [32]. mislabeling or substitution may lead to inadequate nutrient intake, impacting overall health and exacerbating nutritional deficiencies. moreover, introducing fraudulent products into the market erodes consumer trust, hindering their ability to make informed dietary choices. global health is intricately linked to food authenticity and safety. exposure to inauthentic (presumable unsafe) food otherwise known as manipulated food is a threat to public health, as consumers may unknowingly consume substandard or adulterated products, exposing themselves to contaminants [2, 8] allergens, and nutritional deficiencies [1, 33]. furthermore, the health consequences of agri-food fraud can lead to increased healthcare costs, decreased productivity, and economic losses for both consumers and governments since it increases health risks adding to the national burden [34]. this can affect economic development and exacerbate existing health disparities. therefore, comprehending the extent of agri-food fraud and its health implications within the context of african-european trade becomes imperative for safeguarding public health and ensuring the integrity of the food supply chain. to combat the growing threat of agri-food fraud and food safety by extension, the african national regulatory agencies e.g. nigeria (national agency for food and drug administration and control, nafdac), south african (south african health products regulatory authority, sahpra), ethiopia (ethiopian food and drug authority, efda), and cameroon (cameroon food safety framework law) may need to make more provisions for food compliance and inspections. these regulations aim to ensure the safety and authenticity of food products, protect public health, and maintain consumer confidence. as global trade in agriculture commodities and food products expands, the supply chain vulnerability to fraudulent activities becomes increasingly evident. this may be worse partly due to the inadequate food safety laws in african countries or poor enforcement. finally, there is a need for agriculture and food sciences research, 2024, 11(2): 59-64 63 © 2024 by the authors; licensee asian online journal publishing group rapid detection tools to make quick decisions on the field, and a need for supply chain traceability systems [25]. this article has provided relevant information on the extent of food fraud practice across the africa-european trade border and highlighted a potential association with consumers' health in different geographic locations around the globe. 4. conclusion agri-food fraud is a growing syndrome in africa which may unfortunately largely be unchecked. agri-food fraud may be common across the africa-eu borders, with speculated unprecedented implications on consumer health globally. this may partly be due to the absence or inadequate enforcement of food safety-related legislation in some african countries. when speculating that the variety and severity of sicknesses in africa may be associated with fraud, agri-food fraud in africa may require more attention. the need for efficient and effective systems for agri-food fraud prevention, vulnerability assessment, and authorities to track suspicious foods in the market or trade borders and trace them back to their origin/producers cannot be emphasized. there is a need for the producers/industries to devise strategies of not only authenticating their products but also being capable of distinguishing their products in the market from fake copies as a means of protecting their brands and maintaining consumers' trust. safeguarding the genuineness and safety of agri-food commodities or products strengthens consumers’ 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[accessed 2020. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://www.thermofisher.com/us/en/home/industrial/food-beverage/food-beverage-learning-center/food-analytical-testing-information/food-adulteration-analysis-information.html https://www.thermofisher.com/us/en/home/industrial/food-beverage/food-beverage-learning-center/food-analytical-testing-information/food-adulteration-analysis-information.html https://doi.org/10.1016/j.foodchem.2018.10.091 https://doi.org/10.1136/bmjgh-2019-002172 https://doi.org/10.3390/foods12010199 https://www.food.gov.uk/sites/default/files/media/document/the-cost-of-food-crime.pdf 93 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 2, 93-99, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i2.6961 © 2025 by the authors; licensee asian online journal publishing group good agricultural practices in horticultural production under cover in the lvrn, patagonia, argentina lucrecia aviles1 facundo zaragoza2 angel mamani3 omar ariel gajardo4 ( corresponding author) 1,2,3,4complejo universitario regional zona atlantica y sur. universidad nacional del comahue, (8500) viedma, argentina. 1email: malezas@curza.uncoma.edu.ar 2email: facundozaragoza91@gmail.com 3email: angelsfranco@yahoo.com.ar 4email: omararielgajardo@hotmail.com abstract the surface area of covered crops in the lower valley of the negro river (patagonia, argentina) is significant, producing first fruits in summer and leafy vegetables in winter. producers are interested in transitioning to agroecology, moving away from traditional management of horticultural crops. the objective of this work was to characterize vegetable production under cover in the lower valley of the negro river. to achieve this, greenhouses of producers employing both management systems were surveyed. the methodology proposed by sarandon was used, focusing on three dimensions of analysis: economic, social, and environmental. a survey was designed with indicators suggested by experts, and 16 establishments were visited to analyze the structure of the greenhouses, the crops present, and the quality of the soil and water. most producers are beginning their activities with small modules, covering less than 2000 m2, with structures made of treated wood and covered with polyethylene. the technological level is low, requiring training and technical assistance to meet the local demand, with expectations of sustained growth over time. around 10 different vegetables are grown in winter, and between 5 and 7 species in summer, with tomatoes being predominant. since producers in agroecological transition have only recently started, they differ from traditional producers in crop diversity, soil physicochemical and biological conservation, and low dependence on external inputs. keywords: crop diversity, economic dimension, environmental dimension, greenhouse, social dimension, soil quality, sustainability. citation | aviles, l., zaragoza, f., mamani, a., & gajardo, o. a. (2025). good agricultural practices in horticultural production under cover in the lvrn, patagonia, argentina. agriculture and food sciences research, 12(2), 93–99. 10.20448/aesr.v12i2.6961 history: received: 23 june 2025 revised: 25 july 2025 accepted: 29 july 2025 published: 4 august 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 94 2. materials and methods ................................................................................................................................................................... 94 3. results ................................................................................................................................................................................................ 96 4. conclusion ......................................................................................................................................................................................... 99 references .............................................................................................................................................................................................. 99 https://www.doi.org/10.20448/aesr.v12i2.6961 https://orcid.org/0000-0002-3383-1324 https://orcid.org/0009-0009-5170-3906 https://orcid.org/0009-0001-7606-9827 https://orcid.org/0000-0002-4872-9234 mailto:malezas@curza.uncoma.edu.ar mailto:facundozaragoza91@gmail.com mailto:angelsfranco@yahoo.com.ar mailto:omararielgajardo@hotmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ agriculture and food sciences research, 2025, 12(2): 93-99 94 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this research on the implementation of good agricultural practices in greenhouse crops demonstrates the benefits of agroecological management over traditional methods. knowing the installed greenhouse capacity is vital for training producers, promoting crop diversification, improving soil quality, and reducing dependence on inputs, thereby ensuring growth that meets unmet local demand. 1. introduction the lower valley of the negro river (lvrn) covers a total area of 80,000 hectares, of which 18,500 hectares are organized into farms with surface areas ranging from 30 to 120 hectares. these farms have a network of canals to facilitate surface irrigation, utilizing water from the negro river, which is suspended during the winter months for system maintenance. most farms are dedicated to agricultural production, including horticulture, fruit growing, and fodder, as well as livestock activities such as cattle and sheep fattening on irrigated pastures. vegetable production is primarily carried out by small-scale producers who grow diversified crops to supply fresh food to the region. according to sworn crop declarations, 2,500 hectares are cultivated with vegetables, representing between 10% and 14% of the total irrigated area in the lvrn. of these, approximately 1,500 hectares are dedicated to onion production, 300 hectares to squash, and 700 hectares to various other vegetables [1]. most of the horticultural producers who settled in the region started as sharecroppers and now rent land on which they produce. they are compelled to lower costs by reducing labor expenses, and their production is generally family-based. currently, horticultural production is conducted on a small scale, in an intensive and diversified manner, primarily for the domestic market and, to a lesser extent, for the regional markets such as neuquén, bariloche, and comodoro rivadavia. the population is estimated to consist of approximately 150 producing families, mostly tenants who sell around 70% of their produce on the domestic market. although there is limited information on the productive capacity of these horticulturists, they supply fresh vegetables to consumers in the viedma-patagones region [2]. the increase in the surface area of covered crops observed in recent years, with the aim of producing early summer crops and leafy vegetables in winter, indicates the interest of producers in changing traditional management of horticultural crops. this activity intensifies the use of the soil, which means that greater contributions of nutrients, generally of chemical synthesis, must be made, or crops must be rotated by moving the structures to other plots, negatively impacting the sustainability of the system. the greenhouse system is an enclosure delimited by a wooden or metal structure, covered by glass or any transparent plastic material, inside which vegetables and ornamental plants are usually grown during periods when external climatic conditions would not allow obtaining the desired products [3]. already in garcía and quaranta [4], emphasized the importance of having information on the horticultural sector and the territory that enables informed decision-making by different actors (such as the public sector, producer and consumer organizations, etc.) in the green belt of the town of la plata, capital of the province with the largest number of inhabitants in argentina. in the province of río negro, which is among the 10 least populated in the country, there is no systematized information on horticultural production, much less under cover. in this sense, tellería et al. [5] highlighted the social aspect of vegetable production, highlighting the work of women and the precariousness of the production system. zalazar [2] reported on the diversity of horticultural foods offered and quantified the economic impact of lvrn producers in the region. in order to have updated information related to the increase in the surface area of greenhouses, this work proposes updating the information on under-cover production systems, the quality of their soils, and the crops grown, differentiating between producers with conventional management and those in agroecological transition. 2. materials and methods 2.1. study site the lower valley of río negro (lvrn) is located in the se of the province of río negro, argentina (40° south latitude and 63° west longitude), on the southern bank of the homonymous river. the climate in the region is mesothermal semiarid, with an average annual rainfall of 394 mm and an average annual temperature of 14.2 °c. it is a fertile area with fine to medium textured soils. figure 1 illustrates the geographic location of the lvrn in patagonia, argentina. figure 1. geographic location of the lvrn. to characterize the crops under cover, the production systems were studied in three categories: 1. survey of greenhouses: dimensions, construction materials, and crops. 2. analysis of soil and water quality: physical and chemical aspects. 3. survey of producers: environmental, economic, and social dimensions. agriculture and food sciences research, 2025, 12(2): 93-99 95 © 2025 by the authors; licensee asian online journal publishing group 2.2. survey of greenhouses based on geographic information and using google earth®, a total of 31 producers with varying amounts of covered surface area were identified. for the study, representative producers of each type were selected: conventional and agroecological, based on the suggestions of qualified informants. in june and september 2022, 16 establishments of entrepreneurs who produce vegetables in greenhouses were visited: 12 producers who use conventional management with chemical synthesis inputs and 4 who manage production with low dependence on external inputs (agroecological transition). during each visit, in addition to the semi-structured interviews conducted with producers, the dimensions of the production modules (each of the buildings), the materials and construction design, orientation, and protection of the greenhouses and crops present were recorded. 2.3. soil and drilling water analysis for soil characterization, a sample was taken consisting of five subsamples from the first 10 cm of soil in three conventional production greenhouses and three in agroecological transition. a water sample was also collected from the boreholes of four establishments to assess the quality of irrigation water during winter. in soil samples, clay content was determined by the bouyoucos method, ph in a 1:2.5 water solution, electrical conductivity in saturation paste, sodium absorption ratio (sar), total nitrogen by kjeldahl, phosphorus by the olsen method, organic matter by bray-curtis, and potassium by flame spectrophotometry. comparisons between conventional systems and agroecological transition systems were made using anova and lsd mean comparison test (p < 0.05). in june, when producers used drilling water because the irrigation system supplied by the negro river was suspended to clean the canals, the ph was analyzed in water samples, as well as the electrical conductivity, soluble salt content, and sodium content determined by flame spectrophotometry. 2.4. producer surveys semi-structured interviews were conducted with producers, who were grouped according to the production system: conventional and in agroecological transition. the categories, descriptors, and indicators suggested by the experts and collected in the interviews are presented in tables 1, 2 and 3. as a first step, the values of each variable were standardized on the scale recommended by cieza and sarandón [6]. in this work, the scale ranged from 0 to 4, with 0 representing the least sustainable category and 4 the most sustainable, regardless of the original units of each indicator. they were thus converted and expressed in scale values. level 2 was adopted as acceptable for all variables. for example, cultural biodiversity was evaluated with a scale of 5 values (from 0 to 4): 4 indicating six crops; 3 indicating five crops; 2 indicating four crops; 1 indicating three crops; and 0 indicating 1-2 crops. this facilitated comparison between different systems. with the systematized information, the values of the indicators were averaged to determine the value of the corresponding descriptor. the average of the final values was represented in spider web diagrams, as suggested by sarandón et al. [7], to identify strengths and critical points of each system. table 1. categories of analysis for the environmental dimension suggested by local experts. category descriptor indicators soil conservation of physicochemical fertility risk of salinization fertilization management conservation of physical properties management of organic matter tillage practices conservation of biological conditions use of pesticides tillage practices rotations biodiversity conservation of biodiversity use of pesticides crop variability rotations crop diversity water irrigation water quality risk of salinization table 2. categories of analysis for the economic dimension suggested by local experts. category descriptor indicators economic stability productive diversity number of cultivated species land ownership economic benefits economic growth economic growth degree of technical development hybrid seeds type of heating use of venturi use of manometer temperature and humidity recording dependence on external inputs use of synthetic fertilizers use of pesticides table 3. categories of analysis for the social dimension suggested by local experts. category descriptor indicators quality of life basic needs access to health access to education producer satisfaction degree of satisfaction land ownership land ownership line of succession children and family agriculture and food sciences research, 2025, 12(2): 93-99 96 © 2025 by the authors; licensee asian online journal publishing group 3. results 3.1. greenhouse survey greenhouse crops constitute a small part of the productive structure of the lvrn in terms of surface area occupied. with a total of 20 ha out of 2,500 ha dedicated to horticultural production, they practically represent all the vegetables consumed in the region. currently, 31 producers have greenhouses on their farms, totaling 94 modules. forty-one percent of the producers have a single module, 29% have expanded to two or three modules, and the remaining have more than four. notably, two producers have 12 modules on their farms (figure 2a). the increase in the number of modules per producer is due to the experience gained in managing this productive system, which encourages investment in infrastructure. regarding the surface area of the modules, most producers build them 50 or 100 meters long to optimize the use of polyethylene rolls and align with the capacity of pressurized irrigation water pumping systems. greenhouses used for agroecological production tend to be shorter, ranging from 25 to 30 meters, likely due to higher labor demands and the need for improved ventilation to control pests and diseases. concerning width, there is no consistent pattern; modules vary between 9 and 45 meters, with a predominance of 30 meters. the chosen width correlates with the machinery used in the field. all greenhouses feature a gable roof design, with more than half measuring 5 meters at the center and 2.20 meters at the sides, incorporating zenith ventilation to enhance air circulation. additionally, 41% have a central height of 3 meters, which may compromise air exchange due to construction costs and the absence of adequate windbreak curtains. of the 94 covered production modules in the lvrn, 37% have an average surface area of 2000 m2 and 25% have an average surface area of 1000 m2 (figure 2b). in other words, most of the modules are small; however, the number of modules in the establishments differentiates two categories of producers: those who are starting with this activity and have less than 1500 m2 of greenhouse space, and those who have already mastered the system with more than 1 hectare under cover in production (figure 2c). according to hidalgo [8], having very small production units limits access to technology, training, and technical assistance. that is, the costs required to implement new technologies cannot be amortized by small-scale production. figure 2. structural characterization of the lvrn greenhouses. a) number of modules per producer, b) surface area of the modules built, and c) total surface area per producer. the survey was conducted using google earth® for the 31 producers. the structures of the lvrn greenhouses are made of eucalyptus or palm poles that have been treated with sulphate; without this treatment, their lifespan is very short. additionally, to extend the poles' useful life, they are painted with tar on the part that is buried. generally, the thickness of the lateral poles and roof braces causes shading inside, which negatively impacts the crop during periods of high photoluminescent requirements [3]. for this reason, poles with a smaller diameter but made of resistant wood are selected in the region. the covering material used is 150-micron lldpe (linear low-density polyethylene). the incorporation of "long-lasting" additives, which protect it from ultraviolet radiation, and "long-lasting thermal" additives, which make it more opaque to long-wave radiation (thermal pe 0.2 mm = 15-18%), makes it a suitable material for production, especially in the winter months. although the useful life of these polyethylenes is around four years, their low weight, the fact that they are sold in variable widths, and their good light diffusion facilitate their construction without the need for specialized labor (figure 3). the orientation of the modules is mainly parallel to the prevailing winds of the wnw region. to protect the structures from the effects of cold winds, they are built close to windbreaks made of established poplars. since the lvrn has irrigation channels, the modules are built close to the drains (drainage channels) to allow the percolation of irrigation water, thereby preventing waterlogging inside and salinization of the soil. investment in technological equipment is low: they do not have a heating system, automated irrigation, automatic window opening, or high-temperature control that would help the plants develop better and avoid losses due to extreme temperatures. they do not evaluate or control the quality of the soil or irrigation water, which is detrimental to the sustainability of the system. figure 3. internal structure of greenhouses. agriculture and food sciences research, 2025, 12(2): 93-99 97 © 2025 by the authors; licensee asian online journal publishing group 3.2. soil quality plant growth is strongly influenced by soil salinity conditions, and vegetables are no exception [3]. in laboratory studies, no differences were observed between the variables analyzed in the soils of greenhouses where conventional production is carried out compared to those in agroecological transition. the plots where the greenhouses have been installed have a clayey loam texture with high ph values typical of the region. a high salinity (ec) with a high sodium content (sar) is notable, probably associated with the quality of the water used for winter irrigation (table 4). these salinity values are associated with yield losses of around 50%, according to iglesias [3]. the yield is reduced by half in spinach, tomato, and lettuce with ec values of 9, 8, and 5 ms/cm, respectively. regarding fertility parameters, soils in greenhouses undergoing agroecological transition tend to be higher, although they are not yet statistically different from soils under conventional production (table 4). however, the values observed under both greenhouse production systems are higher than those observed by avilés et al. [9] in open-air soils where the nitrogen, organic matter and phosphorus content was 0.20 ± 0.01 g n /100 g, 2.90 ± 0.11 g om/100 g and 27.7 ± 5.9 mg p/kg respectively, evidencing good fertility management within the greenhouse. the potassium contribution should be improved since the content is higher in the field than in greenhouses (632 ± 75 mg k/kg). this deficiency in potassium content and increase in phosphorus content was also observed by silveira and barbazán [10] in greenhouses in uruguay, who suggest that producers review fertilization practices. table 4. physicochemical characterization of greenhouse soils under conventional management and during agroecological transition. soil quality conventional management agroecological management clay (g/100 g) 32.83 ± 9.58 38.73 ± 4.69 ph 8.01 ± 0.15 8.00 ± 0.23 ec (ms/cm) 11.41 ± 3.09 9.99 ± 2.26 sar 14.5 ± 2.1 11.7 ± 0.6 n (g/100g) 0.22 ± 0.00 0.27 ± 0.05 p (mg/kg) 135 ± 33 142 ± 21 k (mg/kg) 254 ± 49 365 ± 67 om (g/100g) 4.30 ± 0.55 5.36 ± 0.49 3.3. irrigation water quality regarding irrigation water, both in conventional production systems and those undergoing agroecological transition, during the summer season (august to april), farms have a water supply from the negro river through a system of high-quality canals (ec 0.4 ms/cm and sar < 1). the irrigation system employs drip hoses with a pressurizing pump and venturi for fertilizer incorporation into the soil, although the smaller systems use a precarious bypass-type fertilization system that involves sucking fertilizer directly with the irrigation pump. during the winter months (may to july), the river water supply is suspended for canal system maintenance, prompting producers with greenhouses to perforate their farms to supply water to crops. studies on some of these boreholes have detected high salt content (ec 6.1 ms/cm and sar 18), which qualifies the water as unsuitable for crop irrigation according to the riverside standard (c6, s3). this issue is exacerbated by the localized irrigation system and its repeated use over time. according to castellón gómez et al. [11], ec values higher than 3 ms/cm and sar higher than 9 in irrigation water are associated with a severe degree of restriction for agricultural production. due to the low annual rainfall in the area (392.2 mm), and particularly during months when irrigation water supply is cut off, where the average rainfall over the last 57 years was 92.2 mm [12], it is not feasible to accumulate rainwater for use during the winter season as proposed by lobos rodríguez, et al. [13] for the o'higgins region in chile. producers should compare yield losses in summer crops due to soil salinization with the gains obtained from winter production and consider the possibility of suspending winter production under cover. they could also evaluate the feasibility of incorporating technology such as reverse osmosis equipment for the winter months. 3.4. survey of producers the results of the interviews were grouped into three dimensions: environmental, economic, and social, according to the indicators suggested by the experts (table 1, 2 and 3),comparing producers who carry out conventional management with those who are in agroecological transition (figure 4). regarding the environmental dimension, it is observed that both production systems manage irrigation similarly, reaching an acceptable level, aided by the contribution of river water during the summer season (figure 4a). a similar pattern occurs with fertilization management: conventional producers supply external nutrients through the fertigation system, optimizing the doses of these compounds with venturi in the drip system during the crop cycle, while those in the agroecological transition stage provide manure and compost during seedbed preparation. producers in the agroecological transition are distinguished by their low use of pesticides (including biopreparations), crop rotations, more conservative tillage practices, and the addition of organic matter. crop diversity, in terms of species and varieties, is enhanced through seed exchanges with agroecological producers from other regions, preserving native germplasm and incorporating aromatic plants to control pests [1]. the analysis of the economic dimension indicates that land ownership and economic growth are adequate and similar in both production systems (figure 4b), because the enterprises in agroecological transition consist of families of conventional producers adopting the new production system based on the capital they already possess. this enables them to gradually enter a niche of specific consumers. they do not depend on external inputs, which enhances their qualifications, and due to consumer demand (market), they have been expanding the diversity and variety of crops. however, both types of producers, possibly due to their low educational level [2] and limited experience in covered systems, present a critical point in terms of the degree of technicalization, as observed by mundo coxca et al. [14]. in the crops under cover of the town of puebla in mexico, they suggest strengthening the link between research and government centers at their different levels with tomato producers through continuous advice and training. in this sense, hernández suárez [15] observed that the federal programs, which were crucial for the establishment of greenhouses and the growth of their number in zacatecas (mexico), were granted to lowincome producers with insufficient capacities to face all the vicissitudes of this type of activity, which is highly uncertain and risky. agriculture and food sciences research, 2025, 12(2): 93-99 98 © 2025 by the authors; licensee asian online journal publishing group since producers in agroecological transition are members of the same families that produce conventionally, the social dimension is similar in most of the evaluated indicators (figure 4c). they live on the same property, in the same home, belong to the same family, have access to the same health system, but are more satisfied with their production due to the response of consumers and because they perceive an improvement in their living conditions. figure 4. comparison of covered production systems in the lvrn: conventional vs. agroecological transition. a) environmental dimension; b) economic dimension; and c) social dimension. 3.5. winter and summer crops although production is traditionally divided into two seasons: winter, for green leafy crops, and summer, for fruit crops, these criteria are becoming less distinct. at the farm level, practices involve cultivating different crops throughout the year. protected cropping is a practice aimed at extending the productive season, which allows for early and late harvests. in the area, the main winter crops are planted in a staggered manner to supply the town with lettuce in its different varieties (curly, head, and butter lettuce), spinach, chard, celery, broccoli, parsley, beetroot, among other minor crops. particularly, producers in agroecological transition add aromatic species that fulfill the double purpose as a crop and pest regulator, such as mint, lavender, marigold, certal, citronella, rosemary, etc. in each module, between 7 and 10 different crops were observed, which implies a complex management by having to regulate the agriculture and food sciences research, 2025, 12(2): 93-99 99 © 2025 by the authors; licensee asian online journal publishing group irrigation and fertilization regime by sectors, although this allows them to offer greater diversity of products to local consumers and expand commercial sales. summer crops are generally less diversified and are sown on a single date to facilitate logistics and management of irrigation and fertilization, compensating for the supply to consumers with field crops. among the crops grown on larger areas in greenhouses, the tomato stands out in its different varieties: pear, platense, red cherry, yellow cherry, and chocolate cherry. bell peppers, peppers, eggplant, green beans, strawberries, broad beans, and cucumbers are also produced, and some farmers even add melon to obtain early fruits. 4. conclusion the surface area of covered crops in the lvrn is significant and responds mainly to the local demand for food. as producers gain experience, they risk adding modules and diversifying production. however, the technological level is still low and requires training and qualified technical assistance to fulfill the still unsatisfied local demand, predicting sustained growth over time. producers in agroecological transition in the lvrn replaced the cultivation system, convinced of the improvement in family health and the social recognition acquired, which encourages other producers who see in this system a healthy, sustainable, and economically viable alternative. references [1] n. seba et al., diversified horticultural production in the lower río negro valley: current socio-productive models and their potential for ecological intensification. buenos aires, argentina: ediciones inta, 2017. 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[14] m. mundo coxca, j. l. jaramillo villanueva, j. m. jimenez, a. macias lópez, and j. ocampo mendoza, "technological characterization of greenhouse tomato production units in puebla," revista mexicana de ciencias agrícolas, vol. 11, no. 5, pp. 979-992, 2020. https://doi.org/10.29312/remexca.v11i5.2010 [15] j. l. hernández suárez, "the development of greenhouses in zacatecas. between boom and abandonment," entreciencias: diálogos en la sociedad del conocimiento, vol. 8, no. 22, 2020. https://doi.org/10.22201/enesl.20078064e.2020.22.70248 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.24215/2346898xe108 https://doi.org/10.24215/16699513e121 https://doi.org/10.29312/remexca.v11i5.2010 https://doi.org/10.22201/enesl.20078064e.2020.22.70248 44 © 2022 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 9, no. 1, 44-49, 2022 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v9i1.3944 © 2022 by the authors; licensee asian online journal publishing group evaluation of cocoyam-soybean flour blends and sensory properties of the amala dumpling ojo, m. a.1 ologunde, m. o.2 alabi, o. d.3 ohijeagbon, o. r.4 ojo, h.5 ( corresponding author) 1,2,4,5department of food science, ladoke akintola university of technology, ogbomoso, oyo state, nigeria. 1email: mayoojo2006@yahoo.com tel: +234(0)8037238432 2email: moologunde@lautech.edu.ng tel: +234(0)803721454 4email: orohijeagbon@lautech.edu.ng tel: +234(0)8132392133 5email: ojohelen278@gmail.com tel: +234(0)8132847556 3department of nutrition and dietetics, ladoke akintola university of technology, ogbomoso, oyo state, nigeria. 3email: ajaladupe@yahoo.co.uk tel: +234(0)9034548501 abstract amala, a staple food among the yoruba people of south west nigeria, is mostly produced from yam. since yam is known to be mostly carbohydrate and soybean is a legume with an appreciable quantity of protein, efforts have been made to prepare amala from the blends of cocoyam and soybean flours. the protein content of the flour blends ranged from 3.73% to 13.81%. in general, there was increase in the concentration of protein with increasing level of soy flour substitution. unlike protein, the crude fibre content of the flour blends decreased with increasing supplementation of soy flour. increase in oil content of the flour samples, with increasing addition of soy flour, predisposes the samples to shorter shell life and off flavor because of liability to rancidity. the lowest bulk density of 0.78 g/cm3 was recorded for sample a (100% cocoyam flour) and hence none of the flour blends could be considered as a complimentary infant substitute. in general, supplementation of cocoyam flour with soy flour enhanced the concentrations of protein, ash and dietary fibre. therefore, it is hoped that consumption of cocoyam amala supplemented with soy flour, a cheap source of plant protein, will not only create dietary diversity, it will make more protein available for the consumers and thus help in alleviating the problem of protein energy malnutrition in developing regions of the world. keywords: cocoyam, soybean, flour blends, elubo, amala dumpling. citation | ojo, m. a.; ologunde, m. o.; alabi, o. d.; ohijeagbon, o. r.; ojo, h. (2022). evaluation of cocoyam-soybean flour blends and sensory properties of the amala dumpling. agriculture and food sciences research, 9(1): 44-49. history: received: 22 february 2022 revised: 20 april 2022 accepted: 6 may 2022 published: 26 may 2022 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. authors’ contributions: all authors contributed equally to the conception and design of the study. competing interests: the authors declare that they have no conflict of interest. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study followed all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 45 2. materials and methods ................................................................................................................................................................... 45 3. results and discussion ................................................................................................................................................................... 46 4. conclusion ......................................................................................................................................................................................... 48 references .............................................................................................................................................................................................. 48 mailto:mayoojo2006@yahoo.com mailto:moologunde@lautech.edu.ng mailto:orohijeagbon@lautech.edu.ng mailto:ojohelen278@gmail.com mailto:ajaladupe@yahoo.co.uk https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v9i1.3944 https://orcid.org/0000-0002-2092-4062 https://orcid.org/0000-0002-2173-501x https://orcid.org/0000-0003-0263-0818 https://orcid.org/0000-0001-6715-0175 agriculture and food sciences research, 2022, 9(1): 44-49 45 © 2022 by the authors; licensee asian online journal publishing group contribution of this paper to the literature: this study reveals the possibility of using the blends of cocoyam and soybean flours to produce amala which hitherto is produced from white yam species. supplementation of cocoyam flour with soybean flour for the production of amala creates dietary diversity, makes nutritionally important nutrient such as protein more available and widen the scope of utilization of cocoyam which is traditionally believed to be a food crop for low-income group. 1. introduction cocoyam (colocasia esculenta) is a well-known food plant that is high in carbohydrates [1]. it is a member of the araceae family [2]. a significant proportion --about 75%, of the world production of cocoyam comes from africa, with nigeria and ghana leading the way. cocoyam is the third most important root and tuber crop in nigeria, following yam and cassava though consumption of cocoyam is assumed to be for low income earners of the society in some communities because it is readily available at a relatively lower price [3, 4]. cocoyam is nutritionally superior to other root and tuber crops [5]. it contains more crude protein than other root and tuber crops, and its starch is highly digestible due to the small size of the starch granules. in addition, it contains calcium, phosphorus, vitamin a, and b-vitamins [6]. cocoyam is an underutilized tropical root plant in nigeria, its utilization is still at the subsistence level, making it a highly neglected crop [7]. in the south eastern part of nigeria, cocoyam flour is commonly used as a thickener in soup preparation. cocoyam has been reported to have a lowering effect on blood glucose levels, confirming its anti-hyperglycemic effect and thus recommended for diabetic patients [8]. the nutritional and chemical compositions of cocoyam indicate that, if fully exploited, it would improve food and nutrition security for people living in the tropics [9]. soybean (glycine max) is a leguminous plant that is widely grown for its edible seed, which has numerous uses. seeds of soybean have an outstanding nutritional and functional food profile. based on their nutrient composition, soy-foods are considered nutritious and healthy [10]. it is the only plant source that contains all of the essential amino acids; the protein content of soybeans is higher than that of other legumes. soy protein is also high in quality and easily digestible. soybeans are high in fat, with the majority of the fatty acids being unsaturated. polyunsaturated (primarily linoleic acid), monounsaturated (oleic acid), and saturated (primarily palmitic acid) fatty acids account for approximately 63%, 23%, and 14%, respectively. soybeans have a high polyunsaturated fat content, which includes alpha-linolenic acid, an essential omega-3 fatty acid. soybeans are one of the few plant foods that contain both essential fatty acids, b-vitamins, calcium, iron, and phosphorus are among the vitamins and minerals found in soybeans [10]. it also contains antioxidants and phytonutrients, like phytosterols and phytoestrogens, which have been linked to a variety of health benefits. soybean is one of the cheapest and richest sources of plant protein that can be used to help millions of people meet their nutritional needs. aside from being a good source of protein, the seed is said to have the highest food value of any plant food consumed worldwide [10]. soybean contains protein (35–45%), oil (15–25%) and carbohydrate content is approximately 33%, out of which 16.6% are soluble sugars [11]. it also contains approximately 19.10% ether extract, 5.71% crude fibre, 5.06% mineral content, and 26.05% nitrogen free extract [12]. amala is a popular yoruba starchy food made by mixing yam flour (elubo) with boiling water and vigorously stirring with a wooden stirring rod until a smooth, consistent paste is formed. continuous and vigorous stirring is required to prevent the formation of lumps. amala is rolled into balls between the fingers and served with a variety of soups and stews that contain fish or meat. it is a meal that can be consumed at any time of day. amala is a popular delicacy in western nigeria and other west african countries. amala is traditionally made mainly from yam flour (elubo) although the use of cassava flour (lafun) and unripe plantain flour (elubo ogede) is not uncommon. yam flour (elubo) is made from yam tubers, primarily white yam (dioscorea rotundata) [13]. efforts have recently been made to produce flour from other roots and tubers, such as potato and cocoyam, that can be used to make amala. furthermore, because amala is a high-carbohydrate food, several studies have been conducted to improve the nutritional quality of this commonly consumed meal. abulude and ojediran [14] supplemented yam flour (elubo) with soybean flour in 2006, and barine and frankline [15] produced composite flour from a blend of yam and african yam beans ((sphenostylis stenocarpa)), both for amala preparation. reports exist in the literature on the use of cocoyam and cocoyam-cowpea flour blends for preparation of amala [16, 17]; paucity of information on the use of cocoyam flour blended with soybean seeds at varying proportions necessitated this study. in this report, amala was made from a blend of cocoyam flour and soy flour, and the amala dumpling made from it was evaluated. this will, hopefully, make a significant contribution to the level of progress being made in using mixtures of local crops to improve nutrition in developing countries; it will also encourage diverse use of locally grown food crops [18]. 2. materials and methods 2.1. materials freshly harvested cocoyam was obtained from ganmo market, ilorin, kwara state, nigeria while soybean was obtained from sabo market, ogbomoso, oyo state, nigeria. other equipment such as cabinet dryer, weighing balance, milling machine, trays and vats were provided by the department of food science, ladoke akintola university of technology, ogbomoso, oyo state, nigeria. 2.2. production of cocoyam flour cocoyam tubers were washed in water to remove dirts. the tubers were weighed, manually peeled with the aid of a stainless-steel knife and sliced into thickness of about 1.0 – 1.5 cm. the sliced cocoyam was blanched at 70 ˚c for 3 minutes and left in blanching water for 12 h while the water cools down to room temperature. the slices were then drained and dried in a cabinet dryer at 55 ˚c until it is properly dried. the slices were then milled using a plate mill. the milled sample was allowed to cool, packaged in a cellophane bag and stored on the laboratory shelf prior to further processing. agriculture and food sciences research, 2022, 9(1): 44-49 46 © 2022 by the authors; licensee asian online journal publishing group 2.3. production of soybean flour the method of udofia, et al. [19] was modified for use. soybean was dried cleaned by winnowing and then washed with clean water. the seeds were then soaked in water for 4 h. the seed coat was removed by rubbing the soaked seeds between the palms of the hands. the decorticated seeds were dried in cabinet dryer at 55 ˚c until properly dried and then milled using a plate mill. the milled sample was packaged in a cellophane bag prior to further processing. 2.4. formulation of flour blends elubo samples were prepared by blending cocoyam and soybean flours in the ratio of 100:0 (a), 95:05 (b), 90:10 (c), 85:15 (d), 80:20 (e), 75:25 (f) and 70:30 (g). the blends were similar to the preparation of julianti, et al. [20]. 2.5. preparation of amala the blend product (elubo) was prepared for consumption by pouring slowly and stirring continuously in boiling water with a wooden rod while being cooked. the stirring was done vigorously to prevent formation of lumps and a smooth consistent paste was formed. the paste was covered and left to steam on the fire for about 5 minutes to cook. it was further stirred, packed and wrapped in sweet prayer (t. daniellie) leave. each of the flour blends was used to prepare amala using this same procedure. 2.6. determination of proximate analysis of cocoyam-soybean flour blends the proximate analysis of the cocoyam-soybean flour blends (samples a, b, c, d, e, f and g) were carried out using standard methods [20]. the proximate parameters determined were total ash, moisture content, crude protein, crude fat (ether extract), crude fibre and total carbohydrate. the total carbohydrate was determined by difference. 2.7. determination of functional properties water and oil absorption capacities were determined using centrifugal method. distilled water was used as a medium for water absorption while refined olein was used for oil absorption [21, 22]. bulk density for each of the flour blends was determined using the method described by ojo and ade-omowaye [23]. 2.8. sensory evaluation sensory evaluation of the amala samples was carried out for taste, color, aroma, texture, and general acceptability. the sensory evaluation was done by 10 panelists who were provided with questionnaire with nine-point hedonic scale. each panelist was requested to assess each coded sample and to record the degree of differences. 2.9. statistical analysis statistical analyses of the data obtained were done using the statistical package for social science (spss version 16.0) [24]. the data were subjected to one-way analysis of variance and the significance difference determined at p>0.05. the means were separated by duncan multiple range test. 3. results and discussion 3.1. proximate composition of cocoyam-soybean flour blends the results of the proximate composition of the composite flour blends of cocoyam and soybean are shown in table 1. the moisture content of flour blends ranged from 5.82 to 7.64% . table 1. proximate composition of cocoyam and soybean flour blends. sample moisture (%) ash (%) cho (%) crude fat (%) crude fibre (%) crude protein (%) calorific value (cal) a 7.64±0.60b 2.94±0.37a 82.18±0.97d 1.78±0.42a 3.02±0.14ab 3.73±0.00a 1624.60±17.54a b 6.32±0.52a 2.97±0.40a 77.35±1.31c 2.97±0.21b 2.48±0.44abc 8.16±0.00a 1619.12±22.53b c 6.69±0.31b 3.03±0.19ab 72.63±0.21b 3.58±0.23c 2.32±0.18a 12.47±0.00a 1590.61±4.90b d 6.32±0.76a 3.11± 0.14ab 73.38±1.13b 4.14±0.08d 2.27±0.26d 9.33±0.00a 1533.11±22.08b e 6.06±0.07a 3.36±0.09ab 73.48±0.15b 5.34±0.21e 2.20±0.11ab 10.05±0.00a 1520.22±1.80c f 6.22±0.16a 3.47±0.11ab 66.44±0.47a 7.75±0.16g 1.82±0.57bc 13.81±0.00a 1515.20±1.94c g 5.82±0.36a 3.58±0.23a 66.74±0.31a 7.46±0.00f 1.75±0.21c 13.81±0.00a 14.99.20±5.13c note: values with different superscript along the same column are significantly different at p>0.05. cho = total carbohydrate; a = 100% cocoyam flour; b = 95% cocoyam flour + 5% soybean flour; c = 90% cocoyam flour + 10% soybean flour; d = 85% cocoyam flour + 15% soybean flour; e = 80% cocoyam flour + 20% soybean flour; f = 75% cocoyam flour + 25% soybean flour; g = 70% cocoyam flour + 30% soybean flour. with sample g having the lowest moisture content while sample a has the highest value. flour samples with high moisture content greater than 12% usually have short shelf stability compared with the ones with lower moisture content of less than 12% [25]. the control sample i.e., 100% cocoyam flour, had the ash content of 2.94%. addition of 5% soybean flour caused sample b to have 2.97% ash. the highest concentration of ash of 3.58% was recorded for sample g with 30% addition of soybean flour. samples c, d, e and f had ash contents of 3.03, 3.11 and 3.36%, respectively. the results showed that there was increase in the ash contents of the flour blends with increasing addition of soybean flour. thus, addition of soybean flour contributed to the increase in the ash contents of the flour blends. in an earlier study, awolu and oseyemi [1] reported increase in ash content in the range of 0.53 – 2.48% on addition of bambara groundnut to cocoyam flour. the significant difference (p>0.05) in the protein content among the flour blends are presented in table 1. the protein content of the flour blends ranged from 3.73% for the control to 13.81% for sample f and g. sample e, containing 20% level of substitution with soy flour had 10.05% protein while sample d had 9.33% protein. in general, agriculture and food sciences research, 2022, 9(1): 44-49 47 © 2022 by the authors; licensee asian online journal publishing group there was increase in the concentration of protein with increasing level of soy flour substitution. increase in oil content of the flour samples predisposes the samples to shorter shell life and off flavor because of liability to rancidity. protein is required for growth, repair and maintenance of fluid and strong immune system [26]. protein also acts as a carrier for other nutrients such as iron, sodium, potassium [27]. foods containing protein of good biological value are of great nutritional importance in developing regions of the world where is prevalence of protein energy malnutrition [19, 28]. in an earlier study, supplementation of cocoyam flour with bambara groundnut flour had been reported to cause increase in protein content [1]. it can therefore be inferred that flour blends of cocoyam and soy flours n the proportion formulated in this study can be used to produce amala dumpling rich in protein. the 100% cocoyam flour contained 1.78% crude oil. the quantity of oil in the flour blends increased with increasing addition of soy flour. this is not unexpected because soy bean seed is an oil bearing leguminous seed. as shown in table 1, 30% level of substitution resulted in 7.75% crude oil content of the flour blend indicating that soybean contributed significantly to the crude oil content of the flour blends. increase in oil content of the flour samples predisposes the samples to shorter shell life and off flavor because of liability to rancidity [29]. sample a contained 3.02% crude fibre. samples b, c and d contained fibre content of 2.48, 2.32 and 2.27%, respectively while sample g had the lowest percentage of 1.75%. unlike protein, the crude fibre content of the flour blends decreased with increasing supplementation of soy flour. crude fiber content of the food product is an index to detect adulteration of flours [30]. dietary fiber has been shown to have a great impact on the health of the consumers. consumption of foods that are rich in dietary fibre is very essential due to its bulk addition to food, prevention of constipation and facilitation of peristalsis [31]. however, the crude fiber content of the flour blends in this study fall below the daily allowance range of 25-50% recommended by american diabetes association [32]. cocoyam had been reported to be a good source of carbohydrate ranging from 72.0 to 87.7% [2]. this is comparable with the range of 66.74 to 82.18% obtained for the flour blends in this study. there was decrease in the total carbohydrate content of the flour blends on increasing addition of soy flour. calorific value ranged from 1499.20 calorie for sample g to 1624.60 calorie for sample a. thus, calorific value decreased with increasing supplementation with soy flour. the energy value of food is very important as it helps to determine the fuel value of food. energy is not a nutrient but is required for metabolic process, physiological functions, heat production, muscular activity, synthesis of new tissues and growth. energy is released from food components by oxidation [33]. 3.2. functional properties of flour blends the functional properties of the flour blends are shown in table 2. water absorption capacity ranged from 107.51 for sample a to 135.23 for sample g. there was significant difference (p> 0.05) among all samples. water absorption capacity is the ability of a product to absorb water [34]. similar observations were observed for oil absorption capacity of the samples. increase in the water absorption capacity with increasing addition of soybean flour might not be unconnected with the increase in the lecithin content of the flour blends occasioned by the addition of soybean flour. addition of lecithin has the propensity to hold cause increase in water absorption capacity. good water absorption capacity of these flour blends makes them ideal for preparation of dumpling such as amala. the highest bulk density of 0.95 g/cm3 was recorded for sample g while sample a has the lowest bulk density of 0.78 g/cm3. in 2015, kiin-kabari, et al. [35] reported bulk density of 0.34 g/ml for whole wheat flour. the high bulk density of these flour blends made them unfit to be used as weaning foods. weaning foods are expected to have bulk density of less than 0.6 g/cm3. low bulk density is of benefit in the formulation of infant complementary foods [36]. table 2. functional properties of cocoyam and soybean flour blends. sample absorption property bulk density oac (%) wac (%) (g/cm3) a 61.37 ± 1.34a 107.51 ± 0.82a 0.78 ± 0.01a b 66.07 ± 2.63b 115.20 ± 2.20b 0.81 ± 0.02ab c 67.44 ± 2.14b 116.75 ± 1.54bc 0.83 ± 0.00b d 67.40 ± 2.90b 122.45 ± 0.06cd 0.87 ± 0.01a e 68.57 ± 3.59bc 126.24 ± 2.19d 0.93 ± 0.02d f 76.49 ± 1.34d 134.70 ± 2.70e 0.93 ± 0.01d g 76.91 ± 3.40d 135.23 ± 2.23e 0.95 ± 0.01d note: values with different superscript along the same column are significantly different at p>0.05. wac = water absorption capacity; oac = oil absorption capacity. a = 100% cocoyam flour; b = 95% cocoyam flour + 5% soybean flour; c = 90% cocoyam flour + 10% soybean flour; d = 85% cocoyam flour + 15% soybean flour; e = 80% cocoyam flour + 20% soybean flour; f = 75% cocoyam flour + 25% soybean flour; g = 70% cocoyam flour + 30% soybean flour. table 3. sensory evaluation of amala from cocoyam and soybean flour blends. sample colour appearance taste flavor texture general acceptability a 6.20±0.27a 6.85±0.13a 6.45±0.26a 5.75±0.18a 6.15±0.25a 6.35±0.25a b 6.90±0.32ab 6.70±0.21a 6.60±0.37a 5.90±0.35a 6.50±0.24a 5.90±0.37b c 6.80±0.34a 6.25±0.32a 6.55±0.39a 6.75±0.33a 6.80±0.24a 6.85±0.36b d 6.95±0.29a 7.05±0.30a 7.40±0.30a 6.55±0.26a 6.35±0.31a 7.30±0.24ab e 6.75±0.33a 6.65±0.35a 6.35±0.41a 6.30±0.36a 6.10±0.30ab 6.20±0.34ab f 5.70±0.31a 5.85±0.29a 5.75±0.26b 5.80±0.28a 5.35±0.31a 5.35±0.30ab g 5.60±0.21ab 6.00±0.23ab 5.69±23ab 5.90±0.28a 5.75±0.34a 5.35±0.23ab note: values with different superscript along the same column are significantly different at p>0.05. a = 100% cocoyam flour; b = 95% cocoyam flour + 5% soybean flour; c = 90% cocoyam flour + 10% soybean flour; d = 85% cocoyam flour + 15% soybean flour; e = 80% cocoyam flour + 20% soybean flour; f = 75% cocoyam flour + 25% soybean flour; g = 70% cocoyam flour + 30% soybean flour. agriculture and food sciences research, 2022, 9(1): 44-49 48 © 2022 by the authors; licensee asian online journal publishing group 3.3. sensory evaluation of amala from flour blends the results obtained for the sensory evaluation of amala dumpling from the blends of cocoyam and soybean flours are shown in table 3. in term of colour appearance, taste and flavour, samples b, c, d and e compared favourably well with sample a (100% cocoyam flour). increased substitution with soybean flour at 30% caused significant difference in the texture of the amala dumpling. amala prepared with 15% level of soybean substitution appeared to be more acceptable to the panelist. there was a significant change in texture with 30% substitution. sample d is more preferable to every other sample when consideration for taste, appearance and colour are of importance. amala prepared from sample d, with 15% soybean flour substitution, is the most preferable amala dumpling. 4. conclusion amala from the blends of cocoyam and soybean was produced and evaluated. the results of the proximate composition of the flour blends showed that supplementation of cocoyam flour with soy flour enhanced the concentrations of protein, ash, oil and dietary fibre. improvement in some functional properties on supplementation with soy flour gives a propensity that the flour blends might be useful for other food preparations such as palp or as an adjunct component of bakery products. encouraging increasing consumption of cocoyam amala supplemented with soy flour, a cheap source of plant protein, will not only create dietary diversity, it will make more protein available for the consumers and thus help in alleviating the problem of protein energy malnutrition particularly in developing regions of the world. references [1] o. o. awolu and g. f. oseyemi, "physicochemical and rheological properties of optimized cocoyam-based composite flour comprising cassava starch," food technology, vol. 20, pp. 65 – 84, 2016.available at: https://doi.org/10.1515/aucft-2016-0016. 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[36] m. akpata and p. akubor, "chemical composition and selected functional properties of sweet orange (citrus sinensis) seed flour," plant foods for human nutrition, vol. 54, pp. 353-362, 1999. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 182 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 182-186, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6250 © 2024 by the authors; licensee asian online journal publishing group bio-geotextiles based acrylonitrile and their evaluation on radish plants (raphanus sativus) alcala-sanchez daniel1 tapia-picazo juan-carlos2 ( corresponding author) 1,2department of chemical and biochemical engineering, national technological institute of mexico / technological institute of aguascalientes, aguascalientes, mexico. 1email: das.oficial@hotmail.com 2email: juan.tp@aguascalientes.tecnm.mx abstract in this study, two innovative biopolymers (biop) were investigated for bio-geotextile (biog) fabrication: an acrylonitrile-2-hidroxyethyl methacrylate-starch terpolymer (st-an), and a composite of acrylonitrile-vinyl acetate-chitosan terpolymer (cs-an) blended with acrylonitrilemethyl acrylate (an-ma) copolymer. using wet-spinning technique, bio-fiber (biof) were produced, characterized and utilized in to construct biog. biof based on st-an and cs-an exhibited linear densities of 10.7 and 19.8 denier, bulk densities of 1.21 and 1.16 g/cm³, tensile strengths of 74.7 mpa and 28.5 mpa, elongation of 10.7% and 11.2%, and moisture retention capacities of 88% and 65%, respectively. the biog made with st-an improved soil moisture retention by up to 130%. radish plants biometric measurement in pots with biog revealed improvements in growth parameters: leaf length increased by 87%, leaf width by 45%, and stem thickness by 142% compared to controls. these findings highlight the potential of bio-based materials to advance sustainable engineering through innovative strategies in synthesis, processing, and application, offering a viable alternative for the partial or complete replacement of plastics associated with microplastic generation and persistent environmental pollution. keywords: acrylonitrile-vinyl acetate-chitosan, acrylonitrile-2-hydroxyethyl methacrylate-starch, bio-geotextile, biopolymer, radish, sustainable engineering, wet-spinning. citation | daniel, a.-s., & juan-carlos, t.-p. (2024). bio-geotextiles based acrylonitrile and their evaluation on radish plants (raphanus sativus). agriculture and food sciences research, 11(2), 182–186. 10.20448/aesr.v11i2.6250 history: received: 1 november 2024 revised: 4 december 2024 accepted: 13 december 2024 published: 26 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: both authors contributed equally to the conception and design of the study. both authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 183 2. materials and methods ................................................................................................................................................................. 183 3. results and discussions ................................................................................................................................................................ 184 4. conclusion ....................................................................................................................................................................................... 185 references ............................................................................................................................................................................................ 185 mailto:das.oficial@hotmail.com mailto:juan.tp@aguascalientes.tecnm.mx https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6250 agriculture and food sciences research, 2024, 11(2): 182-186 183 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study investigates the evaluation of novel polymeric biomaterials within a wet-spinning system to produce fibers for bio-geotextile applications. it examines the physicochemical properties of bio-base fiber and their performance in radish cultivation systems, offering insights for future research on biomaterials in sustainable agricultural practices. 1. introduction microplastics (mps) are particles with sizes between 0.05 and 5 mm [1-5] originating from manufacturing processes or the breakdown of larger plastics. accumulation of mps is more prevalent in soil ecosystems than in aquatic environments, whit agricultural soils showing higher concentrations than urban soils [1]. mps found in the soil ecosystem are mainly sourced from sewage sludge, tire wear, industrial processes, plastic waste, and agricultural films [1, 2]. these particles negatively impact soil properties, nutrient cycling, microbial activity, and plant growth by interfering with roots and nutrient absorption. mps can even enter plant tissues, affecting overall plant health and reducing soil fertility, indirectly hindering crop development. beyond soil and aquatic ecosystems, mps pose significant health risks when they enter the human body via food, air, or direct contact, potentially damaging the digestive, respiratory, and circulatory systems. geotextiles are textile structures with a wide range of engineering applications; for example, they are applied in the soil filtration, drainage, separation, reinforcement, protection and revegetation of eroded areas. due to growing environmental concerns, a shift toward biodegradable geotextiles has emerged. the 90% of geotextiles are made from synthetic polymers such as polyethylene, polyester, and polyamides, which are cost-effective and easy to produce and handle [6]. however, the exposure of these materials to environmental factors can cause their disintegration into mps, resulting in high contamination of the environment. materials of natural origin have been used to replace synthetic materials in the construction of geotextiles. although these materials can be fully environmentally compatible and effective in removing heavy metals, they present some drawbacks, including hygroscopicity, which limits their filtering, drainage, and permeability capabilities, as well as short durability and significant loss of mechanical properties over time [7]. another approach for creating such materials involves the production of fibers from biopolymers [6]. recent advances in biodegradable polymers based on starch (st) [8-13] and chitosan (cs) [14-18] have emerged as key candidates, that can be chemically modified, blended, or copolymerized with other monomers to enhance specific physicochemical properties such as solubility, mechanical strength, viscosity, hydrophilicity, and hygroscopicity [19-27]. these biopolymers are not only biodegradable but also exhibit multifunctional characteristics, such as the adsorption of chemical species [28, 29] and superabsorption of water [22, 30]. these advancements are critical for producing fibers suitable for the generation of geotextiles that are biodegradable, biocompatible, and capable of fulfilling roles that natural fibers cannot. accordingly, our research aims to contribute to the development of new biopolymers that can be utilized in the production of biodegradable and biocompatible fiber to obtain bio-geotextile, with a focus on soil remediation and environmental protection. this study contributes to this field by producing biodegradable fibers from st, cs, and acrylonitrile using a wet-spinning technique. a nonwoven bio-geotextile was then fabricated and evaluated through a simple experiment on the growth and development of radish plants. 2. materials and methods two biopolymers were synthesized for this study: acrylonitrile-2-hydroxyethyl methacrylate-starch (st-an) and acrylonitrile-vinyl acetate-chitosan (cs-an) terpolymers. the synthesis was conducted using aqueous suspension polymerization and a free-radical process, according to previous works [31]. the average molecular weight of st-an and cs-an were determined to be 124 x103 g/mol and 49 x103 g/mol, respectively. the wetspinning process was performed on a laboratory-scale spinning machine, and following the operating conditions listed in table 1 [32]. the resulting fibers were impregnated with a 15% (w/w) aqueous npk-urea nutrient solution (50:50 ratio) at room temperature. after 24 h, the fibers were dried in an oven at 50°c and weighed. table 1. operating conditions for wet-spinning. variable st-an based biog cs-an based biog polymeric solution concentration, % wt* 14 18 blend ratio, (cs-an/an-ma) 0.01 coagulation bath concentration, % wt 30 25 coagulation rate, m/s 1 1 drying temperature, °c 80 70 note: * % wt, weight percent. for soil evaporation and plant biometric measurement, experiments were conducted using recycled pet bottles as pots. the st-an based biof was manually felted to develop networks of mechanically bonded fibers. the interwoven fibers were pressed for 24 h at room temperature. table 2 provides the conditions used to evaluate stan based biog performance. table 2. evaluation conditions for st-an based biog. test code weight, g/cm2 nutrient*, g 1 g200 2 g201 0.28 3 g211 0.025 0.28 note: * grams of npk-urea nutrient per grams of geotextile. biog samples were placed at a depth of 15 mm and covered with soil. radish seedlings, germinated for15 days, were placed into pots containing biog or control soil. experiment was conducted in a greenhouse under controlled conditions (25 ±2 °c during the day and 14 ±2 °c at night). soil moisture content was periodically measured by collection and weighing oven-dried samples. pots were irrigated uniformly, and plant growth was monitored for agriculture and food sciences research, 2024, 11(2): 182-186 184 © 2024 by the authors; licensee asian online journal publishing group seven weeks. two pots were used for each test condition. control pots without biog were fertilized with solid nutrient mixes matching the biog nutrient composition. 3. results and discussions 3.1 characterization of fibers biof were produced using the wet-spinning method and characterized for their morphological and physicomechanical properties. figure 1 and table 3 show the results of biof characterization. microscopic analysis revealed differences in cross-sectional shape: st-an based biof exhibited irregular shapes influenced by diffusion and counter-diffusion mechanism during coagulation, figure 1, while cs-an based biof displayed circular shape. both types of biof showed porous morphologies whit macropores and longitudinal grooves, contributing to their functional properties. the morphology of st-an biof exhibits microstructures resembling lignocellulosic fibers, which are determinant in its mechanical behavior and ability to retain moisture. according to the characterization, no significant differences in biof density were observed. the st-an based biof exhibit a high moisture retention capacity, likely due to the porous surface structure and the affinity with hydroxyl groups (oh) from starch and water. conversely, cs-an based biof demonstrated a lower moisture retention capacity, which can be attributed to the hydrophobic nature of the copolymer used in the blend. interestingly, the cs-an based biof show a greater capacity for nutrient absorption, which can be ascribed to the porous internal structure that allows for the penetration of the npk-urea solution. mechanical testing revealed higher tensile strength for st-an biof (74.7 mpa) compared to cs-an biof (28.5 mpa), indicating that specific interactions in the cs-an/an-ma composite may weaken its intermolecular forces, affecting its structural integrity. a) b) c) d) e) f) figure 1. images of biof. a) and d), cross-sectional shape of st-an and cs-an/an-ma based biof, respectively; b) and e), longitudinal cut of st-an and cs-an/an-ma based biof, respectively; (c) and f), longitudinal cut of st-an and cs-an/an-ma based biof (whit nutrient npk-urea), respectively. table 3. characterization of biof obtained by we-spinning. properties st-an based biof cs-an based biof linear density (monofilament), denier 10.7 19.8 linear density (bundle), denier 1200 2220 nominal diameter, μm 34.0 60.7 density, g/cm3 1.21 1.16 tensile strength (monofilament), mpa 74.7 28.5 elongation, % 10.70 11.19 nutrient impregnation, % 39.8 58.5 moisture retention, % 88 65 3.2 preparation and evaluation of bio-geotextile nonwoven biog constructed from st-an biof was evaluated for its effects on soil moisture retention and radish plant growth. in figure 2, the st-an based biog obtained is shown. a simple experiment was carried out to analyze the effect of geotextiles on radish plant growth and soil moisture retention. in the experiment, radish seedlings with germination time 14 days were used. the effective application of geotextiles can be judged by soil moisture content and biometric parameters of plants. figure 3 show the results of radish plant crops tested with and without the use of geotextiles. agriculture and food sciences research, 2024, 11(2): 182-186 185 © 2024 by the authors; licensee asian online journal publishing group figure 2. nonwoven geotextiles obtained from st-an biof impregnated whit npk-urea nutrient. the results in figure 3 indicate that geotextiles can aid soil moisture retention. this means that the water needed for irrigation can be reduced by using geotextiles. it should be noted that for the soil moisture retention test, all pots in the experiment were irrigated with the same amount of water, which allows for a comparative analysis. in the experiments that have soil with lower and higher percentage of moisture, experiments g201 (without geotextile, moisture retention of 5.48%) and g211 (with geotextile, moisture retention of 14.31%), respectively, a saving of more than 130% in irrigation water consumption can be achieved by using the geotextiles. biometric analysis of radish plants demonstrated substantial growth improvements with biog. compared to control plants (g200), leaf length increased by 87%, leaf width by 45%, and stem thickness by 142%. against g201, which used nutrient-only soil, the improvements were 37%, 25%, and 128%, respectively. these results indicate that nutrient-impregnated biog can serve as a sustainable alternative to conventional fertilization methods, providing controlled nutrient delivery and improving plant growth. figure 3. biometric parameters of radish plants with and without biog 4. conclusion this study successfully developed biodegradable bio-fibers (biof) using acrylonitrile-based terpolymers for the construction of bio-geotextiles (biog). the biog demonstrated performance in soil moisture retention and radish plant growth enhancement. specifically, the use of st-an biog resulted in a 130% improvement in soil moisture retention, significantly reducing water requirements for irrigation. biometric analysis of radish plants highlighted substantial increases in leaf length (87%), leaf width (45%), and stem thickness (142%) compared to control experiments. these findings underscore the potential of bio-based materials as sustainable alternatives to conventional geotextiles, offering multifunctional solutions to address agricultural and environmental challenges. by reducing the reliance on synthetic plastics and mitigating microplastic pollution, biog presents an eco-friendly option for soil remediation and crop productivity. future research will focus on improving the mechanical properties of these fibers and exploring their applications in soil reinforcement and environmental protection. references [1] h. yu, y. zhang, w. tan, and z. zhang, "microplastics as an emerging environmental pollutant in agricultural soils: effects on ecosystems and human health," frontiers in environmental science, vol. 10, p. 855292, 2022. https://doi.org/10.3389/fenvs.2022.855292 [2] s. he, y. wei, c. yang, and z. he, "interactions of microplastics and soil pollutants in soil-plant systems," environmental pollution, vol. 315, p. 120357, 2022. https://doi.org/10.1016/j.envpol.2022.120357 [3] z. zhang, s. zhao, l. chen, c. duan, x. zhang, and l. fang, "a review of microplastics in soil: occurrence, analytical methods, combined contamination and risks," environmental pollution, vol. 306, p. 119374, 2022. https://doi.org/10.1016/j.envpol.2022.119374 https://doi.org/10.3389/fenvs.2022.855292 https://doi.org/10.1016/j.envpol.2022.120357 https://doi.org/10.1016/j.envpol.2022.119374 agriculture and food sciences research, 2024, 11(2): 182-186 186 © 2024 by the authors; 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[32] d. alcalá-sánchez, j.-c. tapia-picazo, a. bonilla-petriciolet, g. luna-bárcenas, j. lópez-romero, and a. álvarez-castillo, "analysis of terpolymerization systems for the development of carbon fiber precursors of pan," international journal of polymer science, vol. 2020, no. 1, p. 8029516, 2020. https://doi.org/10.1155/2020/8029516 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.eti.2022.102408 https://doi.org/10.1016/j.coesh.2022.100344 https://doi.org/10.1016/j.geotexmem.2018.09.006 https://doi.org/10.3390/ma16134849 https://doi.org/10.3144/expresspolymlett.2007.10 https://doi.org/10.1007/s10965-020-02069-6 https://doi.org/10.1016/j.polymdegradstab.2010.05.024 https://doi.org/10.1007/s00289-018-2484-x https://doi.org/10.1007/s11356-014-2946-0 https://doi.org/10.1007/s10924-012-0518-2 https://doi.org/10.1016/j.ijbiomac.2018.04.084 https://doi.org/10.3390/polym14183735 https://doi.org/10.1016/j.polymer.2009.10.004 https://doi.org/10.1016/j.ijbiomac.2017.11.129 https://doi.org/10.1515/polyeng-2013-0105 https://doi.org/10.3390/ijms22094325 https://doi.org/10.1016/j.sajce.2023.12.005 https://doi.org/10.1007/s00289-021-03842-3 https://doi.org/10.1016/j.carbpol.2005.07.016 https://doi.org/10.1002/adv.21943 https://doi.org/10.1080/02757540.2021.1886280 https://doi.org/10.1007/s42452-020-2753-9 https://doi.org/10.1016/j.molliq.2021.115815 https://doi.org/10.1016/j.trgeo.2022.100910 https://doi.org/10.1016/j.reactfunctpolym.2012.09.002 https://doi.org/10.3390/polym13234115 https://doi.org/10.1155/2020/8029516 110 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 110-115, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6179 © 2024 by the authors; licensee asian online journal publishing group design and experimentation of a pressure testing platform for tea tree canopy pengfei wang1 yongguang hu2 decheng liu3 qingmin pan4 wenqiang wu5 ( corresponding author) 1,2,3,4,5jiangsu province, zhenjiang, jiangsu university, china. 1email: jsu.wpf@gmail.com 2email: deerhu@ujs.edu.cn 3email: 30116103@qq.com 4email: 2112116011@stmail.ujs.edu.cn 5email: dilanshengxue@126.com abstract in the mechanized operations of tea gardens, components like cutting blades and suspension systems frequently interact with the tea tree canopy, resulting in complex push, squeeze, and pressure interactions. this makes it challenging to accurately measure the distribution characteristics of the forces involved. the uncertainty in these mechanical properties complicates mechanical design, posture adjustment, and positioning control. to address this, this study designed and developed a testing platform for pressure measurement in the tea tree canopy, aiming to systematically investigate the force characteristics and structural changes of the canopy under different penetration depths. the platform enabling high-precision recording of canopy responses under varying pressure conditions. the experimental results indicated significant differences in pressure values at different depths and locations. at a penetration depth of 1.0 cm, the minimum pressure value was 3.0 n, while the maximum pressure at a depth of 4.0 cm reached 60.0 n. moreover, the pressure exhibited a clear hierarchical distribution with changes in depth and position. the study observed a hierarchical structure within the tea tree canopy, comprise tender leaves, mature leaves, and branches. these findings provide important evidence and data support for the optimization of tea garden management and harvesting machinery design. keywords: pressure testing, tea tree, hilly region, canopy structure of plants, testing platform design, field experiment. citation | wang, p., hu, y., liu, d., pan, q., & wu, w. (2024). design and experimentation of a pressure testing platform for tea tree canopy. agriculture and food sciences research, 11(2), 110–115. 10.20448/aesr.v11i2.6179 history: received: 24 october 2024 revised: 27 november 2024 accepted: 29 november 2024 published: 2 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the regional science foundation projects (grant number: 32460782) and priority academic program development of jiangsu higher education institutions (grant number: papd-2023-87) institutional review board statement: the ethical committee of the jiangsu university, china has granted approval for this study (ref. no. 20241115001). transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 111 2. materials and methods ................................................................................................................................................................. 111 3. result and analysis ........................................................................................................................................................................ 113 4. conclusions ..................................................................................................................................................................................... 115 references ............................................................................................................................................................................................ 115 mailto:jsu.wpf@gmail.com mailto:deerhu@ujs.edu.cn mailto:30116103@qq.com mailto:2112116011@stmail.ujs.edu.cn mailto:dilanshengxue@126.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6179 https://orcid.org/0009-0009-5798-3478 https://orcid.org/0000-0002-3771-9380 agriculture and food sciences research, 2024, 11(2): 110-115 111 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the article presents a device for testing the pressure of the tea canopy layer, which analogizes the pressure test of soil and has found physical stratification in the actual observation of the tea canopy layer. currently, there is no research on measuring the pressure and stratification characteristics of the tea canopy layer. 1. introduction tea tree (camellia sinensis) is a globally significant economic crop, playing a crucial role in the agricultural and economic systems of countries such as china, japan, and india [1]. as the scale of tea cultivation expands and labor costs rise, traditional manual management and harvesting methods can no longer meet the demands for efficiency and scale in modern tea production [2-4]. consequently, the development and optimization of tea garden management and harvesting machinery have become key to advancing the tea industry towards modernization and automation. however, existing tea garden management and harvesting machinery often face a major challenge: the physical damage to the tea tree canopy structure during operation, which adversely affects the health of the trees and the quantity and quality of the tea leaves. therefore, exploring the mechanical properties of the tea tree canopy under mechanical action and optimizing machinery design to minimize damage to the trees has become a core technical issue in the mechanization of the tea industry. the structure of the tea tree canopy not only influences photosynthesis, gas exchange, and moisture management but also directly affects the forces experienced during mechanical harvesting [3]. different tea varieties and cultivation methods result in distinct canopy structures that exhibit varying mechanical properties under external pressure, raising design requirements for tea harvesting machines [5, 6]. during the harvesting process, the canopy experiences mechanical pressures such as compression and bending. accurately designing the harvesting head to adapt to different canopy structures and reduce unnecessary physical damage directly impacts the efficiency and effectiveness of mechanized harvesting [7, 8]. with the advancement of sensor technology and mechanical analysis methods, studying the force response of the tea tree canopy under mechanical pressure has become increasingly feasible [9, 10]. existing studies have shown significant differences in deformation, stress distribution, and physiological responses of plant canopies under varying external forces; particularly, external pressure significantly alters the physiological activities and morphological characteristics of the tea tree canopy. although there are various tea canopy shot detection models available [11, 12]. research on the pressure characteristics of tea tree canopies remains limited, and there is a lack of systematic experimental platforms to dynamically monitor the behavior of tea trees under stress conditions [13, 14]. this study aims to design an experimental platform to simulate the force conditions of the tea tree canopy under different pressure levels, providing a scientific basis for the design of tea harvesting machines. the platform can precisely apply different pressure levels and monitor the stress distribution, morphological deformation, and physiological responses of the canopy in real time. it integrates stress sensors and data acquisition and analysis modules to record the stress and strain conditions of the canopy under mechanical action. this experimental platform not only offers new methods for understanding the pressure characteristics of tea tree canopies but also provides a theoretical foundation for the design and optimization of harvesting machinery. through a systematic study of the response mechanisms of tea tree canopies under various pressure levels, this research will help enhance the operational efficiency of harvesting machines while reducing negative impacts on tea tree growth, thereby promoting the sustainable development of tea industry mechanization and achieving efficient tea garden management and harvesting. 2. materials and methods 2.1. design principle of test platform the dense foliage of the tea tree canopy can be approximated as a compact solid when a large panel is pressed down upon it, resembling compressible soil [15]. this allows for the study of the tea canopy using principles similar to those applied in soil mechanics. the bekker settlement theory is based on statistical analysis of extensive experimental data and incorporates classical theories of soil mechanics. bekker identified essential differences in the settlement processes between off-road vehicles and structures like foundations. structures typically apply static and heavy loads, resulting in slow settlement rates and minimal displacement, while off-road vehicles exert dynamic and lighter loads, causing rapid and significant settlement. the state of the tea tree canopy structure under pressure aligns with bekker's settlement theory. therefore, applying this theory to the pressure characteristics testing of the tea tree canopy aims to analyze the deformation, compression, and mechanical responses of the canopy when subjected to external pressures (such as those from tea harvesting machines or other mechanical operations). this analysis will provide insights into the growth environment of tea and optimize the equipment used, offering foundational theoretical support for the design of the pressure characteristics testing platform for tea tree canopies. a model of soil settlement under pressure is illustrated in figure 1. figure 1. schematic diagram of soil pressure subsidence. note: 1. soil 2. platen. agriculture and food sciences research, 2024, 11(2): 110-115 112 © 2024 by the authors; licensee asian online journal publishing group 2.2. the overall design and function realization of the test platform to meet the agronomic requirements of standardized tea planting models, the main functions and technical parameters of this testing platform are detailed in table 1. the platform is designed with a gantry structure, as shown in figure 2. it is driven by a stepper motor system allowing for three-axis movement: the y-axis for lateral motion mounted on a moving frame; the z-axis installed above the y-axis for vertical movement; and the x-axis, which is fixed between two parallel y-axes to achieve forward and backward movement. the coordinated motion of the three axes ensures precise control of speed and position for the platform. the control system utilizes a plc (programmable logic controller) to send pulse signals to the stepper motor drivers, converting these signals into angular displacement signals that drive each stepper motor for accurate motion control. in the platform design, the z-axis must support equipment weighing up to 30.0 kg, while the xaxis must carry the weights of the y and z axes, along with associated motors and testing equipment. to accommodate the complex terrain of the fields, the platform is equipped with a wheeled moving frame that allows for height adjustments in four directions, ensuring compatibility with uneven tea garden surfaces. the height of the moving frame is set at 1.2 meters to allow the platform to traverse tea ridges during operation. for the canopy compression tests, a membrane-type pressure sensor is chosen to accurately measure the mechanical responses of the tea tree canopy, as shown in figure 3. this sensor offers high precision, sensitivity, and a wide measurement range (0-100 kg), making it suitable for measuring the pressure characteristics of the canopy. its circular membrane design, which can be either of variable cross-section or flat type, provides excellent resistance to lateral loads, ensuring the stability and accuracy of the measurement data. the compact size of the sensor allows for easy miniaturization, making it suitable for installation within the limited space of the canopy testing platform. by providing continuous and stable mechanical parameters, this sensor offers a scientific basis for analyzing the mechanical characteristics of the tea tree canopy, optimizing its structure, and enhancing both the yield and quality of tea leaves. figure 2. side view of the test bench note: 1 z-axis dynamometer, 2 pressure sensors, 3 pressure plates, 4 racks. figure 3. bellows type tension pressure sensor (0-100kg/m8). 2.3. design and implementation of field experiment on test platform the testing platform is illustrated in figure 4, with dimensions of 1.5 m in height, 0.4 m in length, and 1.8 m in width. during the testing process, the measurement depth is first calibrated on the force gauge of the z-axis, after agriculture and food sciences research, 2024, 11(2): 110-115 113 © 2024 by the authors; licensee asian online journal publishing group which the pressure rod is gradually lowered to the specified depths. the collected data is transmitted in real time to a computer for recording. once measurements at each point are completed, the platform is manually moved to the next position for further measurements. to effectively simulate the impact of harvesting machinery on the tea tree canopy, a square wooden board measuring 20 cm × 20 cm is chosen for the pressure application. this size is determined by several considerations. first, it ensures that the pressure coverage area is moderate, effectively simulating the localized force of harvesting machines without interfering with or damaging the natural structure of the canopy due to excessive area. second, the 20 cm × 20 cm board provides high operational convenience and accuracy for positioning, securing, and collecting pressure data. additionally, this size is consistent with commonly used pressure plate dimensions in soil mechanics, enhancing the scientific validity and comparability of the test data. this choice not only meets the experimental design requirements but also provides a solid basis for subsequent analysis of canopy stress and optimization of equipment. to obtain representative pressure data, adult tea trees covering all canopy levels are selected for testing. as shown in figure 5, the platform integrates pressure sensors, a data processing unit, and a recording system, allowing precise capture of pressure changes at different depths. measurements are taken at 12 points across the canopy cross-section to analyze the distribution and patterns of data at varying depths of application. the zero point is established at the position where the sensor first contacts the canopy, resulting in zero pressure. comparative data indicates that the sensor's readings are unstable at depths less than 10.0 mm, while depths exceeding 70.0 mm lead to canopy collapse, which is detrimental to cutting operations. therefore, the final set pressure depths are established at 10.0 mm, 20.0 mm, 30.0 mm, and 40.0 mm to analyze their effects on the pressure characteristics of the zhongcha 108 canopy. figure 4. test bench and test area. figure 5. location diagram of 12 collection points. 3. result and analysis using the testing platform shown in figure 4, pressure measurements of the tea tree canopy were conducted at various compression depths, with results summarized in table 1. analysis of the pressure data at different depths (1.0 cm, 2.0 cm, 3.0 cm, and 4.0 cm) for each measurement point (illustrated in figure 6) revealed a consistent pattern in pressure changes with respect to location and depth, exhibiting significant differences at varying depths. as the compression depth increased, the overall pressure values showed an upward trend, particularly at depths of 3.0 cm and 4.0 cm, where pressure changes were more pronounced. specifically, at a depth of 1.0 cm, the pressure agriculture and food sciences research, 2024, 11(2): 110-115 114 © 2024 by the authors; licensee asian online journal publishing group curve was relatively flat, with small differences in pressure between measurement points, indicating that the upper structure of the canopy was sparse. at a depth of 2.0 cm, pressure gradually increased, especially around measurement points 6 to 8, where a noticeable rise in pressure suggested an increase in canopy density in that region. at depths of 3.0 cm and 4.0 cm, the pressure curve exhibited peaks at measurement points 6 to 8, with values significantly higher than at other points, reflecting a dense middle layer structure in the tea tree canopy. additionally, the pressure peaks concentrated at measurement points 6 to 8, particularly at depths of 3.0 cm and 4.0 cm, where the pressure sharply increased to its maximum. this indicates that the density of the canopy in this area is significantly greater than in other regions, likely corresponding to the distribution of the main leaves and branches of the tea tree. this phenomenon highlights the layered characteristics of the tea tree canopy: the upper layer (1.0 cm and 2.0 cm) is sparse, while the middle layer (3.0 cm and 4.0 cm) has increased density. at measurement points 9 to 12, the pressure gradually decreased with position, especially at deeper measurement points, indicating that the device had penetrated through the denser parts of the canopy into the sparser lower region. overall, the trends in pressure changes with depth and position suggest that the tea tree canopy can be roughly divided into a sparse outer layer, a dense middle layer, and a sparse lower layer, exhibiting significant stratification. in future research, accumulating more pressure data will facilitate a quantitative assessment of the tea tree canopy structure and the establishment of a canopy pressure model. in agricultural machinery operations, real-time monitoring of compression depth and pressure changes can assist in precisely locating equipment within the canopy, supporting accurate pruning and harvesting of tea trees while enhancing operational precision and efficiency. this finding provides essential theoretical and practical support for the application of smart agricultural equipment in tea tree management. the layered structure of the tea tree canopy is distinctly illustrated in figure 7. from top to bottom, the canopy can be divided into layers of tender leaves, mature leaves, fine branches, and old branches, with differences in physiological function and growth state across layers. this stratification is a result of the tea tree’s natural growth characteristics and pruning management, reflecting the process by which tea farmers optimize harvesting efficiency and improve tea quality. the successful design of the pressure testing platform for the tea tree canopy lays a solid foundation for future large-scale experiments. upcoming work can involve accumulating pressure data across broader tests to refine the canopy pressure model, providing precise design guidelines for intelligent harvesting equipment. furthermore, the analysis method based on pressure data demonstrates good versatility, making it applicable to various tea tree varieties and growth environments, thus offering data support for mechanized harvesting under diverse conditions. table 1. corresponding table of pressure/depth of tea canopy. depth/cm pressure/n 1 2 3 4 5 6 7 8 9 10 11 12 1.0 3.0 3.0 3.0 7.0 5.0 10.0 3.0 6.0 10.0 10.0 4.0 4.0 2.0 9.0 8.0 14.0 17.0 18.0 22.0 24.0 15.0 18.0 16.0 10.0 8.0 3.0 13.0 13.0 16.0 21.0 25.0 31.0 35.0 30.0 21.0 24.0 14.0 15.0 4.0 17.0 26.0 21.0 27.0 30.0 43.0 60.0 40.0 30.0 28.0 25.0 18.0 figure 6. measurement results of tea canopy pressure at the same pressure depth. agriculture and food sciences research, 2024, 11(2): 110-115 115 © 2024 by the authors; licensee asian online journal publishing group figure 7. schematic diagram of longitudinal section of tea row. note: 1 tender layer、2 old leaf layer、3 twig layer、4 old branch layer、5 ground、6 twelve pressure sensor and its position. 4. conclusions the pressure collection system designed in this study demonstrates stable performance and effectively gathers pressure data from the tea tree canopy at various compression depths. data analysis reveals the pressure distribution characteristics of the canopy structure at different locations and depths, confirming the non-uniformity of canopy density. notably, localized areas exhibit peak pressure values, indicating a dense structure within the canopy. these findings provide critical references for understanding canopy characteristics and optimizing the design of tea harvesting equipment. the pressure data analysis enables a more accurate assessment of the specific location of machinery within the tea tree canopy, enhancing operational effectiveness. pressure feedback allows the machinery to adjust operations in real time, facilitating more precise execution of harvesting tasks while minimizing disruption to the deeper canopy structure, thus improving positioning efficiency and protecting the healthy growth of the tea trees. this study deepens the understanding of the tea tree canopy structure and provides a scientific basis for the optimized design of intelligent tea harvesting equipment. by analyzing how pressure varies with depth, harvesting machinery can execute tasks more efficiently and intelligently, significantly contributing to the modernization, intelligent transformation, and sustainable development of the tea industry. references [1] g. meirion-griffith and m. spenko, "a modified pressure–sinkage model for small, rigid wheels on deformable terrains," journal of terramechanics, vol. 48, no. 2, pp. 149-155, 2011. https://doi.org/10.1016/j.jterra.2011.01.001 [2] s. ihara, s. furuhata, s. noda, h. nagai, y. kikuchi, and f. tatsuoka, "estimating the compacted dry density of gravelly soil with oversized particles," transportation geotechnics, vol. 49, p. 101379, 2024. https://doi.org/10.1016/j.trgeo.2024.101379 [3] j. c. garcía‐peña, j. g. ramírez‐gil, c. góngora‐canul, l. calderón, j. cuellar, and c. cruz, "multiscale temporal and spatiotemporal analysis of wheat blast infection‐like dynamics using vertical plant stratification, regression and markov chain approaches," plant pathology, vol. 72, no. 9, pp. 1570-1584, 2023. https://doi.org/10.1111/ppa.13789 [4] m. de cristofaro et al., "modeling the excess porewater pressure buildup in pyroclastic soils subjected to cyclic loading," journal of geotechnical and geoenvironmental engineering, vol. 150, no. 12, p. 06024007, 2024. https://doi.org/10.1061/jggefk.gteng-12129 [5] w. wenchao, "reciprocating cutting mechanism and parameter optimization based on load pattern and bending deformation of tea stalks," doctoral dissertation, jiangsu university, zhenjiang jiangsu china, december, 2023. [6] z. zhang, y. lu, m. yang, g. wang, y. zhao, and y. hu, "optimal training strategy for high-performance detection model of multi-cultivar tea shoots based on deep learning methods," scientia horticulturae, vol. 328, p. 112949, 2024. https://doi.org/10.1016/j.scienta.2024.112949 [7] t. jing, z. tang, z. ding, y. liang, m. fang, and t. wang, "paddy soil compaction effect undergoing multi-dimensional dynamic load of combine harvester crawler," agriculture, vol. 14, no. 2, p. 202, 2024. https://doi.org/10.3390/agriculture14020202 [8] u. veyna, s. garcia-nieto, r. simarro, and j. v. salcedo, "quadcopters testing platform for educational environments," sensors, vol. 21, no. 12, p. 4134, 2021. https://doi.org/10.3390/s21124134 [9] w. yan, h. wu, y. qu, and y. yan, "design of hydraulic driving experimental bench for automatic vertical drilling tool," presented at the in journal of physics: conference series (vol. 1314, no. 1, p. 012121). iop publishing, 2019. [10] j. zehui, "optimal parameter design on fracture tea-picking device for famous and high-quality tea," master's thesis, jiangsu university, zhenjiang jiangsu china, september, 2023. [11] z. zhang, y. lu, and y. zhao, "ts-yolo: an all-day and lightweight tea canopy shoots detection model," agronomy, vol. 13, no. 5, p. 1411, 2023. https://doi.org/10.3390/agronomy13051411 [12] d. h. liu, x. xiao, and l. q. yin, "research on adaptation of machine picking in yunnan tea garden," shandong agricultural science, vol. 47, no. 4, pp. 49-51, 2015. https://doi.org/10.14083/j.issn.1001-4942.2015.04.013 [13] m. r. esmaeili, r. khodabakhshian, and m. khojastehpour, "effect of asafoetida extract solution on red apple quality with a focus on the mechanical properties of peduncle, rate fruit drop and orchard pest control," applied fruit science, pp. 1-11, 2024. https://doi.org/10.1007/s10341-024-01205-6 [14] h. adèle, c. buffet, p. d’arras, a. bourmaud, and c. baley, "winter flax: stem structure, fibre properties and reinforcement potential for composite materials," industrial crops and products, vol. 217, p. 118831, 2024. https://doi.org/10.1016/j.indcrop.2024.118831 [15] m. islam, a. g. nizar, and r. stefan, "a case study on advanced cpt data interpretation: from stratification to soil parameters," geotechnical and geological engineering, vol. 4, no. 5, pp. 4087-4113, 2024. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.jterra.2011.01.001 https://doi.org/10.1016/j.trgeo.2024.101379 https://doi.org/10.1111/ppa.13789 https://doi.org/10.1061/jggefk.gteng-12129 https://doi.org/10.1016/j.scienta.2024.112949 https://doi.org/10.3390/agriculture14020202 https://doi.org/10.3390/s21124134 https://doi.org/10.3390/agronomy13051411 https://doi.org/10.14083/j.issn.1001-4942.2015.04.013 https://doi.org/10.1007/s10341-024-01205-6 https://doi.org/10.1016/j.indcrop.2024.118831 131 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 2, 131-139, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i2.7803 © 2025 by the authors; licensee asian online journal publishing group nutrient use efficiency and growth analysis of wheat crop under precision nitrogen management techniques and growth regulators application manisha kapri1 amit kesarwani2 rajeew kumar3 vineeta rathore4 ( corresponding author) 1,2,3,4department of agronomy, college of agriculture, g.b. pant university of agriculture and technology, india. 1email: manishakapri610@gmail.com 2email: getkesar@gmail.com 3email: shuklarajeew@gmail.com 4email: vineetaagron@gmail.com abstract nitrogen occupies a significant position in plant metabolism as an essential component of proteins, which are associated with all vital and metabolic processes in the plant system. nitrogen fertilization in wheat is more complex, and the results are more flexible than in any other field crop. the objective of this study was to determine how nitrogen dose variation interacts with growth regulators and their impact on crop lodging and the yield of wheat. the experiment was conducted during the rabi season 2019-20 to assess these effects, involving various treatment combinations of different nitrogen doses, use of growth regulators, including precision nitrogen management using leaf color charts, greenseeker, and decision-support tools such as nutrient expert. the results showed that precision nitrogen management techniques resulted in higher harvest index and nitrogen use efficiency compared to other treatments. balanced supply of major nutrients, viz., n, p, and k, led to better nutrient uptake compared to treatments where only nitrogenous fertilizer was applied. the use of growth regulators increased the total number of days to achieve physiological maturity, thereby increasing values of growing degree days and heliothermal units. therefore, it is suggested that nitrogenous fertilizer alone should not be applied in wheat crops to avoid soil nutrient imbalance and potential toxicity disruptions in the food cycle. keywords: chlormequat chloride, growth regulators, nitrogen, nutrient expert, nutrient use efficiency, precision nutrient management, tebuconazole. citation | kapri, m., kesarwani, a., kumar, r., & rathore, v. (2025). nutrient use efficiency and growth analysis of wheat crop under precision nitrogen management techniques and growth regulators application. agriculture and food sciences research, 12(2), 131–139. 10.20448/aesr.v12i2.7803 history: received: 17 october 2025 revised: 21 november 2025 accepted: 28 november 2025 published: 5 december 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 132 2. materials and methods ................................................................................................................................................................. 132 3. results and discussion ................................................................................................................................................................. 134 4. conclusion ....................................................................................................................................................................................... 138 references ............................................................................................................................................................................................ 138 mailto:manishakapri610@gmail.com mailto:getkesar@gmail.com mailto:shuklarajeew@gmail.com mailto:vineetaagron@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i2.7803 https://orcid.org/0009-0003-0672-1941 https://orcid.org/0000-0002-6699-9668 https://orcid.org/0000-0001-7082-2868 https://orcid.org/0000-0002-9209-4790 agriculture and food sciences research, 2025, 12(2): 131-139 132 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to the existing literature on the application of nitrogen, along with growth regulators and precision nutrient management. it demonstrates the relevance of balanced nutrients, especially nitrogen application in crops. although the integrated use of growth regulators can also help achieve higher yield targets and improve soil nutrient status. 1. introduction wheat (triticum aestivum l.) is a universal staple food for 40% of the human population in the world and is the second most important cereal after rice [1]. the massive importance of wheat can be understood from the figures of a grown area of 215.48 mha, with an annual production of 731.46 mt and a productivity of 33.9 q/ha during 2018-19 worldwide [2]. in india, a significant portion of total cultivation is devoted to this crop, covering nearly 29.14 million hectares with an annual production of 102.19 million tonnes, carrying an average productivity of 3,506.8 kg/ha in the year 2018-19 [3] but unfortunately, the productivity of bihar and uttar pradesh is 44% and 34%, respectively, which is less than the productivity of punjab [4]. on a deeper level, these yield gaps can be somewhat related to the nutrient (n+p2o5+k2o) consumption per unit of gross cropped area in each individual state. nitrogen is an indispensable protein component required in all dynamic processes of plants. excessive n causes "luxuriant" growth, resulting in the plant being attractive to insects and diseases. the excessive growth can also reduce the strength of stems, which induces lodging at the time of flowering and grain filling. therefore, deciding the optimum dose of nitrogen for crops is a matter of great concern for better yield. on one hand, high-yielding varieties require more inputs like nitrogenous fertilizers for their better performance, but on the other hand, more nitrogen application causes problems like lodging, which reduces their yield [5]. therefore, it becomes quite important to know whether nitrogen management could reduce lodging risk without reducing the yield potential [6]. plant growth regulators are chemical substances that can alter growth and developmental processes, helping to increase yield, improve grain quality, or facilitate harvesting [7]. the chemicals used are most commonly gibberellin inhibitors that block the early stages of gibberellin synthesis, thereby inhibiting cell elongation and growth, which results in shorter, lodging-tolerant plants. some examples include chlormequat chloride, trinexapac-ethyl, ethephon, peclobutrazol, etc. the blanket recommendations of nitrogenous fertilizers for larger areas have helped to serve the purpose of producing optimum yield, but these recommendations are not always helpful in increasing n use efficiency beyond a certain limit [8]. in general, nitrogen use efficiency (nue) or recovery is only 30-50%. the concept of sitespecific nutrient management can help to improve production per unit of nutrient supply. site-specific nutrient management is a method to adjust soil nutrient supply over time and space to match the crop’s requirements through four key principles: right rate, right product, right time, and right place. many instruments, viz. lcc (leaf colour chart), optical sensors, and decision support systems like nutrient expert can be used for this purpose [9]. 2. materials and methods the centre n. e. borlaug crop research centre of govind ballabh pant university of agriculture and technology, pantnagar, is situated at 29°n latitude and 79.50°e longitude, at an elevation of 243.8 meters above mean sea level. the climate of the region is subtropical, with winters that are extremely cold, and the average temperature ranges from 4°c to 35.4°c. the soil at the experimental site is silty clay loam with a neutral ph of 7.4. it contains medium organic carbon content (0.73%), low available nitrogen (217.16 kg/ha), medium available phosphorus (21.4 kg/ha), and potassium (139.1 kg/ha) (figure 1). figure 1. location map of pantnagar, district udham singh nagar, uttarakhand. agriculture and food sciences research, 2025, 12(2): 131-139 133 © 2025 by the authors; licensee asian online journal publishing group the experiment was laid out in a randomized block design with 13 treatments and 3 replications. the treatments included an absolute control, 50% n (recommended dose of n) (75 kg n/ha), 75% n (112.5 kg n/ha), 100% n (150 kg n/ha), 125% n (187.5 kg n/ha), 150% n (225 kg n/ha), 100% rdf (150:60:40 kg n p2o5 k2o / ha). three treatments involved growth regulator spray with chlormequat chloride (lihocin) @ 0.2% of the commercial product dose and tebuconazole (folicur 430 sc) @ 0.1% of the commercial product at the first node and boot leaf stage, along with application of 125% n, 150% n, and 150% rdf in respective treatments. additionally, three precision nutrient management treatments were included. n management by lcc (apply 45kg n/ha if lcc<4 and 35kg n/ha if lcc≥ 4) (135 kg n/ha), n management by green seeker (116.5 kg n/ha) and n management by nutrient expert (125: 71: 89 kg n p2o5 k2o / ha). n was applied in 3 split doses (1/3rd basal + 1/3rd at 1st irrigation + 1/3rd at 2nd irrigation) in only n-containing treatments. full dose of p and k and 1/3rd of n was applied at the time of sowing, and the remaining 2/3rd of n was applied as 1/3rd at first irrigation and 1/3rd at second irrigation. in lcc-based n management, a full dose of p, k, and 1/3rd n was applied as basal, and 1/3rd n at the time of the first irrigation. the readings of lcc were taken at the time of the second irrigation, and 35 kg n/ha was applied as the lcc reading was 4. similarly, in green seeker-based n management treatment, 1/3rd n and full p and k were applied as basal, and 1/3rd at the time of the first irrigation. the remaining n was applied based on green seeker ndvi readings taken at the time of the third irrigation. in the case of nutrient expertbased n management, the fertilizer doses were determined through the ssnm nutrient expert software with a yield target of 8 t/ha. wheat variety wh 1105 was sown at a row spacing of 20 cm on 14th november 2019. other management practices, including irrigation, weeding, and hoeing, were adopted as per the package of practices for the wheat crop. crop growth rate and relative growth rate during different time intervals were calculated using the following formula [10]. cgr = 𝑊2 −w1 𝑇2−𝑇1 where, w2 = dry weight of plant/m2 recorded at time t2. w1 = dry weight of plant/m2 recorded at time t1. rgr = = 𝑙𝑛𝑊2 − lnw1 𝑇2−𝑇1 where, ln = natural log. w2 = dry weight of plant/m2 recorded at time t2. w1 = dry weight of plant/m2 recorded at time t1. the collected grain and straw samples, after harvest, were oven-dried at 70 ± 1 ˚c for 72 hours and ground to estimate the nitrogen content (%) in the samples, which was then converted to kg/ha. nitrogen uptake by grain (kg/ha) = nitrogen conc.in grain (%)×grain yield ( kg/ha ) 100 nitrogen uptake by straw (kg/ha) = nitrogen conc.in straw (%)×straw yield ( kg/ha ) 100 total nitrogen uptake (kg/ha) = nitrogen uptake by grain + nitrogen uptake by straw plant samples were analyzed for phosphorus concentration (%) in grain and straw. plant samples were dried in a dryer at 70 ± 1°c for 72 hours and followed by wet digestion using the vanado-molybdo-phosphoric acid yellow color method [11]. the total phosphorus uptake was calculated as described below. phosphorus uptake by grain (kg/ha) = phosphorus conc.in grain (%)×grain yield ( kg/ha ) 100 phosphorus uptake by straw (kg/ha) = phosphorus conc.in straw (%)×straw yield ( kg/ha ) 100 total phosphorus uptake (kg/ha) = phosphorus uptake by grain + phosphorus uptake by straw. plant samples were analyzed for potassium concentration (%) with the help of a flame photometer, and the total potassium uptake by grain and straw was calculated separately. total potassium uptake (kg/ha) was determined as follows. potassium uptake by grain(kg/ha) = potassium conc.in grain (%)×grain yield ( kg/ha ) 100 potassium uptake by straw (kg/ha) = potassium conc.in straw (%)×straw yield ( kg/ha ) 100 total potassium uptake = phosphorus uptake by grain + phosphorus uptake by straw. partial factor productivity and agronomic efficiency of n were calculated by formula [12]. partial factor productivity (kg/kg n) = 𝐺𝑟𝑎𝑖𝑛𝑦𝑖𝑒𝑙𝑑 (𝑘𝑔/ℎ𝑎) 𝑎𝑚𝑜𝑢𝑛𝑡𝑜𝑓𝑛𝑢𝑡𝑟𝑖𝑒𝑛𝑡𝑎𝑝𝑝𝑙𝑖𝑒𝑑 (𝑘𝑔/ℎ𝑎) agronomic efficiency (%) = 𝑌𝑁−𝑌0 𝑁 ×100 where, 𝑌𝑁 = crop yield (kg/ha) with applied nutrient. 𝑌0= crop yield (kg/ha) without nutrients. n = amount of nutrient applied (kg/ha). weekly weather observations were recorded at the meteorological observatory situated at the research centre, and growing degree days and heliothermal units were calculated using the following formula [13]. gdd (˚c days) = σ t max + t min 2 − base temp. the base temperature for wheat was taken as 5 ˚c. htu = σ gdd x bright (actual) sunshine hrs agriculture and food sciences research, 2025, 12(2): 131-139 134 © 2025 by the authors; licensee asian online journal publishing group observations on different growth and yield parameters were taken, and the data were statistically analyzed with the help of the statistical program opstat. 3. results and discussion 3.1. crop growth rate (g/m2/ day) different treatments had shown no significant effect on cgr values during 90-120 das, while a significant effect was reported during the early stages of the crop (figure 2). cgr increased slowly at early stages of growth and reached the peak at booting to heading stage; thereafter, it declined. the mean value of cgr was lowest in the absolute control during 30-60 das, while achieving the highest value in 100% rdf and 125% n during 60-90 das. these results are in accordance with the results obtained by sugár et al. [14]. this was due to the maximum production of dry matter at vegetative stages and indicates that dry matter production increases throughout the life cycle, but at a slow rate in later stages. crop growth rate (cgr) is an index of agricultural productivity of land in terms of the plant biomass produced per unit area. it represents the net result of photosynthesis, respiration, and canopy area interception [15]. 3.2. relative growth rate (g/g/day) relative growth rate (rgr) is used to express growth in terms of the rate of increase in size per unit of size. there was no significant effect of nitrogen doses on rgr values during 90-120 das, but a significant effect was observed during the initial stages until 90 das (figure 3). the values of rgr were maximum during 6090 das, and after that, rgr values declined. sugár et al. [14] reported similar results. the observed decline in rgr with increasing size is accompanied by systematic changes in physiology, morphology, and allocation [16] often referred to as ontogenetic drift. the rgr values at later stages decrease for several reasons: non-photosynthetic biomass (roots and stems) increases, the top leaves of a plant begin to shade lower leaves, and soil nutrients can become limiting. as rgr is a total function of plant biomass change with time, it indicates that in later stages of plant growth, biomass increases but at a relatively slower rate compared to the initial period. figure 2. effect of different treatments on crop growth rate (g/m2/day) at different intervals. 3.3. harvest index (%) there was no significant difference observed in harvest index due to different treatments (figure 4). the maximum harvest index was recorded from green seeker-based n management, followed by the application of 50% n. while a minimum harvest index was observed from the application of 150% n. a low harvest index means that there is less translocation of assimilates from the source to the sink, which results in less development of seeds. when the harvest index is high, it means that more assimilates were translocated from the source to the grains, which results in improved development and filling [17]. the more nitrogen present, the greater the vegetative growth and development, but only up to a certain limit. beyond that limit, it can cause toxicity to the plant and have adverse effects on yield and yield parameters. therefore, n doses exceeding 125% resulted in a low harvest index [18]. agriculture and food sciences research, 2025, 12(2): 131-139 135 © 2025 by the authors; licensee asian online journal publishing group figure 3. effect of different treatments on the relative growth rate (g/g/day). 3.4. nitrogen content (%) and uptake in grain and straw a gradual and significant increase in nitrogen content in grain and straw was observed with an increase in the dose of nitrogen (table 1). n content in grain varies from 1.18% in the absolute control to 2.01% in 150% rdf + gr. this may be the effect of increased availability of nitrogen to the plants due to higher application amounts. as proper availability of k was present under 150% rdf, and k plays a very important role in the translocation of nutrients in the plants, therefore, n% in grain was higher in this treatment [19]. similarly, maximum n content in straw was observed with the application of 125% n+gr and nutrient expert-based nitrogen management. the application of growth regulators has a positive impact on grain and straw nitrogen content. under similar n application, treatments with growth regulator application showed higher values of grain and straw nitrogen content. belete et al. [20] and singh et al. [21] reported similar results. nutrient expert gave the best results among all precision nitrogen management practices. as nutrients were applied at the right dose and the right time with the help of a software system, the availability of nitrogen content was increased to the plants. figure 4. effect of different treatments on harvest index (%). significant effects of different nitrogen doses and plant growth regulators were observed in the uptake of nitrogen by the crop. n uptake in grain and straw was significantly increased with an increase in the dose of nitrogen. maximum n uptake in grain was obtained from the application of 150% rdf + gr, followed by 100% rdf, while maximum n uptake in straw was recorded in nitrogen management by nutrient expert. eytoo [22] reported similar results. total n uptake is directly related to the grain and straw yield obtained from the particular agriculture and food sciences research, 2025, 12(2): 131-139 136 © 2025 by the authors; licensee asian online journal publishing group treatment and is found to be maximum with the application of 150% rdf + gr. the balanced supply of major nutrients, viz. n, p, and k might have resulted in better development of the active root system, which efficiently helps in better absorption of nutrients from the soil, producing higher biomass and nutrient concentration, resulting in higher nutrient uptake [20]. table 1. effect of different treatments on n content (%) and uptake in grain and straw. sr. no treatment n content (%) n uptake (kg/ha) grain straw grain straw total 1 absolute control 1.18 0.30 28.9 13.9 42.8 2 50% n 1.20 0.38 51.8 29.3 81.1 3 75% n 1.37 0.40 62.9 36.9 99.8 4 100% n 1.77 0.44 82.9 42.3 125.2 5 125% n 1.86 0.46 92.2 43.2 135.4 6 150% n 1.87 0.41 87.2 39.2 126.4 7 100% rdf 1.99 0.49 102.5 47.3 149.9 8 125% n+gr 1.89 0.50 98.8 47.5 146.3 9 150% n+gr 1.90 0.42 93.2 40.3 140.2 10 150%rdf+gr 2.01 0.49 106.1 46.3 152.8 11 lcc 1.66 0.48 79.2 43.5 122.7 12 green seeker 1.62 0.49 76.6 41.2 117.8 13 nutrient expert 1.85 0.50 94.5 48.2 142.7 se(m) ± 0.04 0.01 2.9 2.1 4.2 c.d. (p = 0.05) 0.11 0.03 8.6 6.1 12.4 3.5. phosphorus content (%) and uptake in grain and straw there was a significant difference observed in grain and straw p content due to different nitrogen doses (table 2). the maximum value of grain p content was obtained with the application of 150% rdf + gr, followed by nutrient expert-based n management. however, p content in straw was maximum under both nutrient expert and 150% rdf + gr, followed by 100% rdf. the reason behind the higher p content in these two treatments may be the application of a higher amount of p, which has increased the availability of p to plants. n also plays an important role in the uptake of other nutrients. it encourages the uptake and utilization of other nutrients, including potassium and phosphorus, and controls the overall growth of the plant [23]. treatments in which only n was applied without p and k showed a decrease in p content (%) in grain and straw with an increase in the dose of n. as no p was applied, the application of more n resulted in increased plant growth, and due to the dilution of available phosphorus, less content was observed. total p uptake in plants depends on p content (%) in grain and straw as well as grain and straw yield, and it was found to be maximum with the application of 150% rdf + gr. p uptake in grains ranged from 8.1 kg/ha in the absolute control to 26.3 kg/ha in 150% rdf + gr. p uptake in straw ranged from 6.3 kg/ha in the absolute control to 16.6 kg/ha in nutrient expert-based n management. the amount and timing of application of p in the above two treatments, viz. 150% rdf + gr and nutrient expert played an important role in increasing the availability and uptake of p by the crop. table 2. effect of different treatments on p content (%) and uptake in grain and straw. sr. no. treatment p content (%) p uptake (kg/ha) grain straw grain straw total 1 absolute control 0.33 0.13 8.1 6.3 14.3 2 50% n 0.32 0.13 13.8 9.7 23.5 3 75% n 0.32 0.12 14.7 11.2 26.0 4 100% n 0.31 0.11 14.6 10.3 24.9 5 125% n 0.31 0.11 15.2 10.0 25.3 6 150% n 0.30 0.10 14.0 9.5 23.5 7 100% rdf 0.45 0.16 23.1 15.8 38.9 8 125% n+gr 0.30 0.12 15.9 11.3 27.1 9 150% n+gr 0.29 0.10 14.4 9.8 24.2 10 150%rdf+gr 0.50 0.17 26.3 16.1 42.4 11 lcc 0.44 0.15 21.0 13.5 34.5 12 green seeker 0.42 0.15 20.0 12.8 32.8 13 nutrient expert 0.49 0.17 24.9 16.6 41.4 se(m) ± 0.01 0.01 0.8 0.8 1.1 c.d. (p = 0.05) 0.02 0.02 2.4 2.3 3.3 3.6. potassium content (%) and uptake in grain and straw significant differences were observed in potassium content (%) in grain and straw due to different treatments containing various n doses (table 3). more amount of potassium in grains and straw was obtained from the treatments in which basal application of potassium was done. maximum k content in grains was recorded at 150% rdf + gr, followed by nutrient expert, while maximum k content in straw was obtained at 150% rdf + gr, followed by 100% rdf. remaining precision nitrogen management treatments, viz. lcc and green seeker-based n management have shown better values of k content in both grains and straw compared to other treatments in which only nitrogen was applied. agriculture and food sciences research, 2025, 12(2): 131-139 137 © 2025 by the authors; licensee asian online journal publishing group table 3. effect of different treatments on k content (%) and uptake in grain and straw. sr. no. treatment k content (%) k uptake (kg/ha) grain straw grain straw total 1 absolute control 0.43 0.97 10.6 45.3 55.9 2 50% n 0.42 0.97 18.2 74.2 92.3 3 75% n 0.42 0.96 19.2 87.8 106.9 4 100% n 0.41 0.95 19.4 91.0 110.4 5 125% n 0.40 0.95 20.0 89.6 109.6 6 150% n 0.40 0.93 18.6 88.9 107.5 7 100% rdf 0.63 1.47 32.2 143.1 175.3 8 125% n+gr 0.41 0.97 21.6 91.1 112.7 9 150% n+gr 0.39 0.94 19.3 89.6 108.9 10 150%rdf+gr 0.65 1.53 34.4 144.6 179.0 11 lcc 0.62 1.30 29.3 117.7 147.1 12 green seeker 0.61 1.20 28.8 99.7 128.5 13 nutrient expert 0.64 1.37 32.8 130.7 163.5 se(m) ± 0.01 0.09 1.0 9.8 9.8 c.d. (p = 0.05) 0.03 0.26 2.9 28.9 28.6 significant effects of different treatments were observed in grain and straw potassium (k) uptake. higher values of potassium uptake were observed from the treatments with application of potassium compared to the treatments in which only nitrogen was applied. the highest potassium uptake in grains was recorded at 150% rdf+gr, followed by nutrient expert. the highest potassium uptake in straw was obtained with 150% rdf+gr, followed by 100% rdf. the improved grain and stover (straw) yield also enhanced potassium uptake in the crop, which was also confirmed in the findings of shivay et al. [24]. 3.7. partial factor productivity of nitrogen (kg/kg n) partial factor productivity of n is a useful measure of nitrogen use efficiency as it provides an integrative index that quantifies total economic output relative to the use of nutrient resources in the system [25]. there was a significant difference observed in the partial factor productivity of nitrogen due to different treatments (figure 5). it was observed that with an increase in the level of nitrogen, partial factor productivity decreased. significantly, the maximum (57.6 kg/kg n) and minimum (20.7 kg/kg n) pfp were observed with the application of 50% rdn and 150% rdn, respectively. panayotova and kostadinova [26] reported similar results. precision nutrient management treatments consisting of n management by green seeker and nutrient expert show better partial factor productivity of n compared to the remaining treatments. the fertilizer efficiency increases when suitable sources of nutrients are applied at the exact rate, time, and place, which increases the yield, decreases the loss of nutrients, and improves soil fertility. nitrogen utilization efficiency (nue), or n uptake per unit n applied, is greatest where the yield response to n is highest. therefore, nue is generally greatest with low levels of applied n and decreases as the amount of n applied increases [27, 28]. 3.8. agronomic efficiency of nitrogen (%) agronomic efficiency of nitrogen was significantly affected by different treatments. with an increase in the level of nitrogen, agronomic efficiency decreased (figure 5). maximum (24.9%) and minimum agronomic efficiency (9.8%) were obtained from treatments with application of 50% n and 150% n, respectively. similar results are reported by ayadi et al. [29]. similar to pfp of n, better agronomic efficiency of nitrogen is obtained from precision nutrient management treatments in which n was applied on the basis of nutrient expert and green seeker. figure 5. effect of different treatments on partial factor productivity (kg/kg n) and agronomic efficiency (%). agriculture and food sciences research, 2025, 12(2): 131-139 138 © 2025 by the authors; licensee asian online journal publishing group 3.9. growing degree days (˚c days) and helio thermal units the relationship between temperature and the length of different growth stages of crops can be well understood with the help of growing degree days. the accumulated gdd, taken from the date of sowing to the physiological maturity of wheat for different treatments, is presented in figure 6. wheat crop supplied with higher nitrogen doses accumulated more growing degree days for attaining physiological maturity. the gdd values for different treatments range from 1303 in the absolute control to 1543 in the treatment with application of 150% n with growth regulator. similar results related to the effect of nitrogen doses on gdd are reported by sidhu and raj [30]. due to the application of more nitrogen, the vegetative phase of the crop was enhanced, which resulted in the accumulation of more growing degree days [31]. application of plant growth regulators also increases the gdd accumulation to attain maturity as the total growing period length increases. similarly, the accumulated heliothermal units required to attain physiological maturity differed greatly among different nitrogen doses and growth regulator applications. maximum and minimum values of heliothermal units were recorded from the application of 150% n + gr and absolute control, respectively. the higher values of heliothermal units may be due to the longer duration of the wheat crop in 150% + gr as compared to lower levels of nitrogen doses. the findings confirmed the results of kaur et al. [31]. figure 6. effect of different treatments on growing degree days (˚c days) and helio thermal units. 4. conclusion from the conducted research, it can be concluded that a balanced application of nutrients (npk) is key to achieving better yield of wheat crops in terms of both quality and quantity. the application of only nitrogenous fertilizer should not be recommended, as it leads to a lower harvest index, nitrogen use efficiency, and nutrient uptake in plants. precision nitrogen management techniques have significant potential to increase nitrogen use efficiency and crop yield with reduced use of nitrogenous fertilizer. references [1] a. dwivedi, n. s. rana, r. k. naresh, b. p. dhyani, a. singh, and u. p. shahi, "effect of nutrient management strategies and sources of nutrient on yield and nutrient uptake in wheat under subtropical conditions," bulletin of environment, pharmacology and life sciences, vol. 6, no. 11, pp. 101-107, 2017. 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(2025). freeze assisted-aqueous extraction of rapeseed oil using tween 20. agriculture and food sciences research, 12(1), 31–41. 10.20448/aesr.v12i1.6569 history: received: 28 february 2025 revised: 1 april 2025 accepted: 4 april 2025 published: 9 april 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the research and technology deputy of tarbiat modares university, iran. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: both authors contributed equally to the conception and design of the study. both authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 32 2. materials and methods ................................................................................................................................................................... 33 3. results and discussion ................................................................................................................................................................... 34 4. conclusions ....................................................................................................................................................................................... 40 references .............................................................................................................................................................................................. 40 mailto:ebrahim.ammari.coc@gmail.com mailto:sabbasifood@modares.ac.ir https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6569 https://orcid.org/0000-0003-1503-2227 agriculture and food sciences research, 2025, 12(1): 31-41 32 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the present study, for the first time, examined the capability of tween 20 in recovering oil from crushed rapeseeds under optimized conditions. the findings proved the potential effectiveness of tween 20 under alkaline ph and very low concentrations to recover up to 50% of rapeseed oil. 1. introduction vegetable oils are used in numerous industries, including snacks, cakes, margarine, biscuits, cosmetics, detergents, plastics, etc. with a 42% share of the global market, vegetable oils are among the most important agricultural products (hamm et al. [1]). shahidi [2] noted that rapeseed normally contains 38-44% oil, which can be extracted by mechanical pressing (60% yield) and solvent extraction, primarily using hexane [2]. it is essential that an extraction process is used as part of food processing in order to obtain a particular compound from raw materials [3]. generally speaking, traditional, industrial, and novel methods are all used to extract oil from oilseeds. as part of traditional methods, seeds are prepared and pressed by hand, which involves manual labor. meanwhile, industrial methods utilize chemical and mechanical processes. by using cutting-edge technologies, the quantity and quality of oil extraction can potentially be improved. however, each of these approaches has advantages and disadvantages. therefore, a number of factors are usually considered when choosing the most appropriate method of oil extraction, including the type of seeds, oil content, volume of production, quality of oil required, intended use of the oil, and the availability of appropriate technology [4]. the novel extraction methods, namely: supercritical fluid extraction (sfe), ionic liquid extraction (il), deep eutectic solvents (des), pressurized liquid extraction (ple), supercritical liquid extraction (sle), ultrasound-assisted extraction (uae), microwave-assisted extraction (mae), and aqueous extraction processing (aep) [5-7], can potentially offer less extraction time, use environmentally friendly solvents, reduce solvent consumption, provide full automation, and improve reliability [8, 9]. the aep, as an innovative and sustainable method, is already used to extract edible oils from sesame seeds, babassu fruit, and other agricultural products. a water-based extraction process has a lower environmental impact and energy cost than chemical solvents, such as hexane. an aqueous extraction procedure consists of several steps, including pretreatment (milling), the formation of an emulsion, and the separation of the oil from the emulsion using a variety of methods [10]. three common extraction methods are enzyme-assisted extraction (eaae), ultrasoundassisted extraction (uae), and surfactant-assisted extraction (saaep). in the latter, by using an appropriate surfactant at an optimized level, the oil extraction is conducted [11, 12]. a surfactant molecule has two distinct properties: a hydrophilic (water-loving) head and a hydrophobic (oil-loving) tail. this 'amphiphilic' structure allows it to position itself at the interface between oil and water, reducing the surface tension between the two and enabling them to mix together [13, 14]. the extraction process works by having surfactant molecules gather at the boundary between oil and water, which lowers the surface tension between them. the best results occur when the amount of surfactant reaches the ideal level to form micelles [15]. according to existing reports (mostly non-food), oil removal from various matrices can be achieved by three mechanisms, namely: roll-up or roll-back, snap-off (also known as necking and drawing/emulsification/solubilization), and diffusion or micellar solubilization [13, 14]. in roll-up or roll-back, when surfactant adsorbs at the oil–water interface, it increases the contact angle between the oil and the solid surface and, at the same time, decreases the interfacial tension (ift) between oil and surfactant solution. owing to the ift reduction between the oil and surfactant solution, as well as the solid matrix and surfactant solution, the contact angle of the attached oil droplet increases. in ideal circumstances, the contact angle reaches ∼180°, so that the oil droplet completely detaches from the solid matrix. in contrast to the roll-up mechanism, the snap-off mechanism is applicable when the contact angle is not high enough to detach an oil droplet from the solid surface. therefore, an incomplete detachment of the elongated oil droplets occurs, and the detached oil droplets are then emulsified. finally, in the diffusion or micellar solubilization mechanism, the surfactant micelles dissociate near the oil–water interface and then reform around the oil phase, essentially partitioning the oil into the aqueous phase [13]. based on the origin of surfactants, they can be classified either as natural or synthetic, as well as being classified as food-grade or non-food-grade ones. even though long-chain (or 'extended') non-food-grade surfactants are often very effective in forming emulsions and microemulsions, their toxicity and environmental concerns make them unsuitable for use in food applications [13, 16]. tweens, which are also called polyoxyethylene sorbitan esters (polysorbates), are nonionic, hydrophilic food-grade emulsifiers. sorbitan fatty acid esters are synthesized by adding ethylene oxide. these include tween 20, 40, 60, 65, 80, and 83. their solubility varies; tween 20 is more hydrophilic than tween 60 and tween 80, while tween 65 is the most lipophilic [13]. a yellow liquid made by the ethoxylation of sorbitan monolaurate is known as tween 20 (c₅₈h₁₁₄o₂₆). it is soluble in both water and ethanol. food manufacturers frequently use it in ice cream, baked goods, and salad dressings [17]. in addition, in order to reduce surface tension, tween 20 can also be utilized to extract oil from seeds [14, 18]. by now, several reports have shown the effectiveness of tween 20 on the extraction of oils. in the presence of ethanol, tween 20 formed the largest microemulsion region and extracted 69% of the lipid components from bee propolis [19]. as a result of its use in combination with maceration techniques, tween 20 increased the essential oil yield from rose by 94%, emphasizing its role in increasing cell wall permeability and softening tissues [20]. using tween 20 (0.5%) and heating (60°c for one hour), 80% oil (rich in unsaturated fatty acids and antioxidants) was extracted from oilseeds [21]. similarly, tween 20 (1.2% w/w) at ph 10 was able to extract 76.1% of peanut oil [22]. using tween 20 and span 20 (1%), 80% of the oil was extracted while the bioactive compounds such as phenols and unsaturated fatty acids were preserved [15]. furthermore, cyclic extraction of camellia seed oil was facilitated by tween 20 at 70°c and ph 9.0, whilst interfacial tensions were reduced and overall yields increased [23]. despite its higher hydrophobicity and ability to displace oleosin proteins, tween 20 extracted 60% of walnut oil, whereas with span 20, 91.2% of oil was extracted [24]. lastly, hasanah et al. [25] found that tween 20 (0.5%) in combination with ultrasound treatment recovered 18.54% of rice bran oil [25]. based on these studies, it is evident that tween 20 can potentially improve oil extraction efficiency, quality, and sustainability. considering the potential capability of sae and tween 20, it is evident that tween 20 can reduce interfacial tension and facilitate micelle formation, making it a promising candidate for an efficient and green oil extraction agriculture and food sciences research, 2025, 12(1): 31-41 33 © 2025 by the authors; licensee asian online journal publishing group method. this study aimed to evaluate and optimize the effects of different concentrations of tween 20, along with the rapeseed-to-water ratio, temperature, ph, and freezing-defrosting treatments on the oil extraction yield from crushed rapeseeds. 2. materials and methods 2.1. materials the materials used in this study included tween 20 (thermo fisher scientific, usa), rapeseeds (obtained from oksidaneh oil extraction plant, tehran, iran), hexane (dr. mojallali chemical industries, tehran, iran), sodium hydroxide, and hydrochloric acid (merck chemical industries, germany). double-distilled water was used for all sample preparations. 2.2. methods 2.2.1. determination of selected characteristics of rapeseeds the protein content of rapeseeds was determined via the kjeldahl method using a kjeltec 8400 (foss, sweden). a 0.3 g sample was mixed with 1 g of catalyst and 7 ml of concentrated sulfuric acid. digestion was performed at 360°c for 3 hours. the resulting ammonia was captured in a titration solution and titrated against a standard acid to determine nitrogen content. crude protein was calculated by multiplying the nitrogen content by 6.25 (the standard conversion factor for canola). for ash content measurement, rapeseeds were cleaned, dried, and finely ground. a 2.0 g sample was weighed (vibra ab 323, japan) into a porcelain crucible. the sample was burned in a muffle furnace (heareus, germany) at 500-600°c for several hours. after cooling in a desiccator, the crucible and ash were reweighed. ash content was calculated as the percentage difference between the initial and final weights, based on the dry sample weight [26]. in order to measure the moisture content, 5.0 g of the sample was accurately weighed and placed in the sample chamber of a moisture analyzer (sartorius ma35, germany). during the analysis, the instrument continuously monitored the sample weight until a constant weight was reached, indicating the complete removal of moisture. the instrument then automatically calculated and displayed the moisture content percentage of the sample. the oil of ground rapeseeds was extracted with hexane in a soxhlet apparatus (gbg, iran) at 60°c for 6 hours. the solvent was then removed using a rotary evaporator (ika ks 4000i, germany) under reduced pressure. the oil content was calculated using the equation [27]. 2.2.2. surfactant-assisted aqueous extraction an aqueous method assisted by tween 20 was used to investigate the process of oil extraction from crushed rapeseeds. the seeds were obtained from an oil processing plant and stored at 5°c. prior to the experiment, the seeds were cleaned and then subjected to a heat pre-treatment at temperatures ranging from 170 to 210°c. after cooling, the seeds were ground using a high-speed mill (toos shekan khorasan, iran) to reduce the particle size to < 400 µm. the concentration of tween 20 solution (thermo fisher scientific, usa) ranged from 0.8 to 1.6% (wt). the solution at various ratios (1:4 to 1:14 w/v) was added to the ground seeds. mixtures were homogenized using a vortex mixer (heidolph reax top, germany), and the ph was adjusted (8.0 to 13.0) by adding koh (2n) using a ph meter (metrohm 827, switzerland). the samples were then incubated for 40 min at 50°c while shaking at 165 rpm (ika ks 4000i, germany). they were then centrifuged (sigma 3-30k, germany) at 4000g and 25°c for 15 min to separate the oil, emulsion, and upper oil phases (figure 1a). in order to improve the separation of the free oil, the samples were centrifuged (figure 1b) again (5 min at 25°c, 4000g). lastly, the separated oil was carefully weighed to determine its extraction yield (figure 1). the following equation was used to calculate the free oil yield: free oil content (%) = (𝐴 − 𝐵) (𝐶 × 𝐷) × 100 where: a: weight of the tube with oil (g); b: weight of the empty tube (g); c: weight of the crushed rapeseed (g); d: oil content of the crushed rapeseed (expressed as a decimal fraction). figure 1. demonstration of the separation of a) oil, emulsion, and solid phases and b) emulsion and free oil phases by centrifugation. agriculture and food sciences research, 2025, 12(1): 31-41 34 © 2025 by the authors; licensee asian online journal publishing group 2.2.3. extraction yield optimization (ofat approach) to optimize the oil extraction yield, a one-factor-at-a-time (ofat) approach was employed. the effects of pretreatment temperature (170–210°c), tween 20 concentration (0.8–1.6% wt), ph (8–13), crushed rapeseed: water ratio (1:4 to 1:14 w/v), and thermal pre-treatment time (20–40 min) were investigated. all other factors were held constant during each experiment. for each factor, several levels were tested. for example, when investigating the effect of the crushed rapeseed: water ratio, the following parameters were held constant: tween 20 (1.2% wt), ph (10), pre-treatment temperature (190°c), and pre-treatment time (30 min). in addition, when investigating the effect of tween 20 concentration, the following parameters were held constant: rapeseed: water ratio (1:10), ph (10), pretreatment temperature (190°c), and pre-treatment time (30 min). a similar procedure was followed for the remaining factors, maintaining a similar level of control over the constant variables. 2.2.4. freezing and thawing in order to investigate the effects of freezing and thawing on the yield of oil extraction, upon centrifugation of the samples from the previous step, they were frozen (-18°c for 24 h). then, the effects of rapid (80°c, 10 min) and conventional (25°c, 4 h) thawing methods were tested. as soon as they were thawed, they underwent two additional centrifugations to ensure complete separation of the oil phase from the emulsion layer. the final extraction yield was calculated based on the weight of the extracted oil. 2.2.5. statistical analysis treatments were arranged in a complete factorial random design with three replicates. data analysis was conducted using sas jmp statistical discovery pro 16.0 software, employing anova to assess mean differences. excel software was utilized to draw curves and distinguish their fittings with mathematical models if needed. 3. results and discussion 3.1. chemical composition of rapeseeds the average moisture, protein, oil, and ash contents of rapeseeds, measured in triplicate, were 4.88 ± 0.14%, 24.89 ± 0.02% (on a dry matter basis), 38.11 ± 0.77%, and 3.25 ± 0.01%, respectively. by subtracting the sum of these components from 100, the carbohydrate content was around 28.87 ± 0.06% (dry weight). 3.2. comparing the effects of various factors on oil extraction yield to evaluate the effects of various factors on oil extraction yield from rapeseeds, a one-factor-at-a-time (ofat) approach was used. it was possible to determine the independent effects of each factor using this approach, including the ratio of crushed rapeseeds to water, surfactant concentration, ph, temperature, and pretreatment time. utilizing this method, the optimal conditions for achieving maximum oil extraction yield were identified. 3.3. effect of seed: water ratio (tween 1.2%, ph: 10, pretreatment temp 190°c, 30 min) statistical analysis of the data (figures 1, 2) showed that different seed-to-water ratios had a significant effect on oil recovery (p < 0.05). furthermore, based on the comparison of means, the highest oil recovery (50.96%) was observed at a ratio of 1:10, while the lowest oil recovery was observed at a ratio of 1:4. additionally, there was no significant difference in oil recovery between the 1:4 and 1:6 ratios. it appears that as the oil content of the seeds decreases, the seed-to-water ratio required for optimal oil extraction increases. zhang and wang [22] also reported a direct relationship between oil content and the required seed-to-water ratio, where the total oil content of peanuts was 55%, and the optimal ratio was 1:4. however, in the case of rice bran oil extraction, as its total oil content was low (15%), the optimal seed-to-water ratio was 1.5:10 [22, 28]. figure 2. comparison of the effect of rapeseed: water ratio on oil recovery under constant tween 20 concentration (1.2 wt%) and ph 10. agriculture and food sciences research, 2025, 12(1): 31-41 35 © 2025 by the authors; licensee asian online journal publishing group 3.4. effect of tween 20 conc (seed: water ratio 1:10, ph 10, pretreatment temp 190°c, 30 min) as can be seen (figure 3), tween 20 concentration had a significant effect on oil extraction yield (p<0.05). the highest oil extraction yield (52.23%) was obtained at 1.4% wt of tween 20, while the lowest yield (20%) was attained at 0.8% wt of tween 20. as a result, no significant differences were observed between 1.2% and 1.6% wt concentrations of tween 20. several studies have shown the effects of tween 20 concentration (1– 2% wt) on the enhancement of the oil extraction efficiency through aqueous extraction methods. zhang and wang [22] achieved a 76% oil extraction yield from peanuts using 1.2% of tween 20. surlehan et al. [15] extracted 80% of oil from passion fruit seeds using 1% of tween 20. in the same context, geng et al. [24] reported a 58% oil extraction yield from walnut seeds using 1.7% wt tween 20. surfactants aid in oil removal by disrupting the protein matrix that surrounds oil bodies within oilseeds [29]. additionally, they form channels filled with water between the oil phase and the solid surface [30]. a gel layer occurs at the interface between water and solid, as a result, water molecules penetrate the oil-water interface. surfactantstabilized oil microdroplets undergo extraction by mass transfer of extractant into the droplets, followed by upward movement within the aqueous phase [31]. figure 3. comparison of the effect of tween 20 concentration on the amount of oil extracted from rapeseed (rapeseed: water 1:10 and ph 10). 3.5. effect of ph (seed: water ratio 1:10, tween 1.4%, pretreatment temp 190°c, 30 min) the results of the statistical analysis showed that ph had a significant impact on oil extraction yields (p<0.05). according to figure 4, the maximum yield (54.76%) was obtained at ph 12.0, while the minimum (36.4%) was obtained at ph 11.0. oil extraction yields were not significantly different between ph values of 9.0, 11.0, and 13.0. furthermore, other studies have demonstrated that by using an alkaline ph, the oil extraction yield can be increased. hanmoungjai et al. [28], for example, successfully extracted 80% of the oil from rice bran using an aqueous extraction method at a ph of 12.0 [28]. canola oil extraction with tween 20 is likely to be influenced by ph by reducing protein adsorption at the interface, which destabilizes emulsions and increases oil yield at higher ph levels. with alkaline ph, tween 20 has an advantage over proteins in competition for the interfacial layer, which allows it to adsorb and dominate the interface more rapidly [22]. agriculture and food sciences research, 2025, 12(1): 31-41 36 © 2025 by the authors; licensee asian online journal publishing group figure 4. comparison of the effect of aqueous ph on oil recovery from rapeseed (rapeseed: water ratio 1:10, and tween 20 concentration 1.4% wt). 3.6. effect of pretreatment temperature (seed: water ratio 1:10, tween 1.4%, ph 12, 30 min) regarding the effect of pretreatment temperature on rapeseed oil recovery, the present study found a significant correlation between temperature and extraction yield (p<0.05). according to figure 5, 190°c yielded the highest yield (50.5%), while 120°c yielded the lowest (14.6%). these findings indicate that temperature plays an important role in determining the efficiency of oil extraction. studies on oil extraction from sesame seeds have also demonstrated that higher temperatures resulted in higher yields of oil [32]. the extraction of oil can be significantly enhanced by pretreatment at 180°c. according to jia et al. [33], germinated corn meal was four times more likely to yield oil after pretreatment at this temperature. the same was observed by wang et al. [34] who observed a 40% increase in oil extraction from peanuts after a 180°c pretreatment. oil extraction yields are increased when thermal treatment is conducted at temperatures exceeding 180°c. chemical reaction kinetics is accelerated at higher temperatures, resulting in improved oil absorption. during this process, proteins are hydrated and swollen, which aids in the collection of soluble solids, reduces the formation of emulsions, and increases the amount of free oil recovered. during roasting, proteins become more hydrophilic, resulting in the aggregation and coalescence of oil bodies, as well as cell disruption, which facilitates the oil release [34, 35]. figure 5. comparison of the effect of pretreatment temperature on the oil recovery rate from rapeseed (rapeseed: water ratio 1:10, tween 20 concentration 1.4% wt, ph 12.0, and thermal pretreatment duration 30 min). agriculture and food sciences research, 2025, 12(1): 31-41 37 © 2025 by the authors; licensee asian online journal publishing group 3.7. effect of pretreatment time (seed: water ratio1:10, tween1.4%, ph 12.0, temp 190°c) the present study demonstrated a significant influence (p < 0.05) of pretreatment time on oil extraction yield (figure 6). a maximum yield of 50.4% was obtained after 30 minutes of pretreatment, compared to a minimum of 30.7% at 20 minutes. as a result, thermal pretreatment time was identified as an important factor in optimizing aqueous oil extraction, primarily through its effects on the structure of the cell wall and the subsequent release of oil from the cell. the oil extracted from peanuts and canola seeds was 93% and 95%, respectively, after the seeds were pretreated at 104°c for 35 minutes using extended surfactants [11]. figure 6. comparison of the effect of the duration of thermal pretreatment of rapeseed on the oil extraction yield (rapeseed: water ratio 1:10, tween 20 concentration 1.4%wt, ph 12.0, and temperature of 190°c). 3.8. effect of preheat stage (before or after grinding the seeds) and temperature (170-210°c for 30 min) under constant condition (seed: water ratio 1:10, tween 1.4%, ph 12.0) it can be seen (figure 7) that both the stage of thermal pretreatment (before or after grinding) and the pretreatment temperature independently had a significant impact on the extraction of rapeseed oil yield. in addition, a significant effect (p<0.05) was observed between the pretreatment stage and temperature, indicating that they interact to influence each other. the highest oil extraction yield was obtained at 190°c when heat pretreatment was applied before grinding. this is likely due to the fact that, not only does reducing particle size increase the surface area for oil-solvent interaction, but also applying heat treatment before grinding leads to more extensive cell disruption, thereby facilitating oil release. this finding is in agreement with that reported by sagili et al. [36], where the highest crude oil yield (28% from hemp seeds) was obtained with the smallest particles (0.25-0.5 mm). shejawale et al. [37] also observed an increase in soybean oil extraction when the particle size was reduced, although only up to a certain level (0.239-0.353 mm). in addition, further reductions in particle size (to 0.129 and 0.122 mm) resulted in a reduction in yields as a result of reduced porosity and bed compaction. it is believed that the superior effectiveness of pre-grinding heat treatment is due to the fact that grinding can increase surface area but also cause physical damage to the seed matrix, which entraps oil, preventing its release. alternatively, heat treatment before grinding increases oil accessibility and enhances extraction yields by disrupting the cellular structure more effectively. agriculture and food sciences research, 2025, 12(1): 31-41 38 © 2025 by the authors; licensee asian online journal publishing group figure 7. comparison of the effect of the thermal pretreatment step (190 and 120°c) before grinding rapeseed and the temperature of thermal pretreatment on the oil extraction rate (rapeseed: water ratio 1:10, tween 20 concentration 1.4% wt, ph 12). 3.9. effect of freezing and thawing (seed: water ratio 1:10, tween 1.4%, ph 12, pretreatment temp 190°c, 30 min) the freeze-thaw treatment yielded the second highest oil extraction rate (45.2%). li et al. [38], by optimizing freeze-thaw conditions, recovered 82.28% free oil from soybean during the enzyme-assisted aqueous extraction process [38]. it was reported that freezing destabilized soybean oil emulsions and increased the free oil recovery from 3% to 22% by reducing oil droplet size [39]. zhang et al. [40] also found that freeze-thaw cycles improved the yield and quality of tiger nut oil, with 6-8 cycles being considered optimal. according to ghosh et al. [41], ice crystals formed during freezing disrupt the emulsion structure and encourage oil droplet coalescence. in addition, freezing reduces the emulsifying capacity by affecting the protein's secondary structure, resulting in the release of oil [38]. furthermore, thawing can further disrupt the weakened emulsion network and aggregate the oil droplets, causing the trapped phases to be released. during freeze-thaw cycles, some surfactants also lose their efficacy [42]. in table 1, it can be seen that tween 20 effectively extracts oil from rapeseeds under all tested conditions. in the final separation process of oil and emulsion, ph primarily showed a significant influence. by changing the ph of the emulsion, the emulsion destabilized, thus improving oil recovery. while ph may influence the competition between tween 20 and proteins during oil extraction, its primary function is to break the final emulsion and release the oil from the emulsion. according to hao et al. [43], high ph reduces the stability of oil-in-water emulsions by altering surface tension and surfactant behavior. by altering surface properties and reducing repulsive forces between droplets, cations (na⁺, ca²⁺, mg²⁺) intensify this effect [43]. agriculture and food sciences research, 2025, 12(1): 31-41 39 © 2025 by the authors; licensee asian online journal publishing group table 1. comparison of the effects of thawing conditions, ph, presence/absence of tween 20 (0, 1.4% wt), and thermal pretreatment (25 or 190°c, 30 min) under constant seed: water ratio (1:10) on rapeseed oil extraction rate. treatment variables thawing conditions ambient temp (25°c, 4 h) high temp (80°c, 10 min) tween 20 (%) 0 0 1.4 1.4 1.4 1.4 0 0 0 1.4 1.4 1.4 1.4 0 ph 6.8 6.8 6.8 6.8 12.0 12.0 12.0 6.8 6.8 6.8 6.8 12.0 12.0 12.0 pretreatment temp (°c) 25 190 190 25 25 190 190 25 190 190 25 25 190 190 free oil recovered (%) 3.8e 3.1e 43.3b 26.7d 38.8c 45.2a 3.2e 2.9e 3.4e 44.5b 22.3d 27.5c 49.2a 3.2e agriculture and food sciences research, 2025, 12(1): 31-41 40 © 2025 by the authors; licensee asian online journal publishing group 4. conclusions in this study, tween 20-assisted aqueous extraction was examined as an alternative to traditional solvent-based extraction methods for extracting oil from crushed rapeseeds (canola seeds). based on the results, this method can achieve a notable oil recovery of 50.4% under optimized conditions: a rapeseed-to-water ratio of 1:10, tween 20 concentration of 1.4 wt%, ph 12.0, and a 30-minute thermal pretreatment at 190°c. alkaline ph significantly increased oil recovery as it destabilizes emulsions and allows tween 20 to dominate the oil and water interface. likely, tween 20, by reducing the interfacial tension and disrupting the protein matrix, facilitated the oil release. furthermore, freeze-thaw treatment (freezing at -18°c for 24 hours, followed by rapid thawing at 80°c for 10 minutes) led to a 49% oil recovery rate, offering an alternative approach to thermal pretreatment. in addition, the study emphasizes the importance of thermal pretreatment before grinding in order to achieve better cell disruption and greater oil release compared with post-grinding treatment. although it seems that wet-heat treatment should be much more effective than dry heat, which was tested in this study. references [1] w. hamm, r. j. hamilton, and g. calliauw, edible oil processing, 2nd ed. hoboken, nj, usa: wiley-blackwell, 2013. 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[43] x. hao et al., "role of ph and cations on emulsion formation and stability of crude oils," geoenergy science and engineering, vol. 227, p. 211905, 2023. https://doi.org/10.1016/j.geoen.2023.211905 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.seppur.2022.122470 https://doi.org/10.3390/en13226022 https://doi.org/10.1016/j.foodchem.2024.138934 https://doi.org/10.1007/s13197-021-04981-1 https://doi.org/10.1021/acsfoodscitech.3c00129 https://doi.org/10.1007/s13197-022-05554-6 https://doi.org/10.4028/www.scientific.net/amr.236-238.2598 https://doi.org/10.4028/www.scientific.net/amr.236-238.2598 https://doi.org/10.1007/s11746-008-1199-9 https://doi.org/10.1007/s11746-008-1199-9 https://doi.org/10.1016/j.colsurfa.2005.07.013 https://doi.org/10.1016/j.geoen.2023.211905 1 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 1, 1-14, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i1.5261 © 2024 by the authors; licensee asian online journal publishing group moroccan and spanish agriculture comparative analysis 2008-2021 lahoucine asllam1 ahmed ait bari2 mustapha amzil3 ( corresponding author) 1,2,3laboratory for studies and applied, research economic sciences, faculty of legal, economic and social sciences, ibn-zohr agadir university, morocco. 1email: laho11@gmail.com 2email: a.aitbari@uiz.ac.ma 3email: mustapha.amzil@edu.uiz.ac.ma abstract in the general context of morocco's new development model, where public policies need all their credentials, and more specifically in the context of morocco's new agricultural strategy (green plan), the challenge is to develop modern agriculture with high added value and high productivity, capable of competing with agricultural production in other countries. this article compares the overall characteristics of agricultural production in spain and morocco. to make this comparative analysis, we have focused on a set of agricultural statistical aggregates, such as cultivated area, level of production and yield in relation to each family of agricultural products. these are calculated, using the laspeyres index, over the period 2008 to 2021. in addition, we have chosen 2008 as the base year for plotting the evolution of moroccan agricultural performance against that of spain. the results of our comparative analysis showed that agriculture in spain is more stable than in morocco, particularly in cereals and fresh vegetables. thus, they showed that the majority of spanish agricultural products outperform those of morocco. on the other hand, this comparative analysis showed that moroccan agriculture outperforms spanish agriculture in the production of sheep, apples, fresh peas, carrots, turnips, broad beans, green beans, goat meat, dried lentils and chickpeas, and that moroccan agriculture outperforms spanish agriculture in the production of dates and unshelled peanuts. keywords: agricultural production, plant products, livestock products, laspeyres index, morocco, spain. citation | asllam, l., bari, a. a., & amzil, m. (2024). moroccan and spanish agriculture comparative analysis 2008-2021. agriculture and food sciences research, 11(1), 1–14. 10.20448/aesr.v11i1.5261 history: received: 18 october 2023 revised: 4 december 2023 accepted: 15 december 2023 published: 22 december 2023 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ......................................................................................................................................................................................... 2 2. methodology for comparing agriculture in morocco and spain ............................................................................................. 2 3. comparative analysis of agricultural performance between morocco and spain................................................................ 4 4. conclusion ......................................................................................................................................................................................... 10 references .............................................................................................................................................................................................. 10 appendix ................................................................................................................................................................................................ 10 mailto:laho11@gmail.com mailto:a.aitbari@uiz.ac.ma mailto:mustapha.amzil@edu.uiz.ac.ma https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i1.5261 https://orcid.org/0009-0005-6046-2375 https://orcid.org/0009-0007-9875-1775 agriculture and food sciences research, 2024, 11(1): 1-14 2 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the research makes a comparative analysis between moroccan and spanish agriculture, based on a set of agricultural statistical aggregates for each family of agricultural products, calculated using the laspeyres statistical index for the first time, compared to previous studies between the two countries. 1. introduction the aim of this paper is to compare the overall characteristics of agricultural production between spain and morocco. the comparison of agricultural productivity between two countries is difficult to pin down, given the number of factors that come into play in determining the latter, namely the country's climate and geographical position, the type of farming practiced, the tools and technical arrangements put in place for each type of agricultural product and the area cultivated, as well as its fertility. it should be noted in passing that a country can only be said to be in a better position than another with regard to a given crop or agricultural product by comparing agricultural measurement metrics calculated in a similar way in the two countries in question and possessing the same unit of measurement. this paper would therefore focus on a quantitative analysis, focusing on a set of agricultural statistical aggregates such as cultivated area, level of production and yield in relation to each family of agricultural products. it is therefore a study centered on the desire to position one country in relation to another according to the indicators chosen. the indicators used in this paper are selected on the basis of their informative relevance to what they represent. for example, area under cultivation refers only to the amount of land cultivated for a given agricultural product, not to the type of area under cultivation. so, this indicator says nothing about whether a piece of land is conventionally cultivated or whether the cultivated area was an agricultural greenhouse, but simply indicates the amount of land cultivated for a given product. the first part of this comparative analysis will contain a presentation of agricultural comparison metrics, while the second part will detail the levels of these metrics between morocco and spain over a variable time-span, depending on data availability. 2. methodology for comparing agriculture in morocco and spain 2.1. data structure the comparative analysis of agricultural production in morocco and spain will be carried out between 2008 and 2021. the year 2008, at the start of the gmp (green morocco plan) policy, has been chosen as the base year for tracing the evolution of the agricultural performance of one country in relation to the other. in this study, the agricultural performance of a given country for a given product or group of products is based on the level of production in tons, the harvested area in hectares, and the agricultural yield derived from the division of the two previous metrics [1, 2]. as a result, a given agricultural product or family of products is characterized by three main variables: production, area and yield. to avoid redundancy when comparing the two countries, only production and yield will be retained. the data structure deployed in this study is organized along five dimensions: • the size of agricultural product families. • size of agricultural products. • the dimension of parameters, i.e. production in tons, yield in 100ha/ton and area in hectares. • the geographical dimension, i.e. the realization of these parameters for agricultural products in morocco and spain. • the time dimension, i.e. the achievement of the above in each year between 2008 and 2021, i.e. 14 time points in total. note in passing that the first dimension can itself be broken down into several other dimensions. in our case, we have decided to retain two sub-dimensions of this dimension, namely: • the crop and livestock product dimension. • the dimension that breaks down into • cereals, oilseeds, pulses and sugar • fresh vegetables • fresh fruit • meat • milk 2.2. statistical comparison methodology the dimension of agricultural product families, as well as these sub-dimensions, are distinguished by their ability to synthesize several closely related agricultural products into a single aggregate. this aggregation of agricultural products is made possible by the laspeyres statistical index. in this way, the temporal evolution of the production and yield of several agricultural products can be tracked in a single index, for example, the citrus production index instead of the quantity of oranges produced in tons alone. the choice of weighting within each agricultural product grouping is made on the basis of their respective prices in 2008 (base year) in the geographical area of spain that serves as the base country for the comparison. the synthetic production index vacher [3] is thus constructed as follows: 𝐼𝑧0,𝑡 𝑔 = 𝑁𝑀𝑧0,𝑡 𝐷𝑧0 𝐼𝑧1,𝑡 𝑔 = 𝑁𝑀𝑧1,𝑡 𝐷𝑧0 with: agriculture and food sciences research, 2024, 11(1): 1-14 3 © 2024 by the authors; licensee asian online journal publishing group 𝑁𝑀𝑧1,𝑡 = ∑ 𝑝𝑧0,𝑡0 𝑖 𝐼𝑔 𝑖=1 ∗ 𝑞𝑧1,𝑡 𝑖 𝑁𝑀𝑧0,𝑡 = ∑ 𝑝𝑧0,𝑡0 𝑖 𝐼𝑔 𝑖=1 ∗ 𝑞𝑧0,𝑡 𝑖 𝐷𝑧0 = ∑ 𝑝𝑧0,𝑡0 𝑖 𝐺 𝑖=1 ∗ 𝑞𝑧0,𝑡0 𝑖 𝑧 = (𝑧0, 𝑧1) designates the geographical area vector, 𝑧0 being spain, and 𝑧1 being morocco. 𝑡 = (𝑡0, … , 𝑡14) is the time vector with 𝑡0 equal to 2008 et 𝑡14 equal to 2021. 𝐼𝑔 : represents the number of agricultural products in a given grouping. there are several possible groupings 𝑔 to be explained later [4]. 𝐺: gathering of all agricultural products. 𝑖 : is the agricultural product index within a grouping. 𝑞𝑧0,𝑡0 𝑖 : is the quantity produced in product 𝑖 (or agricultural yield) year-round 𝑡0 in the geographical area𝑧0. 𝑝𝑧0,𝑡0 𝑖 : is the price, in dollars, of the agricultural product 𝑖 year-round 𝑡0 in the geographical area𝑧0. 𝑁𝑀𝑧1,𝑡 : is the total value, in dollars, of morocco's agricultural production, from the𝐼𝑔, in the year 𝑡 in prices of respective agricultural products in spain to the year 2008. 𝑁𝑀𝑧0,𝑡 : is the total value, in dollars, of the agricultural production of spain, the family 𝐼𝑔, in the year 𝑡 in prices of respective agricultural products in spain to the year 2008. 𝐷𝑧0 : is the total value, in dollars, of agricultural production in spain, of all agricultural products, in the year 2008 in prices of the respective agricultural products in spain in the year 2008. so far, we've outlined the method of aggregating several agricultural products into a single indicator to express their common production and yield. now, we will proceed in the same way to express morocco's comparative performance against spain for a single agricultural product or group of products in a single combined index. in particular, this index would show the production and yield advantage of one over the other. the index bernard [5] and bernard [6] would be written as follows: 𝐶𝑡 𝑔 = 𝐼𝑧1,𝑡 𝑔 𝐼𝑧0,𝑡 𝑔 = 𝑁𝑀𝑧1,𝑡 𝑁𝑀𝑧0,𝑡 in particular, 𝐶𝑡0 𝑔 represents the advantage or disadvantage in 2008 of morocco over spain, in terms of production or yield, expressed in prices of spanish agricultural products. table 1. weighting1 of agricultural products retained in each type of grouping. agricultural product groups agricultural products weighting first second third yield production p la n t p ro d u ct s cereals, oilseeds, pulses and sugar crops cereals oats 1 8 wheat 2 61 corn 5 27 barley 2 79 rice 7 10 triticale 1 1 oilseeds unshelled peanuts 12 1 soybeans 2 1 rapeseed 1 1 sunflower seeds 1 14 olives 3 121 legume broad and fava beans, dry 1 1 dry lentils 1 1 chickpeas, dried 2 1 dry peas 1 1 sugar crops sugar, beet 8 6 fresh vegetables2 fresh vegetables fresh garlic 30 7 asparagus 18 3 eggplants 98 5 carrots and turnips 45 5 broad and fava beans, green 24 3 onions, shallots, fresh 56 1 fresh peas 34 6 tomatoes, fresh 121 99 1 lund, p.j. the combination of agricultural output and input price indices, journal of agricultural economics, september 1994. bonnays guy, floch jean-michel l'indice des prix des produits agricoles à la production, méthodologie et séries longues insee, archives et documents n° 122 novembre 1987, 122 pages. bourgey guy l'indice des prix des produits agricoles à la production : méthodologie base 1975 insee, archives et documents n° 33 octobre 1981, 88 pages. 2agreste série d n° 17 avril 1979 l'ippap fruits et légumes en base 1970. agreste données chiffrées n° 73 novembre 1995 15 ans d'indice des prix des fruits et légumes. agreste chiffres et données n° 132 mars 2001 ippap fruits et légumes base 1995. agreste chiffres et données n° 165 février 2005 ippap fruits et légumes base 2000. agreste chiffres et données à paraître fin 2009 ippap fruits et légumes base 2005. fourastier jean (sous la direction de) l'évolution des prix à long terme, puf 1969 (pp 121 à 170 pour les prix agricoles et alimentaires). agriculture and food sciences research, 2024, 11(1): 1-14 4 © 2024 by the authors; licensee asian online journal publishing group agricultural product groups agricultural products weighting first second third yield production fresh fruit3 fruits except citrus, almonds, walnuts and dates red fruit apples 22 11 pears 30 13 apricots 11 3 peaches and nectarines 28 31 plums and sloes 15 4 bananas 53 7 figs 8 1 grapes 8 132 citrus lemons and limes 20 14 oranges 14 33 tangerines, mandarins 13 24 almonds, walnuts and dates unshelled almonds 1 6 unshelled walnuts 10 1 dates 37 1 red berries strawberries 150 18 raspberries 72 2 cherries 15 5 l iv es to ck p ro d u ct s meats meats beef and veal 1 48 sheep meat 1 16 goat meat 1 2 meat chicken 5 40 milk milk raw ewe's milk 1 15 raw goat's milk 1 12 raw cow's milk 7 98 note: the weights reported in this table represent the share of the dollar value of a given agricultural product in the total value of the sum of the values of all the agricultural products included in the analysis, in spain, and in the year 2008. the weighting of each type of agricultural product is therefore calculated according to the following formula: wz0,t0 i = 1000 ∗ pz0,t0 i ∗qz0,t0 i dz0 . they correspond to the share of each agricultural product in the index iz0,t0 g . 2.3. weight of agricultural products in the comparison procedure the rest of the study is based on this table, insofar as all the comparative indices between spain and morocco, 𝐶𝑡 𝑔 , relating to the different groupings, reported in the graphs in the following chapter, have been constructed on the basis of the weightings proposed in it. these weights represent the percentage of each agricultural product in the production and yield index for the base year and the base country, in this case 2008 in spain. in assessing the comparison between the agricultural performances of morocco and spain, we note from table 1 that agricultural products such as grapes, olives, tomatoes, cow's milk, barley and wheat have a considerable influence, as they are the mainstays of agricultural production in spain in 2008, so any evolution in agricultural performance, whether in spain or morocco, is assessed, to a large extent, in the prism of these agricultural products. the approach of establishing weightings to group agricultural products into a single index or several indices is necessary to situate morocco's performance in relation to that of spain in several agricultural product groupings at once4. the synthetic yield index, for its part, has a different weighting structure, with particular emphasis on tomatoes, strawberries, raspberries and eggplants. 3. comparative analysis of agricultural performance between morocco and spain 3.1. comparative analysis of the overall agricultural difference between morocco and spain figure 1 presents a comparative analysis of the overall agricultural difference between morocco and spain, taking into consideration all agricultural products with different weightings; we can see that morocco performs, on average, 32.5% of what spain achieves in agriculture. in terms of annual fluctuations in morocco's advantage over spain, we note some variability around the values of 25% and 37.5% of performance compared to spain. spain's agricultural advantage is defined mainly by those products that have a significant weighting in the agricultural production index, namely grapes, olives, fresh tomatoes, raw cow's milk, barley, wheat, beef, chicken meat, oranges, peaches and nectarines, corn and so on (see table 1). according to the same table, the barley production index in morocco fluctuates considerably compared with that of spain, with a coefficient of variation (cv) of 47.57% in morocco versus 21.46 in spain, indicating that barley production in morocco is twice as unstable as that of spain. we can also cite a basket of products that are important in the comparison, where spain's production is much more stable over time: raw cow's milk, wheat, chicken meat, oranges, corn, strawberries, sheep meat, apples, rice and sunflower seeds, and so on (see table 1). 3agreste série d n° 17 avril 1979 l'ippap fruits et légumes en base 1970. agreste données chiffrées n° 73 novembre 1995 15 ans d'indice des prix des fruits et légumes. agreste chiffres et données n° 132 mars 2001 ippap fruits et légumes base 1995. agreste chiffres et données n° 165 février 2005 ippap fruits et légumes base 2000. agreste chiffres et données à paraître fin 2009 ippap fruits et légumes base 2005. fourastier jean (sous la direction de) l'évolution des prix à long terme, puf 1969 (pp 121 à 170 pour les prix agricoles et alimentaires). 4 eurostat, comparison in real values of the aggregates of esa – 1980. luxembourg, 1983. eurostat, summary and comparison of trends in ec agricultural price indices (output and input) 1985-1993, rapid report on agriculture, forestry and fisheries 19944. luxemburg, 1994. eurostat, agricultural land prices and rents in the eu. 1974-1995, theme 5 series c. luxembourg, 1997. eurostat, manual on the economic accounts for agriculture and forestry eaa/eaf 97 (rev. 1.1), theme 5 series methodology and nomenclatures. luxembourg, 2000. agriculture and food sciences research, 2024, 11(1): 1-14 5 © 2024 by the authors; licensee asian online journal publishing group figure 1. evolution of agricultural performance of morocco compared to spain. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). the agricultural yield index, meanwhile, is largely driven by the following products: strawberries, fresh tomatoes, eggplants, raspberries, onions, bananas, carrots, dates and so on (see table 1). morocco is gradually losing its 20% advantage in 2009 to a -10% disadvantage by 2021. figure 2. evolution of agricultural performance of morocco compared to spain for crops and livestock products. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). 3.2. comparative analysis of morocco and spain by first grouping figure 2 presents the evolution of agricultural performance of morocco compared to spain for crops and livestock products. in fact morocco's production disadvantage for livestock products is relatively stable and decreasing, with a downward trend from -70% in 2008 to around -63% in 2021. morocco's yield performance for livestock products nevertheless remains far behind spain, with a sustainable average disadvantage of -65%. the comparison of agricultural performance in crop products is highly unstable, as it is dictated by agricultural products whose annual production is highly variable depending on rainfall, in morocco's case. as already mentioned in the comparison of overall agricultural performance between the two countries, the cv of the production indices of certain agricultural products determining the comparison are very high for morocco, which has repercussions on the annual agricultural performance of this country in relation to spain. the analysis of yields for plant products is more or less stable, with a downward trend for morocco over time. however, it can be assumed that the two countries have very comparable yields for plant products. agriculture and food sciences research, 2024, 11(1): 1-14 6 © 2024 by the authors; licensee asian online journal publishing group figure 3. evolution of agricultural performance of morocco compared to spain for crops products. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). 3.2.1. plant products figure 3 presents the evolution of agricultural performance of morocco compared to spain for crops products. actually morocco is at a disadvantage to spain in terms of production for the three selected agricultural product groupings. for the cereals, oilseeds, pulses and sugar crops group, morocco outperforms spain by 25%. morocco's performance is highly fluctuating, given the cv it displays in the production index for agricultural products in this family. the main products driving the above comparative production index are: olives, barley, wheat and, to a lesser extent, rice. morocco is at an average disadvantage of -70% compared to spain in fresh vegetable production. this grouping is dominated by fresh tomato production. spain is also making strong annual progress in fresh vegetable production, at a much higher rate than morocco, which explains the downward trend in morocco's comparative performance index compared with spain for this family. in the fresh fruit family, morocco ended 2008 with a disadvantage of -80%, and progressed throughout the analysis to a disadvantage of -70% by 2021.despite the gap in agricultural performance between the two countries in terms of this grouping, there is a clear upward trend for morocco in fresh fruit production over the period 2008-2021. the interpretation of this agricultural family is based on viticulture production (grape production), followed by citrus production (all varieties) and peach and nectarine production. here again, fluctuations in the comparative production index are due to the instability of agricultural production in morocco. in terms of yield, the comparison is a little different, since morocco had a 50% advantage in 2009 in the fresh fruit grouping, then began a consistent loss of yield advantage to spain, ending 2021 with a -10% disadvantage in the same family. for the other two groupings, fresh vegetables and cereals, oilseeds, pulses and sugar crops, the gap between the two countries is clearly visible, but the trend is clearly increasing in morocco's favor. figure 4. evolution of agricultural performance of morocco compared to spain for livestock products. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). agriculture and food sciences research, 2024, 11(1): 1-14 7 © 2024 by the authors; licensee asian online journal publishing group 3.2.2. livestock products figure 4 presente the evolution of agricultural performance of morocco compared to spain for livestock products. in the comparison of animal products, three main products are compared: raw cow's milk for the milk category, and beef and chicken meat for the meat category, as they have the highest weights in the milk and meat production indices respectively. the evolution of the comparative milk production index can be interpreted as follows: morocco's disadvantage is around -70% of spain's performance in the same family. moroccan milk production yields are also stable at under 20% of spanish yields. when it comes to meat, morocco achieves, on average, 50% of spain's meat production index. it is interesting to note that morocco's performance is improving year on year. indeed, morocco's meat production disadvantage in relation to spain has been reduced by 20% between 2008 and 2021. nevertheless, morocco's yield is lower than spain's. in 2008, morocco's disadvantage was 60%, rising to 50% by 2021. 3.3. comparative analysis of morocco and spain by second grouping we'll now take a closer look at another level of grouping, breaking down the components of the groupings analyzed above. figure 5. evolution of agricultural performance of morocco compared to spain for cereals, oilcrops, legumes & sugarcrops. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). 3.3.1. cereals, oilseeds, pulses and sugar crops figure 5 presents the evolution of agricultural performance of morocco compared to spain for cereals, oil crops, legumes & sugar crops. we quickly notice a clear advantage for morocco over spain in terms of the comparative index of legume production, and a gain in advantage in terms of sugar crops over the period 20082021. however, these two groupings do not have a significant weighting in the composition of the comparative index of plant product production. they therefore make only a small contribution to the overall assessment of the two countries' agricultural performance. morocco's advantage in pulses is explained by an average performance advantage of 60% in dry chickpeas compared with spain, and an average advantage of 50% in dry lentils, as well as an advantage of 265.7% in dry broad beans and fava beans, i.e. almost four times spain's performance in this agricultural product. for the other two remaining groupings, cereals and oilseeds, spain produces five times more than morocco, with a much more stable production rate than morocco. the two key products in these two key plant family groupings are, respectively, barley for cereals and olives for oilseeds. in terms of comparative yields, morocco has gained a lot in terms of yields for legumes and oilseeds in the last years of the 2008-2021 period, more precisely, from 2016 onwards. for cereals, spain has twice the yield of morocco. 3.3.2. fresh vegetables figure 6 presents the evolution of agricultural performance of morocco compared to spain for vegetable. the aim of this graph is to show, in more detail, all the products contained in the fresh vegetable family. the products that exceed the 100% threshold in comparative production index (start of advantage gain for morocco) at least once in the period between 2008 and 2021 are: green beans, carrots and turnips, fresh weights and fresh onions/shallots. for other agricultural products, the comparison is of little use given spain's enormous performance advantage. we also note that fresh tomatoes dictate the evolution of the entire fresh vegetable family, given its considerable weighting in the comparative fresh vegetable production index. if morocco gains an advantage between 2008 and 2021, it will necessarily be in one of the above-mentioned products. morocco has achieved a significant rise in onion production, from a -50% disadvantage in 2008 to an advantage of over 100% in 2017, i.e. double. it has also begun a process of regression in fresh pea production, moving from a comparative production advantage of 50% to a disadvantage of -40% by 2021. carrots and turnips are very comparable in terms of production performance for both countries. agriculture and food sciences research, 2024, 11(1): 1-14 8 © 2024 by the authors; licensee asian online journal publishing group figure 6. evolution of agricultural performance of morocco compared to spain for vegetable. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). figure 7. evolution of agricultural performance of morocco compared to spain for fruits. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). 3.3.3. fresh fruit figure 7 presents the evolution of agricultural performance of morocco compared to spain for fruits. the comparative production index for the fresh fruit grouping is largely influenced by citrus. when this grouping is broken down into sub-groupings, as in the figure above, there is a wide gap in performance between the two countries, with spain benefiting in citrus fruits, red fruits and other types of fruit. morocco outperforms spain by an average of 50% in the production of almonds, walnuts and dates. in terms of yield, morocco has achieved a sharp rise since 2017 in almonds, walnuts and dates. for red fruits, it has been steadily declining since 2009, falling from an advantage of 100%, i.e. double spain's yield, to finish on a par with spain in 2021. agriculture and food sciences research, 2024, 11(1): 1-14 9 © 2024 by the authors; licensee asian online journal publishing group figure 8. evolution of agricultural performance of morocco compared to spain for red and white meats. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). 3.3.4. meats figure 8 presents the evolution of agricultural performance of morocco compared to spain for red and white meats. in the interpretation of the meat grouping, morocco was at an average disadvantage of -40% in production performance. this was mainly due to the high weighting of beef in the composition of the meat production index. as a result, it pulls down the comparative meat production index when spain produces large quantities of this livestock product. given this, when we look closely at the products making up this meat production index, we quickly see that morocco has a very interesting advantage in goat and sheep meat production, which, it has to be said, have much lower weightings than chicken and beef in the representativeness of the meat grouping. morocco's average advantage is 177% for goat production and 25% for sheep production. the average disadvantage is -60% for chicken and beef production. morocco also achieves an average advantage of 25% and 50% respectively for sheep and goat meat products, and an average disadvantage of -25% and -50% respectively for beef and chicken meat products. figure 9. evolution of agricultural performance of morocco compared to spain for milk. source: the results of production and yield indices are constructed based on data available on the website of the food and agriculture organization of the united nation (fao). agriculture and food sciences research, 2024, 11(1): 1-14 10 © 2024 by the authors; licensee asian online journal publishing group 3.3.5. milk figure 9 presents the evolution of agricultural performance of morocco compared to spain for milk. morocco is largely dominated by spain in milk production, with an average disadvantage of -67% in raw cow's milk production, -90% in raw goat's milk production and -94% in raw ewe's milk production. this advantage gap is more or less stable from 2008 to 2021 for all types of milk production. the yields of these milk products are not very comparable between the two countries, since morocco's comparative yield index is between 5% and 25% of spain's, with a downward trend in ewe's and goat's milk and an upward trend in cow's milk since 2008. 4. conclusion in general, agriculture in spain is more stable than in morocco, especially for cereals and fresh vegetables. in the majority of agricultural products, spain performs better than morocco, with varying differences depending on the type of agricultural product. to sum up, morocco produces 15% more grapes, 37% more olives, 30% more fresh tomatoes, 34% more raw cow's milk, 27% more barley and 84.5% more wheat than spain. in terms of advantages, morocco surpasses spain in sheep production by 25%, 2% in apples, 20% in fresh weight, 5% in carrots and turnips, 124% in green beans, 177% in goat meat, 265% in dried beans, 238% in figs, 50% in dried lentils, 20% in onions and 60% in dried chickpeas, and far exceeds spain in date and unshelled peanut production. references [1] j. berger, "measurement of agricultural price movements," ministry of economy, finance and planning, national institute of statistics and economic studies, 1995. [2] j. berger, "agricultural prices, observation and measurement – the agricultural product producer price index (ippap), 2000 base methodology," insee methods, vol. 114 pp. 9-30, 2006. [3] j. vacher, "indices statistiques: quels outils pour quelles mesures?," insee premiere, no. 15, 1991. [4] p. j. lund, "the combination of agricultural output and input price indices," journal of agricultural economics, vol. 45, no. 3, pp. 360-368, 1994. [5] p. bernard, "descriptive statistics: new method to understand and succeed," economica, 1996. [6] p. y. bernard, "descriptive statistics: new method to understand and succeed," economica, 1990. appendix appendices: production and yield index statistics tables 2 and 3 present production and yield index statistics for each type of agricultural product in our comparative analysis. table 2. production index statistics for each type of agricultural product. weightings agricultural products statistics 𝑰𝒛𝟎,𝒕 𝒈 (spain) 𝑰𝒛𝟏,𝒕 𝒈 (morocco) 𝑪𝒕 𝒈 15 strawberries average coefficient of variation (cv) 18.9 (17.65) 21.4 (36.29) 117.8 (45.03) trend 491 -512 -5937 minimum maximum 15.0 23.9 17.3 48.0 75.3 290.3 12.1 tomatoes, fresh average cv 13.3 (6.56) 13.2 (12.87) 99.4 (10.77) trend 137 352 1675 minimum maximum 11.9 14.6 9.9 15.5 80.6 121.7 9.8 eggplants average cv 11.4 (10.95) 4.3 (20.96) 37.6 (18.78) trend 273 128 285 minimum maximum 10.0 13.8 2.7 5.2 22.9 48.0 7.2 raspberries average cv 16.8 (43.86) 19.9 (5.12) 139.0 (38.58) trend 1492 172 -10147 minimum maximum 7.2 31.7 18.4 21.7 64.5 258.0 5.6 onions, shallots, fresh average cv 5.9 (7.86) 4.4 (13.05) 71.8 (14.75) trend 17 105 727 minimum maximum 5.3 6.9 2.9 4.9 52.1 84.0 5.3 bananas average cv 5.5 (6.67) 5.1 (6.57) 93.6 (10.18) trend 65 -12 -1326 minimum maximum 4.9 6.1 4.3 5.8 82.6 116.8 4.5 carrots and turnips average cv 4.6 (4.82) 2.4 (25.05) 53.0 (25.79) trend 33 -17 -731 minimum maximum 4.2 5.0 1.4 4.0 29.2 90.3 3.7 dates average cv 3.5 (15.83) 1.3 (13.30) 38.7 (27.60) trend -42 18 764 minimum maximum 2.6 4.5 1.0 1.6 27.3 59.1 3.4 fresh peas average cv 3.6 (7.02) 3.3 (16.44) 91.0 (20.88) trend 41 -70 -3007 minimum maximum 3.1 4.0 2.1 4.1 56.8 127.2 3 fresh garlic average cv 3.3 (7.00) 2.9 (29.64) 90.5 (31.63) trend 32 -43 -2138 minimum maximum 3.0 3.7 1.8 5.2 53.7 165.5 pears average cv 2.7 (6.75) 1.7 (16.29) 63.4 (15.89) trend -29 -15 61 minimum maximum 2.5 3.0 1.3 2.2 50.9 87.5 2.8 peaches and nectarines average cv 2.9 (11.00) 2.2 (14.77) 75.7 (20.65) trend 42 -52 -2936 minimum maximum 2.4 3.6 1.7 2.8 58.0 101.4 2.4 broad and fava beans, green average cv 2.3 (4.59) 3.2 (24.74) 141.1 (26.63) agriculture and food sciences research, 2024, 11(1): 1-14 11 © 2024 by the authors; licensee asian online journal publishing group weightings agricultural products statistics 𝑰𝒛𝟎,𝒕 𝒈 (spain) 𝑰𝒛𝟏,𝒕 𝒈 (morocco) 𝑪𝒕 𝒈 trend 8 -96 -4886 minimum maximum 2.1 2.5 2.0 5.2 87.5 244.9 2.2 apples average cv 2.1 (8.31) 1.6 (15.46) 76.9 (17.03) trend 6 -3 -380 minimum maximum 1.7 2.4 0.9 1.9 41.5 98.1 2 lemons and limes average cv 2.7 (19.45) 1.5 (25.59) 60.3 (42.50) trend 70 -6 -2442 minimum maximum 1.9 3.9 0.8 2.3 33.2 118.3 1.8 asparagus average cv 2.0 (6.13) 3.1 (37.15) 155.2 (36.31) trend -6 -48 -1999 minimum maximum 1.8 2.3 1.1 5.3 54.4 259.0 1.5 cherries average cv 2.0 (12.68) 2.9 (41.13) 142.7 (39.88) trend 14 -5 -1334 minimum maximum 1.5 2.4 1.4 5.1 72.0 274.5 plums and sloes average cv 1.8 (9.98) 1.3 (21.10) 77.5 (25.87) trend -4 39 2436 minimum maximum 1.5 2.0 0.9 1.9 45.8 127.1 1.4 oranges average cv 1.4 (12.47) 1.1 (15.11) 76.4 (19.71) trend 30 10 -946 minimum maximum 1.1 1.7 0.7 1.3 52.3 100.6 1.3 tangerines, mandarins, clementines average cv 1.3 (8.75) 1.2 (21.40) 87.5 (22.75) trend 11 36 2068 minimum maximum 1.2 1.6 0.7 1.5 54.4 122.3 1.2 unshelled peanuts average cv 1.1 (12.81) 0.9 (8.37) 79.6 (18.78) trend -9 16 1347 minimum maximum 0.8 1.3 0.7 1.0 59.7 113.2 1.1 apricots average cv 1.2 (19.88) 1.8 (20.61) 153.1 (40.25) trend 34 -47 -9512 minimum maximum 0.8 1.6 1.1 2.4 80.0 274.3 1 unshelled walnuts average cv 1.0 (11.77) 1.3 (30.30) 125.1 (34.44) trend -17 -4 1765 minimum maximum 0.9 1.3 0.8 2.3 83.6 249.5 0.8 figs average cv 0.9 (22.60) 0.7 (18.63) 80.2 (31.00) trend 34 5 -2184 minimum maximum 0.6 1.2 0.3 0.8 28.3 130.8 grapes average cv 0.9 (12.37) 1.3 (16.28) 138.0 (14.40) trend 14 45 2722 minimum maximum 0.8 1.2 0.9 1.6 100.8 176.8 sugar, beet average cv 0.9 (7.26) 0.6 (8.85) 69.8 (8.12) trend 4 8 544 minimum maximum 0.8 1.0 0.5 0.7 62.5 84.6 0.7 raw cow's milk average cv 0.9 (10.51) 0.1 (8.03) 17.0 (8.12) trend 20 2 -166 minimum maximum 0.7 1.0 0.1 0.2 15.0 20.4 rice average cv 0.8 (3.57) 0.8 (15.69) 99.9 (17.37) trend 0 15 1982 minimum maximum 0.7 0.8 0.7 1.2 83.8 156.7 0.5 corn average cv 0.6 (6.98) 0.0 (29.05) 7.0 (31.78) trend 9 -1 -321 minimum maximum 0.5 0.6 0.0 0.1 3.4 11.5 meat chicken average cv 0.5 (6.72) 0.2 (9.36) 48.4 (13.00) trend 4 -3 -1053 minimum maximum 0.4 0.5 0.2 0.3 36.9 60.0 0.3 olives average cv 0.4 (23.35) 0.2 (15.91) 55.3 (33.52) trend 5 -1 -911 minimum maximum 0.2 0.6 0.1 0.3 34.6 91.7 0.2 wheat average cv 0.2 (18.27) 0.1 (30.08) 57.8 (32.73) trend 4 1 -374 minimum maximum 0.1 0.3 0.1 0.2 21.2 90.1 soybeans average cv 0.2 (11.56) 0.1 (0.00) 35.4 (12.53) trend 4 0 -675 minimum maximum 0.2 0.3 0.1 0.1 30.4 45.1 barley average cv 0.1 (20.56) 0.1 (41.74) 40.5 (44.65) trend 3 1 -387 minimum maximum 0.1 0.2 0.0 0.1 10.3 71.6 chickpeas, dried average cv 0.2 (18.46) 0.1 (32.24) 80.2 (43.48) trend 1 5 794 minimum maximum 0.1 0.2 0.1 0.2 45.1 152.6 0.1 unshelled almonds average cv 0.1 (20.02) 0.2 (11.62) 161.5 (19.27) trend 3 0 -4102 minimum maximum 0.1 0.1 0.1 0.2 108.2 215.0 oats average cv 0.1 (21.34) 0.0 (44.46) 52.5 (44.40) trend 2 -2 -2540 minimum maximum 0.1 0.1 0.0 0.1 13.6 80.2 agriculture and food sciences research, 2024, 11(1): 1-14 12 © 2024 by the authors; licensee asian online journal publishing group weightings agricultural products statistics 𝑰𝒛𝟎,𝒕 𝒈 (spain) 𝑰𝒛𝟏,𝒕 𝒈 (morocco) 𝑪𝒕 𝒈 broad and fava beans, dry average cv 0.1 (14.86) 0.1 (33.56) 55.1 (37.79) trend -1 0 1144 minimum maximum 0.1 0.1 0.0 0.1 28.2 111.9 rape or colza seeds average cv 0.2 (17.37) 0.1 (30.41) 64.3 (30.79) trend 4 4 1504 minimum maximum 0.1 0.2 0.1 0.1 39.0 97.1 sunflower seeds average cv 0.1 (12.22) 0.1 (18.43) 110.9 (23.99) trend 1 2 231 minimum maximum 0.1 0.1 0.1 0.2 73.9 178.1 0.1 raw ewe's milk average cv 0.0 (12.59) 0.0 (9.08) 17.9 (19.50) trend 1 0 -707 minimum maximum 0.0 0.1 0.0 0.0 13.9 23.2 raw goat's milk average cv 0.1 (19.81) 0.0 (9.86) 8.6 (20.12) trend -2 0 206 minimum maximum 0.0 0.1 0.0 0.0 5.6 11.1 dry lentils average cv 0.1 (28.09) 0.1 (42.54) 97.9 (43.50) trend 2 2 -271 minimum maximum 0.1 0.2 0.0 0.2 18.5 147.9 dry peas average cv 0.1 (26.95) 0.0 (25.89) 49.2 (38.37) trend 3 0 -2131 minimum maximum 0.0 0.1 0.0 0.1 17.1 76.6 beef and veal average cv 0.1 (2.72) 0.1 (7.22) 78.1 (5.47) trend 1 2 726 minimum maximum 0.1 0.1 0.1 0.1 70.9 86.4 goat meat average cv 0.0 (4.67) 0.0 (13.43) 159.8 (11.77) trend 0 0 2405 minimum maximum 0.0 0.0 0.0 0.0 141.0 203.8 sheep meat average cv 0.0 (3.21) 0.0 (9.53) 121.3 (9.86) trend 0 0 -443 minimum maximum 0.0 0.0 0.0 0.0 97.4 133.9 table 2 shows production index statistics for each type of agricultural product. table 3. yield index statistics for each type of agricultural product. weightings agricultural products statistics 𝑰𝒛𝟎,𝒕 𝒈 (spain) 𝑰𝒛𝟏,𝒕 𝒈 (morocco) 𝑪𝒕 𝒈 13.2 grapes average cv 13.5 (10.12) 0.8 (12.50) 6.2 (14.46) trend 81 20 114 minimum maximum 11.8 16.6 0.6 1.0 4.6 1.0 12.1 olives average cv 15.0 (24.33) 2.9 (23.35) 20.4 (37.27) trend 281 97 281 minimum maximum 8.3 21.3 1.7 4.1 12.2 4.1 9.9 tomatoes, fresh average cv 11.1 (10.67) 3.2 (6.03) 29.0 (11.87) trend 141 15 -246 minimum maximum 9.2 12.8 3.0 3.5 23.5 3.5 9.8 raw cow's milk average cv 10.8 (7.02) 3.6 (12.90) 33.6 (9.39) trend 178 95 351 minimum maximum 9.8 12.0 2.7 4.0 26.8 4.0 7.9 barley average cv 5.9 (21.46) 1.5 (47.57) 26.7 (54.32) trend 33 -36 -725 minimum maximum 4.1 8.1 0.4 2.7 5.6 2.7 6.1 wheat average cv 6.1 (18.97) 5.0 (33.88) 84.5 (39.44) trend 141 30 -1311 minimum maximum 4.4 7.8 2.3 7.3 31.5 7.3 4.8 beef and veal average cv 4.6 (6.85) 1.7 (16.49) 38.0 (14.10) trend 54 65 960 minimum maximum 4.2 5.2 1.3 2.0 27.3 2.0 4 meat chicken average cv 4.6 (8.68) 1.8 (14.75) 38.7 (8.51) trend 86 54 453 minimum maximum 4.0 5.2 1.4 2.3 32.2 2.3 3.3 oranges average cv 3.1 (9.77) 0.9 (14.97) 28.0 (14.56) trend 40 21 291 minimum maximum 2.6 3.5 0.7 1.1 21.5 1.1 3.1 peaches and nectarines average cv 3.5 (12.94) 0.3 (35.86) 7.8 (36.74) trend 37 20 536 minimum maximum 3.0 4.5 0.2 0.4 4.7 0.4 2.7 corn average cv 3.1 (11.46) 0.1 (57.40) 3.1 (67.82) trend 34 -9 -352 minimum maximum 2.5 3.7 0.0 0.2 0.7 0.2 2.4 tangerines, mandarins average cv 2.4 (21.14) 1.0 (38.31) 42.5 (44.53) trend -7 76 3651 minimum maximum 1.9 4.0 0.4 1.5 15.1 1.5 1.8 strawberries average cv 2.1 (14.53) 1.0 (36.93) 51.5 (47.36) agriculture and food sciences research, 2024, 11(1): 1-14 13 © 2024 by the authors; licensee asian online journal publishing group weightings agricultural products statistics 𝑰𝒛𝟎,𝒕 𝒈 (spain) 𝑰𝒛𝟏,𝒕 𝒈 (morocco) 𝑪𝒕 𝒈 trend 43 -19 -2141 minimum maximum 1.7 2.6 0.7 2.3 35.5 2.3 1.6 sheep meat average cv 1.3 (9.06) 1.6 (15.35) 125.0 (19.43) trend -18 48 5252 minimum maximum 1.2 1.6 1.2 1.8 77.1 1.8 1.5 raw ewe's milk average cv 1.9 (7.33) 0.1 (8.93) 6.6 (9.96) trend 15 0 -48 minimum maximum 1.5 2.0 0.1 0.1 5.7 0.1 1.4 lemons and limes average cv 1.7 (20.65) 0.1 (24.55) 3.7 (17.89) trend 70 3 43 minimum maximum 1.1 2.2 0.0 0.1 2.4 0.1 sunflower seeds average cv 1.4 (13.59) 0.1 (39.72) 3.9 (38.83) trend -10 -2 -136 minimum maximum 1.0 1.8 0.0 0.1 1.8 0.1 1.3 pears average cv 1.0 (18.25) 0.1 (15.17) 9.6 (23.12) trend -39 0 374 minimum maximum 0.8 1.3 0.1 0.1 7.1 0.1 1.2 raw goat's milk average cv 1.2 (5.31) 0.1 (14.73) 9.6 (19.52) trend 7 -1 -100 minimum maximum 1.1 1.3 0.1 0.2 7.8 0.2 1.1 apples average cv 1.0 (9.05) 1.0 (28.26) 102.4 (30.54) trend -6 58 6380 minimum maximum 0.8 1.1 0.7 1.4 61.1 1.4 1 rice average cv 1.3 (11.56) 0.1 (28.10) 6.4 (31.09) trend -18 2 288 minimum maximum 0.9 1.4 0.0 0.1 1.9 0.1 0.8 oats average cv 0.7 (25.31) 0.0 (75.76) 3.5 (77.77) trend 12 -2 -309 minimum maximum 0.5 1.1 0.0 0.1 0.4 0.1 0.7 fresh garlic average cv 1.1 (30.86) 0.1 (36.85) 6.0 (56.13) trend 75 0 -470 minimum maximum 0.7 1.7 0.0 0.1 3.4 0.1 bananas average cv 0.7 (6.41) 0.6 (16.34) 78.1 (13.97) trend 8 19 1668 minimum maximum 0.7 0.8 0.4 0.7 57.8 0.7 0.6 unshelled almonds average cv 0.9 (31.46) 0.4 (19.95) 44.9 (19.96) trend 50 14 -720 minimum maximum 0.5 1.5 0.3 0.6 30.0 0.6 fresh peas average cv 0.8 (19.73) 0.9 (22.69) 121.9 (36.21) trend 32 -33 -8866 minimum maximum 0.6 1.1 0.6 1.2 56.5 1.2 fresh peas average cv 0.5 (19.05) 0.5 (23.98) 98.3 (32.26) trend -18 13 6165 minimum maximum 0.4 0.7 0.3 0.6 47.0 0.6 0.5 eggplants average cv 0.6 (11.27) 0.1 (34.06) 22.4 (31.36) trend 14 7 634 minimum maximum 0.5 0.8 0.1 0.2 9.4 0.2 carrots and turnips average cv 0.5 (4.93) 0.5 (27.92) 105.9 (32.18) trend -1 8 1746 minimum maximum 0.4 0.5 0.3 0.9 57.0 0.9 cherries average cv 0.8 (15.34) 0.1 (37.75) 11.8 (32.43) trend 17 5 410 minimum maximum 0.5 1.0 0.1 0.2 7.4 0.2 0.4 plums and sloes average cv 0.4 (14.44) 0.3 (39.41) 64.5 (51.94) trend -10 21 6780 minimum maximum 0.3 0.5 0.1 0.4 28.0 0.4 0.3 apricots average cv 0.4 (22.44) 0.3 (21.49) 93.3 (45.40) trend 12 -10 -7032 minimum maximum 0.2 0.5 0.2 0.5 51.0 0.5 asparagus average cv 0.4 (13.75) 0.0 (87.59) 2.8 (92.03) trend 10 -2 -557 minimum maximum 0.3 0.4 0.0 0.0 0.0 0.0 broad and fava beans, green average cv 0.2 (15.99) 0.5 (28.81) 224.9 (31.16) trend -5 -19 -3854 minimum maximum 0.2 0.3 0.3 0.8 138.8 0.8 0.2 raspberries average cv 0.5 (70.46) 0.0 (6.84) 1.6 (53.84) trend 77 0 -184 minimum maximum 0.2 1.2 0.0 0.0 0.5 0.0 goat meat average cv 0.2 (8.42) 0.6 (14.51) 277.3 (13.18) trend 1 19 7044 minimum maximum 0.2 0.2 0.4 0.7 211.7 0.7 0.1 unshelled peanuts average cv 0.0 (85.33) 0.2 (13.98) 21497.6 (133.82) trend 0 -4 -2464850 minimum maximum 0.0 0.0 0.2 0.3 1891.3 0.3 agriculture and food sciences research, 2024, 11(1): 1-14 14 © 2024 by the authors; licensee asian online journal publishing group weightings agricultural products statistics 𝑰𝒛𝟎,𝒕 𝒈 (spain) 𝑰𝒛𝟏,𝒕 𝒈 (morocco) 𝑪𝒕 𝒈 dates average cv 0.0 (28.10) 1.1 (19.21) 3461.4 (52.66) trend -1 43 186325 minimum maximum 0.0 0.0 0.7 1.5 1622.4 1.5 soybeans average cv 0.0 (49.21) 0.0 (0.00) 45.2 (70.50) trend 0 0 -5667 minimum maximum 0.0 0.0 0.0 0.0 19.3 0.0 broad and fava beans, dry average cv 0.0 (34.29) 0.2 (38.66) 365.7 (46.84) trend 0 -7 -15285 minimum maximum 0.0 0.1 0.0 0.2 49.5 0.2 figs average cv 0.2 (33.37) 0.5 (23.76) 338.0 (33.03) trend 12 19 -8059 minimum maximum 0.1 0.3 0.3 0.7 131.0 0.7 rape or colza seeds average cv 0.1 (59.85) 0.0 (46.55) 1.9 (53.26) trend 18 0 -162 minimum maximum 0.0 0.3 0.0 0.0 0.9 0.0 dry lentils average cv 0.0 (28.42) 0.1 (50.54) 149.4 (47.76) trend 1 1 -1286 minimum maximum 0.0 0.1 0.0 0.1 6.7 0.1 unshelled walnuts average cv 0.1 (12.68) 0.1 (32.61) 84.3 (28.12) trend 4 4 171 minimum maximum 0.1 0.2 0.1 0.2 49.3 0.2 onions, shallots, fresh average cv 0.1 (27.07) 0.1 (38.26) 124.9 (43.81) trend -2 10 6123 minimum maximum 0.1 0.1 0.1 0.2 40.3 0.2 chickpeas, dried average cv 0.1 (35.34) 0.1 (33.49) 159.9 (38.76) trend 3 3 -4036 minimum maximum 0.1 0.2 0.1 0.2 44.9 0.2 dry peas average cv 0.2 (26.60) 0.0 (44.59) 13.3 (47.76) trend 3 -1 -773 minimum maximum 0.1 0.2 0.0 0.0 2.3 0.0 table 3 shows yield index statistics for each type of agricultural product. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 32 © 2023 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 10, no. 2, 32-36, 2023 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v10i2.5210 © 2023 by the author; licensee asian online journal publishing group on-farm demonstration of improved and drought tolerant maize varieties with associated management practices belay roba1 fistum miruts2 ( corresponding author) 1,2ethiopian institute of agricultural research, melkassa agricultural research center, adama, ethiopia. 1email: robabelay1984@gmail.com 2email: fitsummiruts@gmail.com abstract maize is an important cereal crop in the central rift valley of ethiopia. it plays a greater role in ensuring food security. the study was designed to evaluate and promote melkassa-6q variety with standard check (melkassa-2). the study sites were adama and atjk districts. the districts were selected based on their maize production potential and accessibility of the site for demonstration. training and field days were implemented to facilitate the successful implementation of the activity and create awareness among farmers about the varieties. grain yield was recorded and the varieties were compared to each other based on their yield performance. farmers ‘feedback was also collected and farmers preference ranking of each demonstrated variety was done depending on the farmers’ set criteria for variety evaluation. the mean yield of melkassa-2 (standard check) is about 39 qt/ha across locations whereas the mean yield of melkassa-6q is 36.4 qt/ha which is 2.6 qt below the standard check. in addition to yield, melkassa-2 variety was preferred over melkassa-6q variety in overall farmers’ evaluation criteria for maize variety evaluation. therefore, there should be an adequate quality seed supply of melkassa-2 variety till the best-performing maize variety is released by the research system. keywords: demonstration, drought, evaluation, farmers feedback, grain yield, maize varieties, preference. citation | roba, b., & miruts, f. (2023). on-farm demonstration of improved and drought tolerant maize varieties with associated management practices. agriculture and food sciences research, 10(2), 32–36. 10.20448/aesr.v10i2.5210 history: received: 2 october 2023 revised: 8 november 2023 accepted: 30 november 2023 published: 6 december 2023 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by ethiopian institute of agricultural research (grant number: 62-05). institutional review board statement: the ethical committee of the ethiopian institute of agricultural research, ethiopia has granted approval for this study. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: both authors contributed equally to the conception and design of the study. both authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 33 2. materials and methods ................................................................................................................................................................... 33 3. result and discussion ..................................................................................................................................................................... 35 4. conclusion and recommendation ................................................................................................................................................ 36 references .............................................................................................................................................................................................. 36 mailto:robabelay1984@gmail.com mailto:fitsummiruts@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v10i2.5210 https://orcid.org/0009-0004-2835-8212 https://orcid.org/0000-0003-1390-4669 agriculture and food sciences research, 2023, 10(2): 32-36 33 © 2023 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the uniqueness of the study lies in using newly research released maize varieties. the study also reports the research finding which makes the study original. the study compares two maize varieties across two location in two production season and better varieties discussed as well as suggests to maize producing stakeholders. 1. introduction maize is one of the world's most important crops produced for food. it has the highest average yield per hectare and ranks third in terms of area and total output in the world behind wheat and rice. it is grown in most parts of the world over a wide range of environmental conditions [1]. it is also among the most important and widely grown crops in ethiopia. among cereal crops produced, maize is the most important crop in terms of production and contributes significantly to the national economy of the country [2]. the smallholder farmers that comprise about 80% of ethiopia’s population are both the primary producers and consumers of maize [3]. in the 2017/18 crop production year, around 8.5 million small-scale farmers were engaged in maize production, compared to 6.7 million for teff and 5.3 million for sorghum. cereals contributed 87.48% (about 267,789,764.02 quintals) of the grain production. maize production accounts for 27 percent of total cereal production in ethiopia, with the highest volume of production (about 8.3 million tons), followed by teff and wheat (1.7 million and 1.5 million tons, respectively) [2]. however, the maize production potential was not yet achieved because of limited utilization of improved production technologies. as findings show, the productivity level of maize was 2.2 tons per hectare [4] in 2008/09 . this by far below the world’s average yield which is about 5.21 tons per hectare [5, 6] indicated that maize productivity under farmers’ condition is very low. in addition to the low level of improved production technology utilization, recurrent drought considerably affected the production of the crop [7]. growing varieties that are drought-resistant is thus a possible approach for enhancing food production, particularly in ethiopia's drought-prone regions. access to high-quality maize seed, high-yielding, and drought-tolerant maize varieties influences attempts to attain food security by increasing maize output and productivity. in response to this, 45 varieties of improved maize were released by the national maize research from 1973-2011 in which 11 varieties were recommended for drought-prone areas and the rest 34 varieties were released for high moisture areas with average annual rainfall more than 900mm [5]. despite multiple initiatives to reform smallholder agriculture in general and crops in particular, adoption of improved varieties of major crops such as maize in ethiopia has remained low [8]. aside from the limited adoption of better technologies, primarily seed, other factors contributing to the low productivity level include low yield potential of seed cultivars, low seed quality, irregular rainfall, and inadequate crop management methods [3]. in general, yield gaps are ascribed to a variety of causes such as frequent drought, diminishing soil fertility, poor agronomic practice, limited input utilization, low seed quality, disease, and others [9]. as a result, there is significant potential to increase maize production in the country as a whole and in the study region in particular by utilizing high-yielding varieties and improved management techniques. 1.1. objectives • to evaluate the performance newly released drought tolerant maize varieties with recommended production management practices. • to create awareness, develop confidence among farmers, development agents, agricultural experts and seed producers for further promotion and dissemination. 2. materials and methods 2.1. description of the study area the demonstration was carried out during 2017 and 2018 cropping seasons in adami-tullu jido kombolcha (atjk) and adama districts. figure 1. sketch of study area. agriculture and food sciences research, 2023, 10(2): 32-36 34 © 2023 by the authors; licensee asian online journal publishing group the districts are located in rift valley areas of oromia regional state. the areas have mixed livestock farming system. in the study area, the major crops grown are maize, teff, vegetables and the main livestock kept by the farmers are cattle, sheep and goats. the districts were targeted for the study based on their maize production potential and accessibility of the site for demonstration. the site selection was made in collaboration with the district agricultural and natural resource offices. the above figure 1 illustrates the district where the study is conducted. accordingly, the study is conducted in adama and adami tulu jido kombolcha districts district of east shewa zone, oromia regional state, ethiopia. 2.2. farmers selection and demonstration field establishment the demonstration research activity was conducted in central rift valley of ethiopia, the districts and kebeles were purposefully selected based on their maize production potential and accessibility of the area for the demonstration. a total of 60 maize farmers were participated in the maize demonstration activity in consideration of gender issues (women, men and youth). the demonstration hosting farmers were selected depending on their interest for maize technology, land provision for this pre-extension demonstration, willingness to share experiences for other farmers and their willingness to work on experiment with their full commitment and in close collaboration with researchers, experts and development agents. 2.3. design of the field demonstration activity two quality protein maize varieties released from melkassa agricultural research center; one newly released variety namely, melkassa-6q and melkassa-2 as a standard checks were planted. the varieties were demonstrated side by side in which each variety allocated with 0.25 hectare of land. two maize varieties namely melkass-6q and melkassa-2 were demonstrated side by side. melkassa-2 was the early released maize variety when compared with melkassa-6q. thus, it was used as standard check (control). each farmer allocated 0.25 hectare of land for each maize variety and all the appropriate agronomic management practices were employed equally for each varieties side by side under the close supervision of researchers and development agents. the varieties were replicated across six trial farmers per kebele. planting was done based on the onset of the rainfall of the growing season early whenever there is enough moisture in the soil. each variety was planted on a plot size: 25mx25m, at seeding rate of 25-30kg/ha. a spacing of 75cm*25cm (53,333 plants/ha) (between row and plant) and nitrogin-phosphat-sulfer fertilizer (nps)) at rate of 100kg/ha were equally applied for all of the plots. for weed control twice hand weeding (the first one at 25 to 30 days after sowing and the second at knee height) is while slashing at flowering stage was done. culturally removal of volunteer plants and alternate hosts by hand for insect pest control were performed. finally, the performance of the maize varieties was evaluated against each other which is discussed in detail in the result section. 2.4. monitoring and evaluation the demonstration activity was monitored and evaluated properly starting from the beginning of the implementation to the harvest. all the recommended agronomic management practices were employed properly during the demonstration of the varieties. farmers were advised to prepare their demonstration field in advance. the demonstration farmers were also used the recommended seed rate and recommended fertilizer rate. other agronomic management practices like thinning and hand weeding were also employed at the appropriate time. 2.5. important tools for implementation training, field visit and field days were used as important tools for the implementation of this activity. multidisciplinary research team; crop, extension and socio-economic research team and other stakeholders (offices of agriculture and natural resource) actively participated by sharing their experience and knowledge during the training and field day. participant farmers were selected and orientation on the improved maize technology demonstration was given in cooperation with development agents (das) and subject matter specialist (sms). training materials was prepared and practical trainings were given for participant farmers and extension counterparts (das & sms) on production of improved maize technology. a total of 238 participants (126 farmers, 72 das, and 40 sms trained on maize production and management practices. on top of these 55 participants were participated on different food recipes preparation from maize table 1. from the participants, 37 were farmers (5 male and 32 female) and 18 were das and sms (12 male and 6 female). table 1. the number of participants provided training in the demonstration period (2017-2018). participants type of training provided to the participants total maize production food recipe preparation farmers 126 37 163 das and sms 112 18 130 total 238 55 293 field day was one of the important demonstration implementation tools in this research activity. the demonstrated fields were demonstrated to farmers, development agents, district agricultural sms and other stakeholders to share knowledge and experience about the improved maize technologies and to facilitate for its wider popularization among smallholder farmers. accordingly, the field day was organized in adama and atjk districts with objectives of promoting improved maize production and sharing experiences and knowledge about the variety among the participants. various stakeholders and partners were participated and they visited the maize demonstration field in the area. a total of 436 participants were participated on the field day event. from the participants, 400 were farmers (256 male and 144 female). agriculture and food sciences research, 2023, 10(2): 32-36 35 © 2023 by the authors; licensee asian online journal publishing group 2.6. extension material and media coverages leaflets were produced in local languages (afan oromo and amharic) on common bean production, field managements and, food preparation and utilization. 750 smallholder farmers were provided with the leaflets on field day event. field day was supported by different media coverage like television (oromia broadcasting network) and zonal communication offices. the participants’ feedback about the improved maize technologies was recorded and transmitted to the wider societies through the media (obn) for further awareness creation and promotion. 2.7. data collection the types of data collected were grain yield and farmers’ feedback with regarding to important crop (technology) traits beginning from pre-harvest agronomic practices to post harvest handling. the collected data were analyzed using descriptive statistics and preference ranking based on farmers' criteria. 3. result and discussion 3.1. yield performance of the demonstrated varieties against the standard check the following figure 2 describes the yield performances of the demonstrated varieties across the study site. the yield performance of the improved maize variety (melkassa-6q) was evaluated against the yield performance of the standard check or control which was in the hand of the farmers for relatively many years. thus, melkassa-2 is used as a control in this demonstration research activity. the result shows that the mean yield of melkassa2(control) is greater than the mean yield of melkassa-6q in almost all research locations in 2017 and 2018 growing season this shows the standard check had better yield advantage over the melkassa-6q. figure 2. yield performance of the varieties across location. 3.2. farmers preferences farmers made selection among demonstrated maize varieties depending on their own preference criterion. the criteria used were yield, drought tolerance, market, grain color, food taste and earliness in maturity. the farmers’ preference rank is determined based on the evaluation score weight. the variety with high score weight is most preferred by the farmers and given first rank. accordingly, the result indicated that farmers ranked melkassa-2(control) first and melkassa-6q variety second in both demonstration sites (districts). the variety is ranked first across location, since its mean score weight is 0.78 which is greater than the mean score weight of melkassa-6q which is about 0.64 table 2. to sum up melkassa-2variety is more preferred maize variety than melkass-6 in the research areas. this reveals that melkassa-6q no longer replace the standard check (melkassa-2). table 2. farmers preference ranking for the varieties with respect to different criterion. district varieties yield drought tolerant market grain color food taste earliness total score weight rank atjk melkassa-6q 1 2 2 2 2 2 11 0.61 2 melkassa-2 (check) 3 3 1 1 1 3 12 0.67 1 adama melkassa-6q 2 3 2 2 2 1 12 0.67 2 melkassa-2 (check) 3 3 3 2 2 3 16 0.89 1 mean weight (m-6q) 1.5 2.5 2 2 2 1.5 11.5 0.64 2 mean weight (m-2) 3 3 2 1.5 1.5 3 14 0.78 1 note: preference ranking based on preference scores out of 3 points with 1= low score, 2= moderate score and 3= high score. m-6q= melkassa-6q and m-2= melkassa-2 source: farmers’ feedback. agriculture and food sciences research, 2023, 10(2): 32-36 36 © 2023 by the authors; licensee asian online journal publishing group 4. conclusion and recommendation it is indicated that the standard check (melkassa-2) had shown better performance in grain yield than melkassa-6q. thus, the cultivation of melkassa-2 has been found more productive than cultivating melkassa-6q variety. even though both varieties were found to be suitable in study locations, melkass-6q is no longer replace the standard check (melkassa-2), since it has less grain yield performance and less preferred than melkassa-2 in overall farmers’ preference evaluation criterion. hence, seed producer enterprises, cooperatives or seed producing farmers’ groups and other responsible stakeholder should work for the adequate and quality seed supply of melkassa-2 maize variety till the best performing maize variety that can be replace the existing melkassa-2 variety is released by the national lowland maize research. references [1] n. mandefro, d. tanner, and s. twumasi-afriyie, "enhancing the contribution of maize to food security in ethiopia," in proceedings of the second national maize workshop of ethiopia, 12-16 november 2001, addis ababa, ethiopia. addis ababa, ethiopia: ethiopian agricultural research organization (earo) and international maize and wheat improvement center, 2002. [2] central statistical agency, "agricultural sample survey 2010/11 (september – december 2010) ", report on area and production of major crops, addis ababa, 1: 5–14, 2011. [3] d. alemu, w. mwangi, n. mendefro, and d. spielman, "the maize seed system in ethiopia: challenges and opportunities in drought prone areas," african journal of agricultural research, vol. 3, no. 4, pp. 305-314, 2008. [4] s. rashid, k. getnet, and s. lemma, "maize value chain potential in ethiopia: constraints and opportunities for enhancing the system," gates open research, vol. 3, no. 354, p. 354, 2019. [5] moa, ministry of agriculture, animal and plant health regulatory directorate. addis ababa: the federal democratic republic of ethiopia: crop variety register, 2011. [6] fao, "faostat online database," retrieved: http://faostat.fao.org/. 2011. [7] s. dercon, j. hoddinott, and t. woldehanna, "shocks and consumption in 15 ethiopian villages, 1999–2004," journal of african economies, vol. 14, no. 4, pp. 559-585, 2005. https://doi.org/10.1093/jae/eji022 [8] d. j. spielman, d. byerlee, d. alemu, and d. kelemework, "policies to promote cereal intensification in ethiopia: the search for appropriate public and private roles," food policy, vol. 35, no. 3, pp. 185-194, 2010. https://doi.org/10.1016/j.foodpol.2009.12.002 [9] cimmyt, second semi-annual progress report for the qpm development project for the horn and east africa. addis ababa: csa, 2003. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://faostat.fao.org/ https://doi.org/10.1093/jae/eji022 https://doi.org/10.1016/j.foodpol.2009.12.002 39 © 2022 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 9, no. 1, 39-43, 2022 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v9i1.3865 © 2022 by the authors; licensee asian online journal publishing group influence of packaging material and rates of potassium permanganate on post harvest ripening of african eggplant (solanum macrocarpon) inyang, p.  1,2 bassey, e. m.2 afangide, a.2 ankrumah, e.1 anosike, f.c.1,2 akpaninyang, f3 ndifon, e. m.1 ( corresponding author) 1department of agriculture, alex ekweme federal university, ndufu-alike, nigeria. 2department of crop science and technology, federal university of technology, owerri, nigeria. 3department of crop science, akwa ibom state university, obio akpa, nigeria. 1,2email: paulinyang@yahoo.com 2email: edidiongpaulinyang@gmail.com 2email: afangideakaninyene2000@gmail.com 1email: ankrumahemmanuel@gmail.com 1,2email: francisanosike867@yahoo.com 3email: fidelekan@gmail.com 1email: emndi4nn@yahoo.com abstract climacteric fruits (banana, apple, pear, avocado, garden egg and mango) are actually eaten when ripen; but garden egg is not palatable when ripens and decreases demand. influence of packaging material and rates of potassium permanganate (kmno4) on postharvest ripening of garden egg was evaluated to access the interactive effect of kmno4 and packaging material in delaying ripening. two packaging materials; polythene and jute bags, kmno4 at 0g, 3g, 6g and 9g were used. the experiment was a 2 x 4 factorial laid in completely randomized design (crd) with three replications. eight treatment combinations were randomly distributed. fruits were stored for seven days and were observed every 2 days. result revealed that packaging material influenced postharvest ripening of garden egg with polythene bag reducing the intensity of ripening and significantly reduced shrinkage. similarly, kmno4 at (3g and 9g) significantly delayed ripening intensity at 4 and 6 days of storage. keywords: packaging material, potassium permanganate, postharvest, garden egg, intensity rate, polythene, shrinkage, jute bag. citation | inyang, p.; bassey, e. m.; afangide, a.; ankrumah, e.; anosike, f.c.; akpaninyang, f.; ndifon, e. m. (2022). influence of packaging material and rates of potassium permanganate on post harvest ripening of african eggplant (solanum macrocarpon). agriculture and food sciences research, 9(1): 39-43. history: received: 14 february 2022 revised: 18 march 2022 accepted: 4 april 2022 published: 21 april 2022 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. authors’ contributions: all authors contributed equally to the conception and design of the study. acknowledgment: authors acknowledge the school of agriculture and agricultural technology, federal university of technology, owerri for providing all the necessary materials and support to the full conclusion of this research. competing interests: the authors declare that they have no conflict of interest. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. ethical: this study followed all ethical practices during writing. contents 1. introduction ...................................................................................................................................................................................... 40 2. material and methods ..................................................................................................................................................................... 40 3. results and discussion ................................................................................................................................................................... 40 4. conclusion ......................................................................................................................................................................................... 42 5. recommendation.............................................................................................................................................................................. 43 references .............................................................................................................................................................................................. 43 mailto:paulinyang@yahoo.com mailto:edidiongpaulinyang@gmail.com mailto:afangideakaninyene2000@gmail.com mailto:ankrumahemmanuel@gmail.com mailto:francisanosike867@yahoo.com mailto:fidelekan@gmail.com mailto:emndi4nn@yahoo.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v9i1.3865 https://orcid.org/0000-0002-0561-3792 https://orcid.org/0000-0003-1318-9802 https://orcid.org/0000-0001-6027-4714 agriculture and food sciences research, 2022, 9(1): 39-43 40 © 2022by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study has showed that polythene bags with little kmno4 can extend the shelf -life of garden egg fruit without refrigerator. also maintaining the moisture and appearance of the fruit. there is a dearth of knowledge on controlling garden egg post-harvest losses. 1. introduction eggplant is one of the fruity vegetables in the night shade (solanaceae) family which includes bell pepper, tomatoes and potatoes. eggplants are usually planted as annuals, although some varieties may live into a second season. eggplant can be seen in almost every market in west and central africa, where it is one of the five most important fruity vegetables together with tomatoes, onions, pepper and okra [1]. eggplant is commonly called garden egg. garden egg can be combined with groundnut or groundnut paste to entertain visitors. although far from being nutritional power house, these colourful egg-like fruits provide protein, vitamin and minerals, low in sodium and calories, high in dietary fiber and a good source of potassium [2]. postharvest losses in tropical fruits vary widely from 10 – 80% in both developed and developing countries [3]. it is estimated to range from 10 – 30% per year despite the use of modern storage facilities and techniques [4]. in the tropics, these losses are due to tropical weather, poorly developed infrastructure, poor farm practices, inadequate postharvest handling knowledge, poor storage facilities and epileptic power supply. these losses occur along the supply chain beginning from time of harvest right up to packaging, storage, transportation, retailing and consumption [5]. the average shelf life of garden egg is 3 – 7 days depending on harvesting frequency and conditions [6]. several days after harvest, garden egg change colour from white, cream or stripped green to yellow and or intense orange. at the same time the water content of the fruit also declines causing it to shrivel thus affecting the appearance which changes from a glossy looking skin to a pale/dull appearance. the retail price drops compared to the original value, also the shelf-life reduces making it more susceptible to pest attack. these lead to decrease in its consumption as consumers prefer a freshly looking and unripe fruit causing a loss to the farmer and to the seller respectively. 2. material and methods an experiment was conducted in 2008 at the laboratory of crop science and technology, federal university of technology, owerri to study the influence of packaging materials and rates of potassium permanganate on post harvest ripening of garden egg (solanummacrocarpon). the treatment consisted two (2) types of packaging materials (polythene bag and jute sack bag) and four (4) levels of potassium permanganate (kmno4) (may and baker ltd., dagenham, england) which was packaged in sachet (1 x 1 inch) made from muslin cloth, of which each sachet contained 3g of kmno4 in pellets of 0.3 – 0.5cm in diameter. the levels are 0, 1, 2 and 3 sachet. completely randomized design was used with three (3) replications. garden egg (solanum macrocarpon) variety was randomly picked, weighed (200g) each was packaged into each packaging material. treatments were factorially arranged 2 x 4 x 3 making a total of 24. kmno4 was added to the packaged garden egg as 0 (control), 1 sachet (in the centre of the packaging materials i.e. (polythene and jute sack bags) 2 sachets (one at each end of the packaging materials), 3 sachets (one in the centre and one at each end of the packaging materials). the fruits were stored for seven (7) days and were observed after every two (2) day. data were collected on percentage ripening and intensity of ripening. percentage ripening = number of ripe garden egg in the unit total number of garden egg in the unit x 100 1 intensity of ripening; the following grading system was used to access the intensity of ripening: extremely ripe 4 very ripe 3 moderately ripe 2 slightly ripe 1 not ripe 0 the data collected were subjected to analysis of variance (anova). genstat statistical package 6th edition, where significant difference existed between treatments, least significant difference was used to separate them at 5% probability level. 3. results and discussion the study revealed that packaging materials significantly (p ≤ 0.05) influenced the intensity and rate of garden egg ripening at 4 and 6 days of storage (table 1and table 2). however there was no significant (p < 0.05) difference at 2 days of storage, hence packaging material has little or no effect on ripening. polythene bags at 4 and 6 days (table 1and table 2) of storage significantly (p < 0.05) affected ripening rate and intensity. this can be due to reduced temperature and oxygen in the packaging material. ferris [7] reported that reduced fruit respiration causes decreased ripening and increases storage life. packaging material also significantly (p < 0.05) influences shrinkage after storage (table 3). garden egg packaged or stored in polythene bags did not record any shrinkage, however jute sack recorded some level of shrinkage. this study was in conformity with the work reported by forney and lipton [8] that polymeric films influences the development of chilling injury on fresh produce by creating a modified atmosphere around the commodity and by slowing loss of moisture. intensity and rate of ripening of garden egg stored with different rates of potassium permanganate did not differ statistically throughout the period of the storage (table 4 and table 5). however, there were significant (p ≤ 0.05) effect at 6 days of storage in respect to potassium permanganate on ripening rate. jobling [9] reported that sufficient potassium permanganate should be made available and sprayed over a large surface area for proper reaction with ethylene, hence an increased rate of potassium permanganate may possibly show a significant effect on both ripening intensity and rate. garden egg ripening intensity and rates was not significantly influenced by the interaction between packaging material and potassium permanganate at 2 days of storage (table 6 and table7) in this study, but recorded significantly (p < 0.05) interactive effect between the packaging materials and potassium agriculture and food sciences research, 2022, 9(1): 39-43 41 © 2022by the authors; licensee asian online journal publishing group permanganate on the intensity and rate of garden egg ripening, with garden egg stored in polythene bag treated with 3 and 9 grams of potassium permanganate showed delayed ripening intensity at 4 and 6 days of storage as compared with others. this revealed that garden egg stored on or before 2 days using the packaging materials and treated with potassium permanganate does not influence the fruit ripening. this could be probably due to the fact that no active physiological activities at the early time of fruits under storage condition. this is in line with [10] on their work to evaluate the effects of kmno4 on the extension of postharvest life of 'sunrise golden' papaya reported that at 4 days after bag sealing, carbon dioxide (co2) concentration was higher in bags without kmno4 and was stabled in those treated with varying rates of kmno4. a b c d a b c d garden egg fruits treated with 0g of potassium permanganate garden egg fruits treated with 3g of potassium permanganate garden egg fruits treated with 6g of potassium permanganate garden egg fruits treated with 9g of potassium permanganate figure 1. illustrates garden egg fruits in polythene bag, treated with different rate of kmno4. at 6 days. 3g of kmno4 sealed in polythene bag after 4 and 6 days (figure 1 and figure 2) show significant reduction in the rate and intensity of ripening, and weight loss when compared to that of jute bag. this is because kmno4 is efficient at lower concentrations (1.5g 3g) in polythene bag irrespective of the thickness, in delaying ripening of fruits, extending shelf life and improving textural quality under ambient temperature (26 28 ºc). this agrees with those reported by silva, et al. [10], wabali and esiri [11], azad, et al. [12], anyasi, et al. [13] and wabali, et al. [14] that bags with lower concentration of kmno4 reduces the concentration of co2 after 2 days of storage. the oxidation of ethylene by kmno4 leads to the formation of water (h2o) and co2, but in jute bag the water may have been absorbed by the bag resulting into significant deterioration of garden egg fruit after 2 days and above. a b c d garden egg fruits treated with 0g of potassium permanganate garden egg fruits treated with 3g of potassium permanganate garden egg fruits treated with 6g of potassium permanganate garden egg fruits treated with 9g of potassium permanganate figure 2. illustrates garden egg fruits in jute bag, treated with different rate of kmno4.at 6 days. agriculture and food sciences research, 2022, 9(1): 39-43 42 © 2022by the authors; licensee asian online journal publishing group table 1.effect of packaging materials on intensity of ripening with increasing days to storage. packaging materials days of storage 2 4 6 polythene bag 0.500 0.840 1.525 jute bag 0.833 1.650 2.019 lsd(0.05) ns 0.55 0.359 table 2. effect of packaging materials on the rate of ripening with increasing days of storage. packaging materials days of storage 2 4 6 polythene bag 11.30 23.50 54.20 jute bag 22.20 48.20 69.80 lsd(0.05) ns 16.98 15.58 table 3. effect of packaging materials on shrinkage and weight loss after storage. packaging materials shrinkage weight loss polythene bag 0.000 4.200 jute bag 1.880 7.800 lsd(0.05) 0.280 ns table 4.effect of kmno4 on intensity of ripening with increasing days of storage. packaging materials days of storage 2 4 6 0 0.833 1.500 2.015 3 0.667 1.100 1.545 6 0.500 1.230 1.725 9 0.667 1.050 1.803 lsd(0.05) ns ns ns table 5. effect of kmno4 on the rate of ripening with increasing days of storage. packaging materials days of storage 2 4 6 0 23.40 46.20 80.90 3 14.30 27.00 57.10 6 12.70 41.70 57.50 9 16.70 28.60 52.40 lsd(0.05) ns ns 22.03 table 6. interactive effect of packaging materials and kmno4 on intensity of ripening with increasing days of storage. packaging materials kmno4 (g) days of storage 2 4 6 polythene bag 0 0.667 1.500 2.093 3 0.333 0.667 1.083 6 0.667 0.860 1.600 9 0.333 0.333 1.333 jute bag 0 1.000 1.667 1.947 3 1.000 1.533 2.007 6 0.333 1.610 1.850 9 1.000 1.777 2.273 lsd(0.05) ns 1.030 0.719 table7. interactive effect of packaging materials and kmno4 on the rate of ripening with increasing days of storage. packaging materials kmno4 (g) days of storage 2 4 6 polythene bag 0 21.40 40.00 77.80 3 4.80 11.10 42.90 6 14.30 33.30 48.40 9 4.80 9.50 47.60 jute bag 0 25.40 52.40 84.10 3 23.80 42.90 71.40 6 11.10 50.00 66.70 9 28.60 47.60 57.10 lsd(0.05) ns 33.96 31.15 4. conclusion base on this study, packaging materials influenced post harvest ripening of garden egg, with polythene bag reducing the rate and intensity of ripening and significantly reduce the shrinkage of garden egg fruit. potassium permanganate in this study showed very little effect in its ability to absorb ethylene released from ripening garden egg fruits which was due to the rate or quantity used, however the interaction of polythene bag with 3g and 9g of potassium permanganate delayed the ripening of garden egg. agriculture and food sciences research, 2022, 9(1): 39-43 43 © 2022by the authors; licensee asian online journal publishing group 5. recommendation further studies are thereby recommended to validate these findings and to also identify proper or other suitable package that will absorb moisture in packaging potassium permanganate to avoid dilution. egg fruit borers (darabalaisalis and sceliodeslaisalis) should be effectively controlled on the field as their activities affect the post harvest life of garden egg. references [1] r. r. schippers, african indigenous vegetables: an overview of the cultivated species. chatham uk: natural resources institute/acp. eu. technical centre for agriculture and rural cooperation, 2000. [2] national research council, lost crops of africa: vegetables vol. 2. washington, d.c: the national academies press, 2006. [3] f. m. arshad, a. radam, and z. mohamed, "an economic analysis of the malaysian fruits industry," in proceedings of the international seminar on post-harvest handling & processing of tropical and subtropical fruits. & tfnet general assembly kuala lumpur, malaysia., 2003. [4] j. m. harvey, "reduction of losses in fresh market fruits and vegetables. in: vegetable productivity: the role of vegetable in feeding people and livestock. c.r.w. speeding (1981)," proceedings of a symposium held at a royal geological society, london, 1978, pp. 139 – 172. [5] fao, "post harvest handling losses," presented at the committee on commodity problems: intergovernmental group on bananas and tropical fruits (4th session) guayaquil, ecuador, 2005. [6] d. horna and g. g., "marketing under utilized crops for biodiversity: the case of african garden egg (solanum aethiopicum) in ghana," presented at the 8th international bioecon conference on economic analysis of ecology biodiversity. kings college, cambridge, 2006. [7] s. r. b. ferris, improving storage life of plantain and banana. research guide (62). ibadan: international institute of tropical agriculture, 1997. [8] c. f. forney and w. j. lipton, influence of controlled atmosphere and packaging on chilling sensitivity. in: c. y. wang (editor), chilling injury of horticultural crops. boca raton, fla: crc press, 1990. [9] j. jobling, post harvest ethylene: a critical factor in quality management. australia: sydney post harvest laboratory information sheet, 2000. [10] d. f. p. silva, l. c. c. salomão, d. l. de siqueira, p. r. cecon, and a. rocha, "potassium permanganate effects in postharvest conservation of the papaya cultivar sunrise golden: pesq. agropec. bras," brasília, vol. 44, pp. 669-675, 2009. [11] v. c. wabali and a. esiri, "effect of potassium permanganate on colour and textural characteristics of tomatoes at ambient temperature storage," european journal of agriculture and food sciences, vol. 3, pp. 60-62, 2021.available at: http://dx.doi.org/10.24018/ejfood.2021.3.2.263. [12] m. azad, g. mortuza, a. nahar, s. huq, and a. alam, "effect of potassium permanganate on physicochemical changes and shelf life of mango," the agriculturist, vol. 6, pp. 54–59, 2009. [13] t. anyasi, c. aworh, and o. jideani, "effect of packaging and chemical treatment on storage life and physicochemical attributes of tomato," african journal of biotechnology, vol. 15, pp. 1913–1919, 2016. [14] v. c. wabali, a. esiri, and l. zitte, "a sensory assessment of color and textural quality of refrigerated tomatoes preserved with different concentrations of potassium permanganate," food science & nutrition, vol. 5, pp. 434-438, 2017. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://dx.doi.org/10.24018/ejfood.2021.3.2.263 69 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 69-77, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6820 © 2025 by the authors; licensee asian online journal publishing group assessment of productivity of spring and winter wheat sorts from china and pakistan in high mountain tajikistan kozimamad a abdulamonov1 aslam q qadamov2 aziz ali khan3 roy c. sidle4 ahmad q abdulamonov5 ( corresponding author) 1,5pamir biological institute, nast, 2 kholdorov street, 736000 khorog, tajikistan. 1email: ahmad80@mail.ru 5email: ahmad79.79@mail.ru 2,3,4university of central asia, mountain societies research institute, 155 qimatsho imatshoev street, 736000, khorog, tajikistan. 2email: aslam.qadamov@ucentralasia.org 3email: azizali.khan@ucentralasia.org 4email: ahmad80@mail.ru abstract wheat is a staple crop and critical for food security in the high mountains of tajikistan. despite the local and introduced varieties of wheat, overall yields remain low and crops are vulnerable to diseases and climatic shocks. to improve production, new adapted and disease-resistant crops are crucial. we tested ten spring wheat sorts from pakistan and seven from china with higher yields, disease resistance, and shorter stems compared to local varieties. local varieties performed better than foreign varieties in stem length (77% longer), 25% more grains in the main spike, 45% longer main spike, 4% more spikelets in the main spike, and more productive tillers. foreign cultivars had shorter morphogenic events, 16% higher grain weight in the main spike, 23% higher weight of 1000 grain, and 11% more grains per spikelet. yield components and morpho-physiological traits for chinese cultivars were weakly correlated. the main phenotypic traits affecting crop yield components were general and productive tillering. in contrast to chinese cultivars, the main crop yield component values of local cultivars negatively interacted with most genotypic and phenotypic traits. foreign cultivars were less susceptible to lodging and diseases; however, their shorter stems made them less attractive to local farmers. keywords: breeding parent selection, disease resistance, food security, gorno-badakhshan, grain quality, high-altitude agriculture, lodging resistance, straw yield. citation | abdulamonov, k. a., qadamov, a. q., khan, a. a., sidle, r. c., & abdulamonov, a. q. (2025). assessment of productivity of spring and winter wheat sorts from china and pakistan in high mountain tajikistan. agriculture and food sciences research, 12(1), 69–77. 10.20448/aesr.v12i1.6820 history: received: 28 april 2025 revised: 5 june 2025 accepted: 18 june 2025 published: 25 june 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by aga khan foundation tajikistan within the thrive project, tajikistan (grant number: 122042700045-3). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 70 2. materials and methods ................................................................................................................................................................... 70 3. results ................................................................................................................................................................................................ 72 4. discussion .......................................................................................................................................................................................... 75 5. conclusion ......................................................................................................................................................................................... 76 references .............................................................................................................................................................................................. 77 mailto:ahmad80@mail.ru mailto:ahmad79.79@mail.ru mailto:aslam.qadamov@ucentralasia.org mailto:azizali.khan@ucentralasia.org mailto:ahmad80@mail.ru https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6820 https://orcid.org/0009-0000-6291-7066 https://orcid.org/0009-0005-8571-1585 https://orcid.org/0009-0003-5682-3129 https://orcid.org/0000-0002-5004-4154 https://orcid.org/0009-0009-6621-9455 agriculture and food sciences research, 2025, 12(1): 69-77 70 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature: this is the first wheat trial in gbao that put high-altitude chinese and pakistani lines side-byside with local wheats. earlier tests used only tajik varieties. by using local types as controls, we found foreign lines with valuable traitsearlier ripening, strong disease resistance, and heavier grains. these traits are now being crossed into local wheats, a clear and practical outcome of the study. 1. introduction wheat (triticum aestivum l.), a member of the grass family poaceae [1] is the third most important cereal crop behind rice and maize [2, 3] has profound economic and livelihood benefits [4] and is the most widely grown crop worldwide [5, 6]. the increasing demand for wheat increases its planted area, reaching up to 219 million ha with yields ranging from 732.1 to 760.9 million tons globally [7]. wheat provides about 20% of protein and calories worldwide [8] and its gluten protein is widely used in many foods [9]. wheat is the highest-ranked cereal crop, accounting for 30% of all cereals, and provides staple food for more than 7.9 billion people in up to 43 nations [10]. in developing countries, the demand for wheat is expected to grow by as much as 60% by 2050 [11]. wheat is one of central asia's most consumable cereal crops; almost every meal includes bread made of wheat [12]. in the pamir of tajikistan, located in gorno badakhshan autonomous oblast (gbao), both local and introduced wheat varieties have served as a mainstay of food security and livelihood support for many years, but changes in socio-economic conditions together with climate changes have made wheat less attractive due to declines in yields and disease resistance [13]. lack of knowledge and financial capital prevent local farmers from introducing new varieties that are more resistant to disease and more productive. moreover, this practice requires a preliminary assessment of introduced cultivars aimed at determining the main characteristics of the introduced varieties in the new location. therefore, it is important to identify and select the best wheat varieties that can grow in mountain conditions as well as conduct breeding selection in such environments. the overall goal of this research was to assess foreign wheat sorts from china and pakistan that carried traits such as high yield, disease resistance, and short stems that would not lodge before harvest and compare these to local varieties. ancient wheat varieties have been cultivated by local farmers for centuries, but during the last century were replaced by high-yielding sorts [13] which led to the loss of some varieties. considering the importance of ancient cultivars and to preserve natural agrobiodiversity [14] pamir biological institute (pbi) collected more than 100 local wheat varieties from tajik and afghan parts of badakhshan. the main disadvantage of most local cultivars is their low yields and low baking qualities [15]. pbi with a long history of agricultural research and extension is the only facility in gbao with enough land to test many wheat varieties focusing on developing and preserving local varieties and their gene pools. each year pbi provides small farmers in gbao with top local spring wheat varieties (safedaki ishkashimy, surkhkhusha, bobillo, safedaki bartang, and kilak bartangy) along with the proven lines from afghan badakhshan, such as sadiras, pandaki (facultative), and bludon from takhar province [16]. in recent years, pbi scientists and other tajik breeders have crossed local and foreign wheats at the ishkashim experimental station, producing hybrids that combine valuable traits [13]. surprisingly, results of tested varieties show that no foreign variety surpassed the local lines in a total dry biomass and yield metrics [13]. local wheat varieties have some valuable characteristics such as tolerance to local stresses, yield stability, strong adaptability to high altitudes, and important genetic diversity. many of the local cultivars are ancient wheat. the interest in ancient grains increased during the last decades mainly because of their better nutritional composition [17]. the presence of lipids, trace elements, proteins, and other contents make ancient grains popular among farmers and consumers [18, 19]. another feature of local varieties is the so-called high “straw to grain yield” ratio (the ratio of the height of the cultivar to yield). this is important for local farmers as the wheat straw is usually used as livestock feed during winter [20]. therefore, dwarf and semidwarf cultivars are not popular among local farmers. however, local varieties also have disadvantages such as lodging, susceptibility to diseases, and low baking quality. the low baking qualities relegate these varieties to fodder wheat [21]. to support livelihoods and mitigate food insecurity in these mountainous regions, as well as increase the adaptative capacity of local farmers to climate variability, it is essential to improve the yield and the quality of local cultivars by introducing new varieties for screening and dissemination among local farmers. thus, the main objective of this research was to assess and compare the productivity of foreign spring soft wheat sorts from china and vernalized winter sorts from pakistan to local varieties in the mountain conditions of gbao. in developing countries, people depend on cereal crops as a main nutrient source, therefore production of sufficient grain crops to ensure food security remains significant [22]. recent studies attempted to identify mechanisms to increase yields [9, 23, 24] but wheat yield depends on many factors [25, 26]. thus, the introduction of new cultivars is important because the variety genotype is a key factor affecting yield [27]. on the other hand, only proper selection and introduction of new varieties and their tolerance towards agroecological conditions can guarantee high yields and high-quality grains [28]. therefore, comparative analyses of the performance of introduced varieties in local conditions are important. 2. materials and methods 2.1. site characteristics ishkashim district is situated in the southern part of gbao (figure 1) and is characterized by a continental and arid climate. the summers are warm and dry, and the winters are cold with little snow. annual precipitation is low, only 80-120 mm. precipitation mainly falls during the winter and spring (80%), with negligible rain in summer and autumn. march is the wettest month and august is the driest month of the year. the amount of solid precipitation is quite low, ranging from 10 to 20% of total precipitation. the difference in precipitation between the driest and wettest months is 78 mm [29]. during the growing season (may to september) in this arid area, the relative air humidity is no more than 40%. the average annual temperature in ishkashim is 7.5 °c. july is the warmest month of the year with an average temperature of 19.5 °c, and january is the coldest month with an average temperature of −6.0 °c. the average monthly temperature varies by 25.5 °c throughout the year [30]. agriculture and food sciences research, 2025, 12(1): 69-77 71 © 2025 by the authors; licensee asian online journal publishing group during the research period (2019-2021), no significant differences in mean annual air temperature were observed for the research area (figure 2). precipitation was higher in 2020 (91 mm) compared with the other two years, but the difference was insignificant. figure 1. location of the study area and experimental plot in ishkashim district, gbao, tajikistan. figure 2. the distribution of mean annual air temperature and precipitation within the research period based on data from ishkashim meteorological station. 2.2. experimental design, and agronomic practices the research was conducted at the pbi experimental station in ishkashim, gbao, tajikistan (36°40'42"n, 71°41'22"e) (figure 1). seven breeding sorts (crbw1, crbw2, crbw3, crbw4, crbw5, crbw6, and crbw7) were provided by the agronomy department, university of chengdu, china. ten breeding sorts (trjata, no. 63, no. 76 skd, no. 75, no. 78 skd, no. 105, no. 141, no. 142, no. 144 and no. 147) were provided by the pakistan agriculture research council (parc). the pakistani winter wheat sorts were tested and successfully cultivated in the skardu region (above 2000 m a.s.l.) of gilgit baltistan province, pakistan. as a control local variety, safedaki ishkashimy was compared with the chinese wheat varieties and local varieties joydori and bludon were compared with varieties from pakistan. local varieties were included because they have evolved under the local conditions and are adapted to high altitudes (2000-3250 m a.s.l.). in october 2019 local winter wheats joydori and bludon and ten pakistani lines were sown at the ishkashim experimental site. all plants tillered but later froze because the 2019-2020 winter had little snow cover. in march 2020 some pakistani lines and the two local varieties were vernalized in a freezer (below 0°c for 86 days) and sown in spring. because seeds was limited, the final lines kept only three pakistani lines (no. 75, no. 144, and no. 63) plus joydori and bludon. soil is a sandy loam over pebbles, giving good drainage and high natural fertility [31, 32]. the second-year trail was planted in the third week of april and harvested in late september. before ploughing, we applied 200 kg/ha of nitrogen-phosphorus fertilizer and 100 kg/ha of urea. rows are 50 cm apart, each 1 m strip was separated by 60 cm. we hand-sowed two rows per strip (100 seeds per row, 2 cm between plants, 17 cm between the paired rows). each 0.5 m2 plot was replicated three times, and the replicates were arranged at random. agriculture and food sciences research, 2025, 12(1): 69-77 72 © 2025 by the authors; licensee asian online journal publishing group 2.3. data collection the key growth staged such as germination, tillering, stem elongation, heading, and stage of ripeness (milk and dough stages, and full ripening) were tracked. the time from germination to heading and from heading to ripening was recorded, and some plants showed yellow rust during ear emergence and flowering. at harvest we sampled ten plants per variety and measured plant height, general and productive tillering, length of the main spike, number of spikelets on the main spike, number and weight of grains in the main spike, number of grains per spikelet, grain weight of the main spike, grain weight of one plant, and weight of 1000 grains. each of the 12 traits was tested with a one-way anova in xlstt, with lsd use to separate means. pearson correlations among traits were run in r studio (https://posit.co/download/rstudio-desktop). 3. results 3.1. comparative analyses of spring soft wheat sorts as a widely adapted crop, wheat grows in most environments but thrives in cool environments [33]. the main parameters of the spike that determine productivity (i.e., number of spikelets per spike, number of kernels per spikelet, and weight of individual kernels) are determined genetically but can be affected by environmental conditions [34]. in this study, all relevant parameters related to crop yield were analyzed. the comparative analyses amongst all chinese and local sorts show that the mean plant height of the control variety safedaki ishkashimy ranged from 90.6 to 96.0 cm during the 3-yr period, significantly higher compared to chinese sorts (figure 3a). the height of wheat is an important factor for tajik farmers as the straw is used for winter cattle forage. height of chinese sorts during this period ranged from 52 (crbw-7) to 62 (crbw-6) cm, significantly shorter than controls. all chinese sorts were semidwarf. local varieties surkhkhusha and pandaki showed slightly better results than the control (safedaki ishkoshimi), but these height differences were not statistically significant. pandaki produced the tallest plants of all varieties tested. during the 3-yr period, safedaki ishkashimy (control) had the highest mean of the general tillering, a specialized branching that occurs from the basal node and grows independently of the mother stem [35] (3.5-3.7 pcs), significantly higher than sorts from china (figure 3b). the general tillering of sorts from china in the 3-yr period ranged from 2.4 to 2.9 pcs. local varieties were not significantly different from the control, except in 2020 when surkhkhusha was significantly higher. the mean number of productive tillers (number of tillers that produce spikes and seeds) [36] of the control variety was 3.4 pcs. similar to general tillering, the control variety had a higher number of productive tillers compared to all sorts from china. productive tillering of chinese sorts ranged from 2.3 to 2.7 pcs during the 3-yr period (figure 3c). also, similar to general tillering, local varieties were not significantly different from controls, except in 2020 when surkhkhusa was significantly higher. the mean of the length of the main spike in the control variety was significantly greater than the range reported for chinese types. the spike length of the chinese types ranged from 5.5 to 7.5 cm (figure 3d). local types did not differ significantly from the control, except for surkhkush in 2021 (7.40 cm) and pandaki in 2020 (7.7 cm), which were both shorter than the control. safedaki ishkoshimy, the control, also had significantly more spikelets per main spike than the chinese types. nevertheless, crbw7, crbw6 crbw5 and crbw2 had higher values than the control in 2020 (figure 3e). on average , the chinese types had from 11.2 to 15.1 pcs spikelets per main spike . local varieties surkhkusha and pandaki had significantly fewer spikelets than the control. no significant differences were observed between the control and chinese varieties for the number of grains per spikelet. there were about 2.2 to 3.20 pcs grains per spikelet among the chinese varieties (figure 3f). the sorts from china (crbw7, crbw6, crbw4, crbw5 and crbw2) recorded somewhat higher values than the control in 2029, but the 3-year averages were not significantly different.. the control plants carried about 38–42 grains on the main spike. crbw1 always produced fewer grains than the control. crbw7, crbw6, and crbw5 matched the control in 2019 but fell below the control in 2020 and 2021. crbw3 and crbw2 followed the same pattern. crbw4 was the only line with more grains than the control in 2019 (about 44), yet it dropped below the control in 2020 and showed no difference in 2021. both local sorts equaled the control in 2019 and trailed the control in the next two seasons. the control’s 1000-grain weight ranged from 29.8 to 34.7 g. in 2019, grain weights for crbw1, crbw4, crbw5, and crbw2 were clearly heavier (40–43 g), while crbw7, crbw6, crbw3, and the local varieties showed no statistical difference from the control. local varieties needed the most time to reach heading, about 53–61 days. chinese lines headed sooner at roughly 52–55 days, so they were significantly earlier than the control in 2019 and 2020. in 2021, heading in two of those lines extended to 58–59 days, and the rest matched the control. surkhkhusha reached heading later than the control only in 2019, whereas pandaki was slow every year (65–72 days). the time period from heading to full ripening was shorter for local wheats (51–53 days). most lines mirrored the control in 2019 and 2021, except crbw6, which took slightly longer to ripen in 2019. during the 2020 season all sorts needed 59–62 days from heading to full ripening, outpacing the control. local varieties ripened more rapidly than the control in 2019–2020 but slower in 2021. the germination to ripening period is longer for local varieties compared to foreign sorts (112 and 106 days, respectively; figure 3k). during the 3-year study, this period ranged from 104-113 days. the mean germination to ripening periods for all sorts in 2019 and 2021 were a bit slower compared to the control but not statistically different (table 1). in 2020, all sorts were not significantly different from the control. surkhkhusha was not significantly different from the control in all years, whereas the local variety pandaki had significantly higher values in 2019 and 2020 (109 and 119 days, respectively). agriculture and food sciences research, 2025, 12(1): 69-77 73 © 2025 by the authors; licensee asian online journal publishing group figure 3. main parameters of the chinese and local sorts; a) plant height, b) general tillering, c) productive tillering, d) length of main spike e) number of grains in the main spike. f) number of grains per spikelet, g) number of grains in the main spike h) weight of 1000 grains, i) germination to heading period, j) heading to ripening period, k) germination to ripening period, l) weight of grains in the main spike. there were no differences in the weight of grains in the main spike of the control compared to foreign sorts (figure 3l). grain weights in the control ranged from 1.2 to 1.3 g. some chinese sorts (crbw2, crbw3, crbw4, crbw5, crbw7) showed better performance than the control variety but these changes were not significant (p<0.05). surkhkhush and pandaki had significantly lower weight of grain in the main spike compared to the control. 3.2. comparative analyses of vernalized winter sorts local cultivars were taller than introduced sorts from pakistan. control height ranged between 85.9 to 105.6 cm (figure 4a), generally higher than sorts from pakistan (74.4 to 95 cm), except sort №63 in 2019. the local variety bludon was not significantly different from the control in all years. figure 4. main parameters of the pakistani and local varieties; a) plant height, b) general tillering, c) productive tillering, d) length of the main spike, e) number of kernels in the main spike, f) number of grains per spikelet, g) number of grains in the main spike h) weight of grains in the main spike i) weight of 1000 grains, j) germination to heading period, k) heading to ripening period, and l) germination to ripening period. agriculture and food sciences research, 2025, 12(1): 69-77 74 © 2025 by the authors; licensee asian online journal publishing group the mean value for general tillering in the control was 3.3 pcs., not significantly different from foreign sorts. this value for the sorts from pakistan ranged from 2.2 to 2.7 pcs. (except №144 in 2020) and was not significantly different from the control (figure 4b). local variety bludon had a lower tillering value in 2019 and 2021 (2.2 and 2.6 pcs.) but was not significantly different (3.3 pcs.) from the control in 2020. the mean of productive tillering in the control was 3.2 pcs. for pakistani sorts, productive tillering ranged from 2.2 to 2.6, which was not statistically different than the control (table 1). the local variety bludon had lower values ranging from 2.2 to 2.9 pcs (figure 4c). there was no difference in the length of the main spike among the control and foreign sorts (figure 4d). the mean value of the main spike of the control was 8.3 cm, significantly longer compared to pakistani sorts (6.9 to 8.0) (table 2). local cultivar bludon was not significantly different than the control. the mean number of spikelets in the main spike of the control ranged from 14.5 to 16.2 pcs (figure 4e). all foreign sorts and bludon were not significantly different from the control in all years. no statistically significant differences were found among cultivars. the mean number of grains per spikelet in the control ranged from 2.3 to 2.8 pcs. (figure 4f). sorts №75 and №63 did not differ from the control values in all years. only sort №144 had more grains per spikelet compared to the control, but this was not statistically significant. in 2019 and 2021, local cultivar bludon had a higher value compared to the control, but in 2020 it had significantly lower values. the control had an average of 36.5 grains in the main spike during the 3-year study (figure 4g). sort №75 and bludon were not significantly different from the control. sorts №144 and №63 had significantly higher grains per spikelet in all years except 2019 (№144) and 2020 (№63). the pakistani variety № 63 had the highest mean weight of grains in the main spike (1.6 g) and the local variety bludon had the lowest value (1.2 g), although the difference between the cultivars was not statistically significant (figure 4h). on an annual basis, the highest value was recorded in 2021 (1.7 g) and the lowest in 2020 (1.2 g). the mean weight of 1000 grains in the control ranged from 35.9 to 48.1 g (figure 4i). sorts №75 and №144 had higher values in some years but mean values were not significantly different (table 2). local cultivar bludon had the lowest mean value. the germination to heading period was 65 days for the control, slightly longer but not significantly different from pakistani sorts (59 to 62 days, figure 4j, table 2). local cultivar bludon had the longest germination to heading period (67 days). the control cultivar joydori ripened first (42 days). the pakistani cultivars had a longer heading to ripening period (46-49 days) but this was not significantly different from the control variety (figure 4k). for local cultivar bludon, ripening occurred after 44 days. the germination to ripening period was similar for all varieties (108 to 110 days) (figure 4l). the ripening period varied throughout the 3-year study period. for instance, in 2019 all sorts had significantly lower values (ranged from 98 to 104 days), while in 2020 all sorts and the local cultivar bludon had higher values (ranged from 111 to 112 days) compared to the control. in 2021 all sorts and the local cultivar bludon were not significantly different from the control. 3.3. pearson correlation analyses between the crop parameters the results of spearman’s coefficient of rank correlations between various crop yield parameters of chinese, local, and pakistani cultivars are shown in figure 5. among the phenotypic and genotypic variance affecting the grain yield, the influence of yield components, namely the number of grains per spike, weight of the kernel, and spikes per unit area are significant [37]. we observed a weak correlation between the yield components and morpho-physiological traits for chinese cultivars (figure 5a). the length of the main spike (lms) was negatively correlated with weight of 1000 grains (w1000g), number of grains per spikelet (ngps), and general tillering (gt). lms was positively correlated with the number of spikelets in the main spike (nsms), but had a nonsignificant positive correlation with plant height (ph), number of grains in the main spike (ngms), and weight of grains in the main spike (wgms). among the main phenotypic traits affecting the ngms are gt and productive tillering (pt). the interaction between them is highly significant (p<0.001). a negative correlation was detected for w1000g with morpho-physiological traits, especially with hrp (p<0.01), and grp (p<0.05). however, w100g had a negative non-significant correlation with ngms and nsms. in contrast, nsms was positively correlated with almost all traits, while the level of the interaction between these traits was low. wgms and ngms were also not well correlated with morpho-physiological traits. for local spring wheat cultivars, the highest correlation was detected between the ngps and gt, ngms and wgms, ngms and ph, and between w1000g and pt (figure 5b). the main phenotypic traits affecting the yield components were pt, lms, and gt. in contrast to chinese cultivars, lms, ph, wgms, and ngms values of local cultivars negatively interact with most genotypic and phenotypic traits. the grain yield components wgms and ngms correlated well with ph and lms, though the coefficient of the correlation is not significant (p≥0.05). the height of the plant (ph) of local wheat cultivars had a high negative correlation with most morphophysiological traits (grp, pt, ghp, hrp) and yield trait components (w1000g, ngps). for the pakistani sorts, a high correlation was detected between some phenotypic traits (grp, ghp and lms) and yield components (ngms, wgms) (figure 5c). other phenotypic traits (gt, pt and hrp) showed a negative interaction with most genotypic and phenotypic traits. the 1000 grain weight had a moderate positive correlation with wgms, ph, and hrp, but a non-significant negative correlation with yield components (ngms and ngps). ph and lms interacted significantly with wgms. the main morpho-physiological traits affecting the crop yield components are ph, ghp, and grp. agriculture and food sciences research, 2025, 12(1): 69-77 75 © 2025 by the authors; licensee asian online journal publishing group figure 5. correlation between the main crop yield parameters of chinese (a), local (b), and pakistani (c) cultivars. ph-plant height, ptproductive tillering, nsms-number of spikelets in the main spike, hrpheading to ripping period, grpgemination to heading period, lmslength of the main spike, w1000gweight of 1000 grains, ngmsnumber of grains in the main spike, ngps-number of grains per spikelet, and wgmsweight of grains in the main spike. note: ns p>=0.05; *p<0.05; ==<0.01; and ***<0.001. 4. discussion varieties with long adaptation periods to local stresses favor local cultivars if the high mountain setting is not well suited to newly introduced crops. an ideal cultivar that best fits the local context and contributes to food security in this mountain region must possess ideal traits, including long straw; high numbers and mass of kernels per spike; high numbers of productive tillers; early maturity; and resistance to local environmental shocks and diseases. our findings show that local varieties have some advantages over imported cultivars, especially for crop height, number of spikelets on the main spike, length of the main spike, and number of grains in the main spike. introduced cultivars have higher numbers of grains per spikelet, higher weight of 1000 grains, and matured earlier. these results indicate that despite having longer spikes and longer axes, local cultivars produce small grains diminishing total cultivar yields (table 1). local cultivars produce longer stems and more tillers, which are important for the local farmers because they use wheat straw for cattle fodder during the long winters. the local cultivar safedaki ishkoshimi was 72% higher than the average value of the chinese cultivars. the height of the plants was recognized as a main feature of ancient wheat [14]. on the other hand, tall plants are susceptible to lodging [38] causing decreases in yields because the lodged plant becomes susceptible to diseases. therefore, to keep the stem standing, the spikelets have undergone significant changes during domestication, mainly expressed by changes in their shape and decreases in kernel weight [14]. thus, despite local wheat cultivars producing more grains in the main spike, the weight of grains is lower compared to foreign varieties. local cultivars had longer periods of morphogenic events ranging from 109 to 114 days. chinese cultivars matured somewhat earlier, but results were not statistically significant (table 1). early cultivar maturation is an important trait because of the harsh local climate. the growing season in ishkashim is short, varying from 150 to 155 days, and the sum of effective daily temperatures (>10°c) range from 1000 to 1100°c per year [39]. rain can occur in september making cereal crops vulnerable to microbial diseases, thus farmers prefer to harvest earlier before the onset of september rains. as such, early maturation of the cultivars is significant. in the overall heading stage in 2019 and 2020, five wheat sorts ripened 5 to 11 days earlier compared to the control. in contrast, the heading to ripening period for the local variety (pandaki) was 8 to 13 days later compared to the control. the heading to ripening period in the five sorts in 2020 was 2 to 7 days longer than for the control; however, in other years these differences were not significant. the germination to heading periods for the five sorts were 2 to 7 days shorter than the local variety surkhkusha. the main characteristics of the spike, such as length, number of spikelets and grains in the spike, and size and weight of grains are important to biologists and breeders [40]. local cultivars had higher spikelet counts and higher numbers of grains in the main spike, but the weight of grains per spike was lower than in chinese varieties. thus, total yields of local cultivars were lower. the number of spikelets is generally determined genetically, although they could be influenced by the environment [34]. grain productivity in local wheat varieties is formed based on characteristics such as productive tillering, spike length, number of spikelets, and number of grains in the main spike. in contrast, grain productivity in chinese sorts was based mainly on weight of 1000 grains and a longer heading to ripening period. table 1. analyses of variance of main crop parameters of the local and foreign spring cultivars from china. parameter α f pr > f p-values signification codes plant height 0.05 127.5 0.0001 *** length of the main spike 0.05 15.27 0.0001 *** number of grains per spikelet 0.05 4.39 0.003 ** number of grains in the main spike 0.05 6.81 0.00018 ** number of spikelets in the main spike 0.05 1.78 0.13 ° grain weight of the main spike 0.05 1.75 0.142 ° weight of 1000 grains 0.05 50.38 0.069 . heading to ripening period 0.05 5.27 0.99 ° germination to ripening period 0.05 0.95 0.505 ° germination to heading period 0.05 7.74 0.51 ° general tillering capacity 0.05 9.904 <0.0001 *** productive tillering capacity 0.05 8.02 <0.0001 *** note: computed against model y=mean(y). signification codes: 0 < *** < 0.001 < ** < 0.01 < . < 0.1 < ° <1. agriculture and food sciences research, 2025, 12(1): 69-77 76 © 2025 by the authors; licensee asian online journal publishing group the highest-yield traits, such as number of grains per spike, grain weight of the main spike, and weight of 1000 grains, were found in six chinese lines (crbw1, crbw7, crbw4, crbw5, crbw3 and crbw2). these lines also stood up well to lodging, pests, and diseases and have good baking quality, making them useful parents for crossing with local wheats. the main disadvantage of these sorts in this region is that they have shorter stems with a lower straw-to-grain yield ratio. therefore, they should be used in the breeding and hybridization rather than released directly. during the three-year test, pakistani winter lines (no. 75, no. 144, and no. 63) grew shorter, tillered less, and had shorter main spikes than the local bludon. spikelet and grain counts were mostly similar to the control, except that no. 144 had more grains per spike. all three pakistani lines reached heading 4-10 days sooner and matured 57 days earlier than the control, while bludon reached heading later but ripened at the same time. sort no. 144 and no. 63 also produced more grains per main spike than the control. table 2. analyses of variance of the local and foreign winter sorts from pakistan. parameters α f pr > f p-values signification codes plant height 0.05 4.892 0.019 * length of the main spike 0.05 6.10 0.009 ** number of grains per spikelet 0.05 0.46 0.759 ° number of grains in the main spike 0.05 1.56 0.257 ° number of spikelets in the main spike 0.05 0.367 0.827 ° grain weight of the main spike 0.05 0.64 0.65 ° weight of 1000 grains 0.05 0.61 0.659 ° heading to ripening period 0.05 0.209 0.927 ° germination to ripening period 0.05 0.068 0.990 ° germination to heading period 0.05 2.06 0.161 ° general tillering 0.05 1.79 0.207 ° productive tillering 0.05 1.80 0.204 ° note: computed against model y=mean(y). signification codes: 0 < ** < 0.01 < * < 0.05 < . < 0.1 < ° <1. the pakistani sorts produce smaller grains but are noted for resisting lodging, pest, and diseases, and for their good baking quality (this was not tested but based on the sort characteristics). local wheats lack these strengths, so the pakistani sorts are useful breeding materials and can be included in the hybridization process with local varieties. 5. conclusion comparative analysis of productivity of foreign wheat sorts with local varieties in gbao revealed important insights into the suitability of different sorts for the high mountain region. the research aimed to identify the ideal wheat cultivar that best fits local conditions and contributes to food security in this challenging region. the findings revealed that local varieties had certain advantages over foreign sorts. the local varieties showed better performance in terms of crop height, number of spikelets on the main spike, length of the main spike, and number of grains in the main spike. additionally local cultivars had longer stems and more tillers, which are valuable traits for local farmers who use wheat straw as cattle fodder during the long and harsh winter months in high mountain areas. however, foreign wheat sorts also exhibited favorable characteristics, such as higher number of grains per spikelet, higher weight of 1000 grains, and early maturation. these traits are important and could be beneficial for withstanding the challenging local environment where the growing season is short and unpredictable weather conditions occur; particularly rains in september may make cereal crops susceptible to diseases. early maturation of wheat sorts is desirable in such conditions to avoid the impact of september rainfall; some of the foreign wheat sorts demonstrated this trait. local varieties have longer periods of maturity indicating a longer adaptation period to the local conditions. on the other hand, foreign sorts mature somewhat earlier, though not significantly different from the local varieties. the productivity attributes of local wheat varieties are mainly associated with productive tillering, spike length, number of spikelets, and number of grains in the main spike. in contrast, the productivity attributes of foreign sorts focus on weight of 1000 grains and longer heading to ripening periods for grain productivity. a separate comparison was made with vernalized winter wheat sorts from pakistan during spring sowing under local conditions. the pakistani sorts were generally shorter with lower general and productive tillering and shorter main spikes compared to local variety bludon. however, some pakistani sorts have higher numbers of grains in the main spike and better resistant to lodging, pests, and diseases, making them potential candidates for hybridization with local varieties. in conclusion, the study shows that local wheat varieties in gbao have specific advantages, especially in terms of their adaptability to local stresses and their importance for providing cattle fodder during prolonged harsh winters. however, introduced wheat sorts also have attributes, including higher grain yield, early maturation, and resistance to lodging, pests and diseases. the results suggest that a combination of local and foreign wheat sorts through hybridization could lead to the development of ideal wheat varieties that can thrive in these unique environmental conditions and contribute to food security. further research and breeding efforts should focus on selecting and developing cultivars with the most desirable traits to address the specific challenges of wheat cultivation in this high mountain region. in this study, the genotype 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[42] h. g. gauch jr and r. w. zobel, "identifying mega‐environments and targeting genotypes," crop science, vol. 37, no. 2, pp. 311-326, 1997. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1002/9780470015902.a0003691.pub2 https://www.fao.org/faostat/en/#data https://www.fao.org/3/y4011e/y4011e06.htm http://www.cawater-info.net/books/agro-tajik-2/pages/019.htm http://www.cawater-info.net/books/agro-tajik-2/pages/019.htm 76 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 76-86, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6168 © 2024 by the authors; licensee asian online journal publishing group physical and sensory quality of the fine-flavored and aromatic cocoas growing in different venezuelan regions alvarez, climaco1 perez, elevina2 silva, giovanni3 pavani, angelica4 fernandez, hadith5 ( corresponding author) 1national institute of agricultural research, miranda, venezuela. email: climacoa@hotmail.com 2institute of food science and technology, faculty of sciences, central university of venezuela, caracas, venezuela. email: elevina07@gmail.com 3venezuelan socialist cocoa corporation, caracas, venezuela. email: silvageovannyingucv@gmail.com 4cacao san francisco, cacao share, venezuela. email: angelicapavani@gmail.com 5national institute of agricultural research, aragua, venezuela. email: hadidgizeh@gmail.com abstract this work aimed to determine the cocoa's physical and sensory quality in different venezuelan regions to use in specific flavored chocolate production. 49 samples of cocoa beans gathered in plantations from different regions of venezuela were evaluated using methodologies described to analyze cocoa physicochemical standards. a trained panel selected twenty samples from this lot as standards of fine-flavored cacao. eleven samples with the best sensory attributes of each region were selected from the lot of twenty. these eleven samples showed variability in the physical and physicochemical attributes. the analysis of variance (anova) showed significant differences between aroma and taste attributes in the liquors evaluated. the principal component analysis (pca) expresses the variability of the eleven samples studied for the intensity of the sensory qualities of basic and complementary tastes (54.1%), and aroma (50.4%). besides, the fermentation percent and the external and internal bean characteristics through typification and cut test analysis relevant for its commercialization, the study provides practical information on the sensory attributes that characterize venezuelan fine-flavored cocoas from several regions that are used in the production of chocolate with regional-specific flavors. keywords: attributes, cocoa, fermented, flavor, sensory profiles. citation | climaco, a., elevina, p., giovanni, s., angelica, p., & hadith, f. (2024). physical and sensory quality of the fine-flavored and aromatic cocoas growing in different venezuelan regions. agriculture and food sciences research, 11(2), 76–86. 10.20448/aesr.v11i2.6168 history: received: 14 october 2024 revised: 15 november 2024 accepted: 20 november 2024 published: 29 november 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the national institute of agricultural research, miranda-venezuela, fondo nacional de ciencia y tecnología venezuela (grant number: 201100565) and venezuelan socialist cocoa corporation, caracas-venezuela (grant number: 201100313 2021). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: collected the data, a.c.; contributed data or analysis tools, a.c., p.e., p.a. and s.g.; performed the analysis, a.c., p.e. and p.a.; wrote the paper, a.c. and p.e. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 77 2. materials and methods ................................................................................................................................................................... 77 3. results and discussion ................................................................................................................................................................... 79 4. general conclusions ........................................................................................................................................................................ 86 references .............................................................................................................................................................................................. 86 mailto:climacoa@hotmail.com mailto:elevina07@gmail.com mailto:silvageovannyingucv@gmail.com mailto:angelicapavani@gmail.com mailto:hadidgizeh@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6168 https://orcid.org/0000-0002-0563-9984 https://orcid.org/0000-0001-9788-4946 https://orcid.org/0009-0006-2644-5536 https://orcid.org/0000-0001-6879-5493 https://orcid.org/0000-0001-6313-6413 agriculture and food sciences research, 2024, 11(2): 76-86 77 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the research provides unpublished information on the physical, physicochemical, and sensory characteristics of fine-flavored cocoa beans growing in venezuelan plantations, that are useful for its commercialization and regional flavored-specific chocolate production. 1. introduction good postharvest practices are described as a set of activities that start from the classification and selection of the fruits harvested in their optimal maturation state in the corresponding time. subsequently, the fruit is broken or shelled, the fresh seeds are extracted, followed by the fermentation process under the conditions of the production unit. once the fermentation is finished, the drying, selection and cleaning, packaging, and transport of the dry cocoa bean continue [1]. pérez and silva [2] have defined the post-harvest benefit of cocoa, as any operation to which the agricultural product (cocoa seed) must be subjected to convert it, into a marketable article (beans and primary products). the products obtained must meet physical-chemical, sensory, nutritional, and safety quality criteria, which are determinants in their final acceptance by the consumer [3]. determining the attributes and quality of cocoa and derivatives must be accorded with the export standards and regulations described. usually, these standards and rules depend on the post-harvest and transformation technology used, with a marked influence on the region of origin and type of genetic material [4]. venezuela is characterized by the diversity of cocoa plantations that produce beans classified as "fine flavor and aroma cocoa" with marked genotypic and phenotypic variations depending on the region [5]. cocoa variety and the local harvesting practices (especially the selection of the pods) can greatly affect the quality of the fermented cocoa beans [6, 7]. fermentation constitutes a transcendental stage in the processing or post-harvest benefit of cocoa beans. the characteristic aroma and flavor precursors of chocolate formed during fermentation are enhanced during roasting, causing variations in the chemical composition of the bean. during the process, physical and chemical changes occur in the cocoa beans due to biochemical reactions caused by microbial activity under aerobic and anaerobic conditions in an ecological or introversion succession [2]. this activity induces the development of those precursor chemical compounds that will continue to form during the drying and roasting of the beans and will be responsible for the final flavor of the chocolate. the natural drying stage in the sun completes the oxidative process started during fermentation; playing an important role in reducing the astringency, bitterness, and acidity of the beans. the beans develop a brown coloration, which is the product of the condensation reactions between quinones and proteins after the enzymatic oxidation of the polyphenols (hashim et al., 1999 cited by pérez and silva [2]). hence, the importance of generating the fermentation and drying protocols in the regions, due to the existing diversity between them. subsequent operations after drying before roasting will guarantee a more homogeneous batch of beans according to size and reduce microbiological risks by following in accordance with the demands of the international market. determining the physical characteristics of cocoa will allow it to be typified for a certain market, based on the uniformity in the bean's size, fermentation index, and the number of observable external defects and atypical flavors presented [5]. the classification of the commercial quality of cocoa is primarily due to the physical and aromatic composition of the bean, with the genotype and fermentation being the factors with the highest incidence in the flavor, smell, and color of the end product and the formation of flavor precursors in the chocolate [2, 5, 8]. the objective of the following research was to determine the physical and sensory characteristics of cocoa beans from production units in different venezuelan regions with postharvest protocols that define the chocolate quality. 2. materials and methods to recognize the work of cocoa farmers, it was necessary to organize the national cocoa excellence competition, venezuela 2023 edition, within the framework of the event "international cocoa and chocolate, 2023 (cacao of excellence program) by biodiversity international. a program of training and information activities was established (october-december 2022), directed by a national organization committee (noc) managed by the presidency of the venezuelan socialist cocoa corporation (vscc), national institute of agricultural research(inia), and entities attached to the mppat (ministry of popular power for agricultural production and lands), private entities, and universities (central university of venezuela). 2.1. technical training and call at the national level the period of sample selection was carried out from november to december 2022 by the territorial’s ship, field technicians, and coordinators of the venezuelan socialist cocoa corporation (vscc). these officials, after receiving instruction based on the national competition (online) would be responsible for selecting the best samples fermented and dried cocoa by region and farmer. for the selection and conditioning of the samples, the "technical manual for the selection and sampling of cocoa for quality purposes" directed by a national organization committee (noc) was considered. national training workshops were done so that cocoa producers knew the important parameters for the national bean cocoa quality. 2.2. raw material once the training process for the producers was completed, they were asked for a 10 kg sample of fermented and dried cocoa beans from the november-december 2022 harvest. the manual indicated the procedure for transporting and receiving the sample. each sample received was randomly assigned a five-digit code, which would guarantee the anonymity of the sample throughout the process of evaluation and selection of the best samples for sensory quality. a total of 45 samples were received from the following states: aragua, amazonas, apure, barinas, bolívar, carabobo, yaracuy, portuguesa, la guaira, miranda, monagas, mérida, and sucre. agriculture and food sciences research, 2024, 11(2): 76-86 78 © 2024 by the authors; licensee asian online journal publishing group the samples were stored in adequate conditions to avoid contamination for posterior analysis. from the total population of bean samples, a representative number that met the physical and sensory quality requirements demanded by the norms or standards was preselected as follows: 2.3. physical quality characteristics the protocol of the external analysis beans, dimensions and weight average, the number of beans/100 g sample, and the cut test quality were carried out according to the manual of international standards for the assessment of cocoa quality and flavour [9]; comisión venezolana de normas industriales [10]; comisión venezolana de normas industriales [11] and comisión venezolana de normas industriales [11] in its second revision. 2.4. sensory evaluation of cocoa pastes or liquors the sensory characterization (taste and aroma) was carried out to know the sensory profiles of the materials evaluated. the characterization sessions for sensory profiles were carried out at the sensory quality control laboratory of the venezuelan company intercacao, located in caracas. the protocol used for roasting the beans cocoa, and refining nibs into cocoa liquor was carried out according to international standards for the assessment of cocoa quality and flavour (iscqf) [12] available on website cocoa guide cocoa of excellence. the group of panelists participating as sensory assessors of the cocoa´s pasta was made up of eight (8) trained chocolatiers, who were calibrated for the sensory profile’s characterization of the pasta according to criteria defined in the international standard organization [13] and international standards for the assessment of cocoa quality and flavour (iscqf) [12]. 2.5. recruitment of the sensory assessors the judges (8) were selected from several applications based on an exhaustive review of their curricular synthesis and subsequently submitted to selection according to the criteria defined in international standards for the evaluation of cocoa quality and flavor [12]. the selection of the sensory assessors was carried out in december (2022). a survey was carried out requesting information regarding the age, interest, health, feeding schedules, and availability of the candidates recruited. according to their answers in the survey in the basic taste detection test and odor recognition based on standards international standard organization [13] the members of the group were selected to participate in the evaluation of sensory profiles of cocoa´s pastes. the selected candidate's group had previous experience in the sensory evaluation of food and had been trained in cocoa and chocolate-tasting tests, and they were previously informed about the objectives of the study and the initial considerations for carrying out the training. 2.6. application of the umbral tests of basic tastes and odor detection during the training, the selected judges were instructed on the aspects of sensory physiology and sensory evaluation as a quality control tool for cocoa and chocolate. they were instructed on the aspects and considerations of the lexicon and notes (attributes and sub-attributes) used in cocoa and chocolate. the tests for the detection of basic tastes and odors were carried out on the evaluators to verify their training using the international standard organization [13] standard. the evaluating jury used the forms or formats designed for the training and calibration phases. 2.7. training and calibration the selected judges were trained for six days, and they were to a calibration following the international regulations for the evaluation of the quality and flavor of cocoa [12]. the training was carried out by qualified personnel in the food technology area. the training and calibration activity was gone in december 2023 and january 2024. the calibrations were carried out using different pastes or liquors of different origins obtained from the production units participating in the national contest through the use of the triangle test. 2.8. triangle test triangle taste test is a discriminative method commonly used in sensory research and recommended by international standard organization [13]. the test requires a minimum of at least five participants to state if there is a significant difference. the panel's sensory assessors were presented with three samples, of which one was different and two similar. the two similar samples are made of the same raw material, the same protocol of elaboration, and the same origin. this discriminative test allowed the jury to differentiate between 3 samples, that is, it consisted of simultaneously presenting them with three coded samples, of which two were the same and one was different. the jury had to identify the sample with a different flavor. 2.9. sensory evaluation sections the trained and calibrated judges were summoned to appear at the sensory evaluation laboratory of the inter cacao company, in several sessions to preselect the best samples (11) of cocoa paste or liquor that would go on to the final evaluation by using the aroma and taste profile test. 2.10. final evaluation of taste and aroma profiles: the grand jury (sensory assessors) the judges (8) that made up the final great jury were called to meetings for the sensory evaluation of the best attributes of the preselected cocoa pastes. liquor samples were evaluated, performing three repetitions for each one. the judges panel assigned scores according to the intensity of each attribute on a scale of 0 to 10, with 0 being the absence of the attribute and 10 being the maximum intensity. the judges received a format for the taste and aroma evaluation of cocoa paste based on the attributes and sub-attributes established by the international standards for the assessment of cocoa quality and flavour (iscqf) [12]. thirteen (13) taste attributes for cocoa paste were evaluated: cocoa, chocolate, acid, sweet, astringent, bitter, fruity, nutty, floral, wood, spicy, smoky, and agriculture and food sciences research, 2024, 11(2): 76-86 79 © 2024 by the authors; licensee asian online journal publishing group metallic. twelve (12) aroma descriptors as such as cocoa, chocolate, acid, fruity, nutty, floral, woody, spicy, smoky, metallic, putrid, and moldy were also evaluated. the average values of the ratings assigned by the panelists were presented in radial graphs called "spider webs". 2.11. statistical analysis pearson's linear correlation coefficient was applied for the characteristics of the length, width, and thickness of the cocoa bean samples. a descriptive analysis of the physical characteristics of the quality bean was applied through the cut-test using bar graphs to compare the means between samples according to their attributes. the taste and aroma descriptors were evaluated by a principal component analysis (pca) carried out using the di rienzo [14] student statistical software version. mean flavor profiles were calculated. the comparison of means of the cocoa pastes (anova) was obtained using the tukey test (p≥0.05) according to panelists-attributes interaction. the anova test was carried out with infostat, statistical software updated to version 2020. the graphical representation (radials charts) was carried out in microsoft excel 2010. 3. results and discussion table 1 shows the origin of the eleven (11) cocoa bean samples that presented the best physical and sensory characteristics. table 1. origin of cocoa samples from different regions (states) of venezuela. n º sample 1 2 3 4 5 6 7 8 9 10 11 origin (regions) apure barinas bolívar bolívar yaracuy yaracuy miranda miranda monagas carabobo *clone assigned code 65127 48747 78882 19257 97825 91197 17232 76192 50694 66010 54184 note: *clone reference. 3.1. physical quality characteristics: average dimension beans table 2 summarizes the physical quality requirements demanded for the national and international regulations of the eleven (11) samples from the total selected population. the table indicates the origin place, the random code assigned, and the correlations between the average dimensions (length, width, and thickness) of the cocoa samples selected. strongly positive correlations (0.50-1.00) between length and width beans were observed for pastes from apure (65127), bolívar (78882), carabobo (66010), yaracuy (91197) states, and reference clone (54184). weak or negative correlations for the rest of the cocoa samples were observed. the reflected positive correlation between the length and width of the dry bean is related to the swelling index during the fermentation process. the variable nature of the cocoa lots allows us to infer that there is a high presence of beans with different shapes (oblong, elliptical, or ovoid), which are the product of the existence of different cocoa materials in the plantations, in this case, with the presence of hybrids called trinitarians. no correlation was found between the length and thickness and vice versa, of the cocoa beans, which indicates that there is no effect of the fermentation and drying process on these variables. the results show that the mixtures of beans with variable and intermediate shapes in width and thickness do not reach the swelling process of the seeds during fermentation. when dealing with batches from materials with a homogeneous shape, there is a greater possibility of a positive and significant correlation, such is the case of criollo-type cocoas. 3.2. cut-test, humidity content (%), individual weight and weigh by 100 dry beans (averages), and dry beans counting cut test are an effective way to check for internal defects, and evaluate the level of fermentation, and the health standards in a batch of cacao beans. the cut test is carried out to identify, quantify, and separate broken beans, slaty, moldy, infested by insects, violet or purple beans, partially fermented (partially purple), unfermented (purple and smooth), germinated beans, and well-fermented brown beans [1, 4, 5, 15] . figure 1 shows of important physical attributes of the eleven (11) samples tested. the critical defects referred to the presence of molds, insect infestations, impurities, and black, flat, and double grains, complied with the standards established by comisión venezolana de normas industriales [10] and comisión venezolana de normas industriales [11] and international standards for the assessment of cocoa quality and flavour [9]. table 2. pearson´s lineal correlation between the length, width and thickness of the cocoa beans. average dimensions origin length (cm) width (cm) thickness (cm) apure state (65127) length (cm) 1.00 0.51 0.25 width (cm) 0.51 1.00 -0.15 thickness (cm) 0.25 -0.15 1.00 barinas state (48747) length (cm) 1.00 0.48 0.02 width (cm) 0.60 1.00 0.12 thickness (cm) 0.02 0.35 1.00 bolívar state (78882) length (cm) 1.00 0.60 0.05 width (cm) 0.60 1.00 0.12 thickness (cm) 0.05 0.12 1.00 bolívar state (19257) length (cm) 1.00 0.38 -0.03 width (cm) 0.38 1.00 -0.10 agriculture and food sciences research, 2024, 11(2): 76-86 80 © 2024 by the authors; licensee asian online journal publishing group average dimensions origin length (cm) width (cm) thickness (cm) thickness (cm) -0.03 -0.10 1.00 carabobo state (66010) length (cm) 1.00 0.64 0.16 width (cm) 0.64 1.00 -0.13 thickness (cm) 0.16 -0.13 1.00 average dimensions origin length (cm) width (cm) thickness (cm) miranda state (17232) length (cm) 1.00 0.28 0.06 width (cm) 0.28 1.00 0.35 thickness (cm) 0.06 0.35 1.00 miranda state (76192) length (cm) 1.00 -0.17 -0.16 width (cm) -0.17 1.00 -0.32 thickness (cm) -0.16 -0.32 1.00 monagas state (50694) length (cm) 1.00 0.45 -0.23 width (cm) 0.45 1.00 0.16 thickness (cm) -0.23 0.16 1.00 yaracuy state (97825) length (cm) 1.00 -0.23 -0.06 width (cm) -0.06 1.00 -0.18 thickness (cm) -0.06 -0.18 1.00 yaracuy state (91197) length (cm) 1.00 0.70 0.03 width (cm) 0.70 1.00 0.06 thickness (cm) 0.03 0.06 1.00 reference clon (54184) length (cm) 1.00 0.60 -0.15 width (cm) 0.60 1.00 0.17 thickness (cm) -0.15 0.17 1.00 the moisture content varied in a range of 6.0±0.0 [97825-yaracuy] to 7.5±0.1% (50694-monagas), values that conform to those allowed by the national and international standards by fermented and dry cocoa. the moisture values show the good drying practices used by the producers to obtain dry and microbiologically stable beans (low microbial growth). none of the samples showed moisture values above 8.0 %, which represents a critical value to increase the growth of molds that would cause the deterioration of the coca commercial quality bean [9]. during the inspection phase of the selected samples, an adequate selection and cleaning was observed, registering a low percentage of broken, flat, and multiple beans, which is indicative of good postharvest practices (figure 1). the internal surface of the cotyledons exposed by the longitudinal cut (cut test), allowed them to be classified according to the degree of fermentation into well-fermented beans, partially fermented beans, purple, and slaty beans categorized as non-fermented cocoa [5, 10, 11]. figure 1. physical quality characteristics were obtained through the cut test. agriculture and food sciences research, 2024, 11(2): 76-86 81 © 2024 by the authors; licensee asian online journal publishing group 3.3. insufficiently fermented beans and violet or purple color dried cotyledons insufficiently fermented showed compact surfaces of purple color (light and dark) depending on the type of cocoa. these violet-colored beans result from a minimal fermentation process depending on the fruit's maturity, making the release and oxidation of polyphenolic (taste-responsible) compounds impossible due to poor fermentation. 65127-apure and 66010-carabobo samples presented low percentages of insufficiently fermented beans, with 2.0±1.0% and 3.00±0.6% respectively. apure sample presented a lighter brown tone, representing a characteristic of criollo beans, while the rest consisted of a hybrid or trinitarian-type cocoa population. those with dark breaking usually are trinitarian (criollo moderno). the 97825-yaracuy sample did not show intense purple beans; however, 65127-apure and 50694-monagas samples presented lower and similar values to each other (figure 1). the fermentation method (fermenter type), process time, intervals in the removal of the cocoa mass, and environmental conditions, soil type, are determining factors so that fresh seeds do not reach complete fermentation [5, 8]. these characteristics were grouped by the international standards for the assessment of cocoa quality and flavour [9] into 4 categories according to the internal color of the cotyledons: well fermented (dark to light brown), partially purple or violet, totally purple or violet, and slaty beans. all the analyzed samples showed levels of slaty grains within the limits established by comisión venezolana de normas industriales [10] and comisión venezolana de normas industriales [11]. the results explain the good selection of the fruits in their physiological maturity state carried out by venezuelan cocoa producers. 3.4. brown colored beans (fermented): fermentation index (%) figure 1 shows that the fermentation index varied between 75% and 90% in the samples, which corresponds to the limit’s standard by comisión venezolana de normas industriales [11] for fermented and dry beans. first-class fine cocoa (f1≥74%) was based on samples that exhibited a percentage of 75.0±1.0% (48747-barinas), 78.0±2.0% (17232-miranda), 79.0±2.1% (76192)-miranda, 80.0±1.0% (50694-monagas), and 80.0±2.1% (19257-bolívar) the class extra fine cocoa corresponded from two (2) yaracuy samples: 97825-(82.0±2.5%), and 91197 with 83.0±1.0%. fermented and dry cocoa beans from 65127-apure, 66010-carabobo, and the reference clone (54184) showed fermentation high indices of 90.0±2.0%, 85.0±1.2%, and 80.0% respectively. deep and well-defined grooves and striations inside were present in the brown cotyledons, and their fragile shell was easily detachable, all signs of a well-fermented bean. at the start of the fermentation process, the fresh seed has a coloration that is between light to dark purple and/or white. the cocoa's color will change from light dark brown or reddish at the end of the process [4]. according to the international standards for the assessment of cocoa quality and flavour [9] external and internal evaluations take into account the level of fermentation, the color of cotyledons, and beans with internal infections (molds), and infestation (insects). figure 1 shows that fermentation values correspond to the limit’s standard of comisión venezolana de normas industriales [11] for fully-fermented and dry beans. the color changes that occur inside the grain are due to the oxidation reactions of the anthocyanin compounds, which form brown pigments color. this pigment has a significant on reducing the astringent of the fermented bean (guzman, 2007; gonzález et al., 2012 cited pérez and silva [2]). 3.5. weight of 100 g dry beans, and the count per 100 grams of sample the weight of 100 dry cocoa beans varied in a range from 120.3±1.5 g to 183.0±1.0 g. the highest value was for the 54184-reference clone (183.0±1.0g) followed by the 65127-apure sample (179.1±2.7 g). these values are not in accordance with the commercial cocoas established by comisión venezolana de normas industriales [11] for the f1 class. the weights are equivalent to cocoa that is commercially classified as extra-fine, but the final classification is determined by quality defects and the fermentation index. the amount of edible and usable fat in the bean would be determined by the weight parameter, taking into account the butter, husk, and cotyledon content. the variability of the number of beans/100 g sample varied from 59.00±1.15 to 83.00±1.53 beans/100g. the lowest value in the bean count/100 grams of sample was represented by 65127-apure (59.0±1.15), 19257-bolívar (60.0±2.08), and the 54184-reference clone (62.0±1.0). these characteristics classify them as large beans, categorizing them as a standard bean, with desirable and acceptable sizes for international trade [3]. the individual weight of the dry bean varied from 1.21±0.12 g to 1.80±0,37g, with 65127-apure dry bean being the heaviest (1.80±0.37g), followed by 19257-bolívar (1.76±0.70g) and 50694-monagas (1.60±0.41g). in this context, cocoa beans from venezuela that are produced for commercial purposes have a weight above 1.2 g [16] . this value is considered as a reference for the international market. according to the previous authors, a bean index above 1.2 g is believed to guarantee a higher yield per hectare and is favored by marketers and industrialists. the physical parameters reveal differences that indicate the presence of microbiota or bacterial microflora in the fermentation. the fermentation process could also vary depending on the composition of the pulp (sugar content), moisture content, and the genetics of the cocoa. during the benefit process, the environmental conditions and soil characteristics influence the changes that occur in the cotyledons [8]. therefore, it is important to carry out fermentation protocols that classify cocoa types and establish maximum yields according to their quality. 3.6. results of training the evaluating jury in basic taste tests a group of four (4) women and four men aged 30 and 40 made up the panel of evaluators from the beginning of the study. they received training following international standards described in the methodology. table 3 shows the results obtained (%) by the evaluators during the odor identification and basic taste detection tests. agriculture and food sciences research, 2024, 11(2): 76-86 82 © 2024 by the authors; licensee asian online journal publishing group table 3. results of odor identification and basic taste detection tests, expressed in percentages of right guess. tests (%) *trained panel (jury) *e1 *e2 *e3 *e4 *e5 *e6 *e7 *e8 % total correct answers/group smells 90 92 89 90 88 98 90 90 91.0 basic tastes 100 100 100 80 100 100 100 100 98.0 percentage of correct answers/ jury 95 96 95 85 94 99 95 95 94.0 note: *jury code. the trained panel's evaluation jury reported a mean score of 94% for the two tests conducted (smell and flavor). the lowest value was 85% and the highest was 99% (table 3). the group of evaluators is capable of identifying and distinguishing between various odor and taste substances at concentrations that are set by international standard organization [13] standards. during the training process, 100% of the panelists were able to identify the various cocoa paste samples through the discriminative test (triangle test differentiation). 3.7. variance analysis (anova). panelists · attributes (aroma/taste) interaction table 4 shows the global averages of the intensities reported by the evaluation jury of seven (7) sensory attributes for aroma and ten (10) for taste attributes. the intensity values showed statistically significant differences between the aroma and taste attributes data from cocoa pastes. the results also indicated that there were no significance moldy, putrid, smoky, and metallic defects (p > 0.05). according to the tukey test, there is no significant difference in the cocoa and chocolate attributes between these pastes (p > 0.05). the magnitude of the significance values for the remaining flavor (aroma/taste) attributes are presented in figure 2. the average value for the bitterness and astringent taste remained at intermediate intensities, contributing to the flavor characteristics of the cocoa paste. the figure 2 shows a moderate bitterness and astringency intensity notes, and low floral, nutty, and wood notes. the intensity of the sweet taste was low; however, the acidity of the pastes is influenced by the fruity attribute as a characteristic descriptor of trinitarian cocoas. the commercial cataloging of fine or flavored cocoa in trinitarian hybrid clones reveals similar results from sukha, et al. [17]. the flavor profiles for these local clones revealed that they consistently had higher fruity and acid notes. astringency and bitterness did not vary significantly between the local clones. table 4. differences in the global means of aroma and taste attributes were observed using tukey test. aroma attributes *global means tukey test (p > 0.05) taste attributes *global means tukey test (p > 0.05) cocoa 7.25 a cocoa 7.58 a chocolate 5.41 b fruity 5.07 b fruity 4.44 c chocolate 4.91 c acidity 3.46 d acidity 4.52 d wood 3.17 e bitter 3.92 e nutty 2.62 f astringent 3.46 f floral 2.53 f nutty 3.3 g smoked 0.33 g wood 3.05 h spicy 0.32 g floral 2.47 i moldy 0.26 gh sweet 2.08 j putrid 0.14 hi smoked 0.46 k metallic 0.07 i spicy 0.34 k metallic 0.02 l note: *means with a common letter are not significantly different (p > 0.05). this study found that pastas from different venezuelan regions had significant differences, with genetics and post-harvest handling being the main causes of the variability. the bitter taste of the liqueurs showed acceptable average values, which is an indicator of a higher quality of the liqueur. this attribute has an inverse relationship with the quality of the final product. the greater the bitterness, the lower the organoleptic quality of the paste. besides, one of the reasons why the cocoa processing process is carried out is precisely to minimize the levels of bitterness in the beans, which is related to the content of purines (caffeine and theobromine) [18] . figure 2. global means for the different aromas and tastes. note: means with the same letter are not significantly different (p > 0.05). agriculture and food sciences research, 2024, 11(2): 76-86 83 © 2024 by the authors; licensee asian online journal publishing group figure 3-a revealed the statistically significant differences between the aroma intensities of eight (8) liquors evaluated. these liquors or cocoa pastes presented the best attributes or strong notes of chocolate, fruity and cocoa with an intermediate acidity. the intermediate notes for the nutty attribute only corresponded to the liquors from (7)-miranda (17232), (3)-bolívar (78882), and (2)-barinas (48747) states. the barinas liqueur was characterized by intense chocolate and fruity notes (aromas) and a strong cocoa flavor balanced with a soft nutty aroma. the liquor from (1)-apure state (65127) obtained the best notes of chocolate, fruit, and cocoa in aroma and taste, balanced with low bitterness and sweetness figure 3-b. the evaluation jury designated the intensities of acidity and astringency for this liquor, in intermediate values of 5.00 points with low astringency of 3.30 points on average. in summary, the taste and aroma attributes are statistically significant between the liquors from different areas evaluated (p > 0.05). figure 3. cocoa liqueurs or pastes with the best aroma (a) and taste (b) attributes: 1-(65127-apure), 2(48747-barinas), 3-(78882-bolívar), 4-(19257-bolívar), 5-(97825-yaracuy), 6-(91917-yaracuy), 7-(17232miranda), 9-(50694-monagas), 10-(66010-carabobo), and 11-(54184-reference clone). 3.8. principal component analysis for taste and aroma attributes of cocoa paste (liquors) samples principal component analysis (pca) is a statistical tool that determines the most important taste and aroma attributes that define the profiles of cocoa paste samples, and how they correlate with each other. pca compresses the data based on similarities and differences by reducing dimensions without much loss of information and defining the number of principal components (pcs) [19, 20] . the pca showed the variability of the eleven (11) cocoa materials under study concerning the intensity of the taste attributes, with a 54.1% variation value of the two components (cp1, cp2), as seen in figure 4. figure 4 shows three (3) large groups differentiated mainly by the basic and complementary intensities tastes. the first group (i) conformed by 76192, 50694, and 54184 liqueur, they revealed a greater intensity in the basic tastes of astringency, bitterness, and low intensity in acidity, combined with soft woody notes. the 48747, 19257, 65127, and 66010 codified liquors samples from the second group (ii) showed an intermediate balance in the complementary attributes of fruit, cocoa, and chocolate taste. the liqueur 97825 of group iii presented attributes in the sweet, floral, and nutty flavors combined with very low spicy notes, while 91197 liqueur showed the lowest notes in the previous attributes. the metallic taste was not detected in the liquor 78882; the value of this atypical flavor is not exceeding 2 points. agriculture and food sciences research, 2024, 11(2): 76-86 84 © 2024 by the authors; licensee asian online journal publishing group figure 4. principal component analysis for taste attributes on eleven (11) selected liquors of cocoa. figure 5. principal component analysis for aroma attributes on eleven (11) selected liquors of cocoa. the value of the variability observed in the aroma was 50.4% in the two components of the pca (figure 5). liqueur 1732 from the first group (i) was characterized by an intermediate balance of some more intense aroma attributes (spicy and fruity). liquors 76192, 78882, 97825, and 91197 showed very low values in the intensity of moldy and smoky odors. liqueur 19257 from the second group (ii) showed a notable intensity in the floral aroma agriculture and food sciences research, 2024, 11(2): 76-86 85 © 2024 by the authors; licensee asian online journal publishing group but very low in this attribute in liquors 65127, 78882, and 76192 respectively. liqueur 54184 corresponding to the reference clone showed significant separation within the second group as special attributes liqueur (figure 5). the liquors 66010 and 48747 of the third group (iii) were characterized by presenting strong intensities of cocoa and chocolate aromas combined with high acidity and low fruity and woody notes. while liquors 50694 and 54184 showed very soft notes of chocolate and fruit. 3.9. taste profiles of the eleven cocoa pastes selected (liquors) the intensity values of basic and complementary taste attributes were averaged. the intensity scores (0-10) of the flavor attributes were obtained by the average of each evaluating jury in the sessions. in the basic tastes (cocoa, bitterness, astringency, sweetness, and acidity) a score from 5 to 6 was obtained, in the scale range of 0 to 10 points with an intermediate intensity rating in most of the samples (figure 6). similar results were obtained by rodríguez-silva, et al. [21] on colombian genotypes classified as fine cocoa with flavor and aroma. the dried and fermented cocoa beans, after the roasting showed strong cocoa and chocolate notes. the strong notes of cocoa and chocolate were: 65127-apure, 48747-barinas, 91197-yaracuy, 66010-carabobo, and 54184-clon reference as shown in figure 6 (a, b, f, j, k). the maximum strong cocoa and chocolate intensity (9 points) corresponded to the 19257-bolívar liqueur. figure 6, it was also observed that there is a balanced combination of bitterness, fruity, acidity, and astringency notes in all samples, which can be inferred that the producers applied good postharvest practices (harvest, fermentation, and drying). defect intensity or atypical tastes were very low and did not represent a critical value or risk for the final product. the intensity value in the smoked taste (atypical flavor) ranged from 1 to 2 points, corresponding to 2 samples from the miranda state (17232 and 76192), monagas (50694), and the reference clone (54184). according to the international standards for the assessment of cocoa quality and flavour (iscqf) [12], the previous scores (1-2) are considered low intensity to be categorized as a critical risk. the moldy and metallic notes were absent in the cocoa paste which are considered atypical flavor defects. figure 4 (c-d, e-f, g, h) observed that samples processed in the same state (bolívar, yaracuy, and miranda states) had profiles with different flavor intensities, demonstrating that cocoa type, traditional post-harvest technology, and the agronomic-ecological conditions local have a direct effect on these profiles. the nutty and fruity notes considered complementary attributes were quantified with a maximum value of 6 points. the highest fruity notes (6 points) corresponded to liquors from 65127-apure, 19257-bolívar, 97825yaracuy, 76192-miranda, and 66010-carabobo. the floral notes were very low (2 to 4 points), being the highest for the 97825-yaracuy liqueur. spice notes were perceived with low intensity in all samples. the wood attribute ranged from 3 to 4 points in intensity with intermediate tenors (4 points maximum) for 65127-apure, 48747barinas, 97825yaracuy, and two (2) liquors from miranda state (17232 and 76192). agriculture and food sciences research, 2024, 11(2): 76-86 86 © 2024 by the authors; licensee asian online journal publishing group figure 6. taste profiles of eleven (11) samples of cocoa pastes were selected that describe the basic and complementary attributes according to sensory analysis and origin (venezuelan states): (a) 65127-apure; (b) 48747-barinas; (c) 78882-bolívar; (d) 19257-bolívar; (e) 97825yaracuy; (f) 91197-yaracuy; (g) 17232-miranda; (h) 76192-miranda; (i); 50694-monagas; (j) 66010-carabobo; and (k) 54184-reference clone. 4. general conclusions the analyzed cocoa samples presented a characteristic sensory profile with different intensities, confirming the effect of harvest and postharvest handling in the formation of the chocolate precursors. it is concluded the values obtained in the physical quality and cut-test do not constitute the fingerprint to generate quality profiles in cocoa beans. the majority of producers had complied with good post-harvest practices in the cocoa bean samples, as a requirement for the presentation of excellent quality beans. finally, only eleven (11) samples were for the sensory analysis; since they met the physical quality requirements. selected samples were classified, according to national standards, mostly like “fine or extra-fine cocoa” with the lowest number of defects and atypical flavors. a trained taster's sensorial panel properly evaluated the sensory notes of the samples. the results of the statistical calculation produced by the responses of the panelists indicated that each type of cocoa presented a different sensory profile in its intensity classifying venezuelan cocoa as fine and extra-fine. references [1] t. j. gutiérrez and e. e. pérez, chapter 1. significant quality factors in the chocolate processing: cocoa post-harvest, and in its manufacture. in: chocolate: cocoa byproducts technology, rheology, styling and nutrition, ed. elevina pérez sira. nova publisher. https://novapublishers.com/shop/chocolate-cocoa-byproducts-technology-rheology-styling-and-nutrition/, 2014. [2] e. pérez and n. silva, fine aroma cocoa. in: pérez, e. cañas, i. (eds.), from cocoa to chocolate. a booming industry. madrid, spain: bellisco, 2017. [3] m. j. end and r. r. dand, "caobisco/eca/fcc cocoa beans: quality requirements of the chocolate and cocoa industry," retrieved: http://www.cocoaquality.eu. 2015. [4] c. álvarez, n. liconte, e. pérez, m. lares, and j. perozo, "review of physical, chemical and sensory attributes as indicators of the commercial quality of cocoa [electronic version]," petroglyphs. revista crítica transdisciplinaria, vol. 5, no. 1, pp. 12-25, 2022a. [5] c. álvarez, n. liconte, y. pereira, a. farías, i. buscema, and m. láres, "commercial quality profile of venezuelan cocoa (theobroma cacao l.)," novum scientarum, vol. 3, no. 7, pp. 21-32, 2018. [6] c. álvarez et al., "identification of aromatic compounds in venezuelan criollo cocoa using solid phase microextraction and gas chromatography," vitae, vol. 19, no. 1, pp. s370-s372, 2012. https://doi.org/10.1016/j.recqb.2015.05.003 [7] c. álvarez et al., "the use of near infrared spectroscopy to determine the fat, caffeine, theobromine and (−)-epicatechin contents in unfermented and sun-dried beans of criollo cocoa," journal of near infrared spectroscopy, vol. 20, no. 2, pp. 307-315, 2012. https://doi.org/10.1255/jnirs.990 [8] n. martínez, "evaluation of physical, chemical, organoleptic and yield components of universal and regional cocoa clones (theobroma cacao l.) in the production areas of santander, arauca and huila," master's thesis, national university of colombia, bogotá, 2016. 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[18] c. o. álvarez, e. e. pérez, and m. d. c. lares, "benefit of venezuelan criollo cocoa: variations in proximal composition, methylxanthines and polyphenols," rivar, vol. 9, no. 27, pp. 113-128, 2022. https://doi.org/10.35588/rivar.v9i27.5625 [19] e.-c. shin, b. d. craft, r. b. pegg, r. d. phillips, and r. r. eitenmiller, "chemometric approach to fatty acid profiles in runner-type peanut cultivars by principal component analysis (pca)," food chemistry, vol. 119, no. 3, pp. 1262-1270, 2010. https://doi.org/10.1016/j.foodchem.2009.07.058 [20] j. rodriguez-campos, h. escalona-buendía, i. orozco-avila, e. lugo-cervantes, and m. e. jaramillo-flores, "dynamics of volatile and non-volatile compounds in cocoa (theobroma cacao l.) during fermentation and drying processes using principal components analysis," food research international, vol. 44, no. 1, pp. 250-258, 2011. https://doi.org/10.1016/j.foodres.2010.10.028 [21] l. g. rodríguez-silva, l. f. quintana-fuentes, r. a. coronado-silva, a. garcía-jerez, e. y. báez-daza, and g. a. agudelo-castañeda, "physical and sensory characterization of 24 special cocoa genotypes theobroma cacao," revista udca actualidad & divulgación científica, vol. 26, no. 1, 2023. http://doi.org/10.31910/rudca.v26.n1.2023.2410 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://novapublishers.com/shop/chocolate-cocoa-byproducts-technology-rheology-styling-and-nutrition/ http://www.cocoaquality.eu/ https://doi.org/10.1016/j.recqb.2015.05.003 https://doi.org/10.1255/jnirs.990 https://www.google.com/search?client=firefox-b-d&-q=icsqf https://www.sencamer.gob.ve/ https://www.sencamer.gob.ve/ https://www.iso.org/obp/ui/#iso:std:iso:8586:ed-2:v1:en https://doi.org/10.35588/rivar.v9i27.5625 https://doi.org/10.1016/j.foodchem.2009.07.058 https://doi.org/10.1016/j.foodres.2010.10.028 http://doi.org/10.31910/rudca.v26.n1.2023.2410 203 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 203-221, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6253 © 2024 by the authors; licensee asian online journal publishing group nanotechnology: a double-edged sword for future smart agriculture and phytopathological management in plants dipan adhikari1 parvin khatun2 satyajit koley3 moutushi sen4 sudip kumar ghosh5 ( corresponding author) 1,2,3,4ug and pg department of botany, hooghly mohsin college, hooghly, west bengal (712 101), india. 1email: dipanadhikari@gmail.com 2email: parvin0khatun1996@gmail.com 3email: satyajitkoley87249@gmail.com 4email: moutushisen7@gmail.com 5department of economics, bidhannagar govt. college, sector-1, eb-, salt lake, kolkata, india. 5email: sudipkumarghosh919@gmail.com abstract modern technology that would boost agricultural outputs might therefore preserve the country's sustainable living standards by enhancing food security. with nanotechnology, it’s possible to produce foods of exceptional quality that also could increase the bioavailability of nutrients during the third decade of the twenty-first century which found usage in a variety of industries such as medical science, pharmaceuticals, food, and energy conservation. nanotechnology is the synthesis, designing, characterizing, and utilization of assemblies, tools, and systems via directing the morphology and size variation at nanometer level (1 100 nm). nanochemicals, nanopesticides, and nanofertilizers do increase yielding capacity without damaging agricultural land or irrigation water. nanomaterials (nms) are aimed protect crops from pests, microbial and fungal pathogens thereby lowering nutrient losses. nanotechnology holds the potential to monitor soil quality in agricultural fields and sense crop health. metallic nanoparticles (cu, zn, ni, zn fe, ag, al, ti, and al) can impinge on plant development, metabolism, and stress tolerance. this paper examines the role that nanoparticles (nps) be playing while regulating oxidative stress, ros turnover to mitigate abiotic stress in plants thereby emphasizing the advantages and of nanotechnology for better sustenance of future agriculture practices. keywords: agriculture, food security, nano chemicals, nanotoxicology and nano sensors, oxidative burst, and ros. citation | adhikari, d., khatun, p., koley, s., sen, m., & ghosh, s. k. (2024). nanotechnology: a double-edged sword for future smart agriculture and phytopathological management in plants. agriculture and food sciences research, 11(2), 203–221. 10.20448/aesr.v11i2.6253 history: received: 18 november 2024 revised: 20 december 2024 accepted: 23 december 2024 published: 26 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing grou funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 204 2. the world of nano-agrochemicals ........................................................................................................................................... 204 3. synthesis of nanoparticles ........................................................................................................................................................... 206 4. employment of nanoparticles designed for plant disease management ......................................................................... 207 5. plant disease control and management: applications of nanomaterials ......................................................................... 207 6. bio-efficacy of metallic nps against plant pathogens .......................................................................................................... 207 7. nanoparticles as “strategic weapons” against plant pathogens the mechanism of action ....................................... 210 8. cellular uptake and mechanism of nano-toxicity ................................................................................................................ 210 9. cellular toxicity via dna damage by nanoparticles ........................................................................................................... 211 10. nanomaterials and regulation of oxidative stress .............................................................................................................. 211 11. production and inter switching of reactive oxygen species in microbial cells .......................................................... 212 mailto:dipanadhikari@gmail.com mailto:parvin0khatun1996@gmail.com mailto:satyajitkoley87249@gmail.com mailto:moutushisen7@gmail.com mailto:sudipkumarghosh919@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6253 https://orcid.org/0000-0003-3126-7210 https://orcid.org/0009-0005-8664-4599 https://orcid.org/0009-0007-9339-9370 https://orcid.org/0009-0002-5339-1458 https://orcid.org/0009-0008-9459-1046 agriculture and food sciences research, 2024, 11(2): 203-221 204 © 2024 by the authors; licensee asian online journal publishing group 12. mechanisms of np uptake by higher plants ........................................................................................................................ 212 13. how nanomaterials (nms) would emerge as ‘nano-weapons’ against phytopathogens? ........................................... 213 14. environmental risk assessment of nps ................................................................................................................................. 214 15. bacteria and fungi play crucial roles in promoting plant growth through various mechanisms in natural environment ........................................................................................................................................................................................ 214 16. bacterial nano-toxicity by enps ............................................................................................................................................ 215 17. protozoal nano-toxicity out of enps accumulation ......................................................................................................... 215 18. nano-toxicity of fungi out of enps accumulation ............................................................................................................ 215 19. harmful effects of nanoparticles (nps) on earthworms .................................................................................................... 215 20. biochemical pathways leading to toxicity of soil microflora and macrofauna: oxidative stress and cellular death occurrence after enps exposure ....................................................................................................................................... 215 21. sensing and detection and plant pathogens in agricultural crops through nanomaterials (nms) ......................... 216 22. nanotechnology mediated detection of plant pathogens at early stages of infections ............................................. 216 23. challenges and limitations of nanosensors .......................................................................................................................... 216 24. conclusion and future perspectives for future smart agriculture ................................................................................. 217 references ............................................................................................................................................................................................ 218 contribution of this paper to the literature when applied to plant tissues, nanoparticles can be engineered to enhance resistance to pathogens by inducing the creation of defensive chemicals, which in turn stimulates a plant's inherent defense mechanisms. before being widely used, nanoparticles' potential harm to nontarget creatures, such as helpful bacteria and pollinators, needs to be carefully assessed. 1. introduction nanotechnology is the "synthesis, designing, characterizing, and utilizing of assemblies, tools, and systems" [1] through tailored morphology and size variation on the nanoscale from 1 to 100 nm. the one nanometre is defined as one billionth (10−9) of a metre which is a hint that technology can be used at that size scale. nanoscience and nanotechnologies are deemed novel strategies in developmental study relating to the discovery of miracles and the action of substances on the atomic, molecular, or macromolecular scales, at which their capabilities differ considerably from those at the bulk stage. in this case, the physical, chemical, and biological properties of final materials and the bulk material are very different. changes to properties resulting from working at the nanoscale lead to the introduction of novel materials with interesting architectures, advanced tools and additional beneficial products. increase use of "nanoscience" "nanotechnology" by scientists to create nanomaterials (nms) with a plethora of physical, chemical and biological strategies has contributed to the discipline of "nanoscience" and "nanotechnology" as per zhao, et al. [2].all of these techniques have a few drawbacks regarding isolation and purification of nanoparticles from smilarity micro emulsions and the high use of surfactants. one kind of green technology that is useful is, simple to implement, fast, environmentally friendly, and time–efficient is the development of nanoparticles (nps) from plant cell extraction. figure 1, is showing various ways that eco-friendly formulated nps that can enhance sustainability and the possibilities for agriculture areas, such as improved fertilization, less toxic environmental contaminants, as improved pgrs, and improved pesticides. among other aspects of daily life, nanomaterials (nms) are extensively utilized in industry, medicine, pharmaceuticals, and crop protection for sustainable agriculture. oxidative stress is one type of situation where the excessive creation of reactive oxygen species (ros) affects the cell's redox balance. nms can cause oxidative stress, but they can also ameliorate it, which can lead to phytotoxicity [3]. plant cells often accumulate ros through a variety of mechanisms, including environmental stress [4] and stress from heavy metal build-ups [5-7]. the most frequently reported cases have been biotic stress, salinity stress [8] and physiochemical-cum-allelopathy-induced stress [9]. oxidative stress produces reactive oxygen species (ros), which harm biological materials by altering their structural and functional characteristics and inducing genotoxicity [10] coupled with altered plant metabolism and ultimately triggers cellular death via both short-range and long-range apoptotic signaling cascades [11-13]. (this whole paragraph is rewritten as per instructions) plants frequently produce heat shock proteins (hsps), which function as chaperones and are crucial in granting biotic and abiotic stress resistance, in order to combat different levels of biotic and abiotic stress cum signaling cascades both in cellular and organelles microcosm. additionally, by favorably influencing the antioxidant enzyme systems in plants, hsps usually decrease the levels of reactive oxygen species (ros) and increase membrane stability. in addition, it employs ros as a signal to molecules to stimulate hsp production. hsp also improves plant immunity by accumulating and stabilizing pathogenesis-related (pr) proteins under a variety of biotic stressors[14]. therefore, the purpose behind this study is to provide light on how nms, such as nanopesticides, nanochemicals and nanofertilizers, are used in agricultural soil to promote the safe growth and development of both cash and food crops. this chapter also discusses how nms control oxidative stress in plants and how plants use antioxidants as defense mechanisms [15] to rid of themselves from reactive oxygen species. 2. the world of nano-agrochemicals 2.1. nanopesticides because they keep pests (including rodents, ants, mice, cockroaches, aphids, crickets, and caterpillars) under control and preserve crop growth and development, pesticides are essential to agriculture. the term "nanopesticide" describes pesticides that are applied as agrochemical ingredients (acis) that have two or three dimensions and a nanostructure that ranges from 1 to 200 nm. the technique of enclosing active molecules in a nanoscale protective shell is known as nanoencapsulation (figure 1). effective pest management uses nanoencapsulated pesticides, although residue accumulation must be avoided. various forms of copper oxide-based nanomaterials (cuonps), zinc oxide-based nanopesticides (znonps), magnesium hydroxide-based nanopesticides (mgohnps), and magnesium oxide-based nanopesticides (mgonps) were developed to control pathogenic and agriculture and food sciences research, 2024, 11(2): 203-221 205 © 2024 by the authors; licensee asian online journal publishing group pest attack. manufacturers of pesticides are increasingly using encapsulated nanopesticides. light, humidity, and temperature are examples of environmental factors that can cause them to release the active components in a regulated way [16]. 2.2. nanofungicides the primary reason for the reduction in crop yields and profits is fungal infections. therefore, it is essential to concentrate on creating fungicides that have an inhibitory activity in order to control fungal diseases that are lethal for food crops and to essentially protect and extend the shelf life of harvested commodities. currently, nanofungicides are highly sought after due to their superior solubility, permeability, low dose requirement, and low phytotoxicity.these substances can be used to treat plant diseases and are both safe and environmentally friendly. nps increase a plant's resilience to illness by activating its defenses or rendering microbes incapable of surviving. the antifungal properties of nanoparticles can help in the development of pesticides based on them [17]. due to its many advantages over other nanoparticles, such as cu, zn, au, zno, al2o3, and tio2, numerous researchers have thoroughly studied the manufacturing of silver nanoparticles. nanofungicides that are representative of the environment are favored over non-target areas or organisms that could potentially reduce soil fertility and sustainability either directly or indirectly. 2.3. nanoinsecticides on agricultural land, chemicals called insecticides are being sprayed to either kill insects or prevent them from acting destructively toward crop plants. however, many chemicals are extremely harmful and toxic to biological sources as well as aquatic and terrestrial ecosystems. modern nanoscience research is now creating nanotechnology-based botanical insecticides with active components made from essential oils or plant extracts [18]. for instance, organic extracts from different plant materials like neem leaf (azadirachta indica a. juss) , leaf decoction of citrus limon [(l.) osbeck], extracts of acacia gum (acacia senegal (l.) wild and peels poweder of pomegranate (punica granatum l.) were used in the green synthesis of mgoh [19, 20]. experiment demonstrates that, in addition to their pesticidal activities [21] these botanical nanopesticides have a significant impact on plant growth and development that would boost high-yield organic farming during field trials [22]. figure 1. schematic flow chart of different classes of nanomaterials (nms) shows wider applications for sustainable agriculture. agriculture and food sciences research, 2024, 11(2): 203-221 206 © 2024 by the authors; licensee asian online journal publishing group 3. synthesis of nanoparticles using a variety of processes, including chemical, biological and physical, nanoparticles can be manufactured which have been aimed to be environmentally safe, biodegradable but reproducible in activities, with lower toxicity, higher efficacy and upholding strong antimicrobial, antifungal, antibacterial and antiviral characteristics. these qualities vary depending on the techniques of production (figure 1). the environmentally friendly methods (green synthesis) of producing nanoparticles from plant extracts and with active molecules from biological microorganisms [23] had been a great initiative to its truest sense. these methods are pollutant-free, environmentally beneficial and eventually would produce to little toxic waste [24]. recently, tio2 nps were produced utilizing carica papaya [25]. microorganisms such as bacterial and fungal cells are referred to as "bioreactors" in the context of nanoparticle manufacturing [26]. figure 2 illustrates the synthesis of different various formulations used for types of different nano-particles for commercial uses reported till date. figure 2. synthesis of different types of different nano-particles reported till date. 3.1. biological methods 3.1.1. by plant extracts recently, many ways to synthesizing nanoparticles from plant extracts have been explored. plant-derived nanoparticles are becoming increasingly important in biological applications. copper nanoparticles (cu nps) can be biosynthesized from magnolia sp, syzygium aromaticum and zinziber officinale plant extracts [27]. azadirachta indica and citrus lemon were employed to synthesize gold-based nanoparticles (aunps) and silver-based nanoparticles (agnps) [28]. table 1 illustrates the biosynthesized nanparticles that are currently binging in us employing different plant extracts. table 1. biosynthesized nanparticles by using plant extract. bio-synthesized nanoparticles plant extract plant extract references cuo nps magnoliasp, syzygium aromaticum and zinziber officinale antibacterial yaqub, et al. [29] and yashwant, et al. [30] au nps eclipta alba, nepenthes khasiana antibacterial zhang, et al. [31] and bhau, et al. [32] agno3 nps azadirachta indica, musa acuminate peel antibacterial kumari, et al. [33] and maruthai, et al. [34] tio2 nps carica papaya antifungal saka, et al. [25] fe2o3 nps mentha spicata antibacterial, cytotoxic and anticancerous umar, et al. [35] 3.1.2. by microorganisms verticillium sp., phomasp., fusarium oxysporum, phaenerochaetechrysosporium and aspergilus flavusare some instances of fungi that are used to make silver nanoparticles. (agno3-nps) [36]. however, some microorganisms, such as clostridium versicolor and bacillus subtilis, are additionally employed in the synthesis of silver nanoparticles (agno3-nps) [37]. plant viral capsids are also employed as bio-templates for nanoparticle synthesis, such as tobacco mosaic virus (tmv), which is used to biosynthesise ag and ni nps. 3.2. chemical methods there are several commercial chemical methods for synthesis ofnps that incorporated chemical reduction, microemulsion and electrochemical sapproaches [38]. michael faraday initially discovered the chemical reduction process in 1857. this approach is excellent for producing nanosized copper nanoparticles (cu nps). on the other agriculture and food sciences research, 2024, 11(2): 203-221 207 © 2024 by the authors; licensee asian online journal publishing group hand, the electrochemical approach is utilized to create metal nanoparticles. it is accomplished by sending an electric current between electrodes. 4. employment of nanoparticles designed for plant disease management 4.1. cytoprotectant nanoparticles applied directly to seeds, leaves, or roots, nanoparticles, which have a size range of 10 to 100 nm, have special properties that shield plants against pests and diseases. an extensive investigation has been conducted into the antibacterial, antifungal, and antiviral properties of metal nanoparticles, employing copper, silver, zinc oxide, and titanium dioxide. silver nanoparticles, particularly those produced using "green synthesis," have shown significant antifungal effectiveness against a range of fungal-infections and decreased the risk of viral infection. silver-based nanoparticles provide challenges in terms of manufacturing, toxic interactions against soil-borne, growthpromoting rhizospheric flora. other routinely used metal nanoparticles, such as copper and titanium dioxide, are being investigated for their antibacterial properties. furthermore, chitosan nanoparticles have good biological characteristics and have rendered viral resistance and antibacterial activity against a variety of diseases. chitosan has the ability to function as both solitary nanoparticles and nanocarriers for increased delivery [39]. 4.2. antifungal and fungal-like pathogen killer nanoparticles nanotechnology provides promising options for treating fungal diseases in plants through a variety of ways, including detection, nutrition augmentation, nano-fungicide transporters and direct impacts of nanoparticles (nps) on fungal pathogens. nanoscale nutrients, such as copper, when coupled with other elements, have been proven to increase plant nutrient uptake and immune responses to fungal diseases. standard copper-based fungicides (such copper oxide, copper sulphate, copper oxychloride, dicopper chloride trihydrate, cuprous oxide, copper octanoate, copper sulfide, and copper hydroxide) have been replaced by copper-based nanomaterials, displaying superior efficacy in reducing fungal symptoms and boosting crop output [40]. furthermore, nanomaterials mixed with organic chemicals, such as chitosan-coated iron oxide nps, have demonstrated efficacy in suppressing post-harvest fungal infections on fruits and vegetables [41]. furthermore, nanomaterials mixed with organic chemicals, such as chitosan-coated iron oxide nps, have demonstrated efficacy in suppressing post-harvest fungal infections on fruits and vegetables [41]. nanoencapsulation of fungicides in particles such as mesoporous organosilica has demonstrated enhanced efficiency and stability, resulting in considerable decreases in fungal lesion sizes when compared to established fungicides [42]. these nanotechnology-based techniques provide a secure and more effectual method of addressing fungal diseases in agriculture. 5. plant disease control and management: applications of nanomaterials pesticides and herbicides of various varieties have long been used to control disease. the application of nanoparticles in the management of plant diseases has shown to be highly advantageous in the long run. nanotechnology offers several methods of controlling plant disease. nanotechnology has made significant advances in plant disease management. in the future, it might be used to diagnose diseases caused by bacteria, viruses, fungi, insect pests, pathogenic nematodes and so on. it might well be utilized as a diagnostic tool for diseases caused by bacteria, viruses, fungus and insects. these can be used as biosensors and nano-analytical instruments. nanoparticles have the potential to offer protection against bacteria, viruses, insects and fungi. different metallic nps like, ag, cu, zno and tio2 exhibit strong antibacterial and antifungal activities [43]. different types of nanoparticles (both metallic and green) are used in the management of disease in plants. all things considered, nanoparticles with a variety of modes of action and applications for various pathogens and crops provide potential answers for managing plant diseases brought on by bacteria, fungus, and viruses. 6. bio-efficacy of metallic nps against plant pathogens 6.1. zn nps (zinc nanoparticles) because of their low toxicity, zinc nanoparticles (zn nps) can be used to treat a wide range of illnesses. it possesses viricidal and bactericidal effects. numerous pathogenic fungus, such as mucor plumbeus, botrytis cinerea, and penicillium expansum, have been demonstrated to be effectively combatted by zinc nanoparticles (zn nps) [44]. the antifungal and antibacterial properties of zinc oxide nanoparticles (zno nps) against pathogens including pseudomonas aeruginosa and botrytis cinerea have piqued curiosity. their simple synthesis and photocatalytic actions on plant pathogenic fungi make them useful for disease management and detection [45]. significant antibacterial actions against bacteria, fungus, and spores have been demonstrated by zinc oxide nanoparticles that makes them useful in the treatment of disease. pathogen growth is finally inhibited by these nanoparticles, which also might bring forth oxidative stress, damage pathogen cell structure, and prevent dna replication. thanks to their exceptional catalytic, optical, and physical properties, zinc (zn) nanoparticles—in particular, zno nps—are widely employed in a variety of industries, including food, medicines, chemicals, and agriculture. sclerotinia sclerotiorum, penicillium expansumand staphylococcus aureus are some examples of bacterial and fungal pathogens which are being inhibited to grow vegetatively. they also display concentration-dependent micronutrient releasing characteristics [45]. 6.2. agnps: (silver nanoparticles) silver nps are the most effective ones. it exhibits antibacterial, antiviral, nematocidal and antifungal properties reported till date. silver nanoparticles (ag nps) have been found to have viricidal activity, shielding the faba bean plant from bymv. tomato mosaic virus (tomv) and potato mosaic virus (pmv) resistance had been reported to be effectively provided out of foliar spraying of silver nanoparticles (ag nps). additionally, it had been proved that agnps have a viricidal effect on the banana bunchy top virus (bbtv), as seen by the decreased viral infection in banana plants treated with agnps. it has the ability to inhibit the growth of e. coli, p. aeruginosa, staphylococcus aureus and bacillus subtilis. when applied to staphylococcus aureus it had exhibited strong antibacterial and agriculture and food sciences research, 2024, 11(2): 203-221 208 © 2024 by the authors; licensee asian online journal publishing group antimicrobial properties [46]. according to yakout and mostafa [47] poisonous bacteria like xanthomonas axonopodis and phytoplasma aurantifolia can be identified with the use of silver nanoparticles (agnps), which effectively would change color to brown rapidly from yellow. silver nanoparticles are effective in treating illnesses like sun hemp rosette virus (shrv) in cyamopsis tetragonoloba and late blight in tomatoes because they have shown efficiency against a variety of pathogens, encompassing different genera of viruses and fungi. the unique structure, size, and electrical/optical activity of silver nanoparticles (ag nps) make them interesting antibacterial and antifungal agents. they affect membrane integrity, cause cellular content leakage and do prevent pathogens from producing atp and adhering to cell membranes. furthermore, they oxidize lipids and proteins, cause cellular and ros-mediated cytotoxicity, altering phosphotyrosine levels that would harm mitochondria [48]. ag nps have been shown to effectively suppress the growth of numerous bacterial and fungal infections, including dangerous species like bipolarissorokiniana and magnaporthe grisea. additionally, they reduce the titers of tobacco mosaic virus (tmv) and bean yellow mosaic virus (bymv) in afflicted plants. strong antibacterial action was demonstrated by silver nanoparticles (ag-nps) made from plants such as azadirachta indica (neem, meliaceae) and calligonum comosoum (arta, fire bush, polygonaceae) [49] against plant bacterial and fungal diseases.ag-nps also shown antifungal action against different crop diseases and nematicidal activity against root-knot nematodes, indicating their potential for surface sterilizing of diseased seeds without compromising seed germination. on perennial ryegrass, silver nanoparticles effectively decreased leaf spot and gray leaf spot without causing any phytotoxicity and greatly decreased their antifungal activity. agnps would successfully produce relief following conidial inoculation, reducing diseases that differed significantly between three hours before and twenty-four hours after the treatments. when compared to the water control, the majority of silver formulations administered to plants three hours prior to spore inoculation considerably reduced both phytotoxicity and pathological symptoms. the most successful pre-inoculations were those with agno3 (25 and 50 ppm), ag(p) (200 ppm), and ag(e) (50 ppm), which allowed for less than 7% foliar damage by b. sorokiniana and m. grisea in extremely disease-conducive environmental circumstances, whereas control plants treated with water suffered more than 70% damage. on the other hand, more than 50% of the foliar damage was caused by the inability of delayed treatments of silver preparations 24 hours after spore inoculation to effectively reduce both diseases. silver ions were thought to be very effective after prolonged release because they inhibit microbial respiration and metabolism in addition to causing physical injury inside the cellular microcosm [46]. 6.3. fenps (iron nanoparticles) iron nanoparticles (fe2o3 nps) are less harmful and therefore suitable for everyday use. it is highly reactive and has antiviral properties against tmv. khan, et al. [50] revealed that fe2o3 nps have the capacity to limit the proliferation of phytophthora infestations. iron oxide (fe2o3) nps and magnesium oxide (mgo) nps also demonstrate antibacterial efficacy against various pathogens, with smaller nps showing higher penetration and effectiveness. 6.4. ninps (nickel nanoparticles) cucumber plants treated with ni nps exhibited antiviral action, as well as an increase in leaf count and dry weight. when treating staphylococcus aureus (exhibiting methicillin-resistance) infections, it exhibited antibacterial efficacy. ni nps had been made from an organic extract of the ocimum sanctum plant and exhibit antibacterial effectiveness against bacillus subtilis and e. coli [51]. 6.5. tionps (titanium nanoparticles) it possesses the ability to oxidize biomolecules, which contributes to its significant antiviral activity. it has been found that treating vicia faba l. with tio2 nps reduces viral infection caused by broad bean stain virus (bbsv) [52]. strong antibacterial activity against bacterial phytopathogens and viruses, such as turnip mosaic virus (tumv), had been demonstrated by titanium dioxide (tio2) nanoparticles (nps) produced using sustainable methods (tumv) [53]. 6.6. gold nanoparticles because of their physiochemical characteristics, they have high antibacterial actions. thus, aunps have been employed as biosensor components to diagnose various types and intensity of different plant disease. using the surface plasmone resonance (spr) method, au nps play a crucial role in identifying the pathogen causing wheat kernel bunt disease, as well as late blight of potato and tomato caused by phytophthora infestation using an au npbased lateral flow strip biosensor [54]. au nanoparticles were utilized as the detecting tags for phytopathologists during field trials of cash crops. gold nanoparticles (aunps) have been widely used in numerous detection technologies, including strip-based dna sensors for bacterial and viral infections. they also provide extremely immunospecific sensors for specific infections such as leptosphaeria maculans and pseudocercosporafijiensis [54]. aunps enhance the detection of viruses utilizing dna hybridization assays and label-free colorimetric biosensing techniques, such as cucumber green mottle mosaic virus (cgmmv) [55]. gold nanoparticles exhibit potent antifungal properties against pathogens like aspergillus flavus and aspergillus niger and they have been utilized in combating wheat stem rust and other fungal diseases. 6.7. cunps (copper nanoparticles) it has excellent potential for plant disease control with strong wide spectrum antibacterial actions. fungicides made from cu nps had been having the capability to avert the growth of phytophthora infestations. cu np-based bordeax mixtures lowered the xiphinema index. according to reports, cu nps have had antibacterial activity against xanthomonas compestrispv. vesicatoriain tomatoes [56]. phomopsis sclerotioides, the causal source of cucumber root rot disease, was inhibited most effectively by cuo nps [57]. copper nanoparticles (cunps) are used in nanobiosensors to quantify salicylic acid, which is a molecular marker of biotic stress for several plant diseases such as mungbean leaf spot and bacterial blight disease [58]. owing to the antibacterial characteristics of copper agriculture and food sciences research, 2024, 11(2): 203-221 209 © 2024 by the authors; licensee asian online journal publishing group nanoparticles, the development of bacteria like escherichia coli and staphylococcus aureus as well as fungus like phytophthora infestans had been effectively inhibited. copper nanoparticles (cu nps) exhibit antifungal capabilities, especially when combined with nanochitosan. copper oxide (cuo) nps were mainly explored for managing plant bacteria demonstrating high bactericidal activities against pathogens like escherichia coli and staphylococcus aureus (mrsa, methicillin-resistant) [59]. figure 3 describes the different classes of synthesized nanoparticles using metallic and non-metallic inorganic components. figure 3. classes of synthesized nanoparticles. 6.8. mgnps (magnesium nanoparticles) when it comes to broad-range both gram-positive and gram-negative bactericidal activities, mgnps possess potent antibacterial activities. it has been proven that mgo nps have antibacterial properties against ralstonia solanacearum. mgo nps might have strong antibacterial properties against ralstonia solanacearum-induced tomato bacterial wilt, according to recent studies [60]. 6.9. se and sinps (selenium and silicon nanoparticles) both silicon nanoparticles (si nps) and selenium nanoparticles (se nps) had exhibited strong antibacterial action against a wide range microbial disease [61]. growth and infections plant viruses such as the papaya ringspot virus (prv) and the tomato yellow leaf curl virus (tylcv) had been experimentally inhibited by silicon oxide (sio2) nanoparticles (prsv) [62]. 6.10. pt and pd nps (platinum and paladium nanoparticles) according to chlumsky, et al. [63] platinum nanoparticles (pt nps) and palladium nanoparticles (pd nps) could augur potent antibacterial activities against a variety of food-borne pathogen strains as well as bacteria including escherichia coli and streptococcus mutans. 6.11. al2o3 nps (aluminum oxide nanoparticles) aluminum oxide (al2o3) nps had shown promises in controlling fungal pathogens by stimulating electrostatic attraction with cell membranes, increasing ros production and damaging cellular structures [64]. 6.12. ceo2 nps (cerium oxide nanoparticles) cerium oxide (ceo2) nps exhibit antibacterial properties and have the potential to regulate phytopathogenic activity [65]. 6.13. ni nps (nickel nanoparticles) the antibacterial efficacy of nickel nanoparticles (ni nps) had been comparable against a range of diseases, including viruses [66]. 6.14. chitosan nanoparticles chitosan nanoparticles had shown effectiveness in impeding the spread of viruses within plants and boosting the host's immune response. they could strongly enhance fruit resistance and modulate various physiological processes in plants, thereby aiding in disease management [67]. overall, nanoparticles offer diverse mechanisms for combating plant pathogens, including induction of systemic resistance, inhibition of dna replication, disruption of cell structure and modulation of cellular signalling pathways. their antibacterial qualities make them useful instruments in the creation of successful plant disease management plans. however, the use of cadmium nanoparticles (cd nps) is limited due to their toxicity to humans, animals and plants. agriculture and food sciences research, 2024, 11(2): 203-221 210 © 2024 by the authors; licensee asian online journal publishing group 7. nanoparticles as “strategic weapons” against plant pathogens the mechanism of action nanotechnology presents various promising avenues for managing plant diseases, primarily through the direct application of nanoparticles to foliage, seeds, or soil. nanotechnology is transforming plant disease detection by allowing improved tools to identify pathogens quickly and accurately. nanoparticles, with their small size and distinct features, improve biosensor selectivity, sensitivity and detection limits. when it comes to pathogen detection, metal nanoparticles—such as those made of gold, silver, copper, zinc oxide, and other metals—augur distinct benefits over enzyme-related testing systems. these advantages include their bigger surface area-tovolume ratios and simpler physical sensor-mediated detection processes [68]. 8. cellular uptake and mechanism of nano-toxicity engineered nanoparticles (enps) are absorbed by cells through endocytosis, non-endocytosis, and passive transport [69]. small, positively charged and hydrophobic enps are primarily transported via diffusion, whereas perisorption can occur via intestinal microvilli or enterocyte cells. endocytosis routes, including as receptormediated endocytosis and caveolae/lipid raft-dependent endocytosis, transport enps to lysosomes, where they might induce intracellular toxicity due to metal ions. transporter proteins or ion channels are used to absorb the ions created during enp breakdown. enps can cause toxicity and disturb cytophysiology once they enter inside the cell by causing membrane lipid peroxidations [70] and dna damage plus protein injury [8] and oxidative stress via ros outbursts, followed by other mechanisms that halter the normal physiological states [71]. enps' form, size, and coating composition are examples of their physicochemical characteristics that affect their toxicity. smaller enps may cause more toxicity and particle form also influences nanotoxicity. positively charged enps may be more harmful and bioaccumulative due to their affinity for cellular membranes [72]. dietary exposure to enps might cause cellular stress, affecting eating behavior and lipid peroxidation levels. a multitude of processes, such as direct cellular membrane destruction, disruption of atp generation, and dna replication, are the direct outcomes of nanomaterial toxicity [73]. when organisms consume free nano-ions, morphological changes occur, such as cytoplasm contraction and dna condensation [74]. nano-ions also interact with respiratory enzymes and transport proteins, resulting in proton leakage and affecting cellular activities [75]. nanotoxicity causes oxidative stress by producing reactive oxygen species (ros), which damage dna and cause cell death. the chemical composition of nanomaterials impacts the amount of ros produced. ros predominantly target dna, causing various forms of oxidative dna damage [76]. however, the conversion of less reactive species into more reactive radicals (figure 4) sharply worsens cellular damage. ramos-zúñiga, et al. [77] study the transition of less reactive species into more reactive radicals, particularly the hydroxyl radical, which can cause severe cellular damage ramos-zúñiga, et al. [77]. figure 4 illustrates the detailed processes during nanoparticle toxicity which has a general mechanism. (1) accumulation and dissolution of nanoparticles and entry inside the cell. (2) the concentration of nanoparticles on the cell surface causes depolarization and membrane rupture. (3) nanoparticles’ entry inside the cells causes sustained release metallic ions as a result of np dissolution. the release of ions would produce the following effects: (4) ros ourburst, membrane depolarization via lipid peroxidation and protein oxidation; (5) reduced atp generation; (6) disruption of endomembrane systems and collapse of photosynthesis by chloroplast assembly (7) mitochondrial membrane depolarization leading to apoptosis. figure 4. nanoparticle toxicity has a general mechanism. (1) accumulation and dissolution of nanoparticles and entry inside the cell. (2) the concentration of nanoparticles on the cell surface causes depolarization and membrane rupture. (3) nanoparticles’ entry inside the cells and sustained release metallic ions as a result of np dissolution. the release of ions would produce the following effects: (4) ros ourburst, membrane depolarization via lipid peroxidation and protein oxidation; (5) reduced atp generation; (6) disruption of endomembrane systems and collapse of photosynthesis by chloroplast assembly (7) mitochondrial membrane depolarization leading to apoptosis. agriculture and food sciences research, 2024, 11(2): 203-221 211 © 2024 by the authors; licensee asian online journal publishing group 9. cellular toxicity via dna damage by nanoparticles the dna damage within the microbial cells exposed to agnp or znonp had been probed by electrophoretic analysis dna strand breakage visualized within agarose gels [78]. because they produce reactive oxygen species (ros), zinc oxide nanoparticles (zno nps) had been well known for their antibacterial properties. zno nps caused concentration-dependent toxicity and membrane damage after being extensively absorbed into the bacterial cells. studies on genotoxicity had shown that exposure to zno brought forth substantial dna damage inside bacterial cells. every observation showed that zno nps caused substantial oxidation of proteins, dna damage, and ros over-production along with concurrent thiol group depletion. moreover, zno np exposure was found to downregulate a number of genes linked to the metabolic pathway and dna repair, while simultaneously upregulating the expression of genes connected to the dna damage responses [79]. cui nps generated rosmediated dna damage for transcription suppression, as demonstrated by reporter gene assay, and they also caused ros production in both gram positive and gram negative bacteria. in the presence of amine functional groups from diverse biological molecules, ros most likely would have developed on the surface of cui nanoparticles. additionally, they cause harm to membranes [80]. according to gordon, et al. [81] ag ions would prevent dna replication and deactivate sh-groups in functional proteins. microorganisms' biological activities, including alterations in the structure and function of cell membranes, were known to be impacted by silver. silver also would likely cause the inhibition of the expression of proteins involved in atp generation [82]. it follows that oxidative stress, which is brought on by nanoparticles (nps) might harm dna in microorganisms by resulting in oxidized dna bases, single-strand breaks (ssbs), and double-strand breaks (dsbs). additionally, nps can affect dna repair processes through their effects on gene expression, deregulate dna repair protein functioning, and the deplete nucleotide pool required for dna repair systems. 10. nanomaterials and regulation of oxidative stress since the natural pool of endogenous antioxidants, which may detoxify free radicals in a normal condition, is outweighed by the creation of ros, oxidative stress is a phenomenon that occurs when a cell's redox equilibrium is upset [13]. reactive oxygen species (ros), which are highly reactive free ions or radicals created from molecular oxygen during redox reactions, are able to exist independently and have one or more unpaired electrons in their outer orbit or valency shell. a free radical's odd number of electrons makes it incredibly reactive, short-lived, and unstable. they can steal electrons from other molecules to become stable since they are extremely reactive. the attacked molecule becomes a free radical after losing an electron, which sets off a chain of events that ultimately damage the living cell. the most prevalent ros in plant biological systems are singlet oxygen (1o2), superoxide radical (o2-), hydrogen peroxide (h2o2), and hydroxy radical (oh•). (figure 5). these substances' high reactivity and toxicity make them capable of causing oxidative cell death [83, 84]. the electron transport chain (etc) in mitochondria, the mechler reaction in chloroplasts, and photorespiration (c2 cycle) in peroxisomes are the three subcellular compartments that produce multiple reactive intermediaries promulgating ros outburst [14, 85]. environmental stress and increased ros accumulation are directly connected. when an organism produces too many reactive oxygen species (ros), it damages its physiological capabilities and ultimately leads to cell death. external stimuli such as heat stress, ionizing and non-ionizing radiation, heavy metals, xenobiotic chemicals, uv radiation, and different phytopathogens can upset cellular homeostasis, which in turn can trigger the generation of reactive oxygen species (ros) in plant cells [12]. figure 5. schematical representation of ros generation and their interconversions. agriculture and food sciences research, 2024, 11(2): 203-221 212 © 2024 by the authors; licensee asian online journal publishing group 11. production and inter switching of reactive oxygen species in microbial cells ross have an impact on living cells and tissues that including plants [12, 86] which are both positive and negative [86, 87]. ros must be produced at a low to moderate level in the cellular compartments of aerobic organisms [13]. on contrary, elevated levels ros out of abiotic and biotic stress can generate two distinct groups of reactions out of production of reactive oxygen species (ros: *oh, 1o2 , o. 2) and reactive nitrogen species, (rns: *no, *no2, onoo-, n2o3,hno2, no-, no+, n2o4) (figure 6) which can lead to: lipid peroxidation and protein carbonylation. lipid peroxidation is a process that turns unsaturated lipids into saturated residues, such as hydroperoxide [70, 88]. when proteins are carbonylated, ros can oxidize to the unsaturated nucleus of amino acids, including tyrosine, histidine, tryptophan, phenylalanine, etc.a downstream reaction produces a peroxide product in both cases, which can also progressively break down bimolecular fabrication of living cells. the surplus reactive oxygen species (ros) generated within the cellular microcosm is scavenged by plants' efficient antioxidant defense mechanism under a variety of oxidative stress circumstances.the antioxidant safety net is composed of both enzymatic and non-enzymatic elements [89]. flavonoids, tocopherol, carotenoid, polyamine, alkaloids, and phenolic compounds are examples of non-enzymatic components; glutathione reductase (gr), ascorbate peroxidase (apx), glutathione dismutase (sod), and catalase (cat) are examples of enzymatic components. pizzino, et al. [87] and rudenko, et al. [90] (figure 6). this whole paragraph is rewritten as per instructions. figure 6. an imbalance between ros and antioxidants brought on by abiotic stress causes an oxidative imbalance in live cells' cellular homeostasis. the generation of reactive oxygen species (ros), such as hydroxy radical (•oh), is a key initiator of lipid peroxidation, which degrades lipoprotein and produces a cascade of end products including conjugated dienes and malondialdehyde (mda) [70, 86, 88, 91]. 12. mechanisms of np uptake by higher plants 12.1. passive uptake of nps understanding how nanoparticles (nps) are absorbed by plants is critical for determining their interactions with plant cells and immunological responses. there have been several hypothesized methods for np internalization, including passive uptake processes. depending on their size, surface charge, and hydrophobicity, nps can enter plants passively through diffusion and adsorption [92]. when nps come into contact with a plant's surface, they can disperse through the stomata or cuticle and gather in different plant tissues. their hydrophobicity facilitates passage through the lipophilic cuticle, while stomatal holes serve as entry locations. for example, gold nanoparticles coated with polyvinylpyrrolidone and citrate are passively absorbed by wheat leaves through cuticles [93]. however, additional research is required to understand how foliar nps are translocated. 12.2. active uptake of nps plants may absorb nanoparticles (nps) selectively through active uptake mechanisms, which allows for precise targeting and regulated responses. endocytosis, in which nps are engulfed by the cell membrane to form vesicles, allows plant cells to actively absorb nps. plants can control np absorption and respond to environmental cues thanks to this widely used mechanism. plant cells take up nps more readily when they enter endocytosis pathways mediated by caveolin or clathrin. for example, tobacco protoplasts actively internalize gold nanoparticles via clathrin-dependent pathways [69]. for plant cells to absorb nanoparticles by endocytosis, their surface properties are essential. for instance, triethylene glycol-functionalized silica nps were consumed by tobacco mesophyll protoplasts, while bare, unfunctionalized silica nps were not able to penetrate plant cells [94]. even with these results, there is still a paucity of direct evidence of endocytic np uptake by plant cells, which calls for additional study to comprehend integrated mechanisms of np uptake and translocation within the plant system. agriculture and food sciences research, 2024, 11(2): 203-221 213 © 2024 by the authors; licensee asian online journal publishing group 12.3. uptake and translocation of nps through roots plants are mostly exposed to soil-applied nanoparticles (nps) through their roots, where they interact with root hairs and epidermal cells. plant roots are more likely to attract and retain positively charged nanoparticles (nps) or those that chemically interact with the root cell walls. with the help of transpiration rates, nps can move from inside the root to other plant portions through the xylem and phloem [95]. for example, it has been found that copper nanoparticles migrate with water in the xylem, making it easier for them to go from the roots to the aerial parts of maize. the variety of np absorption mechanisms enhances the potential of these compounds as plant immune modulators by enabling targeted administration and controlled release of bioactive compounds. however, in order to choose application techniques and guarantee effectiveness in agriculture, characteristics such as the physicochemical properties of nps, their intended use, targeted crops, and desired interaction with the plant system are essential [96]. research into np absorption pathways is progressing, and this could lead to the full realization of their potential in sustainable plant disease management. in addition to controlling oxidative stress and maintaining redox homeostasis, nanomaterials can also occasionally cause oxidative stress [97]. owing to these two features, it is critical to comprehend the precise metabolic processes of nms and their interactions with plants. lipid peroxidation and electrolytic leakage were decreased as a result of antioxidants' production squelching the ros level. researchers such as rizwan, et al. [98] discovered that foliar application of si and tio2-nps alleviated oxidative stress in rice plants under cadmium (cd) stress by reducing lipid peroxidation and electrolytic leakage by enhancing the activities of antioxidant enzymes like sod, cat, and apx.given that nanoscience is now being investigated as a discipline connected to redox biology and biochemistry, it is critical to comprehend the exact method by which nanoparticles (nps) avoid ros overproduction mitigating overall normal physiological processes. 13. how nanomaterials (nms) would emerge as ‘nano-weapons’ against phytopathogens? • the special optical characteristics of nanomaterials (nms) enable the very accurate and practical detection of plant pathogens [16, 54, 99]. • nms damage to the membrane transporter and nutrient absorption systems of pathogen, [100]. • nanoparticles generate ros inside the pathogenic cell and developed nanotoxicity which results damage of nucleic acids (dna/rna), uncontrolled cell signalling, cytotoxicity, genotoxicity and pcd [101]. • toxic ions generated as a result of nanotoxicity impair membrane protein activity and cell permeability [102]. • nanoparticles are designed to offer distinct and improved antibiotic activity against plant diseases, depending on their size and dose [46]. • nps as bioactive chemical delivery agents: many bioactive substances, including essential nutrients, plant hormones, rna interference agents, and chemical protectants, may be transported via nanoparticles (nps). this feature highlights how they could completely transform plant genetic engineering, growth regulation, and nutrition management [103, 104]. • nanoparticles can improve nutrient delivery to plants by increasing solubility, stability and bioavailability. nanofertilizers developed for this purpose preserve nutrients from degradation and could release them in a guarded manner, allowing for optimal welcome by the plant tissues. this technique has the potential to reduce nutrient deficits and increase agricultural productivity, hence addressing global food security issues. zinc-based nanofertilizers, for example, stimulate seed germination and growth in a variety of crops, whereas copper oxide nanoparticles efficiently transfer copper to increase crop yields and control plant diseases such as wilt by fusarium sp and bacterial fruit blotch disease (fbd) [105]. despite promising findings in greenhouse conditions, field experiments are required to evaluate their potential in real-world environments. • delivery of plant hormones: plant hormones can be precisely delivered to specific areas of a plant using nanoparticles (nps), which enables the plant to respond to disease or infection [106]. mesoporous silica nps, for example, had been created to supply abscisic acid (aba) to arabidopsis, whilst biogenic iron nps had been utilized to therapeutically deliver salicylic acid (sa) into watermelon plants, for increasing resistance to wilt disease caused by fusariumsp [107]. these nano-enabled delivery systems ensure that plants receive hormone signals at the optimal time, improving stress tolerance and utilization of the chief nutrients. growth hormones, which stimulate root development, shoot growth, and flowering in commercially important plants, can also be distributed using chitosan-based nanocomposites [41]. using nps to deliver hormones offers the chance to control plant development and stress reactions, improving crop resilience and performance. • small interfering (si-rna) rnas delivery through nps: plant cells can be directly exposed to smallinterfering rnas (sirnas), which overwhelmingly mediate rna interference (rnai) responses and confer resistance against pests and diseases, through the use of nanoparticles (nps), an adaptable auto-regulatory delivery system [108]. researchers can regulate the expression of certain genes associated with disease susceptibility, insect resistance, or other agronomically important traits by using this capability, which enables targeted gene silencing. nps increase the effectiveness of gene silencing by protecting sirnas from degradation in the harsh extracellular environment and promoting cellular absorption [109]. compared with naked dsrna, pathogen-specific double-stranded (ds)rna administered by layered double hydroxide nanosheets, for instance, remained stable for as long as 30 days and offered tobacco plants sustained protection against viral infections. this method has the potential to reduce the need for medicinal interventions by conferring resistance to diseases and pests. one useful method for precision farming and developing long-term crop protection strategies is the use of nps as sirna carriers [103]. • chemical cyto-protectants delivery by nps: the innovative application of nanoparticles (nps) in crop delivery for chemical protectants represents a significant advancement in precision farming and disease control. it has been challenging to apply chemical protectants like fungicides and bactericides effectively and efficiently, despite the fact they are essential for shielding crops from phytopathogens. it is possible to agriculture and food sciences research, 2024, 11(2): 203-221 214 © 2024 by the authors; licensee asian online journal publishing group provide these protectants precisely and regulated to particular plant tissues and pathogens by encasing or binding them in nps [110]. for example, prochloraz had been transported via mesoporous silica nanoparticles, providing further defense against rice blast disease (rbd) [111]. similarly, silica nanoparticles loaded with azoxystrobin shown enhanced fungicidal activity against tomato late blight disease (tlbd) [112]. when compared to bulk counterparts, nanoscale fungicides were more effective against germs and prevented disastrous crop illnesses. • weeds and arthropod pests management by nps: several reports established that pests can move across cultivated and noncultivated plants, including weeds, posing issues for disease management. insects play an important role in infection transmission, which complicates the situation. traditional chemical control methods pose environmental problems, emphasizing the necessity for novel approaches such as nanotechnology. nanoformulations show potential in improving pesticide efficiency while decreasing the environmental impact. for example, nanoencapsulation of atrazine boosted herbicidal efficacy tenfold, while light-triggered nanoformulations improved herbicidal effects while reducing toxicity to nontarget plants, such as glycine max [113]. however, optimizing target delivery remains a difficulty, particularly given the similarities between weeds and crops. carbon-based nanomaterials have been investigated for more effective and targeted pesticide delivery. moreover, nanoparticles had shown useful roles in the management of insect pests. for example, encapsulated thiamethoxam has been effective in reducing asian citrus psyllids [114]. eco-friendly pest management solutions are offered by combining bioinsecticides with materials based on nanotechnology. moreover, nanoparticles can be engineered to react to outside stimuli, which enhances their properties. for example, as temperature changed, mesoporous silica nanoparticles boosted insecticidal action against aphids. rna interference technology paired with nanoclay effectively prevented virus transmission via insect vectors [115]. overall, nanotechnology shows potential for transforming agricultural disease and pest management systems. 14. environmental risk assessment of nps the rising use of man-made nanoparticles in the environment highlights the need for rigorous risk assessment and regulatory controls. commercializing nanomaterials for agricultural applications involves a number of obstacles, including production scale, cost reduction and safety. to have a better understanding of the production, toxicity, and field application of nanomaterials in agriculture, more research is required. by producing nanopesticides, nanotechnology has the ability to completely transform agricultural pest management. these nanopesticides have various advantages, including enhanced solubility, reduced toxicity, targeted delivery and phdependent release. nanopesticides are still in the early phases of development, and more study is needed to determine their effectiveness and potential toxicity to soil and the environment, despite their potential advantages. although the use of nanoparticles (nps) is expanding across many industries, there is a risk of environmental pollution from their release. they have the capacity to permanently contaminate the water, soil, and air [71]. among the most popular sources are wastewater treatment facilities, electronics, paints, pigments, coatings, and cosmetics. although nanoparticles can be used to clean up the environment, their extensive their extensive use might effectually jeopardize the ecosystems upon randomization. np dispersion is aided by both nanosensors and nanofertilizers. in general, nanoparticles are challenging to control and contain due to their small size and reactivity. moreover, nps can be designed to release their payload in response to specific environmental conditions or triggers, which reduces the amount of fungicide needed overall and the amount of off-target effects. chemical protectant delivery by nanoparticles (nps) may change agriculture's approach to disease control; however there are safety issues and unintended consequences for other living things and the environment to take into account. all things considered, nanoparticles have shown promise as flexible means of delivering bioactive substances to plants, offering a novel approach to challenges in agriculture. the potential uses of np-mediated smart delivery systems are expanding as nanotechnology research advances, providing new opportunities to increase agricultural output, nutritional value, and sustainability.however, concerns including np biocompatibility, environmental impact assessment, and regulatory factors need to be carefully explored as this technology moves closer to being used in agriculture. 15. bacteria and fungi play crucial roles in promoting plant growth through various mechanisms in natural environment • nutrient cycling: organic debris in the soil is broken down by fungi and bacteria, releasing nutrients that plants can absorb. this cycle of nutrients increases soil fertility and provides essential nutrients for plant development. • nitrogen fixation: certain bacteria build symbiotic relationships with leguminous plants, such rhizobium species, and transform atmospheric nitrogen into a form that plants can utilize. nitrogen is added to the soil through this process, which is necessary for plant growth. • mycorrhizal associations: mycorrhizal fungi create mutualistic relationships with plant roots, extending their reach and increasing nutrient intake, particularly phosphate and micronutrients. this symbiotic interaction can greatly enhance plant growth and health. • disease suppression: by building systemic resistance in plants, manufacturing antibiotics, or engaging in resource competition, beneficial bacteria and fungi in the soil can inhibit the growth of dangerous diseases. this aids in preventing disease in plants. • stress tolerance: certain bacteria and fungi create chemicals that help plants withstand environmental challenges like drought, salt and heavy metal contamination. these bacteria improve plant health and resilience by increasing their stress tolerance. • biofertilizers and biostimulants: in agriculture, several bacteria and fungus serve as biofertilizers and biostimulants. they can boost soil fertility, nitrogen uptake, root growth and plant vigor, resulting in higher crop yields. agriculture and food sciences research, 2024, 11(2): 203-221 215 © 2024 by the authors; licensee asian online journal publishing group • overall, bacteria and fungi have various and crucial roles in encouraging plant development and health, thus they are beneficial. 16. bacterial nano-toxicity by enps bacteria, which are essential for environmental processes such as fermentation and decomposition, are extensively researched in eco-nano-toxicity because of their customization for ecological sustainability. bacillus, pseudomonas, escherichiasp and majority of nitrifying bacteria are typically employed in this research. some engineered nanoparticles (enps) with antibacterial capabilities are included into consumer products and eventually released into the environment. interaction with bacteria has an impact on their growth and ecosystem function. enps such as ag, cuo and carbon-based ones reduce bacterial growth and change enzyme activity, mostly by membrane degradation and oxidative stress generation [116]. studies have shown that enps have a wide range of favourable vis-à-vis toxicological effects upon several beneficial microbiota [117]. 17. protozoal nano-toxicity out of enps accumulation protozoa are single-celled eukaryotic organisms that feed bacteria, thrive in damp environments, and are crucial to the development and maintenance of microbial communities. they are actively researched in econanotoxicity assessments. tetrahymena thermophila is an excellent model for assessing the toxicity of engineered nanoparticles (enps). enps mostly enter protozoa by endocytosis and phagocytosis, generating toxicity by upregulating metallothionein genes, modifying membrane fluidity, causing oxidative stress and lowering viability and motility [118]. protozoa also serve as indicators of enp trophic level transmission in food chains, since studies have shown that cadmium is biomagnified from enp-loaded bacteria to protozoa and beyond. in vivo bioimaging shows enps' trophic transmission across various layers of the food chain. 18. nano-toxicity of fungi out of enps accumulation in a variety of environments, fungi are essential decomposers and are crucial to the nitrogen cycle. understanding the harmful effects of engineered nanoparticles (enps) on fungi is critical to ecosystem health. enp exposure can harm cell membranes, impede development, lower cell density, cause cytoplasm blebbing and impair breakdown activity by influencing lignocellulolytic enzyme synthesis [119]. high doses of enps utilized in toxicity studies may not accurately reflect environmental exposure levels. in natural settings, fungal communities have been constantly exposed to lower concentrations of enps over longer periods of time, emphasizing the need for more representative nano-toxicity studies. 19. harmful effects of nanoparticles (nps) on earthworms although snails and earthworms are beneficial and crucial parts of healthy soil ecosystems, their populations and activities need to be carefully controlled to reduce any potential harm to plant development and agricultural output. earthworms, which belong to the phylum "annelida," are invertebrate animals that live in soil and contribute significantly to soil quality by increasing plant nutrients. the species of earthworms found in the field affect the soil's nutritional value. based on the range of soil types and horizons they inhabit, earthworms are divided into three ecological groups: endogenic species, like aporrectodea caliginosa, which live in organic horizons and dig horizontal burrows; epigeic species, like eisenia fetida, which live on the surface and in litter; and anecic species, like lumbricus terrestris, which live in deep, vertical burrows and consume large amounts of soil. numerous organic acids and enzymes secreted by earthworms interact with soil to promote the growth of bacteria, fungi, actinomycetes, and soil microflora. these microorganisms help fix nitrogen, solubilize nutrients, and increase plant growth and production potential. earthworms are therefore essential to a sustainable ecosystem and are commonly known as "farmers' friends." according to research, earthworms' physiological processes and general environmental activities are negatively impacted by nanoparticles (nps). numerous scientific studies have found detrimental impacts on the digestive and immune systems, species diversity, and the growth, reproduction, and development of earthworms. pheretima posthuma, the most prevalent species of earthworm, is frequently employed as a standard test organism to identify environmental pollution levels [71, 120]. 20. biochemical pathways leading to toxicity of soil microflora and macrofauna: oxidative stress and cellular death occurrence after enps exposure when faced with environmental challenges, reactive oxygen species (ros), which are produced during regular metabolic processes, function as signaling molecules. in rhizospheric microflora and fauna, engineered nanoparticles (enps) can lead to an excess of reactive oxygen species (ros), which can induce oxidative stress, cellular damage, dna fragmentation, dysfunction of proteins and enzymes, and finally cell death. to reduce ros effects, organisms have antioxidant defense mechanisms that include both non-enzymatic (e.g., ascorbate, proline and phenolics) and enzymatic (e.g., catalase, peroxidase and superoxide dismutase) counterparts. however, high enp concentrations can overwhelm these defenses, resulting in oxidative damage to cellular organelles. the efficiency of antioxidant responses to enp-induced oxidative stress varies depending on the rhizospheric species and exposure conditions. non-enzymatic antioxidants including ascorbate, proline and phenolics play important roles in scavenging ros and protecting animals from oxidative damage. to fully comprehend the mechanisms of nanotoxicity in beneficial microflora and fauna, as well as the reactions of antioxidant defense systems to acute and long-term exposure to ambient deposition of manufactured nanoparticles, more research is required. if a virus infects soil it may have a number of regulatory impacts on plant development and health: • nutrient uptake: nanoparticles influence nutrient availability in the soil. if plants are infected with a virus, the nutrition cycle may be disrupted, resulting in deficits in key nutrients required for plant growth. • toxicity: certain nanoparticles (cd, pb, cr and hg based) can be hazardous to plants. if a virus affects the composition or activity of nanoparticles in the soil, the toxicity levels may rise, affecting plant growth. agriculture and food sciences research, 2024, 11(2): 203-221 216 © 2024 by the authors; licensee asian online journal publishing group • microbial interactions: the microbial ecology in the soil, which is essential for the cycling of nutrients and plant health, can be impacted by nanoparticles. unbalances in the soil ecosystem could result from a virus that affects nanoparticles and interferes with these microbial interactions. • plant immunity: nanoparticles have been investigated for their ability to boost plant immunity to diseases. if virus-infected nanoparticles disrupt this function, plants may become more susceptible to disease. • soil structure: nanoparticles can alter soil structure and water retention. a virus-induced change in nanoparticle activity may modify soil characteristics, influencing plant water availability and root growth. • overall, the impact would be determined by the individual virus, the nanoparticles used and their interactions with the soil microflora and plant population ecology khan, et al. [50]. 21. sensing and detection and plant pathogens in agricultural crops through nanomaterials (nms) plant resources are what sustain human civilization. there are three essential needs: shelter, food and fiber. these requirements are increasing every day in line with the global population growth. the yield of crops is greatly hampered by the shrinking agricultural lands brought about by the fast urbanization and industrialization of society. another barrier that results in biotic stress in agricultural crops is plant diseases, which causes the plants to become partially impaired and eventually die. it presents a significant hazard to food production and security. plant infections have wreaked havoc on some major disasters. for instance, phytophthora infestans in ireland produced the irish potato famine in the 1840s, while helminthosporium oryzae in rice caused the great bengal famine in 1943. the majority of research on plant biological stress attempts to reduce appalling yield loss, either straight or circuitously [15]. the administration of previously discussed insecticides, fungicides, pesticides and fertilizers is the primary method used to manage biotic stress in agricultural crops. engineered sensors (figure 7) based on nanomaterials are used to track stress caused by pathogens as well as the environment. researchers discovered that nanosensors worked well to identify fungal pathogens that cause plant illnesses that are soil-borne, air-borne, seed-borne and other. an electrochemical sensor that can identify the causative agent of bacterial specks of solanum lycopersicum (psedumonassyringae dc3000) with a recognition limit scale-up of 214 pm [121]. in order to identify soil-borne pathogens such as ralstoniasolanacearum, which causes bacterial wilt in potato disease, the researchers used au-nps [122]. therefore, remote sensing technology based on nanosensors is a great way to identify, anticipate and control insects, pests and plant diseases in order to safeguard both commercial and agricultural crops and preserve the environment. 22. nanotechnology mediated detection of plant pathogens at early stages of infections researchers found that fungal pathogens that cause soil-borne, air-borne, seed-borne and other plant ailments may be effectively identified using nanosensors. with a recognition limit of 214 pm, an electrochemical sensor can determine the cause of bacterial specks of solanum lycopersicum(psedumonassyringae dc3000) [121]. the researchers employed au-nps to identify soil-borne pathogens, such as ralstonia solanacearum, which causes bacterial wilt disease in potato [122]. in order to protect both commercial and agricultural crops and maintain the environment, remote sensing technology based on nanosensors is an excellent approach to identify, anticipate and control insects, pests and plant diseases. figure 7. flowchart shows the advent of different nanotechnology tools used for pathogen diagnosis. 23. challenges and limitations of nanosensors nanotechnology holds great promise for revolutionizing pest management in agriculture through the development of nanopesticides. numerous benefits are provided by these nanopesticides, including increased agriculture and food sciences research, 2024, 11(2): 203-221 217 © 2024 by the authors; licensee asian online journal publishing group solubility, increased effectiveness with decreased toxicity, targeted administration, and ph-dependent release.nevertheless, study is extremely demanded to determine the efficacy and potential toxicity of nanopesticides to soil and the environment because they are still in the early phases of development, despite their apparent advantages. the precise definition of nanopesticides by regulatory bodies is still pending, and nothing is known about how they affect pesticide resistance. challenges include the lack of regulatory standards, high manufacturing costs and public perception. bridging the knowledge gap among farmers and stakeholders, as well as developing scalable solutions compatible with existing farming practices, is essential for successful implementation. to overcome these obstacles and fully utilize nanotechnology in agriculture, teamwork between companies and researchers is essential. for risk evaluation and regulatory approval, dependable data collection and long-term field experiments are required. additionally, stakeholders should be informed about storage conditions and cytotoxicity concerns associated with certain nanoparticles and efforts should be made to utilize green and natural materials for nanoparticle synthesis to minimize environmental risks. overall, interdisciplinary collaboration and comprehensive research are akin to advancing agricultural nanotechnology responsibly. nanotechnology has potential for sustainable and environmentally friendly agriculture practices by introducing new methods of administering fertilizers, insecticides and other materials. however, current study indicates that nanoparticles could affect soil animals such as earthworms, increasing safety issues in food and agriculture. the environment's growing employment of artificial nanoparticles highlights the significance of stringent risk assessment and regulatory limitations. several obstacles must be overcome in order to commercialize nanomaterials for use in agriculture, including those related to manufacturing scale, cost control, and safety. to fully comprehend the production, toxicity, and practical use of nanomaterials in agriculture, more study is required. nanomaterials (nms) have special qualities that could make them useful in agriculture. these promising qualities include: i) increased bioavailability of the active ingredient; ii) improved pesticide, fungicide, and insecticide solubility; iii) releasing the active ingredient based on ph level; iv) transferring rnai molecules for disease management; v) ros detoxification and oxidative stress regulation in plants; and vi) epigenetic and epigenomic studies of animal stress regulation [123]etc. although nanomaterials (nms) offer many potential uses and benefits in agriculture, particularly in terms of helping crops become more resilient to stress, they also have a number of harmful consequences on our ecosystem and living things [124]. nanoparticles mediated pollution is danger to the natural flora and fauna including beneficial bacteria, fungi and soil-nematodes (soil microflora)by causing long term nanotoxicity in their body, decreasing soil fertility, low level of nutrient solubilization and losing the activities of pgpr in soil [125]. nano-agrochemicals continue to be denied authorization to be commercialized in the market due to their vulnerability towards unfavorable environmental attainments. when it comes to using fungicides, pesticides, herbicides, or other products based on nanoparticles (nps), regulatory bodies or the pharmaceutical industry have failed to give the right ratio of nano substances and their maximum level of effective dose without endangering the environment. furthermore, studies and researches are urgent for eco-friendly use of nanoparticles (nps) in agriculture and its commercialization. additionally, scientists ought to create fresh, workable solutions that our farmers can readily use without interfering with their current methods and practices [126]. in particular, more research using an in-vitro method is still needed to determine the appropriate dose level, establish regulatory requirements, and modify and use crops at the field level when using nms to agriculture [127]. 24. conclusion and future perspectives for future smart agriculture nanotechnology is a vast field of study that combines aspects of environmental biology, ecotoxicology, physics, chemistry, botany, biotechnology, and agricultural science. with atomic sizes ranging from 0.1 to 100 nm, nanomaterials (nms) can have both positive and negative environmental effects. in order to apply nms to their biological roots, researchers should concentrate on using them by highlighting and improving their ecologically favorable properties in the proper amounts. nanomaterials are widely used in agriculture to boost high productivity and sustainability. nms are mostly used as nanopesticides, nanofertilizers, nanoinsecticides, nanofungicides, and nanoherbicides to protect agricultural crops from pests and boost their fertility. this paper discusses these applications. our study also demonstrates how nms help cells defend against redox homeostasis imbalances, which cause ros to be produced in excess and activate the antioxidant defense system. additionally, nms would be improved as a tool for producing heat shock proteins [13] balancing physiological homeostasis, and enhancing epigenetic memory [123] in plants to fight off biotic and abiotic stressors. the type of nms, the dosage required for each type of abiotic stress (drought, cold, salinity, etc.), and their effects on over ten different plant species were all documented. to enhance comprehension, scientific examples are provided on the use of nms in sustainable agriculture, the production of ros and their interconversion, and antioxidant defense mechanisms.despite a number of innovative research initiatives, there are still several gaps in the field of nanoscience. how precisely nanomaterials (nms) regulated and activated the antioxidant system against reactive oxygen species (ros) is still unclear and contradictory, for example. thus, before moving into the new and more sophisticated stage of nm-based agriculture, it is essential to have a precise understanding of how plants and nms interact under different stressful conditions and how this affects long-term exposure. an organization that conducts research on nanotechnology should create suitable rules for the safe handling of nanoparticles (nms), their variety of uses in biological systems, and their appropriate disposal to avoid harm to the environment or public health. furthermore, nms are used as sensitive and efficient pathogen diagnostic tools in the form of mirna-based diagnosis, nano-biosensors, nano-barcoding, and quantum dots, among others. the writers of the review paper anticipate that it will meet the fundamental needs of upcoming students and researchers in nanoscience who want to concentrate on the function and use of nms in agriculture. as nanotechnology research progresses, the potential applications of nanoparticle-mediated smart delivery systems are growing, opening up new avenues to enhance sustainability, nutritional value, and agricultural output. however, issues including np biocompatibility, environmental impact, and regulatory considerations must be appropriately addressed as this strategy becomes more and more viable in agriculture. through the introduction of innovative techniques for applying fertilizers, insecticides, and other items, nanotechnology has the potential to support ecologically friendly agriculture and food sciences research, 2024, 11(2): 203-221 218 © 2024 by the authors; licensee asian online journal publishing group and sustainable agricultural practices. however, recent studies indicate that if nanoparticles are not applied carefully, they may partially damage beneficial microorganisms in the rhizosphere, including earthworms. this may possibly augment raising safety concerns in food and agriculture. regulatory bodies have yet to adequately define nanopesticides and there is a dearth of understanding about their consequences on pesticide resistance. challenges include a lack of regulatory standards, expensive manufacturing costs and public opinion. bridging the knowledge gap between farmers and stakeholders, as well as designing scalable solutions that are compatible with current farming techniques is critical for effective adoption of nano-based strategies for future smart agriculture. to overcome these obstacles and realize the full potential of nanotechnology in agriculture, industry and researchers must work hand-in-hand. risk assessment and regulatory approval depend on long-term field experiments with reliable data. in addition, stakeholders should be informed about the storage conditions and cytotoxicity problems connected with specific nanoparticles and efforts should be made to use green and natural materials for nanoparticle production to reduce environmental dangers. overall, interdisciplinary collaboration and comprehensive research are crucial to developing agricultural nanotechnology ethically. list of abbreviation: nms: nanomaterials; nps: nanoparticles; pgrs: plant growth regulators; ros: reactive oxygen species; cuo: copper oxide; zno: zinc oxide; mgo: magnesium oxide; mgoh: magnesium hydroxide; al2o3: aluminum oxide; tio2: titanium dioxide; au: gold; cuonps: copper oxide-based nanomaterials, znonps : zinc oxide-based nanopesticide, mgohnps: magnesium hydroxide-based nanopesticide, mgonps : magnesium oxide-based 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licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 65-75, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6167 © 2024 by the authors; licensee asian online journal publishing group inventory of the risks of contamination of milk from the neighbourhoods in traditional farms in the poro region in côte d'ivoire frederic harding kouaho1 abdoulaye toure1 abdoulaye soumana gouro2 ( corresponding author) 1,2abdou moumouni university of niamey, bp 10896 niamey, niger. 1email: hardingkouaho@gmail.com 1email: tourabdoulaye@yahoo.fr 1laboratory of biotechnology and valorization of agroresources and natural substances, university peleforo gon coulibaly, bp 1328 korhogo, côte d'ivoire. 2email: abdoulayegouro@yahoo.fr abstract this study, which highlights the situation of dairy farms as well as their development prospects, took place from may to october 2022 in the poro region of côte d'ivoire. and focused on the sociodemographic, zootechnical and health characteristics of 45 traditional farms with a total of 360 dairy cows. trade, the main activity for 51.1% of the owners, followed by agriculture 31.11%, for 88.88% of the indigenous senufo herders and 93.33% are non-indigenous fulani. feed is pasture for 75.6% and 24.44% of farmers providing a feed supplement. the age of first calving is between 3 and 4 years old or 5 to 6 years old. cows that calved the most at the age of 4 years by 46.51% compared to those calved 3 years ago 30.23%, 5 years 19.77% and 6 years 3.49% at the regional level. the frequency of milking per season varies significantly from 0.5 litres/day to 15 litres/day depending on the breeds observed (p<0.05). the main parasitic diseases are trypanosomiasis 28.88%, a bi-infection (safa (foot-and-mouth disease), bovine nodular cutaneous skin disease (bncsd)) 20% and contagious bovine pleuropneumonia (cbpp) 13.30%. for 80% of farmers, the prevalence of clinical mastitis is 60% and 20% estimate the prevalence of subclinical mastitis to be 40%. the milk is harvested by the fulani and 33.33% sell the milk to wholesalers in each department of the poro region. the amount of unsold milk 22.22% filtered for some, boiled (10 to 15 minutes). this study highlights a lack of hygiene during milking and on farms. it is therefore important to carry out mastitis screening actions in traditional dairy farms in the poro region and to structure it. keywords: dairy cow, farm situation, ivory coast, mastitis. citation | kouaho, f. h., toure, a., & gouro, a. s. (2024). inventory of the risks of contamination of milk from the neighbourhoods in traditional farms in the poro region in côte d’ivoire. agriculture and food sciences research, 11(2), 65–75. 10.20448/aesr.v11i2.6167 history: received: 8 october 2024 revised: 11 november 2024 accepted: 18 november 2024 published: 29 november 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by regional center of excellence on pastoral production: milk, meat, hides and skins (rcepp). institutional review board statement: the ethical committee of the university abdou moumouni of niamey, niger has granted approval for this study on 7 november 2021 (ref. no. bp 10896). transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: conceived, planned the experiments, performed the laboratory analyses, collected milk samples, processed cases and prepared the article, f.h.k.; helped with the orientation in the treatment of aspects, contribution to additional reference literature for the improvement of the article and final formatting, a.t.; contributed to the literature and directed the article, a.s.g. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 66 2. materials and methods ................................................................................................................................................................... 66 3. results ................................................................................................................................................................................................ 67 4. discussion .......................................................................................................................................................................................... 73 5. conclusion ......................................................................................................................................................................................... 74 references .............................................................................................................................................................................................. 75 mailto:hardingkouaho@gmail.com mailto:tourabdoulaye@yahoo.fr mailto:abdoulayegouro@yahoo.fr https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6167 https://orcid.org/0009-0007-8882-5419 https://orcid.org/0000-0001-7963-3297 https://orcid.org/0000-0003-0871-6014 agriculture and food sciences research, 2025, 11(2): 65-75 66 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the study provides information on the practices used in traditional dairy farms, to inventory the risks associated with the contamination of milk in the neighborhoods and then provides information on the different diseases on the farms, which is special unlike other studies in this region. it also lays bare the values on the duration of treatment of the herd of the different farms. 1. introduction côte d'ivoire depends on the sahelo-sudan region for its supply of livestock meat [1]. indeed, livestock farming plays a significant role in human nutrition through the production of milk and red meat in particular, and is a source of income for producers and farmers [2]. the deficit in animal protein, particularly in milk and dairy products, is a permanent problem facing côte d'ivoire. its national production covers 51% of national meat consumption, and only 17% of that of milk and dairy products [3]. thus, in order to meet its ever-increasing needs for milk and milk-derived products, the ivorian state has opted for the development of the local milk sector [4]. the state, through donors, has financed various projects, including, among others, the african development bank (bad)-livestock projects phase i and ii, which saw the creation of 12 dairy cattle farms, and the southern dairy project financed by belgian technical cooperation (btc) [5]. with the aim of reducing its dependence on milk and bovine protein from sahelian and european countries [2]which dependence gives rise to products that often do not comply with health requirements. however, despite all efforts, the results of improving milk production are mixed. if the quantity of local milk is insufficient, the hygienic and sanitary quality of the milk produced could be improved in order to make the local sector competitive. in addition, the decade of socio-political crisis has led to the absence of administration in certain areas, particularly the savanes district, and would have led to the destructuring of achievements in cattle breeding [5]. consequently, the dairy potential has been weakened due to the deterioration in the health status of farms, thus causing a resurgence of many diseases, particularly mastitis. it is therefore with the aim of taking stock of the situation with a view to setting up a dairy model in this region considered favorable to livestock farming that this study was established in traditional farms in the poro region of côte d'ivoire. the objective of the work was to take stock of the risks of contamination of neighborhood milk in traditional farms in the poro region of côte d'ivoire by applying survey sheets aimed at assessing the sociological characteristics of the actors, zootechnical characteristics of the farms, the hygiene and health conditions of the farms, as well as the circuit of collection, distribution, processing and trade of neighborhood milk. based on this diagnosis, recommendations for improving dairy farming on farms at each farm visited in the poro region are proposed. 2. materials and methods study area: the region subject to this study is that of poro, it is part of the savannah district. the savannah district is made up of four regions including that of poro which are; the tchologo, bagoué and kabadougou regions with a total area of 62,396 km². figure 1. map of the poro region showing the different departments visited. source: bnetd/cct,2012 p:wgs 1984, zone 30n design and realization:kouaho f.harding et zie mamadou ouattara, 2024 agriculture and food sciences research, 2025, 11(2): 65-75 67 © 2024 by the authors; licensee asian online journal publishing group the poro region is a territorial community in the north of côte d'ivoire whose name comes from the cultural practice of the people of this locality. it is located between 8°30 and 11° north latitude, 5° and 6° west longitude and covers a total area of 13,400 km². the relief of the savannah district is characterized by its horizontal plane made of plains and plateaus, the monotony of which is broken, in places, by the appearance of chains of hills or rocky domes that vary between 400 and 600 m in altitude [6]. on the other hand, the hydrographic network of the poro region is crossed by permanent watercourses such as the bandama blanc, the badénou and the bou [6]. however, the vegetation of this sudanese sector is essentially made up of gallery forests, open forests and savannahs. the savannahs are wooded, shrubby and grassy and as for the sub-sudanese sector, it is defined by a few islands of dense forests, sometimes humid [7]. the poro region includes the departments of korhogo, m'bengué, sinematiali and dikodougou, with the city of korhogo as its capital. in this region, the study was conducted in 45 localities spread across each department of the poro region; we have the villages and the different sub-prefectures figure 1. 2.1. material to facilitate the link between breeders and carry out this survey, it was a question of submitting a request to the regional management of mafr (ministries of animal and fisheries resources) of korhogo and the departmental directors of mafr of the poro region for technical input in the field by their agents. 2.2. methods the study conducted allowed the collection of data on breeding practices, milking management, feed used and the marketing of fresh milk. 2.3. sampling the farms studied are chosen randomly; these farms belong to indigenous people (natives) and non-indigenous people (foreigners), taking into account their availability to participate in the study. the survey covered 45 farms spread across the four (4) departments (korhogo, sinématiali, dikodougou and m'bengué) of the poro region. 2.4. progress of the investigation a survey with a single-pass questionnaire covering farms in the poro region was conducted among breeders and owners. its implementation required a pre-survey among technicians in the poro region in order to select the farms constituting the object of study. the aim was to collect information intended to identify and characterize dairy production systems, the mode of evolution of the farms and the practices of breeders. data were collected on the characteristics of the farms, on the health status of the herd during the study period, on the feeding of the animals, the hygiene conditions on the farm, the conditions of milking milk in order to determine the risk factors for contamination of milk at production. the identification of artisans, the circuit after milking cow's milk and dangerous practices were noted. basically, the questionnaire took into account socio-demographic data, the zootechnical and health characteristics of the farms studied. 2.5. analysis of survey data descriptive statistics were performed for all variables. in order to facilitate data interpretation, qualitative variables were reorganized into dichotomous variables after the fact. the survey forms were entered using the excel 2016 spreadsheet and served as a statistical analysis tool. 95% confidence intervals were calculated for proportions [8]. the difference between variables was considered significant at p < 0.05. the variables of interest are presented in the form of frequency tables. 3. results 3.1. neighborhood milk production system 3.1.1. sociological characteristics the people questioned are mostly breeders (herdsmen) after, owners, as well as people who are both owners and managers in a small proportion. the owners of the farms visited are mostly 88.88% natives of the region are senufo and mostly men. on the other hand, 93.33% of the breeders are non-natives (peuls) who come either from burkina-faso, mali and niger. agriculture, in particular that of cotton, cereals (corn, sorghum, fonio, rice) and peanuts is also an important activity (31.11%) of the breeders after that of trade 51.11%; owners who are mostly illiterate (table 1). it is also observed that the majority of non-natives are breeders, breeders at the same time (farmers and traders) and civil servants who are turned to the activity (figure 2). on the other hand, most of the natives are farmers. figure 2. presentation of the activity and ownership of the owners. agriculture and food sciences research, 2025, 11(2): 65-75 68 © 2024 by the authors; licensee asian online journal publishing group table 1. level of education and activity of owners of cattle farms. study activity none koranic primary secondary superior frequencies breeders 2 2 1 0 0 5 (11.11%) traders 7 6 4 6 0 23 (51.11%) cultivators 3 4 2 5 0 14 (31.11%) officials 0 0 0 2 1 3 (6.66%) total 12 12 7 13 1 45 3.2. zootechnical characteristics of farms 3.2.1. racial composition the survey in the forty-five (45) farms visited in the poro region made it possible to identify five types of breeds (baoulés, méré, n'dama, zébu and métisse); three (03) of which have different phenotypes of cattle which constitute the livestock. and is made up of taurins (bos taurus) (n'dama, baoulé), the méré breed (cross between male zebu and female baoulé), zebus (bos indicus) which is the cross between the zebu ( gobra x goudali ) and crossbreeds resulting from crosses between zebus and taurins or the cross between ( local breeds x exotic breeds ), resulting from artificial insemination were observed in the department of m'bengué [9]. the n'dama is short on legs, a breed with long horns and no hump. the baoulés are small cattle with a massive head and short horns. for the zebu, it is a large, robust animal with a hump and exists in red and light gray colors for the most part. the méré is a small breed with a single color and has a hump without curvature. the mixed breed, for its part, comes from the cross between a local and exotic breed with different colors and a very thick shape presenting a significant milk production than other local breeds. the cattle raised in the region are mostly from other countries (figure 2). 3.2.2. herd composition the number of cows and calves in the parks was 844, the table shows the number of animals by category according to sex and the number of calves by department. the percentage of animals present in each department is 28.43% for korhogo, 32.58% for dikodougou, 29.97% for sinématiali and finally 28.79% for m'bengué. the farms visited have a workforce that varies from 26 to 200 heads of cattle (table 2). 3.2.3. animal feeding the feeding practice observed in the region by the breeders is mainly grazing 75.6% and this, for years. compared to 24.44% of the herdsmen who bring surpluses in terms of food, purchased or produced on site (table 3). the frequency varies significantly according to the farms (p < 0.05). in some places, the herds are led to the pasture every morning by the herdsmen before sunrise then return only at nightfall and others lead to the pasture around 6 am and only return around 11 am or 2 pm (in order to increase their production). during the rainy season, the majority of breeders lead their herds to the classified forests (for those who have the means) to avoid disputes and fines between farmers in case the herd ravages a plantation. the distance traveled for grazing depends on the season. however, several breeders do not use salt crystals (nacl) at the level of the region 88.88%. a : n'dama race b: baoulé race c: zebu breed d: méré breed agriculture and food sciences research, 2025, 11(2): 65-75 69 © 2024 by the authors; licensee asian online journal publishing group e: mixed race figure 3. shows the breeds of cows in the farms visited table 2. cow numbers by department and by category. department staff farms visited dairy cows heifers 1st calving veau farms visited korhogo 12 96 16 22 106 korhogo dikodougou 9 72 23 12 87 dikodougou sinematiali 12 96 33 24 100 sinematiali m'bengue 12 96 17 28 102 m'bengue total 45 360 89 86 395 total table 3. feeding conditions for lactating dairy cows. power condition variables staff numbers of farms (n= 45) % 95% ci origin of the water used at the farm level (here) well 14 31.11 [24.4-38.3%] pond 8 17.78 [6.2-27.8%] river 23 51.11 [36.5-65.5%] salt crystals (nacl) (absent) nacl 40 88.88 [78.5-97.5%] food practice at the herd level (here) pasture 34 75.56 [62.4-87.6%] food supplement (cotton cake, cassava peel, corn bran, rice bran, sorghum, etc.) 11 24.44 [11.6-36.4%] note: ci : confidence interval; n = number of farms where the variables are observed 3.3. management and productivity of cattle farms 3.3.1. stable productivity the results obtained concern productivity, it is based on the data obtained at the level of the questions asked to the breeders of the poro region. however, the data were obtained according to the experience of the herdsmen who made predictions on the average age at first calving and the months of lactation of the calf. the herdsmen interviewed (84.44%) estimate that the primiparous is between 3 years or even 4 years normally and this depends on the feeding of the herd (table 4). because, we could not observe primiparous cows of 5 to 6 years in farms in the poro region see table. this table shows that in the poro region, the cows that have calved the most at 4 years 40/86 are more numerous than those who calved 3 years ago 26/86, 5 years 17/86 and 6 years 3/86 at the level of the region. 3.3.2. milk production most of the time, milking is done once a day early in the morning for each cow. calves are separated from dairy cows, when they return from the pasture to the cattle pen. the majority of shepherds (95.55%) milk for years early in the morning (around 6 am and 7 am). however, they admit that milk production obtained per cow during the dry season remains low compared to that of the rainy season and according to the breeds observed (table 5). the frequency of milk per season varies significantly according to the breeds observed (p < 0.05). the milk obtained remains for the herdsmen after production, the milk is packaged in plastic cans or plates with capacities varying between 2 and 15 liters without being filtered for the most part (table 5). in the presence of the calf, milking is carried out in unhygienic practices. 3.3.3. hygiene on farms among the farms visited during our survey, 24.44% of the farms are mud, 17.77% dry soil and 57.77% wet soil were observed. the frequency of soil scraping per year is 15.55% once a year, 40% 2 times a year and other 20%. however, the presence of fresh stools is 71.11% compared to its absence which is 28.89% observed in the region. agriculture and food sciences research, 2025, 11(2): 65-75 70 © 2024 by the authors; licensee asian online journal publishing group the frequency of the variables varies significantly according to the farms (p < 0.05). no milking hygiene was observed (hand washing before milking, cows' teat cleaned before milking and disinfection using alcohol) (table 6 ). table 4. year of first calving in the different stables. departments cow of 3 years cow of 4 years cow of 5 years cow of 6 years old staff % korhogo 8 12 2 0 22 (25.59) sinematiali 6 10 8 0 24 (27.90) dikodougou 4 4 3 1 12 (13.96) m'bengue 8 14 4 2 28 (32.55) total 26 40 17 3 86 (100) table 5. quantity of milk production by breed according to seasons. quantity of milk produced per season mother n'dama baoule zebus metis dry season 1 to 2.5 liters 1 to 2 liters 0.5 to 1 liter 1.5 to 3 liters 6 to 8 liters breeders' response 8 (17.77%) b 14 (31.11%) a 13 (28.88%) a 8 (17.77%) b 2 (4.44%) c rainy season 1.5 to 4 liters 1.5 to 3 liters 1 to 2 liters 2 to 6 liters 10 to 15 liters breeders' response 11 (24.44%) b 10 (22.22%) b 15 (33.33%) a 7 (15.55%) b 2 (4.44%) c 95% ci dry season 4.5 12.5 10.6 4.5 7 95% ci rainy season 6.4 5.3 13.9 3.3 7 table 6. characteristics of the farms visited. features variables staff numbers of farms (n= 45) % 95% ci physical characteristics of the soil mud 11 24.44 [11.6-36.4%] dry soil 8 17.77 [6.2-27.8%] wet ground 26 57.77 [43-71%] presence of fresh stool presence 32 71.11 [57.8-84.2%] absence 13 28.89 [15-29.3%] frequency of soil scraping 1 time per year 7 15.55 [25.7-54.3%] 2 times a year 18 40 [29.5-58.5%] other 20 44.45 [25.7-54.3%] note: ci: confidence interval; n = number of farms where the variables are observed 3.4. health situation of farms 3.4.1. dominant infections observed in the poro region pleuropneumonia (cbpp), trypanosome, dermatosis (bncsd), safa (foot-and-mouth disease), soumaya-fi (black fever), symptomatic anthrax, hyperkeratosis; are the pathologies encountered in the poro region (north of ivory coast). parasitosis, especially trypanosomiasis (28.88%), is the dominant pathology in the region after the survey. it is followed by a bi-infection safa (foot-and-mouth disease) and bovine nodular skin disease (bnsd), which account for a significant proportion of 20% (figure 4). respiratory diseases, especially contagious bovine pleuropneumonia (cbpp), constitute the third most described pathology (13.30%) in the poro region. this disease manifests itself clinically by dyspnea, contamination due to a heterogeneous assembly of animals and, in terms of lesions, hepatization of the lungs, which is subject to seizure after inspection at the slaughterhouse. on the other hand, a tri-infection (trypanosome, safa and cbpp) was also mentioned (11%) in the farms visited. this is followed by biinfections of 8.88% (cbpp and trypanosome) and 4.44% respectively of (cbpp and safa) followed by (safa and bncsd). dermatosis is a skin disease that can affect the entire body of the animal and has many symptoms (skin spots, redness, sweating, fungus, skin infection, skin discoloration and pimples). a local disease under the name of "soumaya-fi 6.66%" which means "black fever" was mentioned. symptomatic anthrax was reported in the department of sinématiali of 2.22%, caused by ingestion of contaminated food and leads to acute inflammation of the intestinal tract. symptoms include nausea, flesh turns black, loss of appetite, vomiting and fever followed by abdominal pain, vomiting blood and diarrhea. the symptoms described indicate a disease of unexpected onset that affects animals in good condition (well-fleshed bodies). it causes rapid weight loss of the animal. at the advanced stage of the pathology, mortality linked to the disease is low and there is no relapse despite the treatments applied (vaccination and others) on the farms. therefore, on the instructions of the park manager, the breeders slaughter the sick animal because it is difficult to cure. general signs and certain diseases such as bellowing, aggressiveness, food deficiency and hyperkeratosis have been reported by the shepherds of the region. 3.4.2. medical prevention at the level of the farms surveyed during the vaccination period, there were few farms that confirmed vaccinations (15.56%). self-treatments carried out in the farms in the event of illness are (44.88%) (table 7). on the other hand, some breeders questioned (55.12%) say they use private technicians or mafr agents (ministries of animal and fisheries resources) to treat their animals. during the campaign or vaccination period by private veterinarians in the poro region, some breeders say they received their herd vaccination twice (2) in the year, i.e. 35.56%, others often, i.e. 28.88%, and for some once (1) in the year. the prevalence of the variables varies significantly depending on the farms (p<0.05). private breeders are the only ones authorized to carry out treatments on animals in the poro region. however, in the event of a shortage of private technicians, they ask for note: nb: a, b and c proportions of the same column assigned different letters are significantly different. agriculture and food sciences research, 2025, 11(2): 65-75 71 © 2024 by the authors; licensee asian online journal publishing group help from technicians or agents of mafr (ministries of animal and fisheries resources) to carry out the vaccination campaign. 3.4.3. how cows are treated the survey in the forty-five (45) farms visited in the poro region made it possible to identify the methods of treating cows, 18% do the treatment individually at the level of each cow, 82% in groups and none at the drying off is observed. on the other hand, the duration of treatment of sick cows is 88.89% from 2 to 3 days and 11.11% from 1 to 2 weeks table 8. however, the use of appropriate antibiotics was never observed in 51.11% of cases, often in 31;11% and applied in 17.78% in the farms; treatment during milking was never observed in 60% of the farms, often in 28.89% of the farms and applied in 11.11% of the farms. the application of brands on animals was observed in 17.78% of farms, often in 48.89% and never in 33.33% of farms. figure 4. different types of disease encountered in the exploitation of the poro region. note: trypa = trypanosome; cbpp=ppcb = contagious bovine pleuropneumonia; safa = foot and mouth disease; cbnsd=dncb= contagious bovine nodular skin disease; soumaya-fi = black fever nb: safa, soumaya-fi (are vernacular names). table 7. prevention on farms. vaccination variables staff numbers of farms (n=45) prevalence% 95% ci after the vaccination period treatment followed 25 55.55 a [33.3-76.9%] self-medication 20 44.45 b [29.5-58.5%] during the vaccination period often 13 28.88 a [15-29.3%] none 7 15.56 b [4.6-25.4%] 2 times a year 16 35.56 a [21.148.9%] 1 time a year 9 20 b [8.331.7%] note: nb: a and b prevalences of the same column assigned different letters are significantly different. table 8. treatment of cows on farms by department. variables features number of breeders n=45 % duration of treatment 2 to 3 days 40 88.89 1 to 2 weeks 5 11.11 use of appropriate antibiotics never 23 51.11 often 14 31.11 application 8 17.78 treatment of cows during milking never 27 60 often 13 28.89 application 5 11.11 marks on animals never 15 33.33 often 22 48.89 application 8 17.78 3.5. mastitis manifestation 3.5.1. clinical mastitis among the clinical and epidemiological criteria useful for the diagnosis of clinical mastitis , 78% of the farmers in the sample use the criterion of asymmetry of the quarters (figure 5) and 84% use udder deformation. the definition of clinical mastitis also includes the notions of change in udder color and modification of the appearance of the milk for 49% and 41% of cases. for the majority of farmers 36% and 34% of the sample, during clinical mastitis, the udder presents a change in udder color, lesion of the teats hence inflammatory risk while this inflammation was not a criterion for 16% of them. for 80% of farmers after survey, estimate that the prevalence of clinical mastitis is greater than 60%. agriculture and food sciences research, 2025, 11(2): 65-75 72 © 2024 by the authors; licensee asian online journal publishing group 3.5.2. subclinical mastitis subclinical criteria used for the diagnosis of subclinical mastitis, 91% of the sampled farmers use the criterion of quarter asymmetry, 96% use teat lesion and 89% use udder deformation. the definition of subclinical mastitis contains respectively the notions of change in udder colour, change in milk appearance in 75% and 76% of cases (figure 6). for a minority of farmers (8% of the sample), during subclinical mastitis, the udder has a persistent imprint on the skin, while no observation was made at the cmt (california mastitis test) test level and skin mobility 0%. additional examinations can be useful in this case for the diagnosis of mastitis; as the cmt test will be welcome on farms. after investigation, the minority of breeders questioned (20%) believe that the prevalence of subclinical mastitis (inflammations, skin sores, etc.) in dairy farms does not exceed 30%. 3.5.3. review of the first drafts according to the questionnaire results, most of the breeders practice the elimination of the first jets in the farm (100%) in a traditional way (the calf suckles before any milking of the cow), and there is a lack of homogeneity (pus and lump) during milking in the farms visited. however, there is a presence of white color coloration in the milk of 75.56% followed by a liquid viscosity 66.67% and a yellow color in 24.44% followed by a heavy viscosity 33.33% in the farms (table 9). however, milk harvesting is carried out by fulani people who often have family ties with the herdsmen. indeed, 33.33% sell their milk to the main seller who collars motorcyclists (wholesalers) in each department of the poro region (korhogo, dikodougou, m'bengué and sinénatiali) (figure 7). urban and rural consumers are also served by the herdsmen or resellers, i.e. 44.44%. the quantity of unsold milk (22.22%) is filtered for some, boiled (about 10 to 15 minutes) and processed on site, see diagram (figure 8). figure 5. signs of clinical mastitis according to breeders. figure 6. signs of subclinical mastitis according to breeders. note: cmt (california mastitis test). agriculture and food sciences research, 2025, 11(2): 65-75 73 © 2024 by the authors; licensee asian online journal publishing group table 9. analysis of first drafts. variables features staff numbers of farms n= 45 % milk color yellow 11 24.44 white 34 75.56 milk color heavy 15 33.33 liquid 30 66.67 figure 7. distribution of milk in the farms visited. figure 8. distribution diagram and consequences linked to the consumption of local milk from the farms visited. 4. discussion the survey conducted in the poro region highlighted the conditions of cow's milk production and the assessment of the risk of mammary contamination of these. the study showed that the owners of the farms visited are of senufo ethnicity and mainly represented in the poro region by 88.88%. on the other hand, 93.33% of allogeneic (peuls) is the main ethnic group that runs this field of activity. indeed, this high frequency of peuls could be explained by the fact that livestock breeding is the activity that is transmitted from one lineage to another in this ethnic group. however, the major activity of the natives is focused on agriculture. this remark was made by hamadou, et al. [10] in the peri-urban area of bobo-dioulasso in burkina faso where livestock farming is dominated by non-natives 87.13% and a minority is made up of natives 2.69%. agriculture, livestock farming and trade are the activities most practiced by the inhabitants of this area. this could be explained by the fact that its inhabitants have land, but also by the fact that livestock farming is a means of saving agricultural and commercial income, particularly those from the cultivation of corn, cotton, peanuts, etc. this is in correlation with the results of [5] who state that the majority of livestock farmers practice both agriculture and livestock farming. also, the minority of livestock farmers on the farms visited are not illiterate 26.66% and 15.55% of livestock farmers have a primary level. this could be explained by the youth schooling policy implemented by the ivorian government; which was not the case a few years ago for this ethnic group. the fact that these breeders found in breeding a main resource. our results are in the same direction as those of koffi [11]. these authors observe that during their studies, the majority of the breeders met were schooled. as for the characteristics of the farms visited, night parks are used by the majority of breeders and in a traditional way. the latter use small areas which could be explained by the more or less reduced size of the herds which range from 26 to 200 heads. these results are different from those found by assani [12] who notes larger areas used by breeders. the high proportion of the méré breed in the farms of the poro region could be explained on the one hand by the ability of this local breed to resist trypanosomes which is one of the dominant pathologies in this region. other breeds, although in the minority, are also observed in the poro region; zebu and crossbreed breeds are breeds with high milk production observed. these results are consistent with those observed by kouaho, et al. [9] who state that zebu are the least numerous in the agriculture and food sciences research, 2025, 11(2): 65-75 74 © 2024 by the authors; licensee asian online journal publishing group north region. feeding is the pillar of dairy farming. thus, in the study, most breeders use natural fodder 75.6 % as their main feed. this could be explained by the fact that in the extensive system, the herds are mainly fed on natural pasture. however, grazing as the only way to feed livestock is one of the pillars of livestock profitability. this feeding method remains consistent with that observed by marichatou, et al. [13] in extensive production systems. also, most breeders do not use any feed supplement. this could be explained on the one hand by the insufficiency of feed produced by the industry and the high cost of this feed. on the other hand, since milk production is not an objective for most of its owners, the meadows, in addition to their free access, would only serve to cover the needs of the animals. nevertheless, the minority of owners using a supplement composed of rice bran, corn, cottonseed cake and crop residues could be considered as the beginning of intensification. in addition, the majority of breeders water their herds in rivers. this could be explained by the fact that, in the traditional system, animals are generally brought to natural parturitions. thus, during their wanderings, the herd is watered with water from watercourses, including rivers, streams and other natural sources. these results are consistent with the system described by chayer [14]. collecting milk in cans is a characteristic specific to all breeders (collectors) in this region. this preservation technique can affect the appearance of fresh milk. during transport, some natural factors can influence the condition of the milk. in particular, heat and turbidity, which can accelerate the milk fermentation process. this collection system is close to that described by bonfoh, et al. [15] and corniaux, et al. [16]. however, at the level of milk production, the difference between the quantities of milk produced in the farms would be explained by the breed observed at the herd level. indeed, the crossbreed breed and the zebu breed have a low number of suckler cows observed in the region with a high production per season. thus, the average daily production observed after the survey varying from 0.5l/cow to 15l/cow per breed of cows is between that obtained by coulibaly [17] which is 0.88 l/cow for the same rainy season in mali. however, our results are similar to those of bonfoh, et al. [15]. the latter obtained daily quantities varying between 0.5 and 3.5 l / cow and according to the seasons. these results could reflect a better performance of the breeds observed if ever the breeding conditions are taken into account and executed. then, the low production observed in certain farms and the fluctuation noted by seasons could be explained by the feed ration of the herd, the practice of milking, vaccinations in the farms and the hygiene practice implemented. on the other hand, trypanosomiasis and cbpp are among the dominant pathologies in the region which is consistent with those obtained by marichatou, et al. [13] and by coulibaly [17] in mali and burkina faso. this could be explained by the transhumance of animals in these neighboring countries. the lack of private veterinarians and other livestock technical agents leads farmers to resort to vaccination campaigns, carried out by livestock technicians in order to protect their animals. the advance of pathologies, in particular trypanosomiasis, cbpp and foot-and-mouth disease, could affect the productivity of the region. with regard to farmers who take care of their animals themselves, one would be tempted to say that this livestock system consumes very few veterinary drugs. this confirms the results obtained by vias, et al. [18] in the peri-urban area of niamey in niger where the majority of farmers use traditional drugs. in addition, the very insufficient number of private veterinarians, qualified technicians and livestock engineers could explain this observation. a study even in france by pauline [19] among the clinical and epidemiological criteria used for the diagnosis of subclinical mastitis, 78%, 48% and 43% of veterinarians use the increase in the concentration of somatic cells in milk, the absence of damage to the general condition and modification of the milk. and for a minority 27%, the udder does not present an inflamed appearance. which confirms our study, among the clinical and epidemiological criteria used for the diagnosis of subclinical mastitis, 91% of the breeders in the sample use the criterion of asymmetry of the quarters, 96% the lesion of the teat and 89% the deformation of the udder. and 75%, 76% of cases, observe a change in udder color, the modification of the appearance of the milk and an absence of inflammation of the udder. on the other hand, this observation was moderately high for clinical mastitis on both sides. among the clinical and epidemiological criteria useful for the diagnosis of clinical mastitis, 78% of the farmers in the sample use the criterion of asymmetry of the quarters and 84% use the deformation of the udder. the definition of clinical mastitis also includes the notions of change in udder color and modification of the appearance of the milk for 49% and 41% of cases. for the majority of farmers 36% and 34% of the sample, during clinical mastitis, the udder presents a change in the color of the udder, lesion of the teats hence inflammatory risk while this inflammation was not a criterion for 16% of them. here, the minority of farmers surveyed (20%) consider that the prevalence of subclinical mastitis (inflammations, skin sores, etc.) in dairy farms does not exceed 30%. for 80% of them, the prevalence of clinical mastitis is greater than 60%. this is not consistent with the results of pauline [19]. on the other hand, the absence of collectors in some surrounding villages and the distance greater than 30 km could, on the one hand, explain the sale of milk to markets and self-consumption. on the other hand, the cost related to transport could be a brake on milk trade in general. what could explain the prevalence of mastitis of 80%, in traditional farms of the poro region according to the study remains high and close to that conducted in the maghreb and east africa where prevalences can reach 89% [20]. our results are higher than those obtained in local breeds of niger (djelli , azawak , bororo, goudali) with a prevalence of 44.2% [21] as well as the prevalence of 44.71% found in benin in local breeds [22]. 5. conclusion the methodological approach was based on a survey, with attention paid to the description of the actors and practices in traditional dairy farms. the survey was conducted among 45 breeders comprising 844 cattle distributed in the 4 departments of the region. the data were obtained through interviews based on a single-pass questionnaire. the data analysis made it possible to establish descriptive statistics on the data of the sociological and zootechnical characteristics of the farms, the hygienic and sanitary conditions of the farms, as well as the circuit of collection, distribution and sale of unpasteurized milk. from this analysis, remarks can be made. agriculture, an important activity constitutes nearly (31.11%) of breeders after that of trade 51.11% and owners who are mostly illiterate. the animals are taken to pasture for the most part, mainly in the mornings and only return at nightfall. few breeders say they provide a food supplement. the age at first breeding is between 3 to 4 years and 5 to 6 years for some. for milk production, the frequency of milk per season varies significantly depending on the breeds observed; although the quantities remain low in the dry season. the dominant pathologies are respiratory pathologies, namely contagious bovine pleuropneumonia (cbpp) (28.88%), followed by a biagriculture and food sciences research, 2025, 11(2): 65-75 75 © 2024 by the authors; licensee asian online journal publishing group infection safa (foot-and-mouth disease) and bovine nodular skin disease (bnsd), 20%. concerning medical prevention, at the level of the farms surveyed during the vaccination period, there are few farms that confirm vaccinations (15.56%). self-treatments carried out on farms in the event of illness are (44.88%). on the other hand, some breeders (55.12%) say they use private technicians or mafr agents in the event of illness. among the clinical and epidemiological criteria used to diagnose clinical mastitis, 78% of the breeders in the sample use the criterion of asymmetry of the quarters and 84% use udder deformation. and for the subclinical criteria used to diagnose subclinical mastitis, 91% of the breeders in the sample use the criterion of asymmetry of the quarters, 96% use teat lesion. finally, for the collection and distribution of milk, 33.33% sell their milk to the main seller who collars motorcyclists (wholesalers) in each department of the poro region, then urban and rural consumers are also served by herdsmen or resellers, i.e. 44.44%. the quantity of unsold milk (22.22%) is filtered for some, boiled (about 10 to 15 minutes) and transformed on site into milk. references [1] a. azokou, y. achi, and m. koné, "control of livestock ticks in côte d'ivoire using traditional ways," livestock research for rural development, vol. 28, no. 52, 2016. http://www.lrrd.org/lrrd28/4/azok28052.htm [2] b. hafidaha and m. fayza, "contribution to the management of dairy cattle breeding in the walaya of ain defla case of: the sidi belhadj farm," thesis for obtaining the master's degree. department of agronomic sciences, animal production, 2019. 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[22] m. kadja, "study of subclinical mastitis in dairy cattle farms in west africa: case of senegal and benin," doctoral thesis, abomey-calavi university, cotonou, 156, 2010. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. http://www.lrrd.org/lrrd28/4/azok28052.htm https://doi.org/10.1002/(sici)1097-0258(19980430)17:8%3c857::aid-sim777%3e3.0.co;2-e https://ssrn.com/abstract=4860471 https://doi.org/10.19182/remvt.9973 https://doi.org/10.1016/j.landusepol.2015.06.031 187 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 187-194, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6251 © 2024 by the authors; licensee asian online journal publishing group preparation of chemically crosslinked chitosan hydrogels with adipic acid and edac carbodiimide: tailored biomaterials for multifunctional applications talita martins1 ezequiel de souza costa junior2 eduardo henrique martins nunes3 ( corresponding author) 1,3federal university of minas gerais, belo horizonte/mg, brazil. 1email: martins.talita2009@gmail.com 3email: eduardo.nunes@demet.ufmg.br 2federal center for technological education of minas gerais, brazil. 2email: escjr50@cefetmg.br abstract hydrogels are versatile biomaterials with significant potential in agriculture, food preservation, and biomedical applications. hydrogels are three-dimensional networks of hydrophilic polymers that can absorb and retain large amounts of water or biological fluids. among the polymers suitable for hydrogel synthesis, chitosan is particularly notable due to its biocompatibility, biodegradability, and antimicrobial properties. this study investigates chitosan characteristics, focusing on its various properties and how they influence its potential applications in the synthesis of chitosan hydrogels crosslinked with adipic acid using the water-soluble carbodiimide n-(3dimethylaminopropyl)-n'-ethylcarbodiimide hydrochloride (edac) to develop multifunctional films for practical applications. the raw materials were characterized using potentiometric titration, fourier transform infrared spectroscopy (ftir) and x-ray diffraction (xrd). the hydrogels were analyzed for morphology and crystallinity through scanning electron microscopy (sem), ftir and xrd, comparing samples with and without the addition of the carbodiimide crosslinker. quantitative analysis of swelling behavior was also conducted using standard buffer solutions. the results showed that crosslinking with adipic acid enhanced the crystallinity of chitosan, while the inclusion of edac reduced crystallinity and the degree of swelling, indicating improved crosslinking between polymer chains. these changes in structural and functional properties demonstrate the adaptability of these hydrogels for specific applications. this study underscores the potential of chitosan-based hydrogels as advanced biomaterials. keywords: adipic acid, chemical crosslinking, chitosan, edac, hydrogel, biomaterials. citation | martins, t., junior, e. de s. c., & nunes, e. h. m. (2024). preparation of chemically crosslinked chitosan hydrogels with adipic acid and edac carbodiimide: tailored biomaterials for multifunctional applications. agriculture and food sciences research, 11(2), 187–194. 10.20448/aesr.v11i2.6251 history: received: 31 october 2024 revised: 29 november 2024 accepted: 5 december 2024 published: 26 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by fundação de amparo à pesquisa do estado de minas gerais (fapemig) (grant number: bpd-00883-22). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 188 2. experimental ................................................................................................................................................................................... 188 3. results and discussion ................................................................................................................................................................. 189 4. conclusion ....................................................................................................................................................................................... 193 references ............................................................................................................................................................................................ 193 mailto:martins.talita2009@gmail.com mailto:eduardo.nunes@demet.ufmg.br mailto:escjr50@cefetmg.br https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6251 https://orcid.org/0000-0002-0309-9849 https://orcid.org/0000-0003-0925-6400 https://orcid.org/0000-0001-6653-5137 agriculture and food sciences research, 2024, 11(2): 187-194 188 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study is original in its investigation of chitosan hydrogels crosslinked with adipic acid using edac, offering an alternative to commonly used acids. this method enhances properties of chitosan, with potential applications in agriculture and food sciences, particularly in controlled release and preservation technologies. 1. introduction in the field of biomaterials research, the development of novel hydrogels has been a focal point for several years, driven by their potential applications in extending food shelf life, advancing agricultural practices, and supporting biomedical innovations. a critical aspect of these studies lies in the selection of polymers for hydrogel fabrication. this selection must consider the intrinsic properties of the polymers, ensuring they synergistically impart the desired functional characteristics required for the specific target application [1-3]. in the focus of this development, biomaterials for use in a multifunctional application such as and agriculture and food science represent a major area of research today [1]. in these fields, the compatibility between the material and its intended application is crucial for success. hydrogels exhibit properties that make them highly appealing, such as their ability to retain water, hydrophilicity, expandability, selective permeability, soft consistency, and low interfacial tension [4, 5]. chitosan (chi) is one of the most promising polymers for hydrogel applications, as it predominantly consists of d-glucosamine units derived from the n-deacetylation of chitin, a natural component of crustacean and insect exoskeletons [6, 7]. chitosan stands out due to its remarkable properties, including biocompatibility, biodegradability, antimicrobial activity, and high adhesiveness, largely attributed to its polycationic nature [7]. hydrogels are advanced materials characterized by crosslinked polymer networks that form three-dimensional structures [8]. the degree of crosslinking is a critical parameter, as it directly impacts the hydrogel's physicochemical and mechanical properties. subtle variations in crosslinking density can alter key attributes such as swelling behavior, elasticity, permeability, and structural stability [9]. therefore, precise control over the crosslinking process is essential to tailor hydrogels for specific applications, ensuring optimal performance and functionality. crosslinking is a crucial process in hydrogel synthesis, involving the interconnection of polymer chains through reactions with low molecular weight crosslinkers containing at least two functional groups. this process results in a more robust, three-dimensional polymer network, enhancing the structural integrity and mechanical properties of the hydrogel. importantly, crosslinking agents employed for hydrogels designed as biomaterials must be non-toxic to ensure safety and suitability for their intended applications [10, 11]. the objective of this work is to evaluate the feasibility of synthesizing chitosan-based hydrogels chemically crosslinked with adipic acid through a reaction mediated by the water-soluble carbodiimide n-(3dimethylaminopropyl)-n'-ethylcarbodiimide hydrochloride (edac). these hydrogels are designed to function as biomaterials with versatile and multifunctional applications. 2. experimental 2.1. materials chitosan powder (cat. # 419419, sigma-aldrich chemical, milwaukee, wisconsin, usa), high molecular weight, mw= 340.000g/mol, degree of deacetylation, dd= >75%, was used without further purification. adipic acid (vetec química). n-(3-dimethylaminopropyl)-n'-ethylcarbodiimide hydrochloride (edac) (fluka). all salts and reagents used were of analytical degree and milli-q water was used in all solutions (18.0 mω). 2.2. chitosan hydrogels preparation chitosan hydrogels were synthesized by dissolving 0.5 g of chitosan in 50 ml of a 1% (w/w) adipic acid solution under stirring at 60 °c until complete dissolution. gradually 4 ml of naoh (0.1 m) were added with continuous stirring, followed by cooling the solution to 40 °c. to initiate the chemical crosslinking, three formulations were prepared by varying the addition of edac (carbodiimide): sample a (without edac), sample b (0.06% w/v edac), and sample c (0.11% w/v edac). gelation was monitored during the stirring process, and the resulting mixtures were uniformly distributed onto petri dishes. after 7 days at room temperature, the hydrogels were dried at 40 °c for 24 hours. the formed films were washed thoroughly with 5% (w/w) acetic acid, followed by rinsing with milli-q water to remove residual reagents. the films underwent a final drying step in an oven at 40 °c for 24 hours to yield the crosslinked chitosan hydrogels. 2.3. raw materials characterization the degree of deacetylation (dd) of chitosan was performed by the potentiometric titration method in phmeter digimed d-23 which 0.1 g of chitosan were dissolved in 10 ml of standard solution of 0.1 m hcl for protonation of present amino groups. the solution was then introduced in the previously calibrated phmeter and initiated titration with standardized naoh 0.1 m. a typical potentiometric titration curve was obtained [12]. the titration values were applied to equation 1, where m represents the sample mass, na and nb are the concentrations of the acid and base, respectively, and va and vb are their corresponding volumes. the molar masses of the deacetylated and acetylated monomers were considered as 161 g/mol and 204 g/mol, respectively. the calculated degree of deacetylation (dd) was then compared with the reference value provided by the manufacturer. 𝜑 = 𝑁𝐴𝑉𝐴 − 𝑁𝐵𝑉𝐵 / 1000 𝐷𝐷% = (𝜑 / 𝑀 − 161𝜑 204 + 𝜑) 𝑥 100 (1) the analysis of x-ray diffraction (xrd) of the chitosan and adipic acid was performed on shimadzu xrd 7000 from the powdered material with 2θ ranging from 3.0° to 40.0° with a step 0,06°. chitosan was evaluated based on agriculture and food sciences research, 2024, 11(2): 187-194 189 © 2024 by the authors; licensee asian online journal publishing group the monoclinic and orthorhombic structures using it for copper kα radiation6 the areas under the curve were determined using baseline correction and integration, identifying the crystalline fraction (ac) and amorphous fraction (aa). these areas were calculated using the microcal origin® 8.5 software. the degree of crystallinity (gc) was then estimated using equation 2. 𝐺𝐶 = 𝐴𝐶/(𝐴𝐶 + 𝐴𝐴) × 100 (2) ftir spectra of the samples were acquired using a bruker alpha ii ft-ir spectrometer, operating at a resolution of 4 cm⁻¹ with 32 scans accumulated for each measurement. 2.4. hydrogels characterization qualitative visual observations were made taking into account the solubility, miscibility, coloring and phase segregation of the films. the average film thickness was measured with a mitotoyo (â±10μm) micrometer. morphological analysis of the films was performed in scanning electron microscopy (sem), equipment shimadzu ssx-550. the samples were covered with a thin gold layer by "sputtering" and positioned at the the maximum distance of the target, thus preventing damage to the films. the x-ray diffraction (xrd) was performed on shimadzu xrd 7000 equipment with 2θ ranging from 3.0 ° to 40.0 ° with step 0.06°. the degree of crystallinity of the films was calculated using the same methodology as applied to the raw materials. the swelling degree (sd) determination, the films were carefully weighed and transferred to an erlenmeyer flask containing 50 ml of buffer solution at ph 4 and ph 7 (hanna instruments, brazil). the samples were immersed in the solutions for 120 minutes, with swelling measurements taken at regular intervals (30, 60, 90, and 120 minutes) at ambient temperature. the swelling degree was calculated based on the variation between the initial mass (m0) and the final mass (mf), as described in equation 3. 𝑆𝐷 = 𝑀𝑓 𝑥 𝑀0 / 𝑀0 𝑥 100% (3) 3. results and discussion figure 1 shows a typical potentiometric titration curve obtained from the chitosan solution, where the points of inflection correspond to neutralization of the excess hcl solution and neutralization of free amino groups present in the sample respectively. for better observation figure 1 shows the values of the first derivative of chitosan’s potentiometric titration curve. figure 1. potentiometric titration curve of chi and first derivative curve. based on the observed curve and the calculations performed using equation 1, which relies on the volume of naoh consumed at the second inflection point, the molar quantity of -nh₂ groups in the sample was determined. this analysis yielded a degree of deacetylation (dd) of approximately 76% ± 6, aligning closely with the value provided by the manufacturer (>75%) for the raw material. figure 2 shows the diffractogram obtained from the sample of chitosan where two characteristic peaks are observed, the first of high intensity at 19.8° (d = 0.441 nm) and a lower intensity at 37.83° (d = 0.244 nm). the diffractogram highlights the crystalline phase of the sample; however, a broad baseline below the peaks indicates the presence of an amorphous phase in the feedstock. the observed angles in the diffractogram can be attributed to specific crystallographic planes (2 0 0) for the high-intensity (1 2 4) to the low intensity. from the diffractogram of chitosan, it was possible to estimate the crystallinity by 17% [13, 14]. figure 3 shows the diffractogram obtained from the sample of adipic acid where four peaks of highest intensity are observed, the first at 12.9° (d = 0.669 nm), the second high intensity at 21.4° (d = 0.411 nm), the third at 25.7° (d = 0.345 nm) and fourth peak at 31.2° (d = 0.289 nm) after analysis and calculations it was observed that the sample is 100% crystal clear. agriculture and food sciences research, 2024, 11(2): 187-194 190 © 2024 by the authors; licensee asian online journal publishing group figure 2. diffractogram of raw material chitosan. figure 3. diffractogram of the raw material adipic acid. figure 4 shows the ftir spectrum of chitosan, highlighting its characteristic functional groups. key absorption bands include the broad band around 3400 cm⁻¹, attributed to o-h and n-h stretching vibrations, and peaks near 2900 cm⁻¹ associated with c-h stretching. the amide i band, typically found around 1650 cm⁻¹, and the amide ii band, near 1550 cm⁻¹, are indicative of residual acetyl groups. additionally, the peak at approximately 1375 cm⁻¹ corresponds to the c-h bending of the ch3 groups, while the region around 1150-1025 cm⁻¹ reflects the c-o-c stretching of the glycosidic bonds in the polysaccharide backbone. these spectral features confirm the molecular structure and degree of deacetylation of the chitosan sample [15, 16]. agriculture and food sciences research, 2024, 11(2): 187-194 191 © 2024 by the authors; licensee asian online journal publishing group figure 4. ftir spectrum of chitosan showing its characteristic functional groups. figure 5 presents chitosan and adipic acid films in three conditions: without crosslinking (film a), crosslinked with 0.06% edac (film b), and crosslinked with 0.11% edac (film c). all films exhibit a thickness of approximately (75 ± 25μm). figure 5. chitosan films with adipic acid were: (a) 0.0%, (b) 0.6% and (c) 0.11% of edac. a qualitative analysis of the films was performed through visual observation, and the results are summarized in table 1. this analysis provides insights into the physical appearance, homogeneity, transparency, and surface characteristics of the films under different crosslinking conditions. table 1. evaluation by qualitative observation of the films produced. identification of hydrogels segregation opacity coloring flexibility a segregated opaque yellowish flexible b segregated opaque yellowish flexible c segregated transparent portions yellowish flexible figure 6 shows the sem images of the films, revealing dendritic formations resulting from the crystallization process during film formation. these formations are noticeably reduced with the addition of edac, indicating its effect on modifying the film structure. comparing the images of films a, b, and c, film c appears to exhibit a more amorphous structure, likely due to the increased crosslinking in comparison to films a and b. figure 6. images from films of chitosan and adipic acid with (a) 0.0%, (b) 0.6% and (c) 0.11% edac obtained by sem. agriculture and food sciences research, 2024, 11(2): 187-194 192 © 2024 by the authors; licensee asian online journal publishing group in analyzing the diffractogram of the films compared to that of pure chitosan (raw material), it was observed that areas of the peaks representing the chitosan showed a relative decrease mainly after the addition of edac. the relative crystallinity of the films was higher than that of pure chitosan due to the high crystallinity obtained with the addition of adipic acid. after the addition of the activator (edac), there is a reduction in crystallinity from the diffractograms of the films b (48%) and c (45%) compared with the film a (88%) (figure 7), suggesting that the occurrence of crosslinking reduces the mobility of the chains, preventing their crystalline organization. this result was anticipated, as the addition of edac promotes chemical crosslinking, which takes precedence over the ionic crosslinking naturally present in chitosan films when combined with diacids. this shift enhances the structural integrity and reduces the crystallization tendencies of the films [9, 17]. figure 7. diffractogram of the films a (edac 0,00%), b (edac 0,06%) and c (edac 0,11%). figure 8 illustrates the swelling curves of the films at ph 4, while figure 9 shows the swelling degree of the films at ph 7. these figures provide a comparative analysis of the films' responsiveness to different ph conditions, highlighting the influence of ph on their swelling behavior. figure 8. profile of swelling of the films a (edac 0,00%), b (edac 0,06%) and c (edac 0,11%) at ph 4. agriculture and food sciences research, 2024, 11(2): 187-194 193 © 2024 by the authors; licensee asian online journal publishing group figure 9. profile of swelling of the films a (edac 0,00%), b (edac 0,06%) and c (edac 0,11%) at ph 7. the swelling process occurs in three main steps: 1) diffusion of solvent molecules into the polymeric network; 2) relaxation of polymer chains as a result of hydration; and 3) expansion and swelling of the polymeric network, also known as matrix swelling. however, as the degree of crosslinking increases, fewer hydrophilic groups are exposed to water, reducing their ability to diffuse through the polymer network. consequently, this results in a lower swelling degree for films b and c compared to film a, which has a lower degree of crosslinking [18, 19]. 4. conclusion chitosan films were prepared using adipic acid, with or without the addition of the acid activator, a watersoluble carbodiimide. the characterization of the raw material not only confirmed the chemical composition of the material but also allowed for analysis and comparison with the films produced. the film characterization revealed that the use of adipic acid as a crosslinking activator enhances the reactivity of the films due to its functional groups, promoting covalent crosslinking over ionic crosslinking. additionally, changes in the crystallinity pattern of the films were observed. the addition of carbodiimide resulted in an increased amorphous portion of the film, which correlated with a decrease in the degree of swelling. this reduction in swelling is associated with more efficient crosslinking within the films. references [1] m. klein and e. poverenov, "natural biopolymer‐based hydrogels for use in food and agriculture," journal of the science of food and agriculture, vol. 100, no. 6, pp. 2337-2347, 2020. https://doi.org/10.1002/jsfa.10274 [2] h. zhang, f. zhang, and r. yuan, applications of natural polymer-based hydrogels in the food industry. in hydrogels based on natural polymers. amsterdam, the netherlands: elsevier, 2020. 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licensee asian online journal publishing group [14] p. cairns, m. j. miles, v. j. morris, m. j. ridout, g. j. brownsey, and w. t. winter, "x-ray fibre diffraction studies of chitosan and chitosan gels," carbohydrate research, vol. 235, pp. 23-28, 1992. https://doi.org/10.1016/0008-6215(92)80075-c [15] t. martins et al., "novel 3d composites with highly flexible behavior based on chitosan and bioactive glass for biomedical applications," materials chemistry and physics, vol. 189, pp. 1-11, 2017. https://doi.org/10.1016/j.matchemphys.2016.12.012 [16] y. zhang, c. xue, y. xue, r. gao, and x. zhang, "determination of the degree of deacetylation of chitin and chitosan by x-ray powder diffraction," carbohydrate research, vol. 340, no. 11, pp. 1914-1917, 2005. https://doi.org/10.1016/j.carres.2005.05.005 [17] t. phaechamud and j. charoenteeraboon, "antibacterial activity and drug release of chitosan sponge containing doxycycline hyclate," aaps pharmscitech, vol. 9, pp. 829-835, 2008. https://doi.org/10.1208/s12249-008-9117-x [18] t. zhu et al., "recent progress of polysaccharide‐based hydrogel interfaces for wound healing and tissue engineering," advanced materials interfaces, vol. 6, no. 17, p. 1900761, 2019. https://doi.org/10.1002/admi.201900761 [19] o. b. assis, "alteration of the hydrophilic character of chitosan films by hmds plasma treatment," química nova, vol. 33, pp. 603606, 2010. https://doi.org/10.1590/s0100-40422010000300022 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/0008-6215(92)80075-c https://doi.org/10.1016/j.matchemphys.2016.12.012 https://doi.org/10.1016/j.carres.2005.05.005 https://doi.org/10.1208/s12249-008-9117-x https://doi.org/10.1002/admi.201900761 https://doi.org/10.1590/s0100-40422010000300022 52 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 52-58, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.5995 © 2024 by the authors; licensee asian online journal publishing group the grasshopper arcyptera (pararcyptera) meridionalis ikonnikov (insecta, orthoptera) as a possible agricultural pest in temperate asia oxana v. yefremova1 vladimir v. molodtsov2 sergey yu. storozhenko3 anna a. shamychkova4 michael g. sergeev5 ( corresponding author) 1,2,5department of general biology and ecology, novosibirsk state university, novosibirsk, russia. 1email: oxana@fen.nsu.ru 2email: vv@fen.nsu.ru 1,2,4,5laboratory of biogeomodelling and ecoinformatics, novosibirsk state university, novosibirsk, russia. 4email: a.shamychkova@g.nsu.ru 3federal scientific center of the east asia terrestrial biodiversity, far eastern branch of the russian academy of sciences, vladivostok, russia. 3email: storozhenko@biosoil.ru 5institute of systematics and ecology of animals, siberian branch, russian academy of sciences, novosibirsk, russia. 5email: mgs@fen.nsu.ru abstract the goal of the article is to estimate possible changes of the distribution of arcyptera meridionalis in the contemporary and potential future climatic conditions. the data were collected during field trips. two packages to produce the species distribution models, namely maxent and ellipsenm, were used. the generated models predict that the status of a. meridionalis as the important pest will not change significantly in the nearest future, however, in the middle of this century, its distribution patterns will become more complicated. the areas those will be the most suitable for the species will remain mainly in the western and central parts of its range. the harmful activity of a. meridionalis can be especially important due to its early hatching, because hoppers may damage and destroy shoots with first leaves and tillers. the models generated for a. meridionalis look like quite different from the models for other steppe acridids. on the contrary, some resemblance between forecasts for two harmful, but quite different grasshopper species, namely siberian-mongolian a. meridionalis and east-mediterranean a. labiata, are revealed. the models for both species demonstrate some possible significant depletion of the territories with suitable conditions for each one in the second half of the 21st century. keywords: acrididae, climate change, ellipsoid envelope model, forecast, grasslands, maxent, modelling, pest management, plant protection, steppe. citation | yefremova, o. v., molodtsov, v. v., storozhenko, s. y., shamychkova, a. a., & sergeev, m. g. (2024). the grasshopper arcyptera (pararcyptera) meridionalis ikonnikov (insecta, orthoptera) as a possible agricultural pest in temperate asia. agriculture and food sciences research, 11(2), 52–58. 10.20448/aesr.v11i2.5995 history: received: 10 july 2024 revised: 9 september 2024 accepted: 20 september 2024 published: 2 october 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by russian science foundation (grant number: 16-04-00706). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: field collecting of grasshoppers, geographic coordinate determination, specimen identification, o.v.y., v.v.m., s.y.s., a.a.s. and m.g.s.; conceptualization, methodology, supervision, project administration, funding acquisition, writing, m.g.s.; data analysis, s.y.s., o.v.y. and a.a.s.; coordinate checking, map producing, modelling, v.v.m. and m.g.s.; validation, o.v.y. and m.g.s.; general discussion and text editing, s.y.s. and m.g.s. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 53 2. materials and methods ................................................................................................................................................................... 53 3. results ................................................................................................................................................................................................ 54 4. discussion .......................................................................................................................................................................................... 57 references .............................................................................................................................................................................................. 57 mailto:oxana@fen.nsu.ru mailto:vv@fen.nsu.ru mailto:a.shamychkova@g.nsu.ru mailto:storozhenko@biosoil.ru mailto:mgs@fen.nsu.ru https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.5995 https://orcid.org/0000-0002-1562-5252 https://orcid.org/0000-0001-9169-9934 https://orcid.org/0000-0001-9269-4043 https://orcid.org/0009-0001-9458-2712 https://orcid.org/0000-0003-2179-0921 agriculture and food sciences research, 2024, 11(2): 52-58 53 © 2024by the authors; licensee asian online journal publishing group contribution of this paper to the literature this is a first attempt to characterize the main ecologo-geographical peculiarities of the important acridid pest arcyptera meridionalis. the species distribution over the grasslands of south siberia, mongolia and china is described in details, and the several models of this distribution are produced for the contemporary and future climatic conditions. 1. introduction the grasshopper arcyptera meridionalis (figure 1) is the member of the subgenus pararcyptera serg. tarbinsky which includes several important pests, such as a. microptera (fischer de waldheim) and a. labiata (brullè) [1]. all or almost all members of this subgenus prefer grasses and, as result, may significantly damage both different crop fields (e.g. wheat, rye, barley, and oat), pastures, and hayfields, especially in the eurasian steppe and forest-steppe life zones [1, 2]. a. meridionalis per se may be the very serious pest across the eastern parts of the altai-sayan mts. and the southern parts of east siberia, and also in mongolia and the northern parts of china [1, 3-5]. because the species is an early hatching grasshopper, in many regions, the first hoppers can emerge in the end of april and in may and the adults are in june [6]. this means a. meridionalis is able to damage seriously shoots of different crops in the end of spring and in the beginning of summer. the current climate change, mainly associated with global warming, may result not only in species range shifts, but also in alteration of species harmful activities relative to some possible changes in agricultural practices. consequences of such transformations may result, inside the modern range, in both increase of the species devastating effects on agricultural fields, pastures and hayfields, and northward shifts in the species distribution. the goal of this article is to estimate possible changes of the species distribution in the contemporary and potential future climatic conditions. figure 1. arcyptera meridionalis (female) in the typical habitat (dry steppe) of central tuva (republic of tyva, russia) (photo m. g. sergeev). 2. materials and methods 2.1. territory of investigations the southern parts of siberia and the russian far east were studied from 1978 until 2022. in the beginning of the 20th century, forest-steppes, steppes, and semi-deserts covered these vast areas, however, later many habitats were transformed into agricultural lands (fields and pastures) [7]. in the region, mean temperatures of the warmest month vary between 20 °c to 26 °c and the same for the coldest month are from –6 °c to –29 °c. mean annual precipitations vary between 230 to 950 mm. 2.2. field data acquisition the data used were collected during field trips. as a rule, different local habitats, particularly natural and seminatural grasslands and agricultural fields, were surveyed, commonly in the middle of summer. in each habitat, grasshoppers were observed and collected to reveal species richness, and their abundance was normally estimated by one or two methods, namely quantitative sampling during a fixed period of time [8] and/or the standard sweep nettings. geographic coordinates were evaluated by gps (global positioning system)/glonass (globalnaya navigatsionnaya sputnikovaya sistema) hand-held units. the google earth pro (©google 2022) was used to reveal the geographic coordinates for old collections. the main part of studied specimens is in the collections of novosibirsk state university, the institute of systematics and ecology of animals (novosibirsk), and the federal scientific center of the east asia terrestrial biodiversity (vladivostok). the data from different publications [919] and from the collections of the zoological institute (saint petersburg, russia) were also analyzed and used. as a result, our database contains the geographic coordinates of 73 points of the species occurrence. 2.3. species distribution mapping and modelling maps of species distribution were generated on the basis of geographic coordinates with qgis 3.18.3. a lambert conformal conic projection was selected as the basic map. two distinctive packages to produce the species distribution models over its whole range were used. the first one (maxent) is based on the machine learning [20agriculture and food sciences research, 2024, 11(2): 52-58 54 © 2024by the authors; licensee asian online journal publishing group 22]. the second one (ellipsenm in the r environment) is based on producing a multidimensional ellipsoid envelope model of an ecological niche [23]. both are limited by only presence data and depend on the number of localities, selected options of modelling and collection of variables used [20-23]. in the case of the maxent models, the full sets of the so-called standard bioclimatic variables [24, 25] were used. their accuracy was estimated by the area under the receiver operating characteristic curve values (auc) for 25 replicates with cross-validation. the possible effects of the bioclimatic variables were estimated by their predictive contributions and the jackknife tests. the following parameters were used for the maxent models: features — auto, output format — cloglog, regularization multiplier = 1. in the case of the ellipsoid models, the several selected bioclimatic variables, namely the annual mean temperature, maximal temperature of the warmest month, minimal temperature of the coldest month, annual precipitation, precipitation of the warmest quarter, precipitation of the coldest quarter, were used, because this algorithm is very sensitive to correlation between variables. 25 replicates were counted as well, the method was covmat and the level used to produce the ellipsoids was 99%. in all cases, the 30 arcsecond spatial resolution was selected. besides, the predicted averaged bioclimatic variables for 2021–2040 and 2041–2060 [24, 25] based on the global climate model cnrm-esm2-1 [26] and the 3-7.0 shared socioeconomic pathway (high greenhouse gas emissions [27]) were used to produce models for the future. 3. results 3.1. the actual species distribution the taxon was described from the vicinities of qiqihar (now in the heilongjiang province, north-east china) as arcyptera flavicosta meridionalis [9]. later sir b. uvarov [10] described another subspecies of a. flavicosta, namely a. f. sibirica, from the southern parts of yakutia and the baikal region, but, in almost all cases, without exact geographic localities. in the monograph of bey-bienko and mistshenko [28], these subspecies were synonymized. later storozhenko and paik [29] justified its status as the distinct species. the northern boundary of the species range is approximately defined by the southern border of the taiga life zone (about 53°n), and the southern one — by the semi-deserts (about 44°n) in the inner parts of eurasia and by the subtropics (about 39°n) near the pacific coast of the continent (figure 2). at least one insular population of the species was known from the southern parts of republic of sakha (yakutia) [10]. the species range includes the southern parts of central and east siberia (however, in the republic of khakassia and the southern parts of krasnoyarsk krai, a. microptera (fischer de waldheim) occurs) [30] almost all mongolia, except its southern arid parts [13, 15] the steppes and forest-steppes of north and north-east china [31, 32]. it is locally distributed in the southern parts of the russian far east [33, 34] in the northern parts of korean peninsula [14, 29, 35] and in the mountains of the north-eastern parts of the tibetan plateau [36]. over all range, a. meridionalis occurs primarily in semi-arid grasslands, including the mountain ones, because its trophic preferences are commonly limited by grasses (poaceae) [37]. figure 2. the general distribution of arcyptera meridionalis. note: 1 — type locality of a. flavicosta meridionalis; 2 — one of the type localities of a. flavicosta sibirica (with the geographic coordinates); 3 — all other localities. 3.2. the species distribution models the models of the a. meridionalis distribution for contemporary conditions show that the optimal areas for the species are in the steppes of the southern parts of central and east siberia, the northern parts of mongolia and the northwestern parts of inner mongolia (figure 3 and 4). some isolated territories are also in the angara river basin (near irkutsk), the southernmost regions of yakutia, and several areas of the temperate far east, where grasslands are relatively common. besides, the maximum entropy model shows the regions with very suitable conditions for this species outside the actual species range, namely in the mountains of east tien shan, where actually another taxon from this group (a. microptera turanica uv.) occurs. the maximum entropy model is well supported with auc = 0.958 (figure 5). agriculture and food sciences research, 2024, 11(2): 52-58 55 © 2024by the authors; licensee asian online journal publishing group figure 3. predicted probabilities of suitable conditions for arcyptera meridionalis (maxent model, all bioclimatic variables for 1970–2000; point-wise means for 25 replicates with cross-validation). figure 4. predicted probabilities of suitable conditions for arcyptera meridionalis (multidimensional ellipsoid model, selected bioclimatic variables for 1970–2000; point-wise means for 25 replicates). agriculture and food sciences research, 2024, 11(2): 52-58 56 © 2024by the authors; licensee asian online journal publishing group figure 5. reliability test for the arcyptera meridionalis model (all bioclimatic variables for 1970–2000; 25 replicates with cross-validation). several bioclimatic variables significantly contribute to the model. among them are precipitations seasonality (34.5%), annual mean temperatures (16.7), temperature seasonality (15), and annual precipitation (10.9%). the jackknife test enables to add several other variables, such as precipitation of the coldest quarter and minimum temperatures of the coldest month. hence, very low winter temperatures may be very important for the species, especially in south siberia and mongolia where they are often coupled with thin snow cover resulting in deep soil freezing and high levels of acridid egg mortality (cf. [19]). the possible changes in the a. meridionalis distribution based on the actual data concerning its localities and the high greenhouse gas emissions look like relatively moderate (figure 6). in the nearest two decades, the areas with very suitable conditions may remain almost the same. in 2041–2060, the optimal territory may significantly diminish, especially in the angara and selenga rivers' basins and in the far east. the main region of the possible harmful activity will likely remain in tuva in the central part of the altai-sayan mts. and in the adjacent areas of west mongolia. besides, some problems with a. meridionalis will persist in the southeastern parts of transbaikal region (dauria) in russia and in the northwestern parts of inner mongolia in china (figure 6, b). a b figure 6. predicted probabilities of suitable conditions for arcyptera meridionalis (forecasts of all bioclimatic variables for 2021–2040 (a) and 2041–2060 (b) according the global climate model cnrm-esm2-1 and the 3–7.0 shared socioeconomic pathway; point-wise mean for 25 replicates with cross-validation). agriculture and food sciences research, 2024, 11(2): 52-58 57 © 2024by the authors; licensee asian online journal publishing group 4. discussion our models predict that the status of the grasshopper a. meridionalis as the important pest species will not change significantly in the nearest future, however, in the middle of this century, its distribution patterns will become more complicated. the areas those will be the most suitable for the species will remain mainly in the western (tuva and north-west mongolia) and central (the south-eastern parts of transbaikal region and the north-eastern parts of inner mongolia) parts of its range. both territories are characterized by very high activity of herdsmen and density of grazing animals, especially sheep and goats. besides, many plots are ploughed and used as agricultural fields for spring wheat, rye, and barley. in outbreak seasons, a. meridionalis may severely damage local grasslands and fields, particularly in the end of spring and the beginning of summer when young hoppers start to emerge and become active and when, in the region with an extreme continental climate and main precipitations in the second half of warm season, there are a few green plants in the grasslands. such pattern is very typical for the siberian-mongolian steppes, where many grasshopper species are normally very abundant and the main period of their development is the middle and the end of summer [38]. moreover, the possible harmful activity of a. meridionalis can be especially important due to its early hatching, particularly relative to fields of spring wheat and rye, because hoppers may damage and destroy shoots with first leaves and tillers. the models generated for a. meridionalis look like quite different from the models for other steppe orthopteran species, both abundant (oedaleus decorus (germar) [39], bicolorana bicolor (philippi) [40]) and rare (miramiola pusilla (miram)) [41]. the models for these three species are similar, at least for the southern parts of west siberia, and show evident trend: noticeable northward shifts of the areas with very suitable conditions. however, surprisingly, there are some resemblance between forecasts for two related, harmful, but quite different grasshopper species from the subgenus pararcyptera, namely siberian-mongolian a. meridionalis and eastmediterranean a. labiata [42]. the models for both species demonstrate some possible significant depletion of the territories with suitable conditions for each one in the second half of the 21st century. references [1] l. l. mistshenko, orthoptera (saltatoria). in: o. l. kryzhanovsky and e. m. danzig, eds., insects and mites — pests of agriculture crops. leningrad: nauka publ, 1972. 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[41] m. sergeev and v. molodtsov, "new data on distribution of miramiola pusilla (miram, 1927)(orthoptera: tettigoniidae)," far eastern entomologist, vol. 16, no. 496, pp. 16–24, 2024. https://doi.org/10.25221/fee.496.4 [42] b. çıplak and o. uluar, "anatolian short-horned grasshoppers unveiled: integrating biogeography and pest potential," insects, vol. 15, no. 1, p. 55, 2024. https://doi.org/10.3390/insects15010055 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.25221/fee.389.2 https://doi.org/10.1093/jisesa/ieu171 https://doi.org/10.3390/insects12010077 https://doi.org/10.3390/insects13010049 https://doi.org/10.25221/fee.496.4 https://doi.org/10.3390/insects15010055 116 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 116-135, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6226 © 2024 by the authors; licensee asian online journal publishing group effect of salt stress on the growth and development of six species of turf grasses in the eastern region of morocco khadija charif1 maria rimani2 ibtissam mzabri1 said otouya3 noureddine kouddane1 rachid touzani4 larbi rhazi5 alicia ayerdi gotor6 abdelbasset berrichi1 ( corresponding author) 1laboratory, agricultural production improvement, biotechnology and environment, department of biology, faculty of sciences, university of mohammed premier, oujda, morocco. 1email: khadijachariff@gmail.com 1email: btissammzabri@gmail.com 1email: kouddanen@yahoo.fr 1email: abdel20759@yahoo.fr 2laboratory bioresources, biotechnology, ethnopharmacology and health, department of biology, faculty of sciences, university of mohammed premier, oujda, morocco. 2email: rimaniimaria@gmail.com 3national institute of agronomic research, crra oujda, 10 bd mohamed vi, bp 428, 6000oujda, morocco. 3email: otouya1433@gmail.com 4laboratory of applied chemistry and environment, department of chemistry, faculty of sciences, university of mohammed premier, oujda, morocco. 4email: r.touzani@ump.ac.ma 5institut polytechnique unilasalle, université d’artois, ulr 7519, unilasalle, 19 rue pierre waguet, bp 30313, 60026 beauvais, france. 5email: larbi.rhazi@unilasalle.fr 6institut polytechnique unilasalle, aghyle, up 2018.c101, unilasalle, 19 rue pierre waguet, bp 30313, 60026 beauvais, france. 6email: alicia.ayerdi-gotor@unilasalle.fr abstract the turfgrass plays a crucial role in landscaped ecosystems, including sports fields, golf courses, residential lawns, and parks. however, various turfgrass species face diverse abiotic stresses, including salinity as irrigation with fresh water becomes unavailable. these challenging environmental conditions can result in growth inhibition, damage to cellular structure, and metabolic disturbances. therefore, the use of salt-tolerant turfgrass species or varieties proves to be one of the most effective methods to mitigate salinity issues. with this aim, a greenhouse experiment was conducted at the experimental station of the faculty of sciences in oujda. the objective was to investigate the impact of saline stress on the morphological and physiological behavior of six grass species widely used in green spaces in morocco, particularly in the eastern region irrigated with 3 concentrations of saline water. the results revealed that the species a. stolonifera, f. arundinacea, and c. dactylon exhibited some tolerance to salinity, especially at a dose of 3 g/l of nacl, while the species l. perenne was found to be the most sensitive to saline stress, regardless of the applied dose. concentrations of 6 g/l and 9 g/l of nacl led to the decline of all studied species six months after application. keywords: morphological behavior, physiological behavior, proline, salinity, salt-tolerant species, soluble sugars, turfgrass. citation | charif, k., rimani, m., mzabri, i., otouya, s., kouddane, n., touzani, r., rhazi, l., gotor, a. a., & berrichi, a. (2024). effect of salt stress on the growth and development of six species of turf grasses in the eastern region of morocco. agriculture and food sciences research, 11(2), 116–135. 10.20448/aesr.v11i2.6226 history: received: 6 november 2024 revised: 10 december 2024 accepted: 13 december 2024 published: 20 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: the ethical committee of the university of mohammed premier, morocco has granted approval for this study on 26 june 2024. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. https://orcid.org/0000-0001-9445-4131 https://orcid.org/0000-0001-7333-697x https://orcid.org/0000-0001-6489-3109 https://orcid.org/0000-0001-6725-3334 mailto:khadijachariff@gmail.com mailto:btissammzabri@gmail.com mailto:kouddanen@yahoo.fr mailto:abdel20759@yahoo.fr mailto:rimaniimaria@gmail.com mailto:otouya1433@gmail.com mailto:r.touzani@ump.ac.ma mailto:larbi.rhazi@unilasalle.fr mailto:alicia.ayerdi-gotor@unilasalle.fr https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6226 agriculture and food sciences research, 2024, 11(2): 116-135 117 © 2024 by the authors; licensee asian online journal publishing group contents 1. introduction .................................................................................................................................................................................... 117 2. materials and methods ................................................................................................................................................................. 117 3. results .............................................................................................................................................................................................. 120 4. discussion ........................................................................................................................................................................................ 130 5. conclusion ....................................................................................................................................................................................... 132 contribution of this paper to the literature this study contributes to the existing literature by investigating the impact of saline stress on the morphological and physiological behavior of six grass species widely used in green spaces in morocco, particularly in the eastern region irrigated with 3 concentrations of saline water. 1. introduction salinity is a major problem worldwide, affecting around 900 million hectares, i.e. approximately 20% of land area and almost half of irrigated arable land [1]. semi-arid and arid zones are particularly affected by a significant accumulation of salts in the soil [2]. in morocco, soil salinisation is progressing in parallel with the expansion of irrigated areas, impacting around 500,000 hectares of arable land [3]. the harmful consequences of high salinity are felt throughout the life cycle of plants, from inhibition or delay of germination and growth to plant death [4, 5]. in a saline environment, plants must mobilise a series of physiological and biochemical mechanisms to cope with stress. these mechanisms include morphological and anatomical changes, adjustments in water relations and photosynthetic activity, alterations in hormone profiles, redistribution of toxic ions, as well as biochemical adaptations such as the activation of antioxidant metabolism responses [4, 6-9]. excess salts in water can cause damage to turf due to salinity and sodicity [10, 11]. according to qian and mecham [12] long-term irrigation with saline water, over a period of 4 to 33 years, led to an increase in soil electrical conductivity (ec) and sodium absorption rate, as well as elevated ph, sodium, boron and phosphorus levels on golf fairways. turf degradation has also been observed on courses that have been subjected to prolonged irrigation with effluents [12]. many researchers have studied the use of low-quality water for turf irrigation, focusing mainly on natural soils or sand-based artificial substrates [13-15]. leaching plays a crucial role in reducing or preventing salt stress in turf irrigated with salt water [16]. this process involves applying irrigation water volumes greater than the actual water requirements, calculated on the basis of evapotranspiration replenishment, in order to promote continuous vertical movement of salts [17]. choosing turfgrass species suitable for irrigation with saline water is essential to optimise maintenance costs while preserving turf quality [18]. in general, warm-season grasses have higher salt tolerance than cool-season grasses [19]. however, cool-season grasses are more appreciated compared to warm-season ones because these last species express winter dormancy which has as consequence the loss of the green colour during that period. tall fescue (festuca arundinacea (schreb). syn., schedonorus arundinaceus (schreb.) dumort) is the most cultivated cool-season grass in the mediterranean region thanks to its tolerance to the most common abiotic stresses characterizing this climate such as high summer temperatures combined with low or no precipitations, besides the resistance to salinity and the more technical advantage of resisting trampling [20]. literature shows that tall fescue survives with levels of soil salinity up to 10 ds m-¹, making this species moderately tolerant [21-23]. in morocco, grasses are widely used, particularly on football pitches and golf courses, sometimes covering several hundred hectares for a single golf course. these surfaces require large quantities of irrigation water, which is often of poor quality in several regions where these sports and tourist facilities are located. the use of saline water to irrigate lawns is becoming a growing problem [24, 25]. the unavailability of fresh water is forcing lawn managers to explore alternative solutions, such as finding grass species that are more tolerant of salt. the aim of this study is to analyse the impact of salt stress on the morphological and physiological responses of six grass species commonly used in green spaces in morocco, particularly in the eastern region. the aim is to provide information on which species are most resistant to salinity. 2. materials and methods 2.1. plant material the plant material used in this study consisted of seeds from six species of turfgrass, four of which are considered coolseason and two tropical. the selected species are as follows: festuca arundinacea, lolium perenne, poa pratensis, agrostis stolonifera, pennisetum clandestinum, and cynodon dactylon. seedling doses are those recommended for each species [26]. the seeds were donated by the seed producing company ꞌꞌles gazons de france ꞌꞌ, france and by ꞌꞌgolf saidia lacsꞌꞌ, saidia, morocco. 2.2. treatments used to assess the impact of saline stress on turfgrass, the grass, grown from germination and cultivation of the seeds of the studied species, was subjected, starting from january 2019 to three levels of salinity corresponding to three concentrations of nacl in the irrigation water: 3, 6, and 9 g/l. the control was irrigated with tap water from the experimental station, with an electrical conductivity (ec) of 0.7 ms/cm. these treatments will be referred to subsequently as: t0: control, irrigated with tap water. t1: 3 g/l nacl. t2: 6 g/l nacl. t3: 9 g/l nacl. agriculture and food sciences research, 2024, 11(2): 116-135 118 © 2024 by the authors; licensee asian online journal publishing group 2.3. installation and conduct of the experiment this experiment was conducted in a greenhouse at the research experimental station of the faculty of sciences in oujda, morocco. sowing was carried out in october 2018 in cylindrical pots with dimensions of 0.35 m deep by 0.22 m in diameter, filled with a substrate composed of peat and sand in a ratio of 2/3 peat to 1/3 sand by volume, preceded by a drainage layer of 1 cm. light watering was performed once daily to keep the substrate moist with nonsaline irrigation water. starting from january 2019, turfgrass irrigation was carried out based on water requirements, calculated using the reference evapotranspiration (et0) values for the city of oujda [27] and using greenhouse water requirements calculated using the following equation [28]: 𝐸𝑇𝑀𝑠 (𝑚𝑚/𝑚2) = 0.56 ∗ 𝐸𝑇0 + 0.7 (1) where: etms: maximum evapotranspiration under greenhouse conditions. et0: reference evapotranspiration. table 1 shows the quantities of water supplied according to the reference evapotranspiration of the city of oujda [27]. table 1. water quantities supplied for turfgrass species according to the imposed salinity stress based on the reference evapotranspiration of the city of oujda. month daily et0 (mm/day) etms (mm/m2) water quantity (ml/pot) dec -18 1.9 1.8 136 jan -19 1.8 1.7 132 feb -19 2.8 2.3 175 mar-19 3.5 2.7 205 apr -19 4.0 2.9 226 may -19 5.0 3.5 270 jun -19 6.0 4.1 313 jul -19 7.1 4.7 360 irrigations were carried out three times per week to facilitate the watering operation, and the turfgrass species were subjected to this stress for seven months. after establishment, the grass was mowed once a month to a height of 2 cm using a lawn shear. no fertilizers, herbicides, insecticides, or fungicides were applied throughout the duration of the experiment. the average, minimum, and maximum temperatures, as well as the humidity of the greenhouse, were measured daily, and the monthly averages are presented in the (figure 1). figure 1. monthly meteorological data in the greenhouse at the experimental station of the faculty of sciences in oujda for the experimental period from november 2018 to july 2019. 2.4. studied parameters the monitoring of the morpho-physiological behavior of these grasses regarding salt stress was conducted by assessing morphological, physiological, and biochemical parameters used in several studies focusing on plant response to various abiotic stresses. it is worth noting that the initial recordings were conducted after two months from the initiation of the different treatments (from early january to late february). 2.5. morphological parameters 2.5.1. length growth (cm) the length growth of the grass and internodes were evaluated each month using a graduated ruler. agriculture and food sciences research, 2024, 11(2): 116-135 119 © 2024 by the authors; licensee asian online journal publishing group 2.5.2. above-ground dry matter of mowing at the end of each month, the fresh weight (in grams) was determined by weighing the mowing product directly after harvesting. the dry weight (in grams) was determined after drying the mowing product in an oven set at a temperature of 80 °c for 72 hours. the dry matter content (%) is derived from the ratio: 𝐷𝑀 (%) = (𝐷𝑊/𝐹𝑊∗ 100) (2) where: dm is the dry matter; dw is the dry weight; and fw is the fresh weight. 2.5.3. above ground and below-ground biomass at the end of the 6-month experiment, the grass from each pot was unearthed, and the above-ground and belowground parts were separated, cleaned of substrate, dried in an oven at 80°c to constant weight, then weighed. the dry and fresh weights thus determined are expressed in grams. 2.5.4. determination of electrical conductivity the soil electrical conductivity of each sodium chloride concentration was measured at the end of the trial. soil electrical conductivity was measured using a 1:5 dilution of soil: water solution, since this technique enables us to determine the amount of salt present in a soil regardless of its degree of saturation [29]. 2.6. evaluation of the area covered by green grass 2.6.1. coverage rate at the end of each month, photos were taken using a camera to analyze and assess the turf coverage rate. the images were captured in jpeg (joint photographic experts group) format for processing using imagej [26]. 2.7. physiological parameters 2.7.1. chlorophyll pigment total chlorophyll content was measured using the method developed by tran, et al. [30] to asses the impact of saline irrigation on the 6 turf species. 1 g of fresh leaves at the end of the experimental irrigation period were ground and macerated with 10 ml of acetone diluted to 80% in a mortar. to avoid the oxidation, the chlorophyl extracted suspensions were protected from light with aluminum foils once recovered in glass tubes, prior to centrifugation at 2500g for 5 min. the absorbance of recovered surnatant was measured at 645 and 663 nm with a uv-vis spectrophotometer (humterlaba inc. reston, va, usa). the following formulas were used to calculate the total chlorophyll concentration (a+b): [𝐶ℎ𝑙 (𝑎)] = (12.7 × 𝐷𝑂663 − 2.63 × 𝐷𝑂645) × 𝑉/𝑀 (3) [𝐶ℎ𝑙 (𝑏)] = (22.9 × 𝐷𝑂645 − 4.68 × 𝐷𝑂663) × 𝑉/𝑀 (4) [𝐶ℎ𝑙 (𝑎 + 𝑏)] = (7.15 × 𝐷𝑂663 + 18.71 × 𝐷𝑂646) × 𝑉/𝑀 (5) where: chl (a): chlorophyll (a) (mg/g fw); chl (b): chlorophyll (b) (mg/g fw); chl (a+b): total chlorophyll (mg/g fw); v: volume of total extract (l); m: mass of fresh material (g). 2.7.2. relative water content (rwc) the determination of leaf relative water content was performed once a month using the method outlined by barrs [31]. this technique involves cutting the leaves at the base of the lamina and immediately weighing them to obtain their fresh weight (fw). subsequently, these leaves are placed in test tubes filled with distilled water and placed in a cool, shaded area. the leaves are retrieved after 24 hours and blotted dry with absorbent paper to remove surface water. they are then weighed again to obtain the weight at full turgor (ft). finally, the samples are placed in an oven at a temperature of 80°c for 48 hours to determine their dry weight (dw). the following formula by clark and mac-caig (1982), as cited by mouellef [32] is used to calculate the relative water content: 𝑅𝑊𝐶 (%) = [(𝐹𝑊 − 𝐷𝑊) / (𝐹𝑇 − 𝐷𝑊)]∗100 (6) where: rwc: relative water content (%). fw: fresh weight (g). ft: weight at full turgor (g). dw: dry weight (g). 2.8. biochemical parameters 2.8.1. proline proline (pyrrolidine-2-carboxylic acid) is one of the twenty primary amino acids that constitute proteins. it acts as an osmoprotector and an important signaling molecule that accumulates in the plant cytosol, functioning in membrane stabilization and protection. moreover, proline enhances water absorption and reduces the accumulation of toxic ions [33-35]. proline is highly soluble in water, methanol, and benzene and easily oxidized by ninhydrin or tricetohydrindene [36]. the quantification of the proline-ninhydrin reaction is performed using a spectrophotometer. proline reacts with ninhydrin to form a colored complex, and the intensity of the coloration is proportional to the amount of proline in the sample. the quantification method for this amino acid follows the procedure adapted by leport [37]. for the analysis, 100 mg samples are placed in a test tube, and 2 ml of 40% methanol is added. the samples are then heated in a water bath for one hour. after cooling, 1 ml of the extract is mixed with 1 ml of acetic acid and 1 ml of a solution composed of: distilled water (120 ml), acetic acid (300 ml), agriculture and food sciences research, 2024, 11(2): 116-135 120 © 2024 by the authors; licensee asian online journal publishing group orthophosphoric acid (80 ml). and added ninhydrin (25 mg). the optical density is measured using a uv-vis spectrophotometer (iso-ts 3632-1, 2003) at a wavelength of 528 nm. the obtained value is converted into proline concentration using a standard curve established from a series of solutions with known proline concentrations. the contents are expressed in mg of proline per g of fw (fresh weight). 2.8.2. soluble sugars maintaining normal water potential could be attributed to the accumulation of osmotica such as soluble sugars, which enable seedlings to maintain leaf turgor by reducing water potential and thus promoting water absorption despite the presence of salt in the soil. simple sugars (glucose, fructose, and sucrose) are extracted by a solvent capable of solubilizing them and inhibiting enzymatic activities that may degrade them. the assay was conducted referring to the method as reported by sidari, et al. [38]. the principle of the reaction is based on the condensation of neutral oses degradation products by sulfuric acid. the highly concentrated sulfuric acid transforms the oses under heat into furfural derivatives, which give a bluegreen coloration with anthrone. we begin by soaking 100 mg of fresh foliar material in 5 ml of 80% ethanol (v/v) for 24 hours. next, the obtained extract is diluted by mixing it with 80% ethanol at a ratio of 1:10. then, 2 ml of this diluted solution are taken and added to 4 ml of 0.2% anthrone reagent (0.2 g of pure anthrone + 100 ml of sulfuric acid (h2so4)). the mixture, containing both the extract and the reagent, must be kept in ice. subsequently, the tubes are placed in a water bath at 92°c for 8 minutes. after this step, the tubes are allowed to cool for 30 minutes in the dark, and then the optical density is measured at λ= 530 nm using a spectrophotometer uv-vis (humterlaba inc. reston, va, usa). the soluble sugar content of the leaves was calculated with reference to a standard glucose range. 2.9. experimental design the experimental design adopted for this trial is a randomized complete block, with three replicates (figure 2). the design is made up of three blocks corresponding to three replicates, i.e. a total of 12 pots sown for each species studied (number of treatments (4) x number of replicates (3)). figure 2. illustrative photo of the experimental set-up adopted for salt stress (february 2019). 2.10. statistical analysis results were subjected to descriptive statistical analysis and analysis of variance using spss software, version 20. means were compared using tukey's method. 3. results 3.1. effect of salt stress on morphological parameters 3.1.1. internode length the results indicate that salt has a depressive effect on final internode length. the length of the internodes is slightly modified under conditions of moderate stress (3 g/l nacl), particularly for the species a. stolonifera, f. arundinacea and c. dactylon. the sensitivity to salt stress of l. perenne, p. prantensis and p. clandestinum species varies with the amount of nacl applied. on the other hand, intense salt stress (6 g/l and 9 g/l) is extremely destructive from june onwards (5 months after the start of stress application) for l. perenne, p. pratensis and p. clandestinum species, and from july onwards for f. arundinace and c. dactylon species, which have completely suppressed their activity. during the summer, only a. stolonifera maintained a green level of 6 g/l (figure 3). agriculture and food sciences research, 2024, 11(2): 116-135 121 © 2024 by the authors; licensee asian online journal publishing group figure 3. effect of different levels of salinity (nacl) on the monthly average internode length among the six studied species. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). 3.1.2. grass length the results obtained for the length of the produced grass follow the same trend as those recorded for the internode parameter. there is a noticeable reduction for the severe treatments of 6 and 9 g/l for all species and a relative tolerance of the species a. stolonifera, f. arundinacea, and c. dactylon at 3 g/l. here again, only the species c. dactylon was able to maintain length growth at 6 g/l (figure 4). agriculture and food sciences research, 2024, 11(2): 116-135 122 © 2024 by the authors; licensee asian online journal publishing group figure 4. effect of different levels of salinity (nacl) on the monthly average grass length among the six studied species. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). 3.1.3. effect of salt stress on mowing dry matter for all studied species, both the control and lawns irrigated with water containing 3 g/l responded with a higher production of dry matter compared to other treatments. conversely, lawns treated with higher concentrations of nacl (6 g/l and 9 g/l) exhibited low production of dry matter, leading to grass death during the last two months of the trial, except for a. stolonifera at the 6 g/l nacl dose. significant differences were observed between treatments throughout the trial period (figure 5). species a. stolonifera appears resistant to doses of 3 g/l and 6 g/l of nacl, while species f. arundinacea and c. dactylon seem resistant to a dose of 3 g/l. however, species l. perenne, p. prantensis, and p. clandestinum are sensitive to saline stress regardless of the nacl dose applied. agriculture and food sciences research, 2024, 11(2): 116-135 123 © 2024 by the authors; licensee asian online journal publishing group figure 5. effect of different levels of salinity (nacl) on the monthly average dry matter among the six studied species. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). 3.1.4. effect of salt stress on above ground and below-ground biomass the results shown in figure 6 (a-b) demonstrate that the application of severe salt stress leads to a reduction in dry biomass both above and below ground. it is noted that for all species, severe treatments with 6 g/l and 9 g/l of nacl caused increasingly significant reductions in biomass towards the end of the trial. significant differences between treatments were observed for all species (figure 6). agriculture and food sciences research, 2024, 11(2): 116-135 124 © 2024 by the authors; licensee asian online journal publishing group figure 6. effect of different salinity levels (nacl) on the monthly mean above-ground (a) and below-ground (b) biomass for the six species studied. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). 3.1.5. effect of salt stress on substrate electrical conductivity measurements of electrical conductivity (ec) at the end of the trial showed a considerable increase in this parameter for all treatments. the results of the evolution of substrate ec show that the lowest substrate salinization was recorded in the control and the highest in the 9 g/l treatment. the high final electrical conductivities can be explained by the cumulative effect of the different watering solutions, leading to grass death at the end of the trial. results were significant between treatments for all species (figure 7). figure 7. effect of different salinity levels (nacl) on monthly mean electrical conductivity in the six species studied. note: significant differences are indicated as follows: *** very highly significant (p<0.001). 3.1.6. effect of salt stress on ground coverage ratio the ground cover ratio by the turfgrass (figure 8) decreased as the levels of salt stress increased, for all species. this reduction in coverage was observed as early as march for species l. perenne, p. prantensis, and p. clandestinum, particularly at high stress levels (6 and 9 g/l). for the same stress levels, the impact of salinity on this ratio only became apparent from may onwards for species a. stolonifera and c. dactylon, suggesting a relative tolerance for the latter. at 3 g/l, the reduction in the cover ratio was most pronounced for species l. perenne and p. prantensis as early as march. species a. stolonifera and l. perenne continued to provide good ground cover until july, when a decline in this coverage began (figure 9). agriculture and food sciences research, 2024, 11(2): 116-135 125 © 2024 by the authors; licensee asian online journal publishing group figure 8. effect of different salinity levels (nacl) on the monthly average cover rate for the six species studied. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). agriculture and food sciences research, 2024, 11(2): 116-135 126 © 2024 by the authors; licensee asian online journal publishing group figure 9. photos and diagram illustrating the effect of different levels of salinity (nacl) on greenness and turf cover in the six species studied (july 31, 2019) on greenness and turf cover in the six species studied (july 31, 2019); (t0: 0 g/l ; t1: 3 g/l ; t2: 6 g/l ; t3: 9 g/l). 3.2. effect of salt stress on physiological parameters 3.2.1. total chlorophyll overall, increasing salinity tends to decrease the total chlorophyll content for all species. indeed, the moderate level of stress (3 g/l) recorded concentrations exceeding those of the control (0 g/l) for all species. however, the application of high salinity levels (6 g/l and 9 g/l) leads to a decrease in total chlorophyll for all species. significant differences between treatments are observed throughout the trial period (figure 10). agriculture and food sciences research, 2024, 11(2): 116-135 127 © 2024 by the authors; licensee asian online journal publishing group figure 10. effect of different salinity levels (nacl) on monthly mean total chlorophyll (a+b) in the six species studied. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). 3.2.2. impact of salt stress on relative water content the data from figure 11 indicate that the presence of nacl led to a decrease in leaf water content. indeed, for all species, the lowest rates of relative water content were observed in lawns subjected to severe nacl treatments (6 g/l and 9 g/l). significant differences were noted between treatments throughout the trial period (figure 11). agriculture and food sciences research, 2024, 11(2): 116-135 128 © 2024 by the authors; licensee asian online journal publishing group figure 11. effect of different salinity levels (nacl) on monthly mean relative water content in the six species studied. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). 3.3. effect of salt stress on plant biochemical parameters 3.3.1. proline content the effect of nacl on proline content reveals that irrigating grass with high concentrations of nacl induces a significant accumulation of this solute. this accumulation is generally proportional to the nacl concentration in the irrigation water. the highest proline content was recorded for the 9 g/l nacl treatment in all species. there were significant differences between treatments throughout the test period (figure 12). agriculture and food sciences research, 2024, 11(2): 116-135 129 © 2024 by the authors; licensee asian online journal publishing group figure 12. effect of different salinity levels (nacl) on monthly mean leaf proline content in the six species studied. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). 3.3.2. soluble sugar content the analysis of the results presented in figure 13 reveals that the effect of salt stress led to a significant accumulation of soluble sugars. this accumulation increased from february to july. indeed, the highest values for this type of osmoticum were observed under high salinity conditions (6 g/l and 9 g/l nacl) and towards the end of the experimentation period for all species, until the depletion of the grass towards the end of the trial. significant differences exist between treatments throughout the trial period (figure 13). agriculture and food sciences research, 2024, 11(2): 116-135 130 © 2024 by the authors; licensee asian online journal publishing group figure 13. effect of different levels of salinity (nacl) on the monthly average soluble sugar content among the six studied species. note: significant differences are indicated as follows: * significant (p<0.05); ** highly significant (p<0.01); *** very highly significant (p<0.001). 4. discussion the results show that the six turfgrass species studied show contrasting responses to salt stress for most of the parameters analysed. observation of the behavior of grasses under salt stress revealed that irrigation with naclenriched water caused a reduction in morphological parameters, such as the length of blades and internodes. similar reductions were observed for dry matter and aboveand below-ground biomass, with a marked drying out of the grasses when irrigated with solutions containing 6 g/l and 9 g/l nacl, from the fourth month after exposure to the stress, except for agrostis stolonifera, which resisted the 6 g/l nacl concentration. in addition, more pronounced reductions in turf cover were noted under severe stress (6 g/l and 9 g/l nacl), particularly during the last two months. these results are consistent with those of bizhani and salehi [39]; guo, et al. [40] and xu, et al. [41] who reported that leaf elongation rate of tall fescue, shoot growth of seashore paspalum (paspalum vaginatum), shoots and roots of bermuda grass (cynodon dactylon), as well as shoots of kentucky bluegrass (poa pratensis) are significantly inhibited by salt stress. in addition, the cover rate is a key indicator for assessing the visual quality of turf, and in agriculture and food sciences research, 2024, 11(2): 116-135 131 © 2024 by the authors; licensee asian online journal publishing group kentucky bluegrass, this rate decreases significantly under the effect of salinity [42]. previous research has also shown that the main signs of stress in plants subjected to saline irrigation include a stunted appearance, a decrease in root biomass and drying out of the aerial parts, leading to reduced growth [43]. this reduction can be explained by the increase in osmotic pressure in the environment due to nacl, which limits water uptake by the roots. this abiotic stress disrupts turfgrass growth and development by inhibiting water uptake, creating osmotic and ionic imbalances at the root-soil interface [44-46]. under moderate salinity conditions, some salt-tolerant species in cynodon spp., paspalum vaginatum, puccinellia distans and zoysia matrella are able to maintain their shoot growth, on the contrary, sensitive species shoot growth is stopped [13, 24, 47]. lolium perenne ‘belida’, festuca rubra commutata ‘casanova’, poa pratensis ‘evora’, and festuca rubra trichophylla ‘smyrna’, cool-season turfgrasses, millipedes (eremochloa ophiuroides) and when irrigated with saline solutions have maintained the turf quality at the low to moderate salt concentration, whereas at higher concentrations leaf width and length significantly decreased [14, 15]. in addition, all species studied (distichlis spicata, festuca arundinacea, poa pratensis) showed an increase in root/stem ratio at all salinity levels. of these species, only glasswort (distichlis spicata) maintained a relatively stable above-ground biomass despite an increase in soil electrical conductivity (ec) from 2.5 to 25 ds m-¹. an initial increase in root biomass was observed in alkaligrade and glasswort up to 15 ds m-¹, while tall fescue maintained its root mass at the same ec level [24]. a similar stimulation of root growth was observed in a halophytic grass, sporobolus virginicus, when the nacl concentration reached up to 1.0 m [17]. the changes in the morphological characteristics of turfgrass under salt stress are directly linked to physiological changes, in particular the stability of cell membranes and the production of reactive oxygen species (ros). physiological and molecular studies have been carried out to gain a better understanding of the mechanisms by which turfgrass responds to salt stress. it has been reported that malonyldialdehyde (mda) concentration and electrolyte leakage (el) increase under nacl in perennial ryegrass and kentucky bluegrass, indicating damage to cell membranes [42, 48, 49]. under salt stress, the plant first undergoes osmotic stress linked to external salts, then ionic stress when salts accumulate to toxic levels inside the cells, the latter phase being decisive in the genetic differentiation of plants' ability to tolerate salinity [50]. however, some studies suggest that osmotic and ionic stress simultaneously affect plants exposed to salt stress [51-54]. in the same context, smillie and nott [55] and hessini, et al. [56] have shown that the presence of soluble salts in the soil solution lowers its osmotic potential, causing a state of physiological ‘drought’ characterised by a reduction in plant metabolic activity. this drop in osmotic potential limits water uptake by the roots, reducing cell turgor pressure and leading to plasmolysis. however, some plants quickly adjust their osmotic potential in response to the external environment, maintaining a favourable water gradient. inclusive plants store chloride and sodium ions, whereas in exclusive plants these ions are not retained to any great extent and are transported to the roots via the xylem [57]. numerous studies confirm that osmotic adjustment is a central mechanism for the survival and growth of plants subjected to salt stress [58-61]. our observations of physiological and biochemical parameters, including total chlorophyll, relative water content (rwc), proline and soluble sugars, reveal that high concentrations of nacl in the irrigation water led to a significant decrease in chlorophyll pigment content and rwc in all the species studied, particularly at concentrations of 6 g/l and 9 g/l nacl. conversely, an increase in the accumulation of proline and soluble sugars in the leaves was observed as salinity increased. these results agree with those of uddin, et al. [62] who reported a decrease in chlorophyll content and rwc, accompanied by an increase in proline with increasing salinity in paspalum vaginatum, zoysia japonica, zoysia matrella, digitaria didactyla and cynodon dactylon. these observations have also been made for other monocotyledons such as rice, wheat and maize [63-66]. under stress conditions, the decrease in osmotic potential can be due to dehydration processes as well as the accumulation of organic and inorganic solutes [67, 68]. this osmotic adjustment mechanism has been identified as an important tolerance strategy in paspalum vaginatum and agrostis stolonifera [40, 69, 70]. in response to stress, organic solutes such as soluble sugars and proline, as well as inorganic ions such as na+ and k+, play an essential role in the osmotic adjustment of turfgrasses [25, 71]. these mechanisms enable plants to maintain leaf turgidity, thereby reducing water potential and promoting water uptake despite soil salinity [72]. in addition to its role as an osmolyte, proline contributes to the stabilisation of subcellular structures, captures free radicals and acts as a buffer to regulate redox potential during stress situations [73]. the accumulation of soluble sugars, in particular, is a frequent phenomenon in response to salt stress [74]. this accumulation could be linked to enzymatic changes affecting carbohydrate metabolism, as observed in rice plants subjected to salinity, where a drop in fructose-1,6bisphosphatase activity led to an accumulation of sucrose, thereby increasing salt tolerance by increasing cellular osmolarity and carbon reserves [75]. knowledge of the use of osmotic adjustment and its components (organic and inorganic osmolytes) in turf grasses remains limited [76]. in our study, the most marked impact of salt stress on morphophysiological parameters was a significant reduction in species cover during the last three months of the growing season. this reduction in the density of plant cover resulted in a deterioration in the aesthetic quality of the turf. species that demonstrated salt tolerance included agrostis stolonifera, festuca arundinacea and cynodon dactylon, particularly at a concentration of 3 g/l nacl. these species showed an ability to maintain a higher cover rate than the other species. in comparison, lolium perenne proved to be the most sensitive to salt stress conditions, whatever the dose applied. plants react in a variety of ways to salt stress through morphological, physiological and metabolic responses, which can considerably reduce crop yield and quality [77, 78]. plant resistance to salinity results from two complementary processes: avoidance and tolerance. avoidance mechanisms limit the excessive translocation of salts to sensitive leaf tissues, while tolerance mechanisms increase the survival capacity of plants in the presence of salts accumulated in these tissues [79]. this involves various physiological adaptations such as root growth stimulation, osmotic adjustment, ion exclusion and regulation, and ion sequestration and excretion. these resistance mechanisms can coexist within the same species and are common to most salinity-tolerant plant species [78]. agriculture and food sciences research, 2024, 11(2): 116-135 132 © 2024 by the authors; licensee asian online journal publishing group many turfgrass species considered moderately tolerant or tolerant to salt stress often show an increase in root/stem ratio when subjected to salt stress [24]. this is explained by an increase or maintenance of rooting, while shoot growth decreases in response to salinity, which may constitute a tolerance mechanism. salinity resistance within the poaceae family encompasses a wide range of salinity levels, and turfgrass species employ a multitude of resistance mechanisms. these include the maintenance of stomatal conductance, anatomical and morphological adaptations, drought and heat tolerance, signalling cascades, minimisation of na+/cltoxicity, a high k+/na+ ratio, as well as nutrient and osmotic balances, active control of ionic accumulation, and the activation of tolerant genes, accompanied by the production of osmolytes. in response to a limited exposure to salinity levels, salt-tolerant species like halophytic plants and particularly warm season grasses maintain or increase roots growth [24, 77, 80] driving to a higher root biomass which is expected to limit the osmotic stress and salt accumulation in shoots tissues, while storing them in the roots where there are limited negative effects. the 4 studied species bermuda grass, saint augustine grass, seashore paspalum and korean lawngrass irrigated for two years with salty water with an electric conductance of 15 ds m-1compared to the control irrigated with water at 2.5 ds m-1. additionally those species which maintained the roots growth, were also able to maintain the shoot quality [77]. the exclusion of salt ions from shoots, combined with osmotic adjustment, is one of the most widespread mechanisms for avoiding salt stress in c3 and c4 turfgrass species [81]. this process consists of preventing the translocation of na+ to the shoots to minimise the toxic effects associated with salinity [82]. numerous studies have demonstrated a negative correlation between shoot na+ and clconcentrations and the relative salinity tolerance of c3 and c4 turfgrass species [22, 79, 80]. this relationship has been successfully exploited to predict salinity tolerance in cynodon spp. and zoysia spp. [10, 83]. various sodium exclusion mechanisms have been identified, including na+ efflux from roots, na+ compartmentalisation in root cell or mesophyll vacuoles, and control of na+ accumulation and release by xylem parenchyma cells [82]. roots have a developed system to regulate the uptake of water with selected ions from soil solution such as na+, k+ and clwhich are essential elements to maintain the osmotic pressure of plant cells, which is directly linked to the capacity of the plant to absorb water. salt tolerance has been correlated with the capacity to maintain the ratio k+/na+ [79, 80]. this is possible thanks to an increased uptake of k+ by the roots while reducing its cells leakage and reducing the transport of na+ from roots to shoots [80]. paspalum spp, a halophytic grass retrieved in the coasts when exposed to salt present an increased level of na+ of 15-fold compared to non-stressed plants in the shoots, and 25-fold in the roots, associated with a decrease of k+ of 26.02% and 69.68% respectively. calcium ions have probed to play a key role being a secondary messenger for signaling salt stress indirectly activating membrane and tonoplast antiporters [84]. thus, when roots are exposed to nacl there are modifications in cytosolic calcium to increase ca2+ levels. another mechanism of resistance to salt stress is the compartmentalization of na+ ions in organs like vacuole, reducing their concentration in the cytoplasm, thanks to the overactivation of membrane tonoplast, thank to calcium and also some ‘compatible solutes’ such as proline and glycine betaine facilitate this translocation into the vacuole [79, 85]. this process protects the cell structure and the osmotic balance. proline, besides this protecting function, contributes to membrane stability and maintains photosynthetic activity [79, 86]. on the other side, when adding glycine betaine in the solution with nacl, lolium perenne presented a reduced accumulation of na+, a maintained level of the ratio k+/na+ and increased level of enzymes with antioxidant activities like catalases, superoxide dismutases and ascorbate peroxidase which prevents cell membrane damage [87]. 5. conclusion the present study examined the physiological and morphological responses of six turfgrass species (four temperate and two tropical) under conditions of salt stress. the results indicate that the most salinity-tolerant species are a. stolonifera, f. arundinacea and c. dactylon, particularly at a dose of 3 g/l nacl. these species also demonstrated an ability to maintain a higher cover rate than the other species. on the other hand, l. perenne proved to be the most sensitive to salt stress conditions, whatever the dose applied. a. stolonifera seems to be the best adapted to this stress, particularly at the dose of 3 g/l nacl, maintaining a high coverage rate throughout the trial. it is important to note that morphological and physiological traits are strongly affected by severe salt stress conditions (6 g/l and 9 g/l nacl), resulting in zero value records from the fourth month of stress application. overall, these results provide an insight into the morphological and physiological responses of turfgrass species of temperate and tropical origin to high salinity, offering an indication of their level of tolerance. these preliminary data could encourage physiologists and landscape designers to carry out field trials on a regional and national scale. highlights: • a.stolonifera, f. 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licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 1, 15-29, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i1.5359 © 2024 by the authors; licensee asian online journal publishing group inoculating plant growth-promoting bacteria effects on soil hydraulic properties and tomato root development under water stress conditions dragonetti giovanna1 cherradi soumiya2 mapelli francesca3 riva valentina4 choukr-allah redouane5 weldeyohannes amanuel oqbit6 borin sara7 1,2international center for advanced mediterranean agronomic studies, mediterranean agronomic institute of bari, italy. 1email: dragonetti@iamb.it 2email: soumiyacherradihorti@gmail.com 3,4,7department of food, environmental, and nutritional sciences, università degli studi di milano, milan, italy. 3email: francesca.mapelli@unimi.it 4email: valentina.riva@unimi.it 7email: sara.borin@unimi.it 5department of horticulture, hassan ii institute of agronomy and veterinary medicine, agadir, morocco. 5email: redouane53@yahoo.fr 6university of alberta, dept. of renewable resource, alberta, canada. 6email: aweldeyo@ualberta.ca ( corresponding author) abstract the plant growth promoting bacteria (pgpb) harbored in the rhizosphere develop specialized mechanisms that may have a key role to ameliorate soil properties and plant growth under prolonged dry conditions. accordingly, this study aimed to assess the effects of bacterial growth on the soil hydraulic properties and the root response under water stress conditions induced by drip irrigated tomato. at pot scale, a silty soil was inoculated with two pgpb strains (micrococcus yunnanensis m1 and pseudomonas stutzeri sr7-77) to cultivate tomato plants under three different water regimes: full irrigation (100% of pot capacitypc), moderate and severe water stress levels (75 and 50% of pc, respectively). bacterized soil altered the pore size distribution of the rhizosphere compared to no-bacterized soil, increasing root zone plant-available water holding capacity. on the contrary, pgpb occupying the pores reduced the saturated hydraulic conductivity near-saturated soil conditions compared to the uninoculated trial. pgpb shown root surface density (rsd) equal to 0.540 % and 0.355 % to inoculated sr7-77 and m1 tests, respectively and under 50% pc, compared to 0.097% to the uninoculated soil test. soil water potential values, retrieved through soil water retention parameters, were more negative to m1 and sr7-77 strains tests, corresponding to water hold in the pores with smaller radii conferring resistance to the plant following contrasting stresses. the results demonstrated that pgpb elongated continuous transmission pores and bridged with air-filled spaces in stressed periods. keywords: irrigation water regimes, pgpb, saturated hydraulic conductivity, soil water potential, soil water retention. citation | giovanna, d., soumiya, c., francesca, m., valentina, r., redouane, c.-a., oqbit, w. a., & sara, b. (2024). inoculating plant growth-promoting bacteria effects on soil hydraulic properties and tomato root development under water stress conditions. agriculture and food sciences research, 11(1), 15–29. 10.20448/aesr.v11i1.5359 history: received: 23 november 2023 revised: 18 january 2024 accepted: 26 january 2024 published: 6 february 2023 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: carried out the experiment, c.s., r.v. and d.g.; wrote the manuscript with support from b.s., m.f., c-h.a. and w.a.o, d.g.; conceived the idea, b.s., m.f. and d.g.; supervised the project, d.g., b.s and c-h.a. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 16 2. materials and methods ................................................................................................................................................................... 17 3. results and discussion ................................................................................................................................................................... 19 4. conclusions ....................................................................................................................................................................................... 27 references .............................................................................................................................................................................................. 27 mailto:dragonetti@iamb.it mailto:soumiyacherradihorti@gmail.com mailto:francesca.mapelli@unimi.it mailto:valentina.riva@unimi.it mailto:sara.borin@unimi.it mailto:redouane53@yahoo.fr mailto:aweldeyo@ualberta.ca https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i1.5359 agriculture and food sciences research, 2024, 11(1): 15-29 16 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the study delved into aspects of the soil hydraulic properties alteration processes induced by pgpb, not commonly assessed to address the drought. highlighting that are soil water potential and pore size distribution changes to play a pivotal role on the performance improvement of prolonged water stress conditions ensuring agricultural productivity. 1. introduction physical and hydraulic soil characteristics are greatly influenced by microbial community activity [1-4]. the microbe harbored in the rhizosphere assist water uptake under drought stress [5] ensuring crop yield, improving root development system, and reducing pathogen infection and maintaining a sustainable environment [6-9]. among these, the plant growth-promoting bacteria (pgpb) has the role to aid the plants to withstand water stress [10-15]. several studies have shown as the inoculation of pgpb influences plant growth and root development response. for instance, the bacterial inoculation with gluconacetobacter diazotrophicus on red rice plants showed induced a systematic tolerance (ist) response to the drought conditions with an increase of root area [16]. in addition, pgpb inoculation is a practice, especially in the arid and semi-arid irrigated areas, to ameliorate the soil physical and hydraulic properties [4, 17-19]. as a matter of fact, what occurs is the bacteria alter such soil physical and hydraulic properties in the rhizosphere, among them: pore system characteristics, soil water holding capacity and soil hydraulic conductivity. in detail, the bacteria occupying smaller soil pores space making available water at severe stress conditions, modifying its distribution [20] thus, is facilitated root water uptake even though prolonged drought conditions. by assuming soil water potential as one of the soil variables that better may represent the root water uptake behavior, it can be taken into consideration to explore the root water uptake behavior to the inoculated bacteria soil. under drought conditions, what was observed is that the bacteria community alter the range of water potential values (i.e. resulting more negatives) due to osmolytes which stimulated, boost the water holding capacity of dry soils [1]. what commonly happens to inoculated-bacteria soil, is the water content brings towards soil potential values nearly to those most negatives, which corresponds to the water adsorbed and hold in thinner and disconnected pores, making thus difficult the root water uptake under common conditions, since is water not promptly available to the plants. therefore, pgpb might be an alternative strategy to reduce risk of drought, because of despite water held in the smallest pores is become available to the plants when experience water stress conditions. in point of fact, pgpb plays a relevant role in modifying water hold capacity and the soil porous structure, as consequence may ameliorate root water uptake to water scarcity contexts. such results shown that bacteria growth increase air-entry value to coarse-grained soils, while first an increase then a decrease in the air-entry value was observed to finegrained soils compared to uninoculated soil [21]. what was observed is bacterial-induced changes in the hydraulic properties and root growth inducing soil water potential and permeability reduction [22]. moreover, at different levels of soil water storage, the soil and root hydraulic conductivities may increase or decrease accordingly to soil types and agricultural practices. in detail, both two conductivities require to be assessed when it reaches an intermediate soil water status condition [23] being the condition in which it observes a decrease of hydraulic conductivity and limited water root uptake under water stress conditions. since the water root uptake not only depends on the hydraulic conductivity of the roots and of the soil, but also to that occurs at the root-soil interface; its efficient therefore is only occurred whereas mechanisms between soil and root fully satisfy water demand [24] which is not observed under water scarcity conditions. furthermore, soil hydraulic properties and pore characteristics depend on spatio-temporal dynamics induced by differ among changes (i.e. long-term tillage systems, crop rotations) [25]. on the other side, pgpb may cause pores clogging which influence hydraulic conductivity under saturated or partially saturated soil conditions [26-30]. this effect arises from the fact that the bacteria regulate mechanisms between plant roots and soil by producing substances that alter soil structure [4]. in addition, knowing soil hydraulic properties plays a considerable role for estimation of available soil nutrients [31] using pgpb will also contribute to improve nutrient absorption capacity of plant, root biomass and area, as well [32]. in the light of this, pgpb can be considered a purposeful strategy to cope extreme events in agriculture. commonly, inoculation with pgpb adapts the plants to tolerate negative effects induced during drought periods, since they stimulate the root growth inducing the development of the lateral roots and root hair. in particular, the bacteria stimulate the roots to suck water even though water is only available at very high negative potential values, because of they allow to the root system to reach mesopores or micropores. it was observed that the inoculated plants develop lateral roots and root hairs [33, 34] which allow to enhance water stress tolerance [14]. presumably, a good contact between roots and the soil surface was achieved under drought conditions because of bacteria stimulate the formation of extracellular polymeric substances become hydrophobic causing thus an alteration of the soil structure and of the soil hydraulic properties in the rhizosphere zone and influence the plant water use efficiency [19]. knowing that the properties of rhizosphere around the root depend on the capability of a plant to extract water [35, 36] size, shape and continuity of pores [37] becomes therefore relevant to assess water interactions among bacteria-rhizosphere soil. for instance, investigating the pore structural properties of the soil adherent to the root shown the rhizosphere presents different behaviors [37]. in the rhizosphere of wheat, it was obtained higher water content values in comparison to the bulk density [20, 38] reasonably explicable to the effect of the mucilage properties at hygroscopic level. increasing of water content in the rhizosphere was also observed for lupine and especially undergoing drying and wetting cycles [20]. moreover, raddadi, et al. [17] and murgese, et al. [39] shown that bacteria have the ability to produce biosurfactants and bioemulsifers substances contributes to improve soil hydraulic properties in arid soil. among others, murgese, et al. [39] shown as the bacterial consortium of pgpb improved barattiere (cucumis melo l.) physiological response at pot scale, as well as it reduced the use of mineral fertilizer doses. although these studies have demonstrated the effects of exudates in controlling the soil water dynamics in the rhizosphere, it not well-explored how some bio-hydrophysical properties change in the rhizospheric soil and alter pore-size distribution under different water regimes. to understand these behaviors, it becomes thus crucial to characterize soil hydraulic properties to inoculated soils, since they provide insights on the ability of the roots to agriculture and food sciences research, 2024, 11(1): 15-29 17 © 2024 by the authors; licensee asian online journal publishing group uptake water even under severe water conditions. so far, only few studies have been conducted thereon [19, 20, 25]. thus, if the pgpb may or not improve the soil water holding capacity especially under water stress conditions this implies to estimate rhizosphere hydraulic parameters and pore size distribution under vary water irrigation regimes. the fact that the water roots dynamics and their growth strongly are influenced by adequate presence of air-filled pore space and by the stabilization of soil structure, to this study it was therefore characterized the soil properties at the end of tomato pot experiment, assuming that the bacteria would have accomplished their growth. and in doing so, soil water potential data was also retrieved along the irrigation season by using collected soil water content data throughout the experiment. however, because of regulations are not yet well-defined on pgpb use at the open field in most jurisdictions, only trials at pot scale may be carried out since only allow to accurate monitor the effects of pgpb on soil characteristics and plant response, likewise allowing to keep under control the water dynamics reducing thus bacteria concentrations eventually lost by water drainage fluxes [40]. as conclusion, the study here proposed was undertaken to assess the effect of bacteria on soil hydraulic properties, root development and soil pore characteristics of tomato plants under three different water irrigation regimes with the project to consider pgpb as amendment to enhance the soil physical and hydraulic characteristics. accordingly, silt-loam potting soil was inoculated with two pgp b strains: micrococcus yunnanensis (m. yunnanensis m1) and pseudomonas stutzeri (p. stutzeri sr7-77 ps) to determine soil hydraulic parameters at end of pgpb growth, and to estimate root surface density (rsd) in the rhizosphere. 2. materials and methods 2.1. soil hydraulic properties estimation simultaneous measurements of soil water content, θ, and soil pressure head, h, were obtained using a suction table method [41, 42] to determine soil water retention curve; while hydraulic conductivity curve inferring from the evolution of θ and h, and saturated hydraulic conductivity measurements, ks, by darcy’s law [43]. θ(h) and k(h)-k(θ) functions were depicted by using van genuchten mualem (vg-m) semi-empirical model [44]. in detail, θ(h) and ks measurements obtained by laboratory methods [44, 45] allowed to solve vg-m model to the soil water retention curve θ(h) and the hydraulic conductivity k(h)-k(θ). the shape of water retention curves is obtained as follows: mn rs r e )h(1 1 θθ θθ(h) (h)s vg+ = − − = (1a) ( )     n m rs r h 1 1 1 1                 −       − − = (1b) where se is the effective fluid saturation, θr and θs denote the residual and saturated water contents (l3l-3), respectively; and αvg, n and m (m =1-1/n) are empirical shape parameters. a non-linear least-squares curve fitting procedure was used to determine αvg, n and m parameters. while the hydraulic conductivity function is deduced by applying the capillary bundle theory [46, 47] starting from water retention curve and here represented as: ( ) ( ) ( ) ( ) 2 1 0 e e s 0 e e e s e er ds sh 1 ds sh 1 s k sk sk e       ==   (2) in which ks is the saturated hydraulic conductivity, and  is a coefficient which considers the reliance of the tortuosity and the parameters on the water content and being set an optimum average at about 0.5 by mualem [47]. assuming m=1-1/n and applying mualem’s model, van genuchten [48] also found a closed-form analytical solution to eq. 2 to predict kr at a specified volumetric water content: ( ) ( ) ( ) n1-1m 11 2 1 =     −−==  m m ee s e er ss k sk sk (3) where ks is the saturated hydraulic conductivity (lt-1) and τ is the pore-connectivity parameter. the poreconnectivity parameter τ in k(θ) was estimated by mualem [47] and set to 0.5 obtained as an average value from many different soil tests. 2.2. soil pore size distribution (psd) to define soil pore system, it is needed to determine the size distribution of pores because the bacteria function may also depend on their size. several classifications are proposed to define the relation between size of pores and water storage. for instance, greenland [49] classified the classes of pores as follows: i) bonding pores (<0.005 µm) which are fine pores able to aggregate primary particles; ii) residual pores (< 0.5 µm) are those that establish chemical interactions at molecular scale; iii) storage pores (0.5 to 50 µm) defined as the pores that retain water and make it available for plant and microbe community; iv) transmission pores (50 to 500 µm) feed the root growth through the movement of water; v) the pores larger than 50 µm and corresponding to the field capacity, and pores larger than approximately 0.5 µm that could be emptied correspond to the wilting point; v) pores larger than 500 µm are interested to root excavation and water movement. however, the partition of pores between filled air and water and agriculture and food sciences research, 2024, 11(1): 15-29 18 © 2024 by the authors; licensee asian online journal publishing group root growth has not been adequately defined. as well known, to ensure root growth it is necessary to have adequate storage pores (0.5-50 µm) and adequate transmission pores (50-500 µm) in the soil [50]. pore size distribution and water movement at specific water potentials are related through several physical equations and models [51, 52]. in fact, the soil pore size distribution (psd) curve may be derived from soil water retention curve θ = f(h) [53, 54]. this function converts into an equivalent pore size distribution curve based on the young-laplace law and assuming a parallel bundle of cylindrical pores [55, 56]. the shape of the distribution function of pore size can be described as follows: ( ) 0(cm)h ; h 2980 cos4 = gh de   4 ( ) ( ) ( ) ln hd d hf  = where f(h) is the distribution function of pore size versus ln(h), and relate to the equivalent pore diameter (μm), on a log10 scale [57-59] h is the matrix head (cm) (h>0), γ is the water surface tension within the pores (72.8 g s-2), ρ is the water density (0.998 g cm-3), g is the gravitational acceleration (980 cm s-2), and ω is the water contact angle with the soil pores (ω ≈ 0). 2.3. laboratory measurements to estimate soil water retention and hydraulic conductivity properties, 27 undisturbed soil samples were collected at the end of the experiment by using stainless steel cylinders (inner diameter of 7.6 cm and height of 7 cm). afterwards, soil samples were placed in a tank and slowly wetted from bottom with the purpose to reach a saturation condition. water content, θ, corresponding to negative pressure head, h, values were subsequently, measured using a sand-kaolin suction table, whereas saturated hydraulic conductivity, ks, through the constant-head method [45]. in detail, water content, θ, was monitored to the following pressure heads: -1.0, -3.0, -10.0, -15.0, -30.0, -50.0, -80.0, -110.0, -180.0 cm; while to more negative h values: -30 and -120 m was used the pressure plate. at end of the experimental measurements of water retention, the soil cores were oven-dried at 105 °c to determine also bulk density, ρb. as mentioned above, a fitting procedure was used to optimize αvg, n and m water retention parameters. these parameters were used to convert the trend of water contents recorded during the tomato season trial in soil potential by using the eq. 1b. these data were used to explain the bacteria growth and its interactions with rhizosphere soil. 2.4. bacteria inoculation procedure the chosen bacteria for this study were: micrococcus yunnanensis (m1) and pseudomonas stutzeri (sr7-77) strains within the five strains and selected from a previous pot experiment [60]. according to the results obtained from this first trial year, m1 and sr7-77 are the most promised strains by assessing tomato root development, yield, and soil quality. micrococcus yunnanensis strain m1 originally described in 2009 [61] has been isolated from the endosphere of an extremophilic plant and some experimental validation on the strain m1 resistance to ampicillin, rifampicin, chloramphenicol and tetracycline was performed to demonstrate that strain m1 is sensitive to all the tested antibiotics. pseudomonas stutzeri was originally described as bacillus denitrifticans in 1895 [62]. the strain sr7-77 has been isolated from plants. the positive role of p. stutzeri on the plant growth is reported also by bacilio, et al. [63]. they indicated that the use of humic acids and bacterial inoculation mitigated negative effects of salinity gradients in pepper. moreover, different strains belonging to this species were isolated from polluted environment, sometimes described as pollutant degraders [64, 65]. 2.5. pot case study a pot experiment was carried out in a semi-controlled greenhouse of the mediterranean agronomic institute of bari (iamb) located in apulia region, south of italy. the test was carried out between 13 february and 23 july 2018. the pots were filled by a silt-loam soil. soil texture was determined by hydrometer and severs methods. according to the usda textural classification (the united states department of agriculture) the percentage of each soil solid particle class was: 13.75, 16.50 and 69.75% for sand, clay and silt, respectively: the experimental trial was consisted of three irrigation regimes that have been applied to potted tomato plant, which are: (t1) well-watered, with 100% irrigation based on pot capacity, (t2) and (t3) a controlled drought stress cycle with 50% and 75% of pot capacity, besides the 2 tests of bacteria: micrococcus yunnanensis m1 (b1), pseudomonas stutzeri sr7-77 (b2); that have been inoculated in the soil, and control (c) where there is no inoculation of the bacteria. pot capacity (pc) was measured after 48 hours once the pots were saturated and a fraction of delivered water was loosed through the soil gravity force. overall, three water regimes were induced and 2 bacteria (b1and b2) and 1 control (c) treatments were set up getting 9 experiment units. the experimental design adopted was a randomized complete block design with 7 blocks; each block included 9 experimental units with 4 pots each one. the total number of pots used in the experiment was 252 pots. the inoculation was applied once, at the beginning of the campaign, and two weeks after transplantation (13 april 2018). the broth culture of the two inoculated bacteria tests, m1 and sr7-77, has been prepared. the concentration of the solution applied was equal to 10^8 cell/ml. estimating that the plantlet roots occupy in the pot a soil volume of 200 ml. hence 10^8 *200 cells were calculated to be applied to each plant, to reach in the soil surrounding root a concentration of 10^8 cell/g. the 50 ml volume received contained a 10-time concentration of bacterial suspension, so it was diluted with a ratio of 1:9 (50ml of broth culture and 450ml of water). for each plant, 50 ml of the total agriculture and food sciences research, 2024, 11(1): 15-29 19 © 2024 by the authors; licensee asian online journal publishing group diluted bottle solution was poured to the top of the soil next to the tomato plant collar. this inoculation procedure was followed under optimal temperature conditions in order to allow to bacteria strain to colonize the root zone. 2.6. root density measurements 126 root-soil samples were collected and scanned by imagej software. first, the samples were treated by a 5% sodium hexametaphosphate solution to disperse soil attached to the roots, then the roots were washed by hand. afterward, each treated root sample was laid out on a white paper and scanned a 300 dpi resolution. acquired images were analyzed by imagej [66]. to only distinguish live roots achieve, red, green and blue (rgb) color threshold ranges were set between 0-70, 0-60 and 0-20, respectively. root density surface was then obtained with the “analyze particles” command. furthermore, it was selected a measurement range from 10 pixels to the infinity to not include small areas or single pixels that not corresponding to root areas. assuming a cylinder shape, the area measured by the software was multiplied by π to gather the real surface of the roots. while root hair length was obtained with 10 root hair measurements per plant (u.s. national institutes of health, bethesda, maryland, usa). a photographic rank scale between 0 (no root hairs) and 5 (most dense) was chosen to determine root-hair density of plant. 2.7. statistical analysis statistical analyses are performed with minitab 16 statistical software. data are analyzed by two-way anova analysis of variance, using “bacteria” and “water regimes” as factors and block as random factor to assess differences among factors and the interactions between the factors using adjusted sum of squares for tests with an interval of confidence of 95% (p=0.05). 3. results and discussion the experiment units (water regime per treatment) are indicated hereafter with the abbreviations, as follows: micrococcus yunnanensis m1 (b1), pseudomonas stutzeri sr7-77 (b2); and control (c) for treatments, while full (t1), moderate (t2) and severe (t3) for water irrigation regimes. 3.1. soil hydraulic properties characterization the hydraulic parameters of water retention θ(h) and hydraulic conductivity k(h) curves are averaged on 21 replications collected per each experiment unit (exp. unit): t1-b1, b2 and c; t2-b1, b2 and c, t3-b1, b2 and c. the fitted hydraulic parameters n, and αvg, θr and θs are listed in table 1. according to the statistical analysis, there is a highly significant difference between the different bacteria inoculated, between the water regimes and between their interactions too. table 1. soil hydraulic parameters for each experiment unit (treatment x water regime). exp. unit θ0(-) θr(-) α(cm-1) n(-) m=1-1/n ks (cmmin-1) τ t1-b1 0.495a 0.010b 0.310b 1.125a 0.111 0.1789b 0.5 t1-b2 0.511a 0.015b 0.243b 1.140a 0.123 0.1432b 0.5 t1-c 0.497b 0.000a 0.173a 1.146b 0.127 0.2350a 0.5 t2-b1 0.493a 0.020b 0.205b 1.139a 0.122 0.2434b 0.5 t2-b2 0.482a 0.035b 0.212b 1.143a 0.125 0.2271b 0.5 t2-c 0.510b 0.000a 0.193a 1.158b 0.137 0.1959a 0.5 t3-b1 0.489a 0.030b 0.300b 1.140a 0.123 0.2194b 0.5 t3-b2 0.494a 0.040b 0.177b 1.170b 0.146 0.2273b 0.5 t3-c 0.501b 0.000a 0.213a 1.161b 0.138 0.4288a 0.5 note: p ≤ 0.05 were regarded as statistically significant. a, b letters indicate statistical significance. in the table 1, the shape curve parameter, αvg, observed for bacterized test was higher than the control for three water regimes (t1, t2, and t3); while t3-b2 was lower. in other hand, being αvg represents the inflection point of the soil water retention curve (wrc) indicates the amplitude of the water availability range. thus, the greater value of αvg corresponds a lower potential head value which implies an enlargement of the water available range to the inoculated bacteria soil in comparison to the control, since the bacteria activity allowed to occupy smaller pores and increasing the held water. on the contrary, the effect of bacteria growth has not increased saturated water content θs (obtained as average on 21 replicates). it was observed that θs is 49% for bacteria b1 and control c and 52 % for bacteria b2 under full water regime t1; 48% for bacteria t3-b1 and t2-b2. in the case of control test, the θs is 51% and 50% for moderate t2 and severe t3 water stress, respectively. this means the bacteria do not show high significant difference when is reached soil porosity [67]. while to unsaturated soil conditions, bacteria are able to improve the soil water availability because of their capability to bridge air-filled gaps and consequently modify the hydraulic properties. likely, fatty acid produced by pgp b in the silty-loam soils may have influenced the soil hydraulic properties improving the soil water retention. with this regard, it can observe as the soil residual water content θr increased in both under full (t1) and moderate (t2) and severe (t3) water irrigation regimes for bacterized tests (b1 and b2). as shown the table 1, the soil saturated hydraulic conductivity values, ks, highlight the effect of beneficial bacteria showing a main reduction to t1 (full irrigation) and to t3 (severe water stress conditions). to the contrary, by looking at the curve k(h) soil hydraulic conductivity, k, and soil water potential, h, the bacterial growth has caused a different behavior of the soil hydraulic conductivity trend, since to the variable saturated soil condition. over the entire bacteria activity indeed, strong effects of the bacteria on hydraulic conductivity in saturated porous media were founded [30]. based on the fitted soil hydraulic parameters from the measurements of soil water content, θ, and potential, h, couple values, the hydraulic properties were determined to three tests (b1, b2 and c) and under three water regimes (t1, t2 and t3) as depicted in the figure 1 a-c. agriculture and food sciences research, 2024, 11(1): 15-29 20 © 2024 by the authors; licensee asian online journal publishing group figure 1. (a-c) averaged water retention θ(h) and (d-f) soil hydraulic conductivity k(h) curves referred to micrococcus yunnanensis (b1 with red color line), pseudomonas stutzeri (b2 with green color line), and control (c with blue color line) treatments: a) and d) under full water irrigation (t1); b) and e) moderate water irrigation c) and f) severe water irrigation regimes. the full symbols of water retention curves represent the measured couples data θ-h, while the lines refer to that simulated. the figure 1a-c shows as soil water retention θ(h) is quite similar to bacterized and control soils, but with a slight difference. at given soil water content and relative soil water potential under optimal (t1) and water stress (t2 and t3) conditions, it is observed that at partially-saturation conditions changes the shape of curve near to the low potential values (more negative) for the bacterized soil compared to the control. this behavior is induced by a rearrangement of soil structure due to the bacteria growth occupying mesopores and micropores contributed to increase soil water available. in fact, soil water potential, which has been assumed as variable of the root uptake behavior, showed that there is more water but held in the smallest pores, which means yet water available, because the bacteria are able to regulate the mechanisms and overcome a water stress condition stimulating the root system to also search water among the smallest pores. the figure 1 d-f instead, depicts the soil hydraulic conductivity curves trend for all water regimes (t1, t2 and t3) and treatments (b1, b2 and c). under full water regime, t1, the shape of the three water retention curves is quite similar, this is why the water held in the soil always allowed to satisfy the plant water demand in both trials: bacterized and non-bacterized, therefore the bacteria have not induced an evident variation under unsaturated soil conditions during the entire tomato season. pgpb have explicated a high significant under saturated soil conditions, in which the saturated hydraulic conductivity value, ks, is equal to 0.1789 and 0.1432 cmday-1 for b1 and b2, respectively and lower than the control test which provided a value equal to 0.2350 cmday-1. under moderate water stress (t2), the soil hydraulic conductivity curve shows a different behavior. over the entire soil water potential range, it may observe that to the same hydraulic conductivity value corresponds different and more negative potential agriculture and food sciences research, 2024, 11(1): 15-29 21 © 2024 by the authors; licensee asian online journal publishing group values compared to the unbacterized test. during the tomato growing season, water is also supplied according to the 75% of pc, which has shown a curve shape differs compared to the control test. and the explanation can be found in the fact that bacteria had the capability to develop within smallest pores. in the case of severe drought stress (t3), the soil hydraulic conductivity vs. soil potential k(h) shows a decreasing, as shown in the figure 1d-f. by observing the last part of the curve, that is under a soil dry condition water storage is roughly kept to 50% of the pc treatment, b1 has pointed out a reduction of the soil hydraulic conductivity 23%; on the contrary to b2 shown an increasing equal to 23% in comparison to the control c. this can be due to the ability of the bacterized tests to uptake more water that than non-bacterized due to the filamentous development induced by the bacteria. as explained wolf, et al. [67] the capability of bacteria to form filamentous allowed to bridge airfilled pores in water shortage conditions, thus their mobility across a range of pore-size classes facilitated the water uptake under low hydraulic conductivity and lower water potential (i.e. to more negative values). furthermore, bacteria are able to explore micro-habitats at lower water potential (negative values). this is the reason why soil water potential may explain water root uptake behavior. in practical, the root uptake activity depends on how much energy is spent by roots to suck water, and the bacteria stimulated the roots to explore larger range of potential values which means the plant could uptaken water by smallest pores, allowing to reduce water stress experience under t3 (severe irrigation regime). to understand the root uptake behavior induced by pgpb, the soil hydraulic properties were measured under three different water regimes (full, moderate and severe water regimes), and to two bacteria and control treatment at the end of tomato growth season. then, the soil hydraulic parameters were used to retrieve soil water potential data using measured soil water content data throughout the tomato season and the fitted soil hydraulic parameters. the inoculated soil (t1 and t2), shown that soil water potential values increase since bacteria growth stimulate bioclogging and biocementation processes especially in the smaller pores radii. as consequence, increase the number of smaller pores that could have contributed to the small amount of water content, in other words the soil water potential making available that water content that otherwise would be unavailable under control treatment (no-inoculated soil). in addition, bacteria altered pore-size distribution. moreover, differences to the α-parameter, which represent the inverse of the soil air-entry value, was observed among the treatments. the two treatments (t1 and t2) shown slightly different trends to the value of α, which is attributed to the competition occurred between pores bundle that contribute to both the capillary and adsorbed phase because bacteria allowed to involve smaller pores. overall, the changes in the soil water retention trend are attributable to the precipitation-induced pore clogging. to this regard, by considering the soil water retention θ(h) and the hydraulic conductivity k(h) curves representative of root uptake, the pore space geometry is also derived to better understand the behavior of bacteria in the soil [58, 68]. and here in attempts to relate soil hydraulic properties to soil structural under bacteria activity, the soil porous system is derived from soil water retention curve. the pore size distribution (psd) representative of the physical structure of soils is obtained to describe the water flow movement and availability of water in the root zone. in fact, the figure 2a-c shows that at the same water content value, the aggregation of pores is different to the treatment (b1, b2 and c). in the figure 2a, it may observe a shift and increase of the peak of the psd curve under full water regime (t1) showing for b1and b2 that the peak moved from the 0.017 for control test (c) to 0.023 and 0.026 for b1 and b2; in other words, the peak is increased by 26% and 37% in comparison to c, respectively. hence, the presence and mobility of bacteria in soil has permitted to bridge the air-pores, thus are progressively involved different classes of pores making available water to the root uptake even under long dry conditions, as occurred to t3 trial. to t2, the peak of the psd curve is pretty similar to all treatments (b1, b2 and b3) and equal to 0.0200; and it is quite evident that the predominance of bacteria b1 induced to modify the soil pore characteristics. in detail it is observed a peak equal to 0.0286 and 0.019 to b1, which is lower than control (0.022). now, explaining the trend of psd curves in terms of soil water content available in the soil, θ, it is observed that the pore size distribution changed under full water irrigation regime, as shown in the figure 2d. at maximum soil water content θ, the pore size curves shown a different distribution between bacterized and un-bacterized tests. at highest soil water content values, the capability of bacteria to fill pores with water increase compared to c test and under full water regime. on the contrary, t2 regime is not depicted relevant difference between b1, b2 and c (see figure 2e). something quite different is observed for bacterized tests under severe water stress conditions t3, where psd curve obtained to b1 was larger than the other two tests (b2 and c), as illustrated in the figure 2f. agriculture and food sciences research, 2024, 11(1): 15-29 22 © 2024 by the authors; licensee asian online journal publishing group figure 2. (a-c) pore size distribution (psd) vs. pore diameter(de) and (d-f) pore size distribution (psd) vs. soil water content (θ) curves referred to micrococcus yunnanensis (b1 with red color line), pseudomonas stutzeri (b2 with green color line), and control (c with blue color line) treatments: a) and d) under full water irrigation (t1); b) and e) moderate water irrigation c) and f) severe water irrigation regimes. agriculture and food sciences research, 2024, 11(1): 15-29 23 © 2024 by the authors; licensee asian online journal publishing group figure 3. time evolution of (a-c) measured soil water content (θ) in whole tomato season and (d-f) the corresponding soil water matrix potential, h, obtained by convert the water content data based on optimized soil hydraulic parameters curves referred to micrococcus yunnanensis (b1 with red color line), pseudomonas stutzeri (b2 with green color line), and control (c with blue color line) treatments: a) and d) under full water irrigation (t1); b) and e) moderate water irrigation c) and f) severe water irrigation. the figure 3a-c shows the soil water content evolution monitored during the whole tomato season. the trend is very similar between treatments and is observed a difference between water regimes. once these data are converted in the corresponding soil water potential values, the behavior of bacteria is clearer. by looking to the trend of the soil water content overtime to control treatment, similar values are recorded to b1 and b2 too, and for all three water regimes. on the contrary, considering the soil water potential trend and compared to the other two treatments (b1 and b2), the capability of the soil to release water can be only attributed to such classes of pores, especially under severe water regime. this is due to the fact that the root tissues do not lose elasticity under drought periods but rather ensure root-water uptake, thanks to the bacteria activity. as shown the figure 3d-f, soil water potential trends are significantly affected by pore size distribution to all bacteria tests. this effect on soil water potential and pore size distribution (psd) explains how the bacteria improved the plant mechanisms, increasing the tolerance of the plant under water stress levels. to each irrigation event, it is evident as the trend of soil water potentials reached the same values in both of three treatments (b1, b2 and b3). on the contrary, between two irrigation events, that is when evapotraspiration fluxes are trigged due to less water available in the soil and showing water potential values twice higher than those observed to the un-bacterized test in whole the season. agriculture and food sciences research, 2024, 11(1): 15-29 24 © 2024 by the authors; licensee asian online journal publishing group instead, the figure 3e depicts the soil water potential values corresponding to water contents monitored in the soil along the irrigation season and to different bacteria tests under (t2) water regime. the graph shows that the soil water availability goes so far as at lower negative water potential to b1 and b2 compared to the control c. under moderate (t2) and severe (t3) water stress levels, a different shape of pore size distribution curve may be explained by the fact that the bacteria growing in the pores allowed to keep the connection between rhizosphere and root, which is pretty evident to water stress regimes. traditionally, the response of the plant to water stress is to anticipate its vegetative phases in order to reduce the negative effects induced by drought stress conditions [69]. overall, with the presence of bacteria, largely increased water fluxes under water drought conditions. in other words, the capillary force which allows to retain water in pores is altered by the presence of bacteria. in detail, the bacteria lead to a decrease surface tension and contact angle between water surface and particle surface allowing thus to a pore to easily release water [19]. this leads to conclude that the bacteria b2 and b1 may modify the pore system characteristics according to the water regime used. therefore, the characterization of soil hydraulic properties is crucial to understand the bacteria growth behavior. 3.2. water use efficiency according to the results observed, the bacteria inoculated surrounding the soil shown a significant difference to the soil parameters between the two treatments (bacteria and water regime). however, the bacteria influence the soil and in return the plant growth through specialized mechanisms, like the production of plant hormones. moreover, uncontrollable conditions, which affect the plant parameters, such as non-controlled temperature and relative humidity, the nutrient deficit, have also a huge influence in the significance of results. this case is called type 2 error, where we accept the null hypothesis while it is wrong and that could be because of the sample size (in this case study is the number of pots per experimental unit that it’s small), the effect of the treatments is really very low, or the significance threshold is high (p=0.05). figure 4. effects of inoculating pgpb under three water regimes (100% pc, 75% pc and 50% pc) on observed data of wue (bars with the same letter(s) are not significantly different at p ≤ 0.05). in order to explain non-significance difference between the bacteria effect on many parameters, the example of water use efficiency (wue) is taken to show how the comparison between control and bacteria it becomes significant by changing the variability of each test in order to make them comparable. observing the graph of wue data (figure 4); the highest variability was of (sr7-77 strain) bacteria tests represented by the bars of the standard deviation that represent how much the data differs from the mean. agriculture and food sciences research, 2024, 11(1): 15-29 25 © 2024 by the authors; licensee asian online journal publishing group figure 5. effects of inoculating pgpb under three water regimes (100% pc, 75% pc and 50% pc) on soil permeability (bars with the same letter(s) are not significantly different at p ≤ 0.05). according to the statistical analysis, there is a highly significant difference between the different bacteria inoculated (p=0.000) and between the water regimes (p=0.000) and also between their interactions (p=0.000). observing the figure 5, the presence of the beneficial bacteria reduces the soil permeability mainly at full irrigation regime and at severe water stress condition (50% pc). it is known that in rhizosphere, the moisture holding capacity of the soil is improved by the exopolysaccharides (eps) produced by the pgp bacteria. also, it has been proved that the eps production has the ability to improve permeability by increasing soil aggregation and maintains higher water potential around the roots [70]. in addition to the shrink–swell behaviour of eps for different water potentials that affects mean pore size and passage of solutes and colloids of different size is the main cause of the decrease of soil permeability [4]. figure 6. averaged root surface density data under three water regimes (100% pc, 75% pc and 50% pc) (ns: non-significant differences between treatments, bars with the same letter(s) are not significantly different at p ≤ 0.05. agriculture and food sciences research, 2024, 11(1): 15-29 26 © 2024 by the authors; licensee asian online journal publishing group 3.3. tomato root development the results shown in the figure 6 confirm a significant difference in root surface density between bacteria, but not between water regimes or their interaction. therefore, according to the graph, it is obvious that under the two induced deficits irrigation 75% pc and 50% pc levels, the impact of the bacteria (sr7-77 strain) on the root surface density is higher than (m1) bacteria and control (c), confirming what is approved before by patten and glick [71] that the inoculation of various plant species with such bacteria has resulted in increased root growth and formation of lateral roots and root hairs, which affect positively the water acquisition and nutrients uptake, helping plants to cope with water deficit [72] as well as the results of sharafzadeh [73] revealed that p. fluorescens (92rk) increased total root surface area and volume. figure 7. images of scanned roots by imagej software for the three treatments: non-bacterized and two bacterized (m1 and sr7-77) under three water regimes:100%, 75% and 50% of pot capacity (pc). while to full regime irrigation, there is no difference between the effect of two bacteria and control, which can be explained by the efficiency of (b2 sr7-77 strain) under stress conditions. the bars also show standard deviation (sd) values added to show how the data spread around the mean value and how accurately the mean value represents the data. the observed standard deviation of each treatment is quite large, but this doesn’t mean that the data are not reliable because biological measurements are notoriously variable. the figure 7 shows instead, images of scanned roots obtained by imagej software for each treatment. the root images are representative of 126 collected roots samples. the images highlight as m1(b1) and sr7-77 strain (b2) treatment improved the root surface density (rsd) and higher than those obtained to control test and full irrigation regime (100% pc). in detail, rsd was equal to 0.540 % and 0.355 % to m1(b1) and sr7-77 strain (b2), respectively agriculture and food sciences research, 2024, 11(1): 15-29 27 © 2024 by the authors; licensee asian online journal publishing group and under 50% pc compared to 0.097% observed to uninoculated bacteria test, and 0.445 and 0.361 to ps and my, respectively and 0.282% to c, under 100% pc to control test. 4. conclusions prolonged dry conditions reduce plant growth and its development, but an alternative strategy as the pgp b would allow to alter soil water hydraulic properties and enhance the water stress tolerance of plants. therefore, pgpb may be considered as an amendment able to improve the soil hydraulic characteristics with a double benefit: 1) pgpb inoculation extend water stress conditions conferring tolerance to plant; 2) pgpb modify root growth and pores distribution. in term of rhizosphere soil hydraulic properties, micrococcus yunnanensis (m1-b1) and pseudomonas stutzeri (sr777 strain-b2) have altered both pore size distribution and water potential in the rhizosphere. results demonstrated that inoculating bacteria test altered the soil water retention curves in different ways. to m1 and ss7, van genuchten α-parameter (air entry point value of the swrcs) increased. while, the saturated hydraulic conductivity, ks, decreased due to the capability of pgp b to grow within pores. bacteria tended to keep all pores filled by water at high soil water potentials (i.e. less negative values), which is a condition already ensured with a full irrigation regime and where bacteria do not have an evident benefit to enhance the water root efficiency. at partially unsaturated soil conditions, pgp b stimulated the roots development in a way that water may be available even at low water potential values (higher negatives-toward low soil water content values). also, the two pgpb strains shown a change in the soil pore distribution which allowed to connect full and empty pores, an essential condition to reduce the negative effects induced by prolonged water stress conditions. the results indicated these selected pgp strains have the potential to significantly improve pores size distribution (psd) under water stress conditions. the pore size distribution curve shown different trend of pores classes for bacterized and non-bacterized treatments. instead, the soil hydraulic conductivity at soil saturation conditions has been reduced by the bacteria growth inner the pores. moreover, soil water potential variable assumed as representative of the root water uptake efficiency showing a better root growth and uptake because of bacteria allowed a greater stock of water at severe stress conditions in comparison to the control; 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licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 146-155, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6228 © 2024 by the authors; licensee asian online journal publishing group heavy metal accumulation in soil and leafy vegetables irrigated with domestic wastewater in vietnam pham thi thom1 nguyen tuan khoi2 phan le na3 ( corresponding author) 1,2,3bacgiang agriculture and forestry university, vietnam. 1email: thompt87@wru.vn 2email: khoint@bafu.edu.vn 3email: napl@bafu.edu.vn abstract this study was conducted to evaluate the accumulation of heavy metals, including copper (cu), lead (pb), and cadmium (cd), in soil and leafy vegetables irrigated with domestic wastewater. the surveyed vegetables included water spinach (ipomoea aquatica), chinese mustard greens (brassica integrifolia), and malabar spinach (basella alba), grown over three consecutive seasons. results showed that cu accumulation in the vegetables ranged from 4.41 mg/kg to 5.62 mg/kg, pb from 0.11 mg/kg to 0.13 mg/kg, and cd from 0.02 mg/kg to 0.03 mg/kg. all values were below the safety limits set by vietnam’s national technical regulation (qcvn), with thresholds of 30 mg/kg for cu, 0.3 mg/kg for pb, and 0.2 mg/kg for cd. similarly, heavy metal accumulation in the soil remained within the permissible limits under qcvn standards. the analysis further indicated that seasonal variations had minimal impact on heavy metal concentrations, suggesting that consistent wastewater management practices contributed to maintaining soil and crop safety. in addition, the research highlighted that water spinach accumulated slightly higher levels of heavy metals compared to the other vegetables, likely due to its faster growth rate and higher water absorption capacity. despite this, the concentrations remained well within safety thresholds, affirming the viability of these crops for safe consumption. the study concludes that using domestic wastewater for vegetable irrigation does not lead to heavy metal accumulation beyond safe thresholds, supporting sustainable and safe agricultural practices. however, continued monitoring and appropriate wastewater treatment remain essential to ensure long-term safety, particularly as environmental and anthropogenic factors evolve. keywords: domestic wastewater irrigation, environmental health risks, food safety and heavy metals, heavy metal contamination, leafy vegetable safety in vietnam, soil and plant health risks. citation | thom, p. t., khoi, n. t., & na, p. l. (2024). heavy metal accumulation in soil and leafy vegetables irrigated with domestic wastewater in vietnam. agriculture and food sciences research, 11(2), 146–155. 10.20448/aesr.v11i2.6228 history: received: 11 november 2024 revised: 9 december 2024 accepted: 12 december 2024 published: 20 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 147 2. content and research methodology .......................................................................................................................................... 147 3. research results ............................................................................................................................................................................ 148 4. conclusion ....................................................................................................................................................................................... 153 references ............................................................................................................................................................................................ 153 mailto:thompt87@wru.vn mailto:khoint@bafu.edu.vn mailto:napl@bafu.edu.vn https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6228 https://orcid.org/0009-0008-3017-8738 https://orcid.org/0000-0002-5920-1486 https://orcid.org/0009-0002-5198-7485 agriculture and food sciences research, 2024, 11(2): 146-155 147 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to sustainable agriculture by demonstrating that domestic wastewater can be safely used for irrigating leafy vegetables without exceeding heavy metal safety thresholds. it highlights the potential for resource optimization, provides baseline data for safe wastewater reuse practices, and underscores the importance of monitoring to ensure long-term environmental and food safety. 1. introduction in the context of freshwater scarcity, the reuse of domestic wastewater in agriculture has emerged as a potential solution to meet irrigation demands and alleviate pressure on freshwater resources [1]. however, domestic wastewater may contain heavy metals such as copper (cu), lead (pb), and cadmium (cd) [2] which are elements capable of accumulating in soil and crops [3] posing potential risks to agricultural product quality and human health if they exceed safe limits [4]. these heavy metals can be toxic in the environment, reducing crop yields and posing health risks if they enter the food chain [5]. leafy vegetables, which are often consumed directly [6] present a particular concern regarding heavy metal accumulation from soil and irrigation water, necessitating rigorous assessment [7]. this study aims to evaluate the accumulation levels of cu, pb, and cd in soil and leafy vegetables irrigated with domestic wastewater over three consecutive growing seasons. the selected vegetables include water spinach (ipomoea aquatica), chinese mustard greens (brassica integrifolia), and malabar spinach (basella alba). the accumulation levels of cu, pb, and cd in both the vegetables and soil will be compared to the safety limits set by vietnam’s national technical regulation (qcvn), thereby assessing the safety and feasibility of applying domestic wastewater in agriculture. 2. content and research methodology 2.1. research content this study assesses the accumulation of heavy metals in leafy vegetables and soil when irrigated with domestic wastewater. the vegetables examined include water spinach (ipomoea aquatica), chinese mustard greens (brassica integrifolia), and malabar spinach (basella alba). domestic wastewater is collected from ponds that receive discharge from surrounding households and then used to irrigate the vegetables in the experiment. the objective is to determine the heavy metal accumulation in both vegetables and soil over different planting seasons. 2.2. research methodology 2.2.1. experimental design subjects: three types of leafy vegetables: water spinach, chinese mustard greens, and malabar spinach. experimental period: conducted over three planting seasons: season 1: starting june 1, 2022. season 2: starting august 10, 2022. season 3: starting october 15, 2022. replications: each vegetable type is grown in three replicates (3 plots per vegetable). experimental plots are arranged randomly. 2.2.2. sample collection and analysis after each planting season, samples of both vegetables and soil are collected from the experimental plots. random samples are taken from the three replicates of each vegetable type to calculate the mean values of the parameters. sample collection and preparation for vegetables follow tcvn 9016:2011. soil sampling, preservation, and preparation adhere to standards: tcvn 7538-2:2005 (iso 10381-2:2002). heavy metal content in vegetable and soil samples is analyzed using atomic absorption spectroscopy (aas) as per tcvn 6496:2009. 2.2.3. data analysis method data are analyzed by calculating the mean values of parameters across replicates for each vegetable and season. spss statistical software is used to assess the differences in heavy metal content between vegetable types and planting seasons. table 1 presents the analysis results of domestic wastewater samples utilized for vegetable irrigation. table 1. analysis results of domestic wastewater samples used for vegetable irrigation. indicator parameter qcvn 08-mt:2015/btnmt temperature (°c) 24.42 ± 0.37 ph 7.52 ± 0.14 5.5-9 total suspended solids (tss) (mg/l) 482.6 ± 14.3 50 electrical conductivity (ec) (ms/cm) 0.59 ± 0.01 ammonium (nh₄⁺-n) (mg/l) 5.12 ± 0.14 0.9 phosphate (po₄³⁻-p) (mg/l) 1.58 ± 0.07 0.3 dissolved oxygen (do) (mg/l) 3.08 ± 0.17 ≥ 4 turbidity (ntu) 79.2 ± 2.72 cadmium (cd) (mg/l) 0.032 ± 0.0005 0.01 copper (cu) (mg/l) 0.019 ± 0.0005 0.5 lead (pb) (mg/l) 0.03± 0.0006 0.05 note: qcvn: vietnam’s national technical regulation. btnmt: ministry of natural resources and environment. agriculture and food sciences research, 2024, 11(2): 146-155 148 © 2024 by the authors; licensee asian online journal publishing group 3. research results the wastewater analysis indicates several parameters exceeding the acceptable limits set by the ministry of natural resources and environment (qcvn 08-mt:2015/btnmt), meaning they do not meet the required safety standards for irrigation. specifically, tss is at 482.6 mg/l, far above the permissible 50 mg/l. nh₄⁺-n is measured at 5.12 mg/l, surpassing the limit of 0.9 mg/l, and po₄³⁻-p is 1.58 mg/l, exceeding the allowed 0.3 mg/l. additionally, cd at 0.032 mg/l is above the limit of 0.01 mg/l. do at 3.08 mg/l falls below the required minimum of ≥4 mg/l. in contrast, pb at 0.03 mg/l and cu at 0.019 mg/l are within safe limits, meeting the standard requirements. these results highlight the need for treatment to reduce tss, ammonium, phosphate, and cadmium levels to comply with safety standards before irrigation use. table 2 illustrates the impact of domestic wastewater irrigation on copper (cu) accumulation in leafy vegetables. table 2. impact of domestic wastewater on cu accumulation in leafy vegetables. vegetable varieties the first season the second season the third season average regulations according to qcvn water spinach 5.57 ± 0.26 5.36 ± 0.27 5.11 ± 0.1 5.35± 0.28 30 chinese mustard greens 5.9 ± 0.28 5.6 ± 0.34 5.57 ± 0.21 5.62 ± 0.25 malabar spinach 4.43 ± 0.11 4.31 ± 0.22 4.48 ± 0.22 4.41 ± 0.18 note: (unit: mg/kg dry vegetable). qcvn: vietnam’s national technical regulation. excessive copper (cu) accumulation in leafy vegetables poses serious health risks due to its potential toxicity [8]. while cu is an essential micronutrient in small amounts, high concentrations can disrupt cellular processes by generating reactive oxygen species (ros), damaging plant tissues and reducing crop yield [9]. in humans, chronic exposure to elevated cu levels through food consumption may lead to symptoms such as gastrointestinal distress, liver dysfunction, and, in severe cases, liver diseases [10]. studies show that leafy vegetables like spinach can absorb substantial amounts of cu when grown in contaminated soils or irrigated with cu-laden wastewater [11]. in regions with high levels of cu contamination, concentrations in vegetables often exceed safe limits set by the world health organization (who), thus increasing the risk of bioaccumulation and adverse health effects [12]. this issue is especially concerning in areas where untreated wastewater is used for irrigation, emphasizing the need for stringent monitoring and improved agricultural practices to prevent cu-related health hazards in the food supply [13]. the study on copper (cu) accumulation in three types of leafy vegetables—water spinach (ipomoea aquatica), chinese mustard greens (brassica integrifolia), and malabar spinach (basella alba)—was conducted over three growing seasons with these vegetables irrigated using domestic wastewater. results indicate that cu accumulation in all these vegetables remained below the permissible limits set by the national technical regulation on heavy metal limits in food (qcvn), which specifies a maximum threshold for copper at 30 mg/kg. figure 1 compares copper (cu) accumulation in leafy vegetables irrigated with domestic wastewater against vietnamese safety standards. figure 1. comparison of cu accumulation in leafy vegetables with vietnamese standards. note: qcvn: vietnamese national technical regulations. specifically, over the three growing seasons, water spinach exhibited an average cu accumulation of 5.35 ± 0.28 mg/kg, chinese mustard greens 5.62 ± 0.25 mg/kg, and malabar spinach 4.41 ± 0.18 mg/kg. in individual seasons, cu accumulation in water spinach ranged from 5.11 mg/kg to 5.57 mg/kg, in chinese mustard greens from 5.57 mg/kg to 5.9 mg/kg, and in malabar spinach from 4.31 mg/kg to 4.48 mg/kg. chinese mustard greens showed the highest cu accumulation, especially in the first season at 5.9 mg/kg, while malabar spinach consistently had the lowest accumulation across all seasons, with an average of only 4.41 mg/kg. agriculture and food sciences research, 2024, 11(2): 146-155 149 © 2024 by the authors; licensee asian online journal publishing group although there are some differences in cu accumulation among the vegetable types, the variation is not substantial. chinese mustard greens tend to absorb more cu compared to water spinach and malabar spinach, but all three vegetables maintained safe cu accumulation levels, significantly below the maximum threshold of 30 mg/kg set by qcvn. this indicates that, under experimental conditions, the use of domestic wastewater for irrigating leafy vegetables does not lead to a significant increase in copper accumulation that would pose health risks to consumers. table 3. presents the impact of domestic wastewater on pb accumulation in leafy vegetables. vegetable varieties the first season the second season the third season average regulations according to qcvn water spinach 0.12 ± 0.006 0.13 ± 0.006 0.12 ± 0.01 0.12± 0.007 0.3 chinese mustard greens 0.12 ± 0.006 0.13 ± 0.006 0.15 ± 0.006 0.13± 0.014 malabar spinach 0.12 ± 0.006 0.11 ± 0.006 0.11± 0.006 0.11± 0.007 note: (unit: mg/kg dry vegetable). qcvn: vietnam’s national technical regulation. lead (pb) accumulation in leafy vegetables and domestic wastewater poses significant health and environmental risks [14]. pb is a toxic heavy metal with no known biological function in plants or humans [15]. its presence in soil and water sources, often due to industrial activities and urban runoff, leads to absorption by plants, especially leafy vegetables, through both roots and foliage exposed to contaminated water and atmospheric particles [16]. when pb concentrations exceed the world health organization’s safety limits, its consumption through contaminated vegetables can result in serious health consequences, including neurological damage, kidney dysfunction, and developmental issues, particularly in young children [1]. in domestic wastewater, pb can accumulate from various sources, including plumbing, industrial discharge, and household products [2]. when this wastewater is used for irrigation without adequate treatment, it increases pb levels in the soil, which plants readily absorb [3]. this cycle not only impacts food safety but also threatens soil health, as pb can persist in the environment, leading to long-term ecological contamination [17]. these risks highlight the critical need for managing pb in wastewater and ensuring it does not enter the agricultural food chain [18]. table 3 assessed the accumulation of lead (pb) in leafy vegetables, including water spinach, chinese mustard greens, and malabar spinach, when irrigated with domestic wastewater over three growing seasons. the results indicate that pb levels in all vegetable samples remained within the safety limits set by the national technical regulation (qcvn) for heavy metal limits in food, with a maximum threshold for pb at 0.3 mg/kg. figure 2 compares pb accumulation in leafy vegetables irrigated with domestic wastewater to vietnamese safety standards. figure 2. comparison of pb accumulation in leafy vegetables with vietnamese standards. the pb content in water spinach fluctuated slightly across the seasons, with values of 0.12 ± 0.006 mg/kg in the first and third seasons and 0.13 ± 0.006 mg/kg in the second season, averaging 0.12 ± 0.007 mg/kg. chinese mustard greens exhibited slightly higher pb accumulation than water spinach, ranging from 0.12 ± 0.006 mg/kg in the first season, 0.13 ± 0.006 mg/kg in the second, and peaking at 0.15 ± 0.006 mg/kg in the third season, with an average of 0.13 ± 0.014 mg/kg. for malabar spinach, pb levels were more stable and lower than in the other vegetables, ranging from 0.11 ± 0.006 mg/kg in the second and third seasons to 0.12 ± 0.006 mg/kg in the first season, averaging 0.11 ± 0.007 mg/kg. a comparison across the three vegetables shows that chinese mustard greens had the highest pb accumulation, particularly in the third season at 0.15 mg/kg, while malabar spinach had the lowest accumulation across all agriculture and food sciences research, 2024, 11(2): 146-155 150 © 2024 by the authors; licensee asian online journal publishing group seasons. however, these differences are minimal, and all values are well below the maximum threshold of 0.3 mg/kg set by qcvn. these results indicate that, although domestic wastewater may contain pb, the accumulation levels in leafy vegetables grown under the study conditions remain safe and do not exceed permissible limits. table 4 demonstrates the impact of domestic wastewater irrigation on cd accumulation in leafy vegetables. table 4. impact of domestic wastewater on cd accumulation in leafy vegetables. vegetable varieties the first season the second season the third season average regulations according to qcvn water spinach 0.03 ± 0.006 0.03 ± 0.006 0.03 ± 0.006 0.03 ± 0.005 0.2 chinese mustard greens 0.03 ± 0.006 0.03 ± 0.006 0.03 ± 0.006 0.03 ± 0.005 malabar spinach 0 0.03 ± 0.006 0.03 ± 0.006 0.02 ± 0.004 note: (unit: mg/kg dry vegetable). qcvn: vietnam’s national technical regulation. the accumulation of cadmium (cd) in leafy vegetables and domestic wastewater poses serious health hazards to humans [19]. cd is a toxic heavy metal that, unlike essential nutrients, serves no beneficial role in biological systems [20]. when cd enters the food chain through contaminated leafy vegetables, it can lead to severe health issues [21]. long-term ingestion of cd-contaminated foods can result in kidney damage, osteoporosis, and even cancer due to cd's high toxicity and tendency to accumulate in the body over time [22]. in domestic wastewater, cd often originates from industrial waste, batteries, and household products [23]. when such wastewater is used for irrigation, cd can be transferred to the soil and then taken up by plants, especially leafy vegetables, which are prone to heavy metal accumulation [24]. this cycle not only increases cd concentrations in edible crops but also creates persistent contamination in agricultural soil [25]. the risks associated with cd accumulation underline the necessity for effective wastewater treatment and regular monitoring to protect public health and ensure safe food production [26]. in the study on cadmium (cd) accumulation in three types of leafy vegetables—water spinach, chinese mustard greens, and malabar spinach—irrigated with domestic wastewater over three growing seasons, results consistently showed that cd accumulation levels remained within the safe limits set by the national technical regulation (qcvn), which specifies a maximum threshold for cd at 0.3 mg/kg. figure 3 compares cd accumulation in leafy vegetables irrigated with domestic wastewater against vietnamese safety standards. figure 3. comparison of cd accumulation in leafy vegetables with vietnamese standards. water spinach showed stable cd accumulation across the three seasons, with values of 0.03 ± 0.006 mg/kg in each season and an average of 0.03 ± 0.005 mg/kg. similarly, chinese mustard greens also exhibited steady cd levels, ranging around 0.03 ± 0.006 mg/kg in each season, averaging 0.03 ± 0.005 mg/kg. malabar spinach, however, showed a slight variation, with no detectable cd accumulation in the first season (0 mg/kg), increasing to 0.03 ± 0.006 mg/kg in the second and third seasons, resulting in a three-season average of 0.02 ± 0.004 mg/kg. comparing the vegetables, both water spinach and chinese mustard greens showed similar cd accumulation across all seasons, while malabar spinach had significantly lower levels in the first season. nonetheless, this difference holds little significance for food safety, as all values were well below the qcvn safety threshold of 0.3 mg/kg. table 5. shows the impact of domestic wastewater irrigation on cu accumulation in soil. vegetable varieties the first season the second season the third season average regulations according to qcvn water spinach 23.62 ± 1.13 24.15 ± 0.49 23.81 ± 1.09 23.86 ± 0.85 100 chinese mustard greens 23.62 ± 1.13 24.08 ± 0.43 23.88 ± 0.98 23.86 ± 0.81 malabar spinach 23.69 ± 0.49 24.21 ± 0.38 23.83 ± 0.46 23.91 ± 0.45 note: (unit: mg/kg). qcvn: vietnam’s national technical regulation. agriculture and food sciences research, 2024, 11(2): 146-155 151 © 2024 by the authors; licensee asian online journal publishing group copper (cu) accumulation in agricultural soil poses significant risks to both plant health and human safety. in soil, excessive cu levels can lead to phytotoxicity, disrupting key plant processes like photosynthesis and enzyme activity, ultimately reducing crop yield and quality [27]. cu’s high affinity for binding to soil particles also means it can persist in the environment, gradually building up to toxic levels in areas subjected to repeated cu exposure from fertilizers or wastewater irrigation [28]. for plants, cu toxicity often manifests in stunted growth, leaf discoloration, and reduced root development, as high cu concentrations interfere with nutrient absorption [29]. when leafy vegetables are grown in cucontaminated soil, they tend to absorb and accumulate cu in their tissues, which can be transferred to humans through consumption [30]. long-term exposure to elevated cu levels in food has been associated with gastrointestinal distress, liver toxicity, and, in severe cases, neurological impacts [31]. these potential health effects highlight the need for careful cu management in agricultural soils to protect both environmental and public health [32]. table 5 presents data on the impact of domestic wastewater on copper (cu) accumulation in soil used to grow leafy vegetables, including water spinach, chinese mustard greens, and malabar spinach, over three planting seasons. the results indicate that cu accumulation in soil ranged from 23.62 ± 1.13 mg/kg to 24.21 ± 0.38 mg/kg, with average values between 23.86 ± 0.85 mg/kg and 23.91 ± 0.45 mg/kg. compared to the vietnamese national technical regulation (qcvn), which sets the maximum allowable cu level in soil at 100 mg/kg, all cu accumulation values in the soil for the three vegetables remained well below this limit, with averages significantly lower than the regulated threshold. figure 4 illustrates cu accumulation in soil irrigated with domestic wastewater compared to qcvn standards. figure 4. cu accumulation in soil with domestic wastewater irrigation compared to qcvn. note: qcvn: vietnam’s national technical regulation. cu accumulation showed minor seasonal fluctuations, with standard deviations ranging from 0.38 mg/kg to 1.13 mg/kg, indicating stability in the accumulation process. malabar spinach showed the least variation, while water spinach and chinese mustard greens exhibited slightly higher fluctuations but remained within safe levels. these results demonstrate that using domestic wastewater for soil irrigation does not lead to excessive cu accumulation, ensuring safety for the soil ecosystem and crop health. table 6 details the impact of domestic wastewater irrigation on pb accumulation in soil. table 6. impact of domestic wastewater on pb accumulation in soil. vegetable varieties the first season the second season the third season average regulations according to qcvn water spinach 1.9 ± 0.07 1.86± 0.11 1.92± 0.04 1.89 ± 0.07 70 chinese mustard greens 1.93 ± 0.16 1.62 ± 0.17 1.76 ± 0.2 1.77 ± 0.2 malabar spinach 1.82 ± 0.12 1.8 ± 0.08 1.88 ± 0.13 1.83 ± 0.11 note: (unit: mg/kg). qcvn: vietnam’s national technical regulation. agriculture and food sciences research, 2024, 11(2): 146-155 152 © 2024 by the authors; licensee asian online journal publishing group lead (pb) accumulation in agricultural soil, particularly from the use of untreated domestic wastewater for irrigation, presents severe risks to both crops and human health [33]. pb is a highly toxic heavy metal with no essential role in biological processes, and its persistence in soil means it can build up over time, especially when wastewater containing pb residues is repeatedly used for irrigation [33]. in soil, pb binds tightly to particles, making it challenging to remove and allowing it to be absorbed by plant roots [34]. when vegetables absorb pb from contaminated soil, they become vectors of pb exposure for humans, who may ingest the metal through their diet [35]. ingesting pb-contaminated vegetables can lead to a range of health problems, from developmental delays and neurological damage in children to kidney dysfunction and cardiovascular issues in adults [36]. because pb is not readily excreted from the body, it accumulates over time, causing long-term health impacts, particularly when exposure occurs regularly [37]. this risk underlines the importance of proper wastewater treatment and soil monitoring to ensure safe levels of heavy metals in agricultural soils and prevent pb from entering the food chain [38]. pb accumulation in the soil ranged from 1.62 mg/kg to 1.93 mg/kg, with an average of 1.89 ± 0.07 mg/kg for water spinach, 1.77 ± 0.2 mg/kg for chinese mustard greens, and 1.83 ± 0.11 mg/kg for malabar spinach. compared to the vietnamese national technical regulation (qcvn) limit of 70 mg/kg for pb in soil, all pb accumulation values were significantly below the safety threshold. figure 5 illustrates pb accumulation in soil irrigated with domestic wastewater compared to qcvn standards. figure 5. pb accumulation in soil with domestic wastewater irrigation compared to qcvn. note: qcvn: vietnam’s national technical regulation. seasonal fluctuations in pb levels were minimal, with chinese mustard greens showing the most variation and malabar spinach the most stability. these results indicate that the use of domestic wastewater for irrigation does not result in significant pb accumulation in soil, keeping pb content well within the safe limits set by qcvn. thus, domestic wastewater can be safely utilized in agricultural production without posing a risk of excessive pb accumulation. table 7 presents the impact of domestic wastewater irrigation on cd accumulation in soil. table 7. impact of domestic wastewater on cd accumulation in soil. vegetable varieties the first season the second season the third season average regulations according to qcvn water spinach 0.1 ± 0.06 0.1 ± 0.06 0.2 ± 0.06 0.13 ± 0.05 1.5 chinese mustard greens 0.2 ± 0.06 0.2 ± 0.06 0.2 ± 0.06 0.15 ± 0.07 malabar spinach 0.2 ± 0.06 0.1 ± 0.06 0.2 ± 0.06 0.17 ± 0.07 note: (unit: mg/kg). qcvn: vietnam’s national technical regulation. cadmium (cd) accumulation in soil used for vegetable cultivation presents significant health and environmental risks, particularly when domestic wastewater is used for irrigation [39]. cd is a toxic heavy metal with no beneficial role in plants or animals and is known for its persistence and bioaccumulation potential in soil [40]. over time, soils irrigated with cd-contaminated water sources, such as untreated wastewater, can accumulate cd to levels that impact both plant health and the safety of food produced in these soils [41]. agriculture and food sciences research, 2024, 11(2): 146-155 153 © 2024 by the authors; licensee asian online journal publishing group in plants, cd disrupts growth by interfering with essential nutrient absorption and generating reactive oxygen species (ros), which damage cellular structures [42]. leafy vegetables, in particular, tend to absorb and concentrate cd in their tissues, making them significant sources of cd exposure when consumed [43]. chronic ingestion of cd through contaminated vegetables can lead to serious health issues in humans, including kidney damage, bone demineralization, and increased cancer risk [44]. the persistence of cd in soils further compounds these risks, as it accumulates over time, presenting a continuous hazard for crops and consumers alike [45]. cd levels ranged from 0.1 mg/kg to 0.2 mg/kg, with an average of 0.13 ± 0.05 mg/kg for water spinach, 0.15 ± 0.07 mg/kg for chinese mustard greens, and 0.17 ± 0.07 mg/kg for malabar spinach. compared to the vietnamese national technical regulation (qcvn), which sets a maximum permissible limit of 1.5 mg/kg for cd in soil, all cd accumulation values remained well below this threshold. seasonal variations in cd content were minimal, indicating stable accumulation levels in soil irrigated with domestic wastewater. figure 6 depicts cd accumulation in soil irrigated with domestic wastewater compared to qcvn standards. figure 6. cd accumulation in soil with domestic wastewater irrigation compared to qcvn. note: qcvn: vietnam’s national technical regulation. these results demonstrate that using domestic wastewater for irrigation does not lead to excessive cd accumulation in soil, ensuring no harm to the soil environment or crops. therefore, domestic wastewater use in agriculture can be considered sustainable and safe concerning heavy metal accumulation in soil. 4. conclusion the research results indicate that the accumulation levels of cu, pb, and cd in soil and leafy vegetables irrigated with domestic wastewater are within the safety limits specified by vietnam's national technical regulation (qcvn). specifically, cu accumulation in vegetables ranged from 4.41 mg/kg to 5.62 mg/kg, pb from 0.11 mg/kg to 0.13 mg/kg, and cd from 0.02 mg/kg to 0.03 mg/kg, all significantly below the qcvn safety thresholds of 30 mg/kg for cu, 0.3 mg/kg for pb, and 0.2 mg/kg for cd. similarly, in soil, cu accumulation ranged from 23.86 mg/kg to 23.91 mg/kg, pb from 1.77 mg/kg to 1.89 mg/kg, and cd from 0.13 mg/kg to 0.17 mg/kg, all well within the regulatory limits established by qcvn. these findings demonstrate that using domestic wastewater for leafy vegetable irrigation does not lead to excessive heavy metal accumulation in vegetables or soil, ensuring safety for crops and soil ecosystems. this study thus confirms the feasibility of reusing domestic wastewater in agriculture. utilizing domestic wastewater not only helps reduce the pressure on freshwater resources but also maintains sustainability and safety in agricultural production. references [1] h. waheed, n. ilyas, n. iqbal raja, t. mahmood, and z. ali, "heavy metal phyto-accumulation in leafy vegetables irrigated with municipal wastewater and human health risk repercussions," international journal of phytoremediation, vol. 21, no. 2, pp. 170-179, 2019. https://doi.org/10.1080/15226514.2018.1540547 [2] h. kim, t. jang, s. kim, and s. park, "impact of domestic wastewater irrigation on heavy metal contamination in soil and vegetables," environmental earth sciences, vol. 73, pp. 2377-2383, 2015. https://doi.org/10.1007/s12665-014-3581-2 [3] s. anwar, m. f. nawaz, s. gul, m. rizwan, s. ali, and a. kareem, "uptake and distribution of minerals and heavy metals in commonly grown leafy vegetable species irrigated with sewage water," environmental monitoring and assessment, vol. 188, pp. 1-9, 2016. https://doi.org/10.1007/s10661-016-5560-4 [4] m. k. souri, n. alipanahi, and g. tohidloo, "heavy metal content of some leafy vegetable crops grown with waste water in southern suburb of tehran-iran," vegetable science, vol. 43, no. 2, pp. 156-162, 2016. https://doi.org/10.1080/15226514.2018.1540547 https://doi.org/10.1007/s12665-014-3581-2 https://doi.org/10.1007/s10661-016-5560-4 agriculture and food sciences research, 2024, 11(2): 146-155 154 © 2024 by the authors; 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[44] m. m. ngugi, h. i. gitari, c. muii, and j. p. gweyi-onyango, "cadmium mobility, uptake, and accumulation in spinach, kale, and amaranths vegetables as influenced by silicon fertilization," bioremediation journal, vol. 26, no. 2, pp. 113-127, 2022. https://doi.org/10.1080/10889868.2021.1924111 [45] q. xiao, s. wang, and y. chi, "accumulation and chemical forms of cadmium in tissues of different vegetable crops," agronomy, vol. 13, no. 3, p. 680, 2023. https://doi.org/10.3390/agronomy13030680 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.ecoenv.2015.05.019 https://doi.org/10.1016/0048-9697(90)90300-j https://doi.org/10.1016/j.envpol.2020.114677 https://doi.org/10.3390/life12030339 https://doi.org/10.1080/10889868.2021.1924111 https://doi.org/10.3390/agronomy13030680 25 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 25-30, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6568 © 2025 by the authors; licensee asian online journal publishing group application of shiitake and maitake extracts to wheat flour and their effect on technological and baking properties tatiana bojnanska1 matej cech2 anna kolesarova3 jozef bojnansky4 ( corresponding author) 1,2,3institute of food sciences, slovak university of agriculture in nitra, tr. a. hlinku 2, 949 76 nitra, slovak republic. 1email: tatiana.bojnanska@uniag.sk 2email: matej.cech@uniag.sk 3email: anna.kolesarova@uniag.sk 4institute of accounting and informatics, slovak university of agriculture in nitra, tr. a. hlinku 2, 949 76 nitra, slovak republic. 4email: jozef.bojnansky@uniag.sk abstract shiitake and maitake extracts were added to composite flours in the amounts of 5%, 7.5%, and 10%, which subsequently changed the basic technological properties of the composite flours. compared to control flour, wet gluten content decreased only slightly, but gluten swelling and gluten extensibility decreased significantly. falling number, zeleny sedimentation volume, and crude protein also decreased. the experimental loaves were baked from composite flours and evaluated by volscan. the weight of the experimental loaves decreased in proportion to the amount of addition of medicinal mushroom extract. the volume and specific volume of the experimental loaf decreased with the addition of maitake extract in proportion to its amount. the addition of shiitake extracts in the amount of 5% increased the volume and specific volume of the experimental loaf; however, higher additions (7.5%, 10%) reduced the evaluated parameters in the case of shiitake extract. nevertheless, all additions were technologically acceptable, and the differences in technological quality of experimental loaves compared to control loaves were not significant. these types of designed foods with nutritional benefits have potential for producers and are desirable to consumers. keywords: composite flours, mushroom extracts, gluten, falling number, zeleny index, baking test, quality evaluation, volscan. citation | bojnanska, t., cech, m., kolesarova, a., & bojnansky, j. (2025). application of shiitake and maitake extracts to wheat flour and their effect on technological and baking properties. agriculture and food sciences research, 12(1), 25–30. 10.20448/aesr.v12i1.6568 history: received: 27 february 2025 revised: 31 march 2025 accepted: 2 april 2025 published: 9 april 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by slovak research and development agency (grant number: apvv-21-0206). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 26 2. materials and methods ................................................................................................................................................................... 26 3. results and discussion ................................................................................................................................................................... 27 4. conclusion ......................................................................................................................................................................................... 29 references .............................................................................................................................................................................................. 29 mailto:tatiana.bojnanska@uniag.sk mailto:matej.cech@uniag.sk mailto:anna.kolesarova@uniag.sk mailto:jozef.bojnansky@uniag.sk https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6568 https://orcid.org/0000-0001-6862-8337 https://orcid.org/0000-0003-3260-2447 https://orcid.org/0000-0001-6438-0039 https://orcid.org/0009-0009-1609-1166 agriculture and food sciences research, 2025, 12(1): 25-30 26 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature the research provides information on the changes caused by the addition of non-baking ingredients to wheat flours intended to produce bread. the presented additives are exceptional in their nutritional benefits, and the results obtained are original regarding the possibilities of their application to make bakery products more attractive. 1. introduction cereals are key plant resources that have provided human nutrition for thousands of years. wheat and rye, as the basic cereals of europe, are irreplaceable from the point of view of the production of bread and pastries as basic foods. despite the fact that the consumption of bread and pastries in slovakia has been decreasing for more than 30 years, in 2022 it was at the level of 62 kg per person per year [1], which is definitely not low. such consumption is significant enough to call attention to the nutritional benefits of consuming these basic foods. the main raw material used worldwide for bread production is wheat flour, which is characterized mainly by a high proportion of starch and a low proportion of proteins that are deficient in some essential amino acids, but with an exceptional ability to capture fermentation gases formed during the fermentation process, thanks to gluten [2, 3]. efforts to improve the nutritional quality of bread are made by partially replacing wheat with non-bakery raw materials, which are valued for their content of biologically active ingredients that traditional breads do not contain. this produces a variety of products with beneficial health effects [4-9]. however, in composite flours prepared by combining wheat flour and flours from non-bakery raw materials, or directly by specific valuable ingredients, a lower proportion of gluten causes problems related to the technological properties of the flour, dough, and finished product. the gluten-free addition causes the wheat dough to be diluted and subsequently weakened; however, the effect of the addition can be very specific [10-13]. a balance must be struck between nutritional benefits and technological requirements. the group of non-bakery raw materials in which nutritional benefits can be assumed includes medicinal mushrooms, which exhibit a wide range of pharmacological activities, including antiallergic, antibacterial, antifungal, anti-inflammatory, antioxidant, antiviral, cytotoxic, immunomodulatory, antidepressant, antihyperlipidemic, antidiabetic, digestive, hepatoprotective, neuroprotective, nephroprotective, and osteoprotective [14]. the authors summarized known findings and compared clinical studies aimed at confirming the positive effects of the consumption of medicinal mushrooms. in shiitake (lentinula edodes), an important compound with immunomodulatory effects is lentinan, β-1,3-dglucan. polyphenol-rich mushroom extract also shows high free radical scavenging [15-18]. the antioxidant effect of shiitake is caused by ergothioneine (2-mercaptohistidine trimethylbetaine), whose significant increase in blood within two hours after mushroom consumption was demonstrated by weigand-heller et al. [19]. as a potential raw material, shiitake is interesting because of its content in therapeutic compounds that have antitumor, antifungal, antibacterial, anti-inflammatory, hypocholesterolemic, antihypertensive, hypoglycemic, and antioxidant effects [18]. the potential positive effects of maitake extract (grifola frondosa) on the human body can be summarized in the benefits related to its antioxidant mechanisms [20], which are conditioned by the content of valuable bioactive components [21]. moreover, the content of polysaccharides, especially β-glucans and heteroglycans, as well as other bioactive macromolecules, is potentially useful in terms of antitumor and immunomodulatory effects [22, 23], prebiotic effects [24], or effects on visceral leishmaniasis [25]. the aim of the study was to verify the possibilities of adding extracts from medicinal mushrooms, shiitake and maitake, to wheat flour, and subsequently to evaluate the basic technological properties of composite flours and the resulting experimental breads. 2. materials and methods 2.1. materials for the preparation of composite flours, commercial wheat flour (mlyn grznár, veľké hoste, slovakia) was used, the quality of which is defined by the following parameters: energy value (1464 kj), carbohydrate content (71 g/100 g) including sugars (1.59 g/100 g), fiber content (3.3 g/100 g), protein content (11 g/100 g), fat content (1.3 g/100 g) including saturated fat (0.3 g/100 g), salt (0.01 g/100 g), and ash content (0.65 %). as part of the presented research, two types of non-bakery additives were added to wheat flour: maitake extract and shiitake extract. the extracts were obtained from natural field (xi’an natural field bio-technique co., ltd., xi’an, china), from the source one pharma s.r.o, janka kráľa 5, 974 01 banská bystrica, slovakia. formulations using the evaluated composite flour (experimental design) are listed in table 1. table 1. experimental design. wheat flour t650 shiitake extract maitake extract identification of the samples 100 % 0 % 0 % wf 95 % 5 % 5 % sh_5 ma_5 92.5 % 7.5 % 7.5 % sh_7.5 ma_7.5 80 % 10 % 10 % sh_10 ma_10 2.2. methods as part of the evaluation of the technological quality of composite flours, the following parameters were determined: moisture by gravimetric method in % (icc standard 110/1, 1976), gluten content g30 (iso 214151:2006), plus the swelling of gluten t30 (cm), and the extensibility of gluten q30 (ml), sedimentation index by zeleny in ml (stn iso 5529), falling number in seconds (icc 107/1, falling number 1500, fa perten instruments, ab), crude protein (kjeldahl, n x 5,7), starch content by ewers (iso 10520:1997), conversion factor 1.898, ash content in % (icc standard 104/1) and titration acidity of flour (mmol/kg). agriculture and food sciences research, 2025, 12(1): 25-30 27 © 2025 by the authors; licensee asian online journal publishing group experimental breads were baked from composite flours (table 1) by the processing procedure described by bojňanská et al. [26] and analysed using a volscan profiler volume analyser (stable mycrosystems, surrey, uk). the following parameters were evaluated: weight of the bread (g), bread volume (ml), specific volume (ml/g), volume yield (ml/100 g flour), and aspect ratio of a middle slice. 2.3. statistical analysis all analyses were performed in triplicate, and average values were calculated. xlstat 2020.5.1 [27] together with microsoft excel 365 [28] was used as the statistical and data analysis software (anova and a duncan’s multiple range test). 3. results and discussion the addition of non-bakery raw materials generally worsens the technological parameters of composite flours, which has also been confirmed in the case of the addition of maitake and shiitake extracts. the amount of wet gluten, its ductility, and swelling decreased in proportion to the amount of extracts added (figure 1). these findings are not surprising, as the change in protein-protein fraction ratios in composite flours affects the rheological properties of the dough [8, 26, 29]. the main problem in applying non-wheat flour to dough is the disturbance of the starch– gluten matrix and the dilution of gluten, which reduces its ability to retain gases produced during dough leavening [30-32]. in addition to the decrease in wet gluten content, which is understandable, the extensibility of gluten decreased significantly. nevertheless, the values observed in table 1 can be characterized as adequate, indicating the usability of such composite flours. other parameters evaluating the technological quality of flours were also influenced by the addition of extracts. compared to control wheat flour, flours with added extracts had demonstrably higher enzymatic activity and lower sedimentation index (zi) values. figure 1. gluten content and its properties in composite flours. note: legend: g30 wet gluten content after 30 minutes of resting (%); t30 the swelling of gluten (cm); q30 the extensibility of gluten (ml). * means significantly different samples compared to control (p≤0.05). starch content did not change significantly, but crude protein content was lower. the addition of shiitake extract reduced the ash content. interestingly, the addition of maitake significantly increased the acidity of the composite flour, up to 23% with just a 5% addition compared to control flour. this finding does not have to be negative, as the higher acidity of the flour can positively influence the sensory evaluation of the final products. in bread and pastries, it is the cause of a fuller and better perceived taste [33-36]. figure 2 shows comparisons expressed as a percentage, representing 100% of the value of a given property in control wheat flour (wf). figure 2. technological quality of composite flours with the addition of 5 % of extracts. note: legend: n x 5,7 crude protein content compared with control (100 %); starch starch content compared with control (100 %); ash ash content compared with control (100 %); fn – falling number compared with control (100 %); zi – sedimentation test compared with control (100 %); ta titration acidity compared with control (100 %). *means significantly different samples compared to control (p≤0.05). agriculture and food sciences research, 2025, 12(1): 25-30 28 © 2025 by the authors; licensee asian online journal publishing group the indirect indicators of bakery quality evaluated indicated a potentially negative impact of the additions of medicinal mushroom extracts, although most of the evaluated indicators did not show changes at a level that would predict that it is unsuitable to use the composite flours under consideration. on the contrary, in the case of an addition of 5%, we can expect good results from baking experiments. more interesting, more informative, and more convincing in terms of quality, usable results in practice were found within the baking experiment, which is considered a direct method of determining bakery quality. when preparing the dough, it is important to add a suitable amount of water, which will ensure the formation of dough with optimal consistency and affect the total weight of the dough being processed. the final weight of the bread is subsequently affected by baking losses. in the case of the evaluated experimental bread, their weight decreased in proportion to the amount of additions. figure 3 shows comparisons of important bread quality parameters, expressed as a percentage, representing 100% of the value of a given property in control wheat flour (wf). figure 3. qualitative parameters of experimental breads. note: *means significantly different samples compared to control (p≤0.05). the bread volume can clearly be considered the most important quality parameter. it was very interesting to find that the addition of shiitake extract in the amount of 5% had a positive effect on the bread volume, and in the experimental bread, the bread volume was higher than in the control breads. higher additions of shiitake extract already reduced the bread volume in comparison with the control. in the case of all additions (5%, 7.5%, and 10%) of maitake extract, the properties of the experimental breads were worse than in the control breads (figure 3). this is also documented by the scans obtained by volscan (figure 4a) and the photo documentation of the experimental bread compared to the control (figure 4b), on which the differences in the volume, shape, and porosity of the breads are very clearly visible. the volume and other characteristics of experimental loaves are key in evaluating their quality, but trade-offs must be sought in the case of value-added foods where the expected nutritional benefits of non-bakery additives are pursued. we believe that bread's slightly reduced level of technological quality is significantly outweighed by its nutritional benefits. sensory evaluation is also considered an important part of a comprehensive assessment of innovative products, as it ultimately determines whether a product will be suitable for consumption. figure 4. a, b. scans of experimental bread and photos of experimental bread. agriculture and food sciences research, 2025, 12(1): 25-30 29 © 2025 by the authors; licensee asian online journal publishing group in general, value-added foods are rated worse than standard ones, especially in relation to the additives used and their amounts [26, 36-39]. when applying dried shiitake powder at levels of 5% and 10%, the results found by van toan and thu [36] were comparable to or slightly better than the evaluators' overall preference compared to control pastries. pastries with the addition of 15% were no longer acceptable to evaluators. other authors [33] found significantly lower bread preferences, in which wheat flour was replaced by shiitake powder as early as 5%. in most cases, non-bakery ingredients worsen the technological parameters of bread, which is also associated with impaired perception by consumers [40-42]. in the case of composite flours and experimental bread evaluated by us, it can be stated that the additions of maitake and shiitake extracts influenced the technological properties of both composite flours and breads; however, the found values can be considered acceptable and balanced with the expected nutritional benefits. the method of adapting and targeted design of staple foods to the intended nutritional benefits leads to healthier foods and potential improvements in consumer health. 4. conclusion due to the addition of extracts of medicinal mushrooms shiitake and maitake, the basic technological properties of composite flours have changed. in proportion to the amount of the addition, wet gluten content decreased, and its swelling and extensibility decreased. the value of the falling number also decreased, so composite flours have higher enzymatic activity, but all values complied with legislative requirements. the value of the zeleny index, crude protein content, and, in the case of composite flour with added shiitake, ash content also fell. its acidity did not change, unlike composite flour with the addition of maitake, in which the acidity increased significantly compared to both control flour and composite flours with the addition of shiitake. in relation to the results of the bakery experiment, it was found that the weight of the loaves decreased in proportion to the amount of addition of medicinal mushroom extracts. the volume and specific volume of the loaves decreased with the addition of maitake extract in proportion to its height. the addition of shiitake extracts in the amount of 5% resulted in an increase in the volume and specific volume of the experimental loaf (by 4.7% and 8.5% respectively), so that with such an addition, apart from the nutritional benefits, there was also an improvement in the technological quality of the bread. higher additions of extracts reduced the evaluated parameters. nevertheless, all additions were technologically acceptable. verified composite flours and breads with nutritional benefits have potential for manufacturers and are desirable for consumers. references [1] datacube, "statistical office of slovak republic," 2023. retrieved: https://datacube.statistics.sk/. 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licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 43-51, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.5870 © 2024 by the authors; licensee asian online journal publishing group evaluation of the mineral and antinutritional composition of bottled and sterilized tigernut (cyperus esculentus) milk chinasa okorie1 gregory ikechukwu onwuka2 nneoma elechi obasi3 ( corresponding author) 1,2,3department of food science and technology, michael okpara university of agriculture umudike, p.m.b 7267 umuahia, abia state, nigeria. 1email: okorie.chinasa@mouau.edu.ng 2email: onwuka.gregory@mouau.edu.ng 3email: obasi.nneoma@mouau.edu.ng abstract nowadays, beverages are not just considered as thirst quenchers; specific functionality is the major ingredient expected in beverages, which has becomes a lifestyle. in recent years, these initiations have led to newly developed products in the beverages sector. companies are learning the consumption styles of various individuals and making novel beverages through blends of milk and energy drinks. tigernut milk is among the most valued plant-based beverages, obtained from the aqueous extract of tigernuts tubers. this study investigated the mineral and antinutritional components of bottled and sterilized tigernut milk. two varieties of tigernut tuber (yellow and brown) were processed using three methods (boiling, soaking and malting), and the milk was bottled and further sterilized. the results obtained showed that iron recorded the least value at (1.73mg and 2.24mg) for yellow and brown tigernut tubers; alsotigernut tubers were low in some antinutrients except flavonoids. the result for the tigernut milk elucidated that processing and sterilization had little effect on the mineral content of tigernut milk. the study also showed that the pre-processing treatment applied (soaking, boiling and malting) to the tigernut either increased or caused a decline in the mineral content of the tigernut milk, whereas, processing and sterilization affected the antinutrient content of tigernut milk. all processing methods showed a significant difference (p<0.05.) as there was a decrease in the phytate and oxalate content. the study highlighted the possibility of processing tigernut tubers into milk extract that can be bottled and sterilized for a period of time. keywords: tigernut, minerals, antinutrients, plant-based beverages, processing. citation | okorie, c., onwuka, g. i., & obasi, n. e. (2024). evaluation of the mineral and antinutritional composition of bottled and sterilized tigernut (cyperus esculentus) milk. agriculture and food sciences research, 11(2), 43–51. 10.20448/aesr.v11i2.5870 history: received: 30 may 2024 revised: 15 july 2024 accepted: 29 july 2024 published: 5 august 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 44 2. materials and methods ................................................................................................................................................................... 44 3. results and discussion ................................................................................................................................................................... 46 4. conclusion ......................................................................................................................................................................................... 50 references .............................................................................................................................................................................................. 50 mailto:okorie.chinasa@mouau.edu.ng mailto:onwuka.gregory@mouau.edu.ng mailto:obasi.nneoma@mouau.edu.ng https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.5870 https://orcid.org/0000-0003-2455-0122 https://orcid.org/0009-0005-8484-4570 https://orcid.org/0009-0002-2413-5164 agriculture and food sciences research, 2024, 11(2): 43-51 44 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this research study provides information on the heat penetration study used to determine the sterilization temperature suitable for thermal processing of tigernut milk, which is unique, compared with existing studies. it also gives data on the effect of different processing methods on the nutrient composition of bottled and sterilized tigernut milk. 1. introduction 1.1. background of study recently in research studies, there has been emphasis on new food product development that is required to meet the present demands of consumers. the upsurge in urban development has increased these demands; which makes studies in functional beverages the recent trend. beverages are no longer used to quench thirst; it is required that the functionality in these drinks are considered before consumption, and various beverage producing companies are using the perceptions of various persons as a vital tool in promoting innovative beverage combinations. therefore, the growing interest on the need to promote healthy living by people has created a prospect for plant-based beverages to thrive. an example of such functional requirement is milk alternatives, which is used to mitigate issues of lactose intolerance, calorie concern, cow milk allergy and prevalence of hypercholesterolemia [1]. ethnic origin is responsible for lactose intolerance. for instance, adult white north europeans, australasians and north americans record the lowest rates, ranging from 5 % in the british population to 17% in northern france and finland. in, africa, south america and asia, over 50 % of the population experience lactase non-persistence, and almost 100 % in some asian countries [2]. hence, there is a great need to explore the potentials of plant based milk alternatives. additionally, plant sources are good functional food and nutraceuticals due to healthy components such as vitamins minerals and antioxidants present in them das, et al. [3]. non-dairy products that are commonly used are soy milk, peanut milk, rice milk, safflower milk, and tigernut milk[4-9]. tigernut milk is one of the most consumed plant-based beverages, obtained during aqueous extraction from tigernuts tubers (cyperus esculentus) [10]. they are rich in minerals[11, 12] yet they are vastly underutilized. tigernut is widely cultivated in nigeria, so exploring both its economic and nutritional potential is crucial. the findings of this research will provide reference data on tigernut production at a commercial scale, thereby diversifying its use and increasing production. thus, food professionals, prospective researchers, and food industries will find the data obtained in this study highly valuable. 2. materials and methods 2.1. sources of raw materials brown and yellow varieties of tigernut tubers were purchased from farmers in jalingo, taraba state, nigeria. a 26mm crown corks manufactured by pellconi company in italy was successfully delivered through a courier servicedalsey hillblom lynn (dhl) and used for capping. the automated machine used for capping was purchased from canada and shipped to nigeria, the manufacturing company is officine pesce bottling system italy with the model number pg 93/c. vitamilk bottles were washed, sterilized and reused for bottling. 2.2. sample preparation tigernut extract used for the study was extracted from tigernut tubers (plates 1-3). the tigernut tubers were sorted to remove defective tubers. matured tigernut tubers selected were weighed and portioned, the tubers were also washed to remove sands and debris prior to use. different processing methods (boiling, malting and soaking) were used before the extraction process was carried out. 2.2.1. boiling method the method described by asante, et al. [13] with slight modification was used. about 200g of tigernut tubers was weighed out and boiled at the temperature of 1000c for 5 minutes before extraction. omniblend v model tm800 was used to reduce the particle size of the boiled tubers, then milled into paste. after the blending process, the slurry was filtered with a clean, damp muslin cloth. distilled water (500 ml) was used together for the blending and slurring process. before bottling and capping process, the filtrate from the extract was boiled at 70 0 c for 15 minutes to avoid curdling (figure 1). 2.2.2. malting method the method of ndubuisi [14] with modification was used. about 200g of tigernut tubers was weighed out then submerged in water (600ml) for a period of 24 hours before draining. during soaking, the tigernut tubers were washed and drained at the interval of 6 hours before placing them in a jute bag and kept at room temperature. water was sprinkled on the tubers twice a day. at the fifth day, germination occurred, and the tubers gave out shoots and roots. the germinated tubers were washed/devegetated. omniblend v model tm 800 was used to reduce the particle size of the malted tubers, and then milled into paste. after the blending process, the slurry was filtered with a clean, damp muslin cloth. distilled water (500 ml) was used together for the blending and slurring process. before bottling and capping process, the filtrate from the extract was boiled at 70 0 c for 15 minutes to avoid curdling (figure 1). 2.2.3. soaking method the method according to ndubuisi [14] with modification was used a weighed portion of tigernut tubers (200g) was cleaned with water, drained, and immersed in a 600ml of water for a period of12 hours. then the tubers were drained. omniblend v model tm 800 was used to reduce the particle size of the soaked tubers, then milled into paste. after the blending process, the slurry was filtered with a clean, damp muslin cloth. distilled water (500 agriculture and food sciences research, 2024, 11(2): 43-51 45 © 2024 by the authors; licensee asian online journal publishing group ml) was used together for the blending and slurring process. before bottling and capping process, the filtrate from the extract was boiled at 70 0 c for 15 minutes to avoid curdling (figure 1). 2.3. heat penetration studies heat penetration study was done on the bottled tigernut milk to determine the appropriate sterilization condition to ensure that the indicator organism (clostridium botilinum) was eliminated and nutrients retained. the goal was to achieve an f0 (f zero) in the range of 12-15 minutes which is recommended for low acid canned foods. in carrying out this process, the sterile bottles were hot filled with the tigernut milk then capped before inserting a thermocouple at the center of the can to evaluate the temperature of the milk as sterilization is being carried out (plates 4-6). it should be noted that the inserted thermocouple must be well sealed with thermostable gum to avoid leakage during sterilization. the instrumented bottle was placed in a retort autoclave along with other capped bottled tigernut milk. the thermocouple captured the temperature of the tigernut in the bottle every two minutes. the appropriate sterilization for the tigernut milk was achieved after 12.82 minutes. the temperature profile for the calculation of f0 using modified trapezium approach is illustrated in table 1. figure 1. a flow chart illustrating the processing, production and sterilization of tigernut milk. varieties of tigernut tubers plate 1: yellow tigernut plate 2: brown tigernut plate 3: malted tigernut heat penetration/ bottling process plate 4: penetration studies plate 5: bottling process plate 6: bottled tigernut milk agriculture and food sciences research, 2024, 11(2): 43-51 46 © 2024 by the authors; licensee asian online journal publishing group table 1.temperature and time profile for the sterilization of the tigernut milk. time 2 (min) temperature (oc) l area under the curve 0 108.4 0.054954 0.409767 2 116.5 0.354813 0.743859 4 116.9 0.389045 0.825561 6 117.4 0.436516 0.915146 8 117.8 0.47863 0.979817 10 118 0.501187 1.025995 12 118.2 0.524807 1.155765 14 119 0.630957 1.261915 16 119 0.630957 1.291651 18 119.2 0.660693 1.352524 20 119.4 0.691831 1.399777 119.5 0.707946 1.466523 fo for tigernut 12.8283 2.4. analysis 2.4.1.determination of mineral content sodium (na), magnesium (mg), iron (fe), calcium (ca), potassium (k), and copper (cu) were analyzed using the method as described by aoac [15]. 2.5. determination of phytochemical properties the method as described by harborne [16] was used to determine alkaloids while the method of amadi, et al. [17] was used to determine tannins and phytate. also the method described by vichapong, et al. [18] was used to determine flavonoid, and oxalate was determined using the method described by savage, et al. [19]. 2.6. statistical analysis the mean and standard deviation of the data obtained from the study was calculated and analyzed with single factor analysis of variance (anova) spss version 22.0. the significant difference between mean values was determined using the duncan new multiple range test. minitab version 19.0 was used to show interaction. 3. results and discussion 3.1. mineral composition the mineral composition of the tigernut tubers are shown on table 2. the mineral content of the tigernut tubers ranged from 1.7mg/100g to 526.25mg/100g. iron recorded the least value at 1.73mg and 2.24mg for yellow and brown variety respectively. while potassium recorded the highest value 460mg-526.25mg for yellow and brown variety. there was no significant difference p<0.05 in the copper content in both varieties. also no significance difference p<0.05 was recorded for the sodium content in both variety. the calcium content is lower than values obtained by arafat, et al. [20] whose study revealed that tigernut tubers have high calcium (152mg) but low values of iron (2mg), and copper (1.30mg) mineral contents. the values of calcium found in the tigernut, are adequate for teeth and bone development in infants. magnesium aids enzyme activities, provides bone strength, support nerve and heart functions. according to ahmed and hussein [21]tigernut tubers contain small quantities of iron, phosphorus, manganese, zinc and copper but have a high content of sodium and calcium. tigernuts could contribute sufficient magnesium to the daily requirements of growing children. oladele and aina [22] reported the mineral element of tigernut, their result showed that potassium had a value of (216 mg/l00g), sodium had a value of (245 mg/l00g) and calcium had a value of (155 mg / l0g) these values are lower than values obtained in this study except for calcium which is slightly higher. although it has been known that tigernut is an underutilized tuber, this study has shown that tigernuts contain essential minerals. from the result of the study, it can be concluded without doubts that tigernut is a good source of some minerals, and having these minerals present; it could serve as an important dietary supplement. table 2.mineral composition content of tigernut tubers. micronutrient yellow variety brown variety calcium (mg/100 g) 79.92**±0.01 119.70**±0.00 magnesium (mg/100 g) 121.21**±0.06 169.41**±0.00 iron (mg/100 g) 1.73*±0.09 2.24*±0.09 sodium (mg/100 g) 395.00ns±17.68 392.50ns±0.00 potassium (mg/100 g) 460.00**±0.00 526.25**±8.84 copper (mg/100 g) 17.84ns±0.81 15.57ns±0.81 note: values are mean ± standard deviation of duplicate determinations. **: means placed in the same row are significantly different at (p<0.05). *: means placed in the same row are significantly different at (p<0.01). ns: means placed in the same row are not significantly different at (p<0.05). 3.2. antinutrient composition of tigernut tubers table 3 shows the antinutrient composition of tigernut tubers. the values recorded ranged from 0.12mg to 19.70mg.the yellow variety recorded the least value of 0.12mg for oxalate and phytic acid content and there was no significant difference p<0.05 between with phytic acid content of the yellow and brown varieties. the result obtained showed that tigernut tubers are low in some antinutrients except flavonoids. even though alkaloids are known for their toxicity, not all are toxic. they are also responsible for the inhibition of some mammalian enzyme activities such as phophodiesterase, and prolonging the action of cyclic adenosine monophosphate (camp). they also affect stimulating hormones such as thyroid and glucagons. some forms of alkaloids have also been shown by agriculture and food sciences research, 2024, 11(2): 43-51 47 © 2024 by the authors; licensee asian online journal publishing group scientist to be carcinogenic [23]. however, antinutrients content obtained from this study were relatively low when compared to the antinutrients content reported for nuts such as peanuts [24]. when phytates are present in a biological systems it may chelate divalent metals such as magnesium and calcium, or inhibit the absorption of essential minerals in the intestinal tract[25] thus reducing its bioavailability [26]. phytates chelate with mineral elements therefore having an effects on the utilization of those minerals. they also react with residues of proteins. tannins and sometimes oxalates, also bind to proteins thereby causing difficulty in protein digestion. oxalates can expunge calcium in the form of calcium oxalate in the blood which may result to the damage of the organ [27]. table 3.antinutrient composition of tigernut tubers. antinutrient yellow variety brown variety alkaloid (mg/100 g) 1.37**±0.04 2.27**±0.07 phytic acid (mg/100 g) 0.12ns±0.00 0.14ns±0.01 oxalate (mg/100 g) 0.12**±0.01 0.22**±0.01 tannin (mg/100 g) 0.99ns±0.08 1.24ns±0.08 flavonoid (mg/100 g) 19.70*±0.00 18.45*±0.35 note: values are mean ± standard deviation of duplicate determinations. **: means placed in the same row are significantly different at (p<0.05). *: means placed in the same row are significantly different at (p<0.01). ns: means placed in the same row are not significantly different at (p<0.05). agriculture and food sciences research, 2024, 11(2): 43-51 48 © 2024 by the authors; licensee asian online journal publishing group table 4. mineral composition of tigernut milk. pre-processing method variety post-packaging calcium (mg/100 g) magnesium (mg/100 g) iron (mg/100 g ) sodium (mg/100 g) potassium (mg/100 g) copper (mg/100 g) malted yellow fresh 78.90d±0.08 71.86b±0.04 0.76f±0.00 212.50c±7.07 136.25c±1.77 14.43bcd±0.81 malted yellow sterilized 79.07d±0.07 47.82e±0.25 0.70f±0.09 136.25g±5.30 103.75h±1.77 17.28a±1.61 malted brown fresh 78.61e±0.12 71.59b±0.00 1.26cd±0.00 192.50e±3.54 83.75i±1.77 12.73d±1.61 malted brown sterilized 39.84f±0.11 48.50c±0.56 1.21d±0.08 190.00e±3.54 118.75de±1.77 13.29cd±0.81 soaked yellow fresh 38.70g±0.06 46.87f±0.03 0.94e±0.07 215.00c±3.53 111.25fg±1.77 15.57abc±0.81 soaked yellow sterilized 39.76f±0.00 48.23cde±0.00 0.83ef±0.09 107.50h±3.53 116.25ef±1.77 16.14ab±1.60 soaked brown fresh 39.66f±0.17 72.32a±0.09 1.27cd±0.00 201.25d±5.30 110.00g±0.00 14.43bcd±0.81 soaked brown sterilized 39.77f±0.00 48.27cd±0.04 1.27cd±0.00 133.75g±1.77 77.50j±3.53 15.57abc±0.81 boiled yellow fresh 80.04a±0.01 48.56c±0.02 1.61a±0.09 257.50b±3.54 131.25c±1.77 17.84a±0.80 boiled yellow sterilized 79.45c±0.04 48.17cde±0.00 1.52b±0.00 208.75cd±1.77 123.75d±1.77 16.14ab±1.60 boiled brown fresh 78.98d±0.00 47.94de±0.05 1.39bc±0.00 160.00f±3.53 220.00a±3.54 6.48e±0.80 boiled brown sterilized 79.64b±0.06 24.19g±0.07 1.21d±0.09 268.75a±1.77 191.25b±5.30 8.75e±0.81 note: values are mean ± standard deviation of duplicate determinations. means placed in the same column with different superscripts are significantly different at (p<0.05). a represents: highest significant mean among the treatments b represents: second highest significant mean among treatments c represents third highest significant mean among treatments d –i represents: significant mean after a b, c j represents: least significant mean among treatments agriculture and food sciences research, 2024, 11(2): 43-51 49 © 2024 by the authors; licensee asian online journal publishing group 3.3. mineral composition of tigernut milk the mineral composition of tigernut milk is presented in table 4. the result recorded ranged from 39.70mg to 80.04mg for the calcium content, magnesium content ranged from 24.19mg to 72.32mg while the iron content recorded 0.70mg to 1.61mg. the result of the sodium content was 133.75mg to 268.75mg, while potassium ranged from 77.50mg to 220mg and copper ranged from 6.48mg to 17.84mg. the boiled brown variety had the least value for copper. sodium, calcium, and potassium are still the major constituents of tigernuts, the result recorded showed all samples were significantly different p<0.05. processing and sterilization had little effect on the mineral content of tigernut milk. although there was a decline on the calcium content of the malted brown which recorded 78.61mg for freshly produced tigernut milk and 39.84mg for the sterilized tigernut milk. however, there was a significant increase on the potassium content. also there was a significant difference p<0.05 on the sodium composition for the boiled samples. the disparity within the findings may be as a result of the type of soil used for planting, variations in the mineral composition of soil, and location where the crop was planted [28]. a number of studies have proven that mineral and acceptability of tigernut milk is usually affected by the planting period and location of the tigernut tubers [29]. in addition, variation in the mineral content of the samples could be attributed to the different processing methods (soaking, boiling and malting) used in this study. onyeka [30] reported that mineral elements are inorganic that are found in minute quantity and play vital roles in human nutrition and their deficiency may affect the body functions. this study showed that the pre-treatment applied (soaking, boiling and malting) to the tigernut either improved or caused a decline on the mineral content of the tigernut milk. 3.4 antinutrient composition of tigernut milk the result for the antinutrient composition of the tigernut milk is shown in table 5. results obtained showed that processing and sterilization affected the antinutrient content of tigernut milk. this could be attributed to the fact that processing generally reduces the antinutrient content of tigernut milk [31]. soaking serves as a medium for the removal or reduction of possible antinutrient present, such as tannins and oxalates, phytic acids, polyphenols which could be released in to the water and discarded during draining process [32]. similarly onwuka [33] reported a decrease in tannin content of vegetable cowpea (vigna unguiculata) and pigeon pea (cajanus cajan) with a rise in soaking time. values obtained for tannins in this study was lower than 2.02% obtained by adekanmi, et al. [34] for soaked tigernut at room temperature but higher than 0.07% and 0.15% reported for brown millet and soybean reported by liu [35]. the effect of soaking on the antinutrient composition could be as a result of leaching during soaking period due to the solubility of tannin in water. soaking is a crucial step during malting in the production of vegetable milk because it tenderizes plant materials to help facilitate milling and extraction, makes milk easily digestible and palatable, and removes anti nutritional factors [3436].tannins inhibit the activities of enzymes such as amylase, trypsin and lipase by forming insoluble complexes with protein [37] and divalent ions such as zinc and iron thereby inhibiting their absorption 2+ 2+ in the body [38]. also the malting process decreased the tannin contents as a result of the activity of enzyme phenyl oxidase [39] microbial activities could also result to the decrease especially the phytic acid due to increased microbial activities during the malting process. all the processing methods used in this study significantly affected the phytate content. according to khan, et al. [40] the higher the moisture content, the higher the phytate loss. it has been reported the malting as a pre-processing method is known to be the most effective method of reducing phytates [41]. a broad range of microflora has been known to possess phytate activity that may be responsible for the reduction of phytate content [42]. the oxalate content was significantly reduced. similarly, obasi and wogu [43] recorded a significant decline in the oxalate content of yellow maize soaked in water for a period of 12 hours. the alkaloid content also reduced, although the different processing methods had effect on the alkaloid, sterilization had more effect in reducing the alkaloid content. the low alkaloid content obtained in this study agrees with studies reported by yadesa and biadge [44] on the effect of some traditional processing methods on nutritional composition and alkaloid content of lupin bean. the concentration of alkaloids, phytate, tannin, hydrogen cyanide, oxalate and saponin reduces after fermentation [45]. tannin, saponin and oxalate can be reduced by soaking [46]. table 5.antinutrient composition of tigernut milk. pre-processing method variety postpackaging alkaloid (%) phytic acid (%) oxalate (mg/100 g ) tannin (%) flavonoid (%) malted yellow fresh 1.05f±0.03 0.040b±0.000 0.025ab±0.007 0.650de±0.00 18.70a±0.00 malted yellow sterilized 0.94g±0.01 0.055a±0.007 0.010c±0.000 0.595de±0.08 14.70ef±0.71 malted brown fresh 1.81c±0.02 0.050ab±0.000 0.020b±0.000 0.815bc±0.08 15.45de±0.35 malted brown sterilized 1.29d±0.04 0.040b±0.001 0.020b±0.000 0.705cd±0.08 10.70h±0.71 soaked yellow fresh 1.20e±0.02 0.055a±0.007 0.010c±0.000 0.880b±0.00 17.70ab±0.00 soaked yellow sterilized 1.01f±0.03 0.060a±0.000 0.010c±0.000 0.805bc±0.08 16.70bc±0.71 soaked brown fresh 2.08a±0.03 0.055a±0.007 0.030a±0.000 0.990a±0.00 16.45cd±0.35 soaked brown sterilized 1.93b±0.04 0.060a±0.000 0.010c±0.000 0.825b±0.08 11.95g±0.35 boiled yellow fresh 0.16i±0.00 0.050ab±0.000 0.010c±0.000 0.220g±0.00 17.70ab±0.00 boiled yellow sterilized 0.09j±0.01 0.050ab±0.000 0.010c±0.000 0.220g±0.00 17.70ab±0.00 boiled brown fresh 1.03f±0.03 0.060a±0.000 0.025ab±0.007 0.540ef±0.00 17.45bc±0.35 boiled brown sterilized 0.65h±0.01 0.050ab±0.000 0.020b±0.000 0.440f±0.00 13.95f±1.06 note: values are mean ± standard deviation of duplicate determinations. means placed in the same column with different superscripts are significantly different at (p<0.05). a represents: highest significant mean among the treatments b represents: second highest significant mean among treatments c represents third highest significant mean among treatments d –i represents: significant mean after a b, c j represents: least significant mean among treatments agriculture and food sciences research, 2024, 11(2): 43-51 50 © 2024 by the authors; licensee asian online journal publishing group also, it has been reported by researchers that hydrothermal treatment, fermentation and some processing methods have the ability to reduce this antinutrients [47].the flavonoid contents recorded shows that tigernut can be used as a functional food as it is rich in flavonoid. administration of tigernut extract on wistar rats resulted in delayed manifestation of diarrhoea symptoms which suggests that anti-diarrhea property of tigernut milk is linked to the presence of flavonoids and tannins [21]. the availability of numerous phytochemicals with antioxidant activity agrees that tigernut milk are beneficial to health [48-50]. due to the chemical composition of tigernut it is suggested that tigernut milk can be used as functional oluwadunsin, et al. 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(2025). fermentation characteristics of yeasts isolated from apple and kiwi. agriculture and food sciences research, 12(2), 116–122. 10.20448/aesr.v12i2.7530 history: received: 21 august 2025 revised: 16 september 2025 accepted: 26 september 2025 published: 10 october 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. contents 1. introduction .................................................................................................................................................................................... 117 2. materials and methods ................................................................................................................................................................. 117 3. results .............................................................................................................................................................................................. 118 4. discussion ........................................................................................................................................................................................ 120 5. conclusion ....................................................................................................................................................................................... 121 references ............................................................................................................................................................................................ 121 mailto:hiroko.sk@yamanashi.ac.jp https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i2.7530 https://orcid.org/0000-0003-2426-7554 agriculture and food sciences research, 2025, 12(2): 116-122 117 © 2025 by the author; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to existing literature by investigating the fermentation characteristics of two natural yeasts. it provides new insights into the effects of temperature, sugar type, and sugar concentration on the fermentation capacity of these yeasts, thereby deepening our understanding of bread making and sake brewing. 1. introduction alcohol production is primarily carried out through fermentation using saccharomyces cerevisiae, although other natural yeasts are also commonly utilized. for instance, torulaspora delbrueckii is used in beer production [1]. additionally, metschnikowia pulcherrima is applied in wine production [2], schizosaccharomyces pombe in rum production, and kluyveromyces marxianus in the fermentation of whey from cheese production to produce vodka and gin [3]. in recent years, bread production with natural yeast has garnered considerable interest [4]. natural yeast is a type of fungus that thrives under diverse environmental conditions. unlike bread made with commercial dry yeast, bread produced using natural yeast has a complex taste, as different yeasts impart unique aromas and flavors [5]. currently, natural yeast bread made with strains isolated from dried grapes [6], sudachi petals, and fruits and flowers collected in tokachi region, hokkaido, japan, is commercially available [7]. moreover, bread has been successfully produced using lachancea fermentati, lachancea kluyveri, and torulaspora species isolated from rose cultivars grown in fukuyama city, hiroshima, japan [8]. saccharomyces rouxii, saccharomyces bisporus, and saccharomyces exigus have been isolated from grape juice, pineapple juice, and rice obtained from local markets in savar, bangladesh [9]. in bread production, yeast fermentation generates carbon dioxide, which causes wheat gluten to expand, and differences in yeast fermentation capacity directly affect bread quality. commercial yeast has a strong fermentation capacity, resulting in a softer texture. in contrast, natural yeast is less stable, ferments slowly, and generally shows weaker activity [10]. therefore, when producing bread with natural yeast, it is essential to adjust the sugar type and concentration as well as temperature to stabilize fermentation and improve efficiency. for example, bread can be successfully produced with hanseniaspora meyeri when monosaccharides such as glucose and fructose are provided at fermentation temperatures of 25–28°c [11]. kluyveromyces delphensis ferments dextrose and sucrose but not fructose or lactose, as demonstrated by sugar-specific fermentation tests [12]. furthermore, s. cerevisiae has an optimum growth temperature of 25–30°c and a fermentation temperature of 45°c; it can utilize glucose, fructose, and sucrose but not in the presence of maltose and amylose [13]. its ethanol production capacity is maximized at a sugar concentration of 125 g/l rather than 100 g/l. however, ethanol productivity decreases at concentrations above 150 g/l [14], suggesting that both sugar type and concentration substantially influence the fermentation efficiency of yeast. as different natural yeasts respond differently during fermentation, appropriate conditions should be selected for each yeast to ensure successful bread production. in this study, i focused on metschnikowia pulcherrima [15], isolated from fermented apple syrup, and candida apicola [16], isolated from fermented kiwi syrup. metschnikowia pulcherrima has previously been isolated from the spontaneous fermentation of tannat and marselan grape musts in concordia (entre rios, argentina) [17], cabernet sauvignon wine in ningxia, china [18], and marastina wine [19]. candida apicola has been isolated from naturally fermented alorena green table olives [20], wine yeasts that ferment grapes with high sugar content [21], and cachaca (sugarcane wine) [22, 23]. here, the effects of fermentation temperature as well as the sugar type and concentration were assessed to determine the optimal fermentation conditions required by each species in bread production. the results of this study may have potential applications in the production of alcoholic beverages. 2. materials and methods metschnikowia pulcherrima (nrbc0863) was purchased from the nite biological resource center, and c. apicola was isolated from kiwi syrup (accession number lc878464, dna data bank of japan). to prepare each yeast solution, yeast monocultured on agar medium was inoculated into a liquid medium containing 0.25% yeast extract, 0.50% peptone, and 0.10% glucose (for m. pulcherrima) or 5.0% glucose (for c. apicola). the inoculum was incubated at 30 °c for approximately 1 week. after incubation, the medium was removed using centrifugation (890 × g, 5 min, 25 °c), and the yeast was weighed. subsequently, the yeast was suspended in a 1% glucose solution to obtain a yeast solution with a final concentration of 0.78%–3.9%. 2.1. examination of the optimal fermentation temperature for each yeast yeast solution (3.0 g) and strong flour (3.0 g; tomizawa shouten inc., tokyo, japan) were mixed in a test tube (1.5 cm φ × 18 cm) and placed in a hot water bath adjusted to 20–35 °c in a low-temperature cooker (anova; axia international co., ltd., tokyo, japan). the increase in dough height was measured every hour for 5 hours and calculated as the increase in dough height per 0.1 g of yeast. 2.2. investigation of the optimal sugar type for each yeast yeast solution (3.0 g), strong flour (3.0 g), and sugar (0.15 g; glucose, sucrose, starch, fructose, and maltose) were mixed in a test tube (1.5 cm φ × 18 cm) and placed in a hot water bath at 25°c (m. pulcherrima) and 35°c (c. apicola) for 12 hours, and the fermentation capacity was calculated as the increase in dough height per 0.1 g of yeast. 2.3. examination of the effect of sugar concentration yeast solution (3.0 g), strong flour (3.0 g), and sugar (0.03–0.15 g) were mixed in a test tube (1.5 φ × 18 cm) and placed in a hot water bath at 25°c (m. pulcherrima) and 35°c (c. apicola) for 12 hours. the fermentation capacity was calculated as the increase in dough height per 0.1 g of yeast. 2.4. statistical analysis data were obtained in triplicate using the same sample based on fisher's three principles. the mean differences between groups were assessed using t-tests, and those among three groups were assessed using a one-way analysis agriculture and food sciences research, 2025, 12(2): 116-122 118 © 2025 by the author; licensee asian online journal publishing group of variance. all statistical analyses were performed using microsoft excel (microsoft corporation, redmond, wa). the significance level was set at p < 0.05. 3. results 3.1. examination of the optimal fermentation temperature for each yeast figure 1 shows the effect of different temperatures on m. pulcherrima fermentation. at 35°c, no change was observed in dough height after 1 hour; however, the dough height increased to 1.3 cm after 5 hours (p < 0.05). at 30°c, the height increased to 1.6 cm after 5 hours (p < 0.05). at 25°c, no change was observed in dough height after 2 hours; however, the dough height increased to 1.7 cm after 5 hours (p < 0.05). at 20°c, no change was observed in dough height after 1 hour, but the height slightly increased to 0.16 cm after 2 hours (p > 0.05). thereafter, no changes were observed up to 5 hours. figure 1. effect of temperature on metschnikowia pulcherrima fermentation. ● indicates 35°c, ■ indicates 30°c, ◆ indicates 25°c, and ▲ indicates 20°c. note: the fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as an integrated value. measurements were obtained in triplicate. the error bars indicate the standard deviation. figure 2 shows the effect of temperature on c. apicola fermentation. at 35°c, no change was observed in dough height after 1 hour; however, the dough height increased to 2.0 cm after 5 hours (p < 0.05). at 30°c, the height increased to 1.9 cm after 5 hours (p < 0.05). at 25°c, no change was observed in dough height up to 4 hours, and the height slightly increased to 0.49 cm after 5 hours (p > 0.05). at 20°c, no change was observed up to 2 hours; nevertheless, the height slightly increased to 0.25 cm after 3 hours (p > 0.05) and remained unchanged for up to 5 hours (p > 0.05). figure 2. effect of temperature on candida apicola fermentation. ● indicates 35 °c, ■ indicates 30 °c, ◆ indicates 25 °c, and ▲ indicates 20 °c. note: the fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. measurements were obtained in triplicate. the error bars indicate the standard deviation. agriculture and food sciences research, 2025, 12(2): 116-122 119 © 2025 by the author; licensee asian online journal publishing group 3.2. investigation of the optimal sugar type for each yeast figure 3 shows the effects of various sugars on m. pulcherrima fermentation. the addition of glucose increased the dough height to 8.1 cm after 12 h (p < 0.05). the addition of sucrose did not alter the dough height until 3 h, but it increased to 0.17 cm after 4 h (p < 0.05). thereafter, no change was observed in dough height until 6 h (p > 0.05), followed by an increase to 0.34 cm after 7 h (p < 0.05). no further change was observed until 12 h (p > 0.05). with starch addition, the height increased to 3.2 cm after 8 h (p < 0.05), then slightly increased to 3.3 cm after 12 h (p > 0.05). with fructose addition, the height increased to 6.7 cm after 11 h (p < 0.05), but slightly decreased after 12 h (p > 0.05). with maltose addition, the height increased to 2.5 cm after 7 h (p < 0.05); however, no change was observed after 8 h (p > 0.05). a slight increase to 2.9 cm was observed at 9 h (p > 0.05). no further change was observed until 12 h. figure 3. effect of various sugars on metschnikowia pulcherrima fermentation. ● indicates glucose, ■ indicates sucrose, ◆ indicates starch, ▲ indicates fructose, and 〇 indicates maltose. note: the fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. measurements were obtained in triplicate. the error bars indicate the standard deviation. figure 4 shows the effects of various sugars on c. apicola fermentation. the addition of glucose increased the dough height to 4.9 cm after 10 h (p < 0.05); however, no significant change was observed after 11 h (p > 0.05), followed by a slight increase to 5.2 cm after 12 h (p > 0.05). sucrose increased the height slightly to 1.6 cm after 2 h (p > 0.05); the height remained unchanged until 3 h (p > 0.05), but increased to 7.7 cm after 12 h (p < 0.05). with starch addition, the height increased to 0.25 cm after 2 h (p < 0.05), remained unchanged until 6 h (p > 0.05), and then increased to 0.81 cm after 8 h (p < 0.05). thereafter, it remained unchanged until 11 h (p > 0.05) and increased to 1.3 cm after 12 h (p < 0.05). with fructose addition, no change was observed until 4 h (p > 0.05), and the height increased to 1.7 cm after 9 h (p < 0.05). thereafter, no change was observed until 11 h (p > 0.05). subsequently, the height increased to 2.2 cm after 12 h (p < 0.05). with maltose addition, the height increased to 0.50 cm after 2 h (p < 0.05), but did not change until 6 h (p > 0.05). thereafter, it increased slightly to 1.2 cm after 9 h (p > 0.05), remained unchanged until 10 h (p > 0.05), and increased to 2.0 cm after 12 h (p < 0.05). figure 4. effect of various sugars on candida apicola fermentation. ● indicates glucose, ■ indicates sucrose, ◆ indicates starch, ▲ indicates fructose, and 〇 indicates maltose. note: the fermentation capacity is calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. measurements were obtained in triplicate. the error bars indicate the standard deviation. agriculture and food sciences research, 2025, 12(2): 116-122 120 © 2025 by the author; licensee asian online journal publishing group 3.3. examination of the effect of sugar concentration figure 5 shows the effect of glucose concentration on m. pulcherrima fermentation. when 1.0% glucose was added, no change was observed in dough height after 1 hour, but the dough height increased to 6.9 cm after 11 hours (p < 0.05) and remained unchanged until 12 hours (p > 0.05). at a concentration of 2.0%, no change was observed in dough height after 1 hour; however, the height increased to 7.3 cm after 12 hours (p < 0.05). at a concentration of 3.0%, no change was observed after 1 hour (p > 0.05), but after 12 hours, the height increased to 8.7 cm (p < 0.05). at a concentration of 4.0%, the height slightly increased to 0.12 cm after 3 hours (p > 0.05), followed by an increase to 8.7 cm after 12 hours (p < 0.05). following the addition of 5.0% glucose, the height increased to 10 cm after 12 hours (p < 0.05). figure 5. effect of glucose concentration on metschnikowia pulcherrima fermentation. ● indicates 1.0%, ■ indicates 2.0%, ◆ indicates 3.0%, ▲ indicates 4.0%, and 〇 indicates 5.0%. note: the fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. measurements were obtained in triplicate. the error bars indicate the standard deviation. figure 6 shows the effect of sugar concentration on c. apicola fermentation. when sucrose was added at a concentration of 1.0%, no change in dough height was observed after 12 hours. furthermore, at 2.0%, no change was observed in dough height up to 2 hours, but it increased slightly to 0.57 cm after 3 hours (p > 0.05) and remained unchanged after 12 hours (p < 0.05). at 3.0%, no change was observed after 1 hour, but the height increased slightly to 0.71 cm after 3 hours (p > 0.05) and remained unchanged until 12 hours (p > 0.05). similarly, no change was observed in dough height at 4.0% until 1 hour; however, after an increase in dough height to 0.71 cm after 2 hours (p < 0.05), no further change was observed until 4 hours (p > 0.05). after 10 hours, the height increased to 6.0 cm (p < 0.05), and no change was observed after 12 hours (p > 0.05). at 5.0%, the height increased to 3.3 cm after 3 hours (p < 0.05), but no change was observed after 4 hours (p > 0.05). thereafter, it increased to 11 cm after 12 hours (p < 0.05). figure 6. effect of sucrose concentration on candida apicola fermentation. ● indicates 1.0%, ■ indicates 2.0%, ◆ indicates 3.0%, ▲ indicates 4.0%, and 〇 indicates 5.0%. note: the fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. measurements were obtained in triplicate. the error bars indicate the standard deviation. 4. discussion 4.1. examination of the optimal fermentation temperature for each yeast in this study, the optimal temperature, sugar type, and concentration required for fermentation using m. pulcherrima and c. apicola as natural yeasts in bread production were investigated. after 5 hours of fermentation, m. pulcherrima and c. apicola showed high fermentation capacities at 25°c and 35°c, respectively; however, after 5 hours, they exhibited relatively high fermentation capacities at 30°c and 35°c, respectively. metschnikowia pulcherrima agriculture and food sciences research, 2025, 12(2): 116-122 121 © 2025 by the author; licensee asian online journal publishing group grows efficiently at temperatures in the range of 15–20°c [2]; whereas, c. apicola grows efficiently at 25–37 °c [12], indicating that it does not grow well in low-temperature environments. metschnikowia pulcherrima has been isolated as a dominant fermentation species during the fermentation of fiano di avellino grapes at 9 °c in wine production [24], suggesting that it can survive at low temperatures and show high fermentation potential. in addition, c. apicola can survive at low temperatures while marginally maintaining its fermentation potential. detailed reports on the optimal fermentation temperature of c. apicola are lacking. as a species similar to c. apicola, c. tropicalis has been reported to be heat-resistant and capable of producing ethanol from xylose [25]. candida species may have a high fermentation capacity at relatively high temperatures, suggesting that it is difficult for c. apicola to ferment substrates at lower temperatures. 4.2. investigation of the optimal sugar type for each yeast in this study, m. pulcherrima and c. apicola showed high fermentation capacities following the addition of glucose and sucrose, respectively. glucose, a monosaccharide, is used by yeast in ethanol fermentation. in a previous study, 31 types of s. cerevisiae isolates from palm wine were provided with glucose, galactose, fructose, sucrose, maltose, trehalose, and raffinose, either in combination or individually, to ferment a substrate, and glucose was used for fermentation in all tests [26]. among the aforementioned sugars, glucose, galactose, and fructose are monosaccharides; sucrose, maltose, and trehalose are disaccharides; and raffinose is the only trisaccharide. as these disaccharides and the trisaccharide contain glucose, they can be fermented if the yeast produces enzymes that can break them down into glucose. reportedly, the strength of sucrase activity varies among different species of s. cerevisiae [27]. hence, the type of sugar available and the fermentation rate may differ among different yeast species. in this study, the fermentation capacity of m. pulcherrima for sucrose was weaker than that for maltose and starch (figure 3). metschnikowia pulcherrima uses glucose and not sucrose in the fermentation of soy whey and produces ethanol as a product [28-30], consistent with the findings of the present study, suggesting that m. pulcherrima does not produce sucrase or possesses weak sucrase activity. alcohol fermentation has been reported to be slow with maltose [31], and as starch is maltose-bound, m. pulcherrima may have weak maltase and amylase activities. candida apicola has weak maltase or amylase activity (figure 4) and shows high fermentation efficiency for sucrose. in fermentation capacity tests by sugar type, c. apicola was reportedly able to ferment sucrose but not fructose [12]. similar results were obtained in this study. hence, c. apicola has high sucrase activity; however, it was also able to ferment the dough with fructose in this study. the glucose and fructose produced from sucrose through sucrase were used for fermentation. however, the slow onset of fermentation of glucose and fructose suggests that sucrose may promote fermentation by c. apicola. in addition, as c. apicola has been reported to tolerate high ethanol concentrations [21], it is possible that ethanol produced through fermentation promoted the fermentation of glucose and fructose. however, baking bread was the focus of this study, and therefore, the fermentation mechanism of these yeasts in baking is a subject for future research. 4.3. examination of the effect of sugar concentration metschnikowia pulcherrima and c. apicola showed the maximum fermentation efficiency at a sugar concentration of 5.0%. during fermentation, the fermentation capacity of c. apicola was affected by sugar concentration. in a previous study, fermentation by s. cerevisiae increased ethanol concentration with increasing glucose concentrations from 2.0% to 8.0% [32]. furthermore, as starmerella zemplinina (synonym candida zemplinina) has been reported to grow faster when glucose is added at concentrations of 2.0% and 20% [21], fermentation efficiency may have increased with increasing glucose concentrations up to 20%. in contrast, the fermentation efficiency of hanseniaspora guilliermondii reportedly decreases at sugar concentrations >300 g/l [14]. these results indicate that the effect of sugar concentration on fermentation varies depending on the yeast type. the yeasts investigated in this study showed the highest fermentation efficiency at a 5.0% sugar concentration, and it is highly probable that the efficiency can be increased by increasing the sugar concentration. however, as a considerable increase in sugar concentration significantly affects the flavor of bread, the effect of sugar concentration on flavor must be considered. this study has some limitations. as both wheat and water are used in bread production, the effects of wheat and the ions in water should be considered. however, these factors were not examined in this study; 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content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1094/cchem-03-17-0044-r https://doi.org/10.47371/mycosci.2021.10.003 https://doi.org/10.3390/applmicrobiol2030040 https://doi.org/10.20582/nfcc.42.0_45 https://doi.org/10.1002/jib.536 https://doi.org/10.1007/s00203-023-03646-1 https://doi.org/10.3389/fmicb.2023.1253969 https://doi.org/10.1016/j.foodres.2024.114072 https://doi.org/10.1016/j.ijfoodmicro.2010.11.006 https://doi.org/10.1016/j.ijfoodmicro.2009.01.024 https://doi.org/10.1023/b:wibi.0000013286.30695.4e https://doi.org/10.1007/s11274-004-4490-4 https://doi.org/10.3390/foods12030526 https://doi.org/10.1080/17597269.2018.1564480 https://doi.org/10.1007/s11274-020-02990-4 https://doi.org/10.1007/s00253-016-7449-0 https://doi.org/10.1016/j.fm.2018.07.016 https://doi.org/10.1007/s00217-020-03665-y https://doi.org/10.1007/s00217-020-03665-y https://doi.org/10.1002/jsfa.7253 https://doi.org/10.1007/s00217-020-03632-7 https://doi.org/10.1007/s10973-020-09629-4 1 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 1-11, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6455 © 2025 by the authors; licensee asian online journal publishing group chemical analysis of cocoa butters and risk assessment of production processes in five localities in cameroon nelly ndee mougang1 stephano tambo tene2 ronice zokou3 hermann arantes foffe kohole4 alix ntongme mboukap5 arnellie archelle foudjin abaoabo6 elsa nguepi solefack7 hilaire macaire womeni8 1,2,3,4,5,6,7,8research unit of biochemistry of medicinal plants, food sciences and nutrition, department of biochemistry, faculty of science, university of dschang, dschang, cameroon. 1email: nellyndee2001@gmail.com 2email: stambotene@yahoo.fr 3email: zokouronice@gmail.com 4email: hermannarantes@gmail.com 5email: mboukapalix@gmail.com 6email: foudjinarchelle@gmail.com 7email: elsanguepi12@gmail.com 8email: womeni@yahoo.fr ( corresponding author) abstract the quality of the cocoa product depends on the post-harvest treatment processes and the extraction method used; therefore, it is important to control the conditions applied to obtain a product of recommended quality. this study aimed to identify the critical stages in the post-harvest treatment and extraction of butter and to propose solutions for controlling them to obtain a product of good chemical quality. a survey of post-harvest treatments and extraction methods for cocoa butter (theobroma cacao l.) was carried out, after which the critical control points of the process were determined. samples of butter were collected from producers in various localities, and chemical analyses of fat were conducted. three production methods were identified: artisanal, semi-industrial, and industrial. the producers of this semi-finished product were mainly women aged between 40 and 50 with no education. the cocoa butters marketed were generally of poor chemical quality. oils from yaoundé producers were overall the most chemically acceptable, while those from douala were less so. these data suggest that producers should take these factors into account when post-harvesting cocoa beans and extracting the fat to guarantee a more stable, high-quality product. keywords: artisanal extraction, chemical quality, cocoa beans, cocoa butter, post-harvest treatments, theobroma cacao l. citation | mougang, n. n., tene, s. t., zokou, r., kohole, h. a. f., mboukap, a. n., abaoabo, a. a. f., solefack, e. n., & womeni, h. m. (2025). chemical analysis of cocoa butters and risk assessment of production processes in five localities in cameroon. agriculture and food sciences research, 12(1), 1–11. 10.20448/aesr.v12i1.6455 history: received: 26 december 2024 revised: 7 february 2025 accepted: 18 february 2025 published: 28 february 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ......................................................................................................................................................................................... 2 2. materials and methods ...................................................................................................................................................................... 2 3. results ................................................................................................................................................................................................... 4 4. discussion .......................................................................................................................................................................................... 10 5. conclusion ......................................................................................................................................................................................... 11 references .............................................................................................................................................................................................. 11 mailto:nellyndee2001@gmail.com mailto:stambotene@yahoo.fr mailto:zokouronice@gmail.com mailto:hermannarantes@gmail.com mailto:mboukapalix@gmail.com mailto:foudjinarchelle@gmail.com mailto:elsanguepi12@gmail.com mailto:womeni@yahoo.fr https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i1.6455 https://orcid.org/0000-0002-2454-5983 https://orcid.org/0000-0002-1117-997x https://orcid.org/0009-0009-9971-1542 https://orcid.org/0009-0005-3444-283x https://orcid.org/0009-0009-2771-1527 https://orcid.org/0009-0006-4162-8998 https://orcid.org/0000-0001-6893-2865 agriculture and food sciences research, 2025, 12(1): 1-11 2 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this work has made it possible to identify the post-harvest treatments that can influence the quality of the final product from cocoa beans and to propose solutions using the haccp tool to limit losses in order to improve the cost of the products. 1. introduction the cocoa sector directly or indirectly supports thousands of families in cameroon. cocoa is the leading export commodity (58.70%), according to an [1]. fermented and dried cocoa provide the country's second-largest source of foreign currency (12% of exports) behind hydrocarbons (39.60%) and is grown in seven regions, of which the centre and south-west are the two main production areas, with 50.36% and 31.54%, respectively [1]. the selection of cocoa quality and its properties (flavor, nutritional composition, and suitability for processing) are linked to numerous parameters such as variety, growing conditions, climate, soil, and agronomic practices [2]. cocoa is exported in raw form (unprocessed form) but also undergoes artisanal processing locally. it is the raw material used to make certain food products (chocolate, cocoa paste, cocoa powder, cocoa butter, cocoa fondue, cakes, and cookies), as well as many other pharmaceutical and cosmetic products such as lotions, lip balms, and creams [3, 4]. the use of cocoa involves a number of stages, including bean selection, drying, and fermentation. these stages can have an impact on the final quality of the cocoa, particularly its flavor and the quality of by-products such as cocoa butter [5]. cocoa butter, also known as 'theobroma oil', is an edible vegetable fat obtained from cocoa beans and has a sweet chocolate flavor. it is highly prized for its melting characteristics, remaining solid at room temperature or below but melting just below body temperature [6]. cocoa butter contains antioxidants that prevent rancidity and give it a shelf life of two to five years. processing cocoa butter to remove flavor and color also removes antioxidants, increasing its susceptibility to rancidity [6]. cocoa butter is obtained using a variety of processes, from traditional to modern. its quality can therefore be influenced by the quality of the cocoa beans used and the extraction method chosen [7]. modern methods include hydraulic presses, centrifugation, and solvent-assisted extraction [7-9]. the so-called artisanal methods include cold pressing, which has a low yield, the use of jute sacks, and thermal degreasing, which is very widespread [10]. thermal extraction, although leading to the formation and release of flavors, also leads to lipid oxidation, which can significantly affect the final quality of the butter [11]. lipid oxidation products responsible for the formation of rancid odors are involved in the genesis of cardiovascular disease and are carcinogenic [12]. in developing countries, particularly in cameroon, small-scale production and marketing of cocoa butter remain booming businesses. small-scale producers do not master the quality criteria for the cocoa beans they use, and there is significant variability in post-harvest treatments, as well as variations in the implementation of traditional cocoa butter extraction processes. these factors lead to inconsistencies in the quality of marketed cocoa butter, thereby reducing its marketability and competitiveness compared to products from industrial production. in light of the above, the aim of this work is to determine the critical stages in the post-harvest treatment and extraction processes for cocoa butter that influence the chemical quality of cocoa butter marketed in several localities in cameroon. 2. materials and methods 2.1. materials 2.1.1. survey data collection material the collection of information on the various stages of cocoa butter processing and production, with a view to identifying the risky or critical stages, was carried out using a survey form. the target population consisted of butter producers, and the aim was to highlight the post-harvest processing methods, the extraction of cocoa butter, and the characteristics of the final butter. 2.1.2. plant material the cocoa pods used in this study consisted of cocoa butter collected from 25 producers in douala (littoralcameroon), limbé (south-west cameroon), yaoundé (central cameroon), bertoua (east cameroon), and kékem (west cameroon), then transported to the laboratory for various chemical analyses. figure 1 shows a map of the cocoa butter samples collected from the different collection areas. figure 1. cocoa butter collected. 2.2. methods 2.2.1. study area the study was conducted in the five leading cocoa butter-producing localities across five regions of cameroon. the sample collection locations are detailed in table 1. agriculture and food sciences research, 2025, 12(1): 1-11 3 © 2025 by the authors; licensee asian online journal publishing group table 1. geographical localisation of studies area. locality region douala littoral limbé south-west yaoundé centre bertoua east kékem west 2.2.2. size of the study population this work was carried out among cocoa butter producers in the aforementioned localities. to do this, 25 producers were surveyed in the five localities, and the distribution by region was done randomly and depended on the density of activity in the area. 2.2.3. survey of post-harvest cocoa treatment and butter extraction processes a survey of the production process was carried out, during which information on post-harvest cocoa processing methods and the characteristics of the butter obtained were collected using a survey form. producers were met in the towns of douala, yaoundé, kékem, limbé, and bertoua. they answered the questions on the form, and their answers were recorded on it. data collection was prospective and semi-administered. 2.2.4. sampling method, frequency and size a random sampling plan was adopted in this study. the different cocoa butter samples were collected under aseptic conditions using single-use sterile gloves. tins of butter were taken at random, and those sampled were tightly packed to prevent oxidation by atmospheric oxygen. the cocoa nut butters were collected directly at the end of each extraction process and transported on ice to the laboratory. the proximate mass per box of butter was 250 g. table 2 shows the number of samples collected according to locality. table 2. number of samples collected by locality. locality number of samples collected douala 10.00 limbé 1.00 yaoundé 6.00 bertoua 5.00 kékem 3.00 2.2.5. transport of samples to the laboratory once they had been sampled, all the samples, previously packed in the containers used by the producers, were inserted into sterile stomacher brand bags (seward limited, england) and carefully labeled with an individual identification number. the samples were transferred to the laboratory after being placed in a cool box containing labsolute reusable ice packs (ref: 827.100001.49), which kept the temperature down to around 4°c to slow down the oxidation of the lipids within 24 hours. 2.2.6. determination of the chemical quality of marketed butters the quality of marketed butters was determined by means of lipid alteration quality indices. 2.2.6.1. acid index the acid value was determined using the titimetric standard method of afnor [13]. specifically, 5 g of cocoa butter was weighed and introduced into a 250 ml conical flask, to which 100 ml of 95% ethanol and two drops of 1% phenolphthalein were added. the mixture was vortexed (perkin elmer, usa) and then titrated with a 0.1 n potassium hydroxide ethanolic solution until the indicator turned pink (the pink color persisted). the results were expressed as a percentage of oleic acid according to equation 1: 𝐴𝑐𝑖𝑑 𝑣𝑎𝑙𝑢𝑒 (% 𝑜𝑓 𝑜𝑙𝑒𝑖𝑐 𝑎𝑐𝑖𝑑) = acid value ( mg g kohof fatty acid)∗282∗100 56.1∗1000 (1) 2.2.6.2. determination of the peroxide value the peroxide value was determined using the standard spectrophotometric method of idf [14]. to do this, 50 mg of the butter sample was mixed with 9.8 ml of a chloroform-methanol mixture (7:3), and the mixture was vortexed (perkin elmer, usa) for 10 seconds. next, 50 μl of a 30% aqueous ammonium thiocyanate solution was added, and the mixture was stirred again before the addition of 50 μl of an aqueous iron ii chloride solution, followed by further stirring. after 5 minutes of incubation at 25°c, the absorbance of the reaction mixture was read at 500 nm against a blank containing all the reagents except the oil (perkin elmer brand spectrophotometer, lambda 35, usa). the procedure took place in a dimly lit chamber and was completed within a maximum of 8 minutes per sample run. the peroxide value, expressed in ppm, was calculated according to equation 2: 𝑃𝑒𝑟𝑜𝑥𝑖𝑑𝑒 𝑣𝑎𝑙𝑢𝑒 (𝑝𝑝𝑚) = (as−ab)∗m 55.84∗w (2) as = absorbance of the sample; ab = absorbance of the blank; m = slope, obtained from the calibration curve (38.46); w = mass of the sample in g; 55.84 = molar mass of iron ii. agriculture and food sciences research, 2025, 12(1): 1-11 4 © 2025 by the authors; licensee asian online journal publishing group 2.2.6.3. determination of the thiobarbituric acid index (tba) the thiobarbituric acid index, expressed as malondialdehyde equivalent or mda (mg/kg of sample), was assessed by reading the absorbance of sr-tba on a visible spectrophotometer, following the methodology of draper and hadley [15]. in a 10 ml test tube, we weighed out 1 g of each butter sample and then added 1 ml of 0.1% trichloroacetic acid. the mixture was vigorously vortexed (perkin elmer, usa), and 1 ml of 0.375% thiobarbituric acid, 1 ml of 15% trichloroacetic acid, and 1 ml of 0.25 m hydrochloric acid were added. the mixture was again stirred and incubated in a water bath (heraeus) at 95°c for 30 minutes. at the end of this step, the tubes were cooled in an ice bath, and the absorbance was read at 532 nm against a blank on the aqueous phase. the thiobarbituric acid index was calculated as follows (equation 3): 𝐼𝑛𝑑𝑖𝑐𝑒 𝑇𝐵𝐴 (𝑝𝑝𝑚) = 𝐴𝑏𝑠∗𝑉𝑇𝐶𝐴∗0.02∗𝑀 1.56∗𝑚 (3) abs = absorbance of sample; vtca = volume of trichloroacetic acid solutions; m = molar mass of malondialdehyde (72 g/mol); m = mass of sample. 2.2.6.4. determination of the iodine value (ii) the iodine value was determined according to the titimetric standard protocol of afnor [13]. to this end, 0.2 g of fat was introduced into a flask, followed by 15 ml of carbon tetrachloride (ccl4) and 25 ml of wijs' reagent. the flask was capped and gently shaken, then placed in a dark place for 60 minutes before the addition of 20 ml of 10% aqueous potassium iodide solution and 150 ml of distilled water. using the starch solution (5 drops of 1% starch paste) as a color indicator, the solution was titrated with a 0.1 n sodium thiosulfate solution. titration continued until, after vigorous shaking, the blue color no longer appeared. the iodine index, expressed in g i2/100 g fat, was determined according to equation 4. 𝐼𝑜𝑑𝑖𝑛𝑒 𝑖𝑛𝑑𝑒𝑥 (𝑔 𝐼2/100𝑔) = (𝑉𝑜−𝑉1)∗12.69∗𝑇 𝑚 (4) t = exact titre of the sodium thiosulphate solution; m = mass in grams of the test sample; vo = volume of the thiosulphate solution for the blank (ml); v1 = volume of the thiosulphate solution for the sample. 2.2.7. determination of critical control points for the post-harvest cocoa treatment process and the artisanal cocoa butter extraction method the haccp: hazard analysis critical control point recommended by the fao: food and agricultural organization and adopted by the codex alimentarius [16] was used to identify the critical control points (ccps) for the cocoa post-harvest treatment process and the artisanal cocoa butter extraction method. this method is based on the 7 principles of haccp and consists of 12 steps, 8 of which were carried out. • set up the haccp team. • describe the product. • determine the intended use of the product. • draw up a flow chart. • confirm the flow chart on site. • list all the potential hazards associated with each stage, carry out a risk analysis and define the measures needed to control the hazards identified. • determine the critical control points (ccps) using the decision tree. • set critical thresholds for each critical control point (ccp). 2.3. statistical analysis the survey results were analyzed using spss version 18.0 software. the tables were generated using excel 2013. excel 2013 was used to express the results of the chemical analysis of the various butters in the form of means and standard deviations, and the analysis of variance (anova) was carried out using minitab version 18.1 (ibm statistics). means were compared using the fisher test, and the confidence level was set at 5%. 3. results 3.1. survey of cocoa post-harvest treatment and butter extraction processes 3.1.1. socio-demographic characteristics of cocoa butter producers the socio-demographic characteristics of cocoa butter producers are presented in table 3. this table indicates that the majority of producers are women (59.30%), with most being aged between 40 and 50 (37.00%). the predominant religion is animism (63.00%). additionally, the majority of producers reside in douala (37.00%), followed by yaoundé (25.90%), and generally have primary education (37.00%). 3.1.2. post-harvest treatment of cocoa and extraction of cocoa butter information on post-harvest treatments and cocoa butter extraction methods used by the surveyed producers is provided in table 4. it shows that several producers (44.44%) did not remove the pods and, therefore, did not carry out post-harvest treatments, but instead purchased cocoa beans that were already dry. the majority of producers (55.56%) surveyed conducted post-harvest treatments. furthermore, the time intervals between harvesting and shelling, as well as between shelling and fermentation, were one hour for most of the surveyed producers. additionally, the majority of producers (88%) fermented their cocoa for 7 to 9 days, and almost all the surveyed producers (93.33%) used bag fermentation. agriculture and food sciences research, 2025, 12(1): 1-11 5 © 2025 by the authors; licensee asian online journal publishing group table 3. socio-demographic characteristics of cocoa butter producers. characteristics modalities frequencies producers (%) gender female 16 59.3 male 11 40.7 age (years) [20-30] 3 11.1 [30-40] 7 25.9 [40-50] 10 37 [50-60] 3 11.1 >60 4 14.8 religion animist 17 63 christian 10 37 city of residence bertoua 6 22.2 douala 10 37 kékem 3 11.1 limbé 1 3.7 yaoundé 7 25.9 education level no education 3 11.10 basic school education 5 18.5 primary education 10 37 high school education 9 33.3 university level 0.00 0.00 with regard to the type, time, and place of drying, it emerged that solar drying was the only method used by the producers surveyed. moreover, the majority (66.67%) dried for a period of between 5 and 8 days. producers dried cocoa beans on black tarpaulins (53.33%) and cemented areas (46.67%). before being processed, dried beans could be kept for 5 days by the majority of producers surveyed (59.99%). with regard to the cocoa butter extraction method, it emerged that it was primarily produced using the artisanal method (66.70%), followed by the semi-industrial method (29.60%) and finally the industrial method (3.70%). extraction generally took between 4 and 7 hours (55.60%), with a yield of 0.25 liters of cocoa butter per kilogram of beans. the majority of producers kept the butter obtained for more than 24 months (29.60%). table 4. post-harvest treatments and cocoa butter extraction methods. production stages characteristics modalities frequencies percentages (%) debossing debossing yes 15.00 55.56 no 12.00 44.44 time between harvest and shucking (hour) <1h 1.00 6.67 1h 11.00 73.33 >1h 3.00 20.00 fermentation time between shelling and fermentation (hour) 1h 15.00 100.00 fermentation time (hour) [4-6] 3.00 20.00 [7-9] 12.00 80.00 fermentation containers bag 14.00 93.33 heap 1.00 6.67 drying heat source sun 15.00 100.00 drying surface black tarpaulin 8.00 53.33 cemented area 7.00 46.67 drying time (days) [0-4] 3.00 20.00 [5-8] 10.00 66.67 [9-12] 2.00 13.33 bean storage time (days) 2 1.00 6.67 3 1.00 6.67 4 4.00 26.67 5 9.00 59.99 extraction methods artisanal 18.00 66.70 semi-industrial 8.00 29.60 industrial 1.00 3.70 extraction time (hours) [0-3] 11.00 40.70 [4-7] 15.00 55.60 [7-10] 1.00 3.70 extraction yield (l/kg of beans) 0,25 27.00 100.00 shelf life (months) [0-6] 5.00 18.00 [12-18] 4.00 14.80 [18-24] 3.00 11.10 [6-12] 7.00 25.90 more than 24 8.00 29.60 in order to identify the best method for extracting cocoa fats, we cross-referenced the duration and frequency of use of each method by the different producers. this analysis, presented in table 5, shows that the traditional method takes longer (4 to 7 hours) than the other methods used by the producers surveyed (semi-industrial and industrial methods). agriculture and food sciences research, 2025, 12(1): 1-11 6 © 2025 by the authors; licensee asian online journal publishing group table 5. production time according to cocoa butter extraction method. production time (hours) total [0-3] [4-7] [7-10] method used artisanal 7.40% (2) 55.60% (15) 3.70% (1) 66.70% (18) industrial 3.70% (1) 0.00% (0) 0.00% (0) 3.70% (1) semi-industrial 29.60% (8) 0.00% (0) 0.00% (0) 29.60% (8) note: numbers in brackets represent frequencies. 3.1.3. cocoa butter extraction diagram figure 2 shows the cocoa butter production diagrams for the industrial, artisanal, and semi-industrial methods. in the industrial method, the beans were cleaned to remove stones, pieces of wood, crabs, broken pieces, and ferromagnetic materials, then dried at 110°c for 3 minutes. this was followed by hulling, roasting at 120°c for 50 to 60 minutes before cooling to 65°c, and then grinding, pressing, filtering, and crystallizing the butter at 20°c. the semi-industrial method began with sorting; then the beans were roasted in an aluminum pot, crushed, winnowed, and pressed at temperatures of around 40°c. in the artisanal method, unlike the semi-industrial method, the beans were not pressed but crushed and then brought to a boil in an aluminum pot with 1.5 to 2 liters of water for 1 kg of paste. the butter was then collected on the surface of the dough, and the rest of the dough was soaked in a large quantity of water (3 l/kg of dough) for 6 to 8 hours. the butter collected on the surface was mixed with the previous batch and then heated. figure 2. different cocoa butter production diagram. 3.2. analysis of the chemical quality of marketed cocoa butters the table 6 presents the values of the thiobarbituric acid, peroxide, acid, and iodine indices determined for each of the cocoa butters collected during the survey in the towns of douala, limbé, yaoundé, kékem, and bertoua. cocoa butter sample 11 (limbé) showed the highest thiobarbituric acid index (3.95±0.39 meq mda/kg), while the lowest was noted with sample 10 from douala (1.09±0.02 meq mda/kg). a significant difference (p˂0.05) was observed between samples from different localities and between samples from the same locality. the evaluation of the peroxide value revealed that sample 8 (douala) of cocoa butter had the highest lipid peroxidation value (20.45±2.67 meq o2/kg), while sample 6 (douala) had the lowest value (3.05±0.00 meq o2/kg). overall, the samples from kékem were the least oxidized. in terms of acid value, sample 10 (douala) showed the highest value (16.97±0.23% oleic acid), while number agriculture and food sciences research, 2025, 12(1): 1-11 7 © 2025 by the authors; licensee asian online journal publishing group 5 from the same town showed the lowest value (0.50±0.00% oleic acid). a significant difference (p˂0.05) was observed between the different samples regardless of locality. evaluation of the degree of unsaturation showed that this parameter varied from 23.21±2.31 g i2/100 g to 51.28±0.00 g i2/100 g, and a significant difference (p˂0.05) was observed between the different samples. table 6. chemical quality of marketed cocoa butters. city of origine samples thiobarbyturic acid value (meq mda/kg) peroxide value (meq o₂/kg) acid value (% oleic acid) iodine value (g i/100 g) douala 1 3.90±0.23b 17.81±1.12a 7.44±0.20e 35.40±0.50e 2 2.48±0.19b 17.27±0.70c 4.00±1.23f 35.10±0.29e 3 1.75±0.33b 15.34±3.60d 12.64±0.24c 27.81±0.87g 4 1.36±0.00j 5.46±0.49f 2.17±0.68g 26.17±1.12h 5 3.91±0.11g 8.55±1.02a 10.19±3.13d 23.21±2.31j 6 3.21±0.22l 3.05±0.00b 0.84±0.39k 25.06±0.44i 7 1.37±0.31k 4.00±0.26f 0.50±0.00l 35.10±0.93e 8 1.16±0.14a 20.45±2.67g 7.33±0.00e 45.63±0.22b 9 1.13±0.02h 7.41±1.02g 4.10±0.44f 38.87±0.35d 10 1.09±0.02i 6.27±0.64g 16.97±0.23a 40.08±0.27d limbé yaoundé 11 3.95±0.39i 6.81±1.27a 4.03±0.54f 42.28±0.57c 12 1.66±0.02c 12.37±2.11d 14.92±0.00b 51.28±0.00a 13 2.52±0.47g 8.78±0.80c 1.91±0.41h 26.51±0.03g 14 1.58±0.16j 5.71±1.18e 0.54±0.00k 33.93±0.39e 15 3.01±0.06k 4.46±0.75b 6.63±0.78e 32.99±2.69f 16 1.36±0.04g 8.30±0.01f 2.46±0.42j 39.54±1.18d 17 1.24±0.20d 11.45±1.48f 1.38±0.03j 35.99±0.48e bertoua 18 2.26±0.17l 3.21±0.50c 0.79±0.37j 33.63±2.69e 19 1.09±0.01e 10.53±2.84g 3.01±0.42f 32.92±1.30f 20 3.68±0.01d 11.16±2.63a 2.89±0.33g 33.05±1.12f 21 2.05±0.18f 9.48±0.67c 4.23±0.39f 24.50±0.46i 22 3.23±0.29k 4.24±0.19b 1.58±0.00h 38.98±0.42d kékem 23 1.67±0.17m 3.31±0.46c 4.10±0.21f 34.12±0.12e 24 2.18±0.07m 3.33±0.78c 1.87±0.47h 26.80±1.11g 25 1.79±0.12m 3.25±0.22c 1.69±0.00h 36.18±2.76h note: values (n =3) with different superscript (a,b,c,d,e,f,g,h,i,j,k,l,m) letters in the same column are significantly different (p < 0.05) according to fisher post hoc test. in order to better highlight the impact of the collection area on the chemical characteristics of the cocoa butters, an overall average for each index according to the cities was calculated. table 7 shows that the collection area has a significant influence (p < 0.05) on the various quality indices evaluated. butters from limbé had the highest thiobarbituric acid index (3.93 meq mda/kg), while those from yaoundé and kekem were statistically similar. the peroxide value was highest in the butters from douala (10.56 meq o2/kg), while those from limbé, yaoundé, and bertoua were statistically similar. butters from the town of kekem, on the other hand, showed the lowest value for this index. the acid value ranged from 2.50% oleic acid (bertoua) to 6.57% oleic acid (douala). no significant difference (p > 0.05) was observed between the samples from limbé and yaoundé and those from bertoua and kekem. the iodine index ranged from 42.28 g i2/100 g (limbé) to 29.03 g i2/100 g (kekem). the samples from bertoua and kekem were significantly (p < 0.05) similar. table 7. quality of butter sold by town. locality thiobarbyturic acid value (meq mda/kg) peroxide value (meq o2/kg) acid value (% oleic acid) iodine value (g i2/100 g) douala 2,13±0.16c 10.56±1.56a 6.57±0.66a 33.24±0.73b limbé 3.93±0.39a 6.81±1.27b 4.03±0.54b 42.28±0.59a yaoundé 1.90±0.16d 8.52±1.06b 4.64±0.27b 36.71±0.79b bertoua 2.47±0.13b 7.73±1.37b 2.50±0.31c 32.61±1.20c kékem 1.88±0.12d 3.33±0.49c 2.55±0.23c 29.03±1.33c note: values (n =3) with different superscript (a,b,c,d) letters in the same column are significantly different (p < 0.05) according to fisher post hoc test. 3.3. determining the critical points for mastering the cocoa post-harvest treatment process and the artisanal cocoa butter extraction method 3.3.1. post-harvest treatment process 3.3.1.1. description of the post-harvest treatment of the product dry cocoa beans (theobroma cacao l.) harvested from pods, debulked, then fermented in bags and sun-dried on cemented areas or black tarpaulins were used as the study matrix. the dried beans are either used by the local people themselves or placed into jute bags and sold to cameroonian industries or exported to other industries abroad. this raw material is used as a basic ingredient in the small-scale and industrial production of cocoa butter, chocolate, and cocoa powder. obtaining dry beans involves a series of operations summarized in figure 3. agriculture and food sciences research, 2025, 12(1): 1-11 8 © 2025 by the authors; licensee asian online journal publishing group figure 3. diagram of operations for post-harvest treatment of cocoa beans. 3.3.1.2. risk analysis table 8 summarises the stages in the cocoa post-harvest treatment process, as well as the associated chemical risks and the preventive measures to deal with them. it should be noted that the shelling, fermentation, drying and storage stages are conducive to the presence of free fatty acids and lipid oxidation (this information have been obtained during survey). table 8. risk analysis of post-harvest treatment of cocoa beans. step associated chemical risks preventive measures debossing presence of damaged beans. clubs will be used instead of sharp instruments fermentation presence of slaty and mouldy beans ensure adequate fermentation time drying presence of mouldy beans dry on cemented areas or black tarpaulins, avoid slow drying, stir the beans 5 to 10 times a day, and cover the beans overnight to prevent moisture build-up. presence of biological contaminants sort beans when collecting or displaying them storage presence of mouldy beans avoid storing in damp conditions and stacking bags more than 5 m high, and sort the beans. presence of moth-eaten beans insect repellent, bag maintenance, avoiding dust and breakage high content of free fatty acids in butter avoid storage in damp conditions, avoid breaking the beans 3.3.1.3. determination of critical control points (ccps) the determination of critical control points for the control of post-harvest cocoa processing, preventive measures, and threshold points to be respected was carried out and presented in table 9. an analysis of this table shows that fermentation and drying, in particular, require precise and controlled conditions to prevent the risk of free fatty acids and lipid oxidation. table 9. identification of critical points for the control of post-harvest treatment of cocoa beans. critical treatment preventive measures threshold points fermentation respect the fermentation time 6 to 8 days brewing the beans after 24 hours, then every 48 hours drying dry on suitable surfaces cemented areas and/or black tarpaulins avoid slow drying 4 to 6 days stir the beans 5 to 10 times daily cover the beans overnight to prevent moisture build-up sort beans when collecting or displaying them remove all defective beans (flat, caked, broken, mouldy, etc.)., attacked by insects and sprouted beans) and contaminants storage avoid storage in humid conditions relative air humidity ≤ 70%. avoid stacking bags too high height ≤ 5 m sorting the beans eliminate all defective beans (flat, agglomerated, broken, mouldy, attacked by insects and sprouted beans) insect repellent, bag maintenance, avoiding dust and breakage new bags, soft handling agriculture and food sciences research, 2025, 12(1): 1-11 9 © 2025 by the authors; licensee asian online journal publishing group 3.3.2. artisanal cocoa butter extraction 3.3.2.1. description of the product preparation the cocoa butter (theobroma cacao) is extracted using a traditional method that involves roasting the beans, grinding, and heating the paste to collect the butter. the cocoa butter (theobroma cacao) is marketed by local companies, which either export it or use it to produce local chocolate. it is also produced by local people who sell it and use it therapeutically to treat ailments such as rheumatism, burns, scars, and other skin defects, including stretch marks. the diagram of artisanal cocoa butter production is summarized in figure 4. figure 4. diagram of artisanal cocoa butter production. 3.3.2.2. risk analysis the risk analysis of the artisanal cocoa butter extraction process is summarized in table 10. it shows that several stages are conducive to the risk of butter oxidation and acidity. these stages, from roasting to the conservation of the butter, are crucial for maintaining the chemical quality of the oils. table 10. risk analysis of artisanal cocoa butter extraction. step associated risks preventive measures collection of dried beans collection of defective beans only healthy beans will be collected sorting omission of defective beans defective beans will be carefully removed grilling butter oxidation respect the roasting time and temperature winnowing presence of high levels of hulls careful winnowing collecting the butter in the boiling dough butter oxidation check the heating time of the paste heating the butter high water content, butter oxidation check the heating time storing the butter rancidity of butter store in an opaque tin with a wide opening, away from light agriculture and food sciences research, 2025, 12(1): 1-11 10 © 2025 by the authors; licensee asian online journal publishing group 3.3.2.3. determination of critical control points (ccp) table 11 presents the critical control points for the artisanal method of extracting cocoa butter, the preventive measures, and the threshold points to be respected. it shows that grilling, boiling the paste, and heating the butter, in particular, require precise and controlled conditions, with heating times of 30 minutes, 30 minutes, and 15 minutes, respectively, in order to prevent the risk of oxidation and acidity of the butter. table 11. critical points for mastering the artisanal extraction of cocoa butter. critical treatment preventive measures threshold points grilling respecting the time and temperature of grilling ≤ 30 min boiling the dough check heating time and temperature ≤ 30 min heating the butter check heating time and temperature ≤ 15 min 4. discussion the socio-demographic data revealed that most of the cocoa butter producers were women (59.30%). this could be explained by the fact that the artisanal cocoa butter production process, which was the most represented, can be likened to culinary preparation. indeed, from an early age, young african girls learn to cook, more by immersion than by explicit learning [17]. the majority of the producers (37.00%) were aged between 40 and 50, and most of them were animists (62.00%). people in this age group are more attached to old practices than the younger generation [17], hence their preference for work in the fields, artisanal extraction, and the animist religion. the majority of growers lived in douala (37.00%) because it is one of cameroon's major economic zones and is suitable for growing this plant. in addition, the high population density in this metropolis makes this activity more lucrative. the most common level of education was primary school (37.00%). growers do not have a very high level of education, so they have turned to an income-generating activity that only requires practical skills [18]. post-harvest processing includes three main stages: shelling, fermentation, and drying. some of the producers surveyed (44.44%) purchased dry beans to produce their cocoa butter, as they felt they needed to produce large quantities to satisfy the high demand. as a result, post-harvest processing was seen as a waste of time and energy, leading to a loss of financial income. the most commonly used fermentation time (7-9 days) is closest to the recommended conditions of 6 to 8 days [19]. fermentation time varies according to climatic conditions, fruit maturity, and the mass to be fermented [20]. the most commonly used fermentation method (in bags) is not the one recommended, although it is easier to implement given the relatively small quantity of cocoa to be fermented [20]. the heat source used by all the producers was the sun, as it was the cheapest and easiest to use. drying was carried out for 5 to 8 days by the majority of producers; this range is close to that recommended [21]. the drying time varied according to local conditions and/or the possible use of an artificial dryer, with the ideal being 6-10 days for sun drying [21]. the drying surfaces used were primarily cemented areas and black tarpaulins, which corroborates the work of barel [3] and lainé [22], who also reported the use of these drying methods in their research. most cocoa butter is then extracted using the artisanal method. this can be attributed to the ease of its implementation, its relatively low cost, the accessibility of the necessary tools, and its yield, which is practically equal to that of the other methods when performed correctly [23]. the artisanal method is the most time-consuming and labor-intensive compared to the semi-industrial method [24]. analysis of the chemical quality of the cocoa butters marketed revealed that these butters were of poor quality. indeed, 72% of the samples have an acid value higher than the codex alimentarius standard for cocoa butter (≤ 1.75% oleic acid), and 40% of the cocoa butter samples have an iodine value outside the normal range defined by the codex alimentarius, which is 32 to 41 g i₂/100 g [16]. this may be due to poor execution of post-harvest treatments, the production method used, or even inadequate preservation of the butter, which promotes oxidation and rancidity [25]. in fact, during the bean drying stage, prolonged exposure of the almonds to the sun is responsible for the oxidation of the fat, leading to significant production of peroxides that destabilize compounds with a high melting point and low stability [26]. similarly, roasting and heating steps are likely to alter the quality of the product obtained, as reported by żyżelewicz, et al. [11] in their work. the haccp carried out on all stages of the post-harvest treatment process reveals fermentation and drying as critical points for the control of this process. the primary objective of post-harvest treatments is to transform the fruit borne by the cocoa tree into a marketable product. to this end, the simplest succession of treatments is as follows: picking, extraction of the beans (shelling), and drying. all these operations affect the characteristics of the beans and are highly interdependent. they, therefore, form a chain that determines the quality of the final product [21]. fermentation is one of the main post-harvest activities in the production of cocoa beans [27], but it remains a complex process that is difficult to control, especially in the case of spontaneous or natural fermentation of cocoa beans, where microbial contamination occurs in a haphazard and random manner [28]. the drying process, on the other hand, must be carried out carefully to ensure that the beans have been properly prepared for storage and transport without being contaminated by mold, bacteria, polycyclic aromatic hydrocarbons (pahs), and other contaminants. drying must be complete, and moisture content must be reduced to less than 8% [29]. the haccp carried out on the artisanal cocoa butter extraction method revealed that the roasting, paste boiling, and butter heating stages are critical points for controlling this process. the time and temperature of roasting contribute to many physical and chemical changes, such as the intensification of the brown color and the development of the characteristic cocoa aroma [30]. on the other hand, the roasting process can contribute to the alteration of the composition of bioactive compounds, leading to changes in lipid degradation due to oxidation [11]. boiling the paste and heating the butter are done at relatively high temperatures that can alter the quality of the cocoa butter obtained if the time taken for these operations is too long [31]. these results are similar to those obtained by womeni, et al. [32], who carried out the haccp on the industrial method of extracting cocoa butter and identified roasting as a critical control point. according to womeni, et al. [32], the cooking time and drying temperature of shea seeds caused biochemical changes affecting the quality of the butter. agriculture and food sciences research, 2025, 12(1): 1-11 11 © 2025 by the authors; licensee asian online journal publishing group 5. conclusion at the end of this study, which focused on the analysis of post-harvest cocoa treatments, cocoa butter extraction, and the identification of the critical stages of the two processes (haccp), using a survey of producers, it emerged that butter extraction methods differ according to the producers. however, post-harvest treatments are generally standard, and the cocoa butters marketed are typically of poor quality. with regard to the critical stages, fermentation and drying were identified for post-harvest treatments, while roasting the beans, boiling the paste, and heating the butter were noted for the artisanal butter extraction method. in view of the above, compliance with production conditions and haccp rules would result in better quality butter and more competitive products in the market. references [1] oncc, "order establishing the terms and conditions for marketing cocoa beans, n° 00026. " 2017. [2] m. crafack et al., "impact of post-harvest practices on cocoa (theobroma cacao l.) flavour quality," food research international, vol. 54, no. 1, pp. 150-158, 2013. [3] m. barel, "from cocoa to chocolate, the epic tale of a delicacy," quae, 2009. [4] k. davison, p. howe, and j. buckley, "potential implications of dose and diet for the effects of cocoa flavanols on cardiometabolic function," the journal of nutritional biochemistry, vol. 64, pp. 1-10, 2019. [5] a. aprotosoaie, s. luca, a. miron, and a. trifan, "the sensory impact of cocoa fermentation on chocolate quality," foods, vol. 5, no. 3, pp. 1-13, 2016. [6] b. encylopaedia, cocoa butter consulted online at cocoa butter | definition, characteristics, & uses |. britannica, 2023. [7] d. ghosh, a. konar, and s. sircar, "solvent extraction of cocoa butter: effect of some process parameters on extraction yield and oil quality," journal of food process engineering, vol. 35, no. 2, pp. 204-222, 2012. 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[15] h. h. draper and m. hadley, "malondialdehyde determination as index of lipid peroxidation," methods enzymology, vol. 186, pp. 421431, 1990. [16] c. alimentarius, "codex standard for cocoa butter," codex-stan, pp. 86-1981, 2001. [17] p. erny, "ethnology of education," vol. 1. paris: harmattan, 1991, p. 163. [18] t. amraoui, "african artisans: social study and organization of work. i. social and legal status based on latin inscriptions from algeria," antiquités africaines. l’afrique du nord de la protohistoire à la conquête arabe, vol. 52, pp. 59-80, 2016. https://doi.org/10.4000/antafr.728 [19] s. s. thompson, k. b. miller, and a. s. lopez, "cocoa and coffee food microbiology fundamentals and frontiers." washington, d.c: asm press, 2001, pp. 721-733. [20] c. amani, "cocoa: obtaining quality beans," agroivoire, 2014, p. 23. [21] m. end and r. dand, "cocoa beans: quality requirements of the chocolate and cocoa industry," eca-caobisco-fcc cocoa research fund, 2015. [22] k. lainé, "survey on cultural practices in cocoa plantations in ivory coast," rapport de projet paccc/icco/industrie sur l’amélioration de la qualité du cacao en côte d’ivoire, vol. 27, 2001. [23] b. njonga, "cameroon: the manufacture of cocoa butter. the voice of the peasant. consulted on march 18, 2023 on," retrieved: https://www.lavoixdupaysan.net/cameroun-la-fabrication-du-beurre-de-cacao/. 2018. [24] agriculture au cameroun, "cameroon: how to make cocoa butter agriculture in cameroon," retrieved: https://www.agricultureaucameroun.net/cameroun-comment-fabriquer-du-beurre-de-cacao/. 2019. [25] m.-e. cuvelier and m.-n. maillard, "stability of edible oils during storage," ocl oilseeds and fats crops and lipids, vol. 19, no. 2, pp. 125-132, 2012. https://doi.org/10.1051/ocl.2012.0440 [26] h. m. womeni, r. ndjouenkeu, c. kapseu, f. t. mbiapo, m. linder, and j. fanni, "application of the drying-frying process to shea almonds: influence on the composition of unsaponifiable matter in the butter," oléagineux, corps gras, lipides, vol. 14, no. 6, pp. 366370, 2007. http://dx.doi.org/10.1051/ocl.2007.014 [27] r. f. schwan and a. e. wheals, "the microbiology of cocoa fermentation and its role in chocolate quality," critical reviews in food science and nutrition, vol. 44, no. 4, pp. 205-221, 2004. https://doi.org/10.1080/10408690490464104 [28] b. koffi, j. diopoh, t. karou, j. nemlin, g. ouattara, and s. t. guehi, "fermentations and marketability of cocoa beans," agronomie africaine, vol. 25, no. 2, pp. 159-170, 2013. [29] w. amoah-awua, "methods of cocoa fermentation and drying," cocoa and coffee fermentations, pp. 71-128, 2015. [30] s. t. guehi, s. dabonne, l. ban-koffi, d. k. kedjebo, and g. i. b. zahouli, "effect of turning beans and fermentation method on the acidity and physical quality of raw cocoa beans," journal of food science and technology, vol. 2, no. 3, pp. 163-171, 2010. [31] h. a. kohole, "influence of drying and cooking processes of peanut seeds (arachis hypogaea) on the quality of lipids and some functional properties of proteins," thesis university of dschang, 2015. [32] h. womeni, r. ndjouenkeu, c. kapseu, f. tchouanguep mbiapo, m. parmentier, and j. fanni, "effect of cooking and drying of shea nuts (butyrospermum parkii (g. don) kotschy) on butter quality," tropicultura, vol. 24, no. 3, pp. 175-182, 2006. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.foodres.2014.04.053 https://doi.org/10.1002/jpln.201200054 https://doi.org/10.4000/antafr.728 https://www.lavoixdupaysan.net/cameroun-la-fabrication-du-beurre-de-cacao/ https://www.agricultureaucameroun.net/cameroun-comment-fabriquer-du-beurre-de-cacao/ https://doi.org/10.1051/ocl.2012.0440 http://dx.doi.org/10.1051/ocl.2007.014 https://doi.org/10.1080/10408690490464104 110 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 2, 110-115, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i2.7404 © 2025 by the authors; licensee asian online journal publishing group enhancing land use efficiency for tea cultivation: a case study in quang binh district, ha giang province phan thi thanh huyen1 dinh hong duyen2 nguyen thi hue3 nguyen thi thao4 truong quang ngan5 ( corresponding author) 1,2vietnam national university of agriculture, vietnam. 1email: ptthuyen@vnua.edu.vm 2email: dhduyen@vnua.edu.vn 3,4hanoi university of natural resources and environment, vietnam. 3email: nguyenhuebn87@gmail.com 4email: thaotnmtmt@gmail.com 5nghe an university, vietnam. 5email: ngantq@nau.edu.vn abstract the study aims to evaluate the efficiency of tea land use in quang binh district across three aspects: economic, social, and environmental. the study utilized random survey data from 60 tea-growing households, combined with both quantitative and qualitative analysis. findings indicated that the total tea cultivation area remained relatively stable between 2020 and 2024; however, economic efficiency sharply declined in 2024 due to disruptions in export markets and a significant drop in tea prices. tea cultivation continues to be highly accepted by local populations, especially among ethnic minorities, owing to its compatibility with traditional practices and its role in ensuring employment and land tenure stability. additionally, tea cultivation contributes to increasing land cover, maintaining and protecting soil quality, and reducing land degradation. nonetheless, the declining wages of daily labor and limited market connectivity threaten the long-term sustainability of economic, social, and environmental aspects of tea cultivation. to enhance effective land use, a comprehensive approach is necessary, which includes rational land planning, adoption of sustainable farming techniques, increased application of organic and biological inputs, and strengthened training and technology transfer. these measures support long-term economic and environmental sustainability in tea-growing regions, while also increasing production and improving soil health. keywords: economy, efficiency, environment, society, tea. citation | huyen, p. t. t., duyen, d. h., hue, n. t., thao, n. t., & ngan, t. q. (2025). enhancing land use efficiency for tea cultivation: a case study in quang binh district, ha giang province. agriculture and food sciences research, 12(2), 110–115. 10.20448/aesr.v12i2.7404 history: received: 1 august 2025 revised: 3 september 2025 accepted: 9 september 2025 published: 12 september 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 111 2. methods ........................................................................................................................................................................................... 111 3. empirical results ........................................................................................................................................................................... 111 4. conclusion ....................................................................................................................................................................................... 114 references ............................................................................................................................................................................................ 115 mailto:ptthuyen@vnua.edu.vm mailto:dhduyen@vnua.edu.vn mailto:nguyenhuebn87@gmail.com mailto:thaotnmtmt@gmail.com mailto:ngantq@nau.edu.vn https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i2.7404 https://orcid.org/0000-0002-1990-2261 https://orcid.org/0000-0001-7846-0179 https://orcid.org/0000-0002-5615-6595 https://orcid.org/0009-0008-6368-6771 https://orcid.org/0009-0008-2892-6514 agriculture and food sciences research, 2025, 12(2): 110-115 111 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study contributes to the existing literature by providing new empirical evidence on tea land use efficiency in quang binh. the primary contribution of the paper is the finding that economic, social, and environmental outcomes vary under market and labor challenges. this study documents integrated solutions to improve sustainable land use for tea. 1. introduction tea (camellia sinensis) is one of the most consumed beverages in the world, with significant economic, cultural, and nutritional value [1-3]. vietnam has unique natural advantages for tea development, with a total cultivation area of approximately 122.4 thousand hectares, producing over 1.12 million tonnes of fresh tea buds annually [4]. in the north, where the majority of the country's tea regions are concentrated, tea production not only provides hundreds of thousands of households with a substantial source of income, but it also plays a crucial role in developing commodity agriculture and stabilizing livelihoods. tea also promotes the preservation of indigenous culture, the growth of agricultural tourism, and the export of high-value agricultural products. improving the use of agricultural land plays a key role in better organizing farming areas. one way to assess how efficiently land is being used is through comparative analysis. this helps identify which natural conditions are most suitable for different types of agricultural production, such as crop farming or livestock raising [5]. previous studies also emphasize that land use efficiency is a composite indicator, reflecting the coordination among natural, technical, economic, and social factors in agricultural production [6]. agricultural land use efficiency can be assessed through various methods, such as output per unit area, output per worker, or the total value of agricultural production. among these, measuring output per unit area is the simplest and most widely used [7]. the effective use of arable land resources is essential for maintaining social stability and promoting economic development [8]. the economic, social, and environmental aspects of land use efficiency must be considered to produce sustainable agriculture [9]. economic efficiency in agriculture is demonstrated through both quality and quantity indicators. solving land use issues today requires accounting for and evaluating the specific conditions of each cultivated plot [5]. social efficiency is the extent to which agricultural activities contribute to human development, social stability, and improved quality of life in rural areas. phuong et al. [10] stated that social efficiency was measured by the number and value of working days for each land use type. since different crops have varying growth stages, the required working days also differ. environmental efficiency is the extent to which agricultural activities maintain, protect, and improve the quality of the ecological environment, including soil, water, air, biodiversity, and rural ecosystems. environmental efficiency is the extent to which agricultural activities maintain, protect, and improve the quality of the ecological environment, including soil, water, air, biodiversity, and rural ecosystems. quang binh district, located in the southwest of ha giang province, has an area of 780.60 km² and a population of 65,337 people, mainly ethnic minorities such as ha giang’s provincial statistical office [11]. the primary source of income for the populace is agricultural output, with tea being the main crop and occupying the second-largest area in the province after bac quang district. although quang binh's tea area has shown improvement recently, poor product value and weak competition have kept the effectiveness of tea land usage constrained. tea revenue experienced a significant decline due to a steep decrease in tea prices and challenges in exportation in 2024. thus, assessing the efficiency of tea cultivation in quang binh district is vital for appropriately reorganizing production areas and optimizing land potential in line with the specific natural conditions. moreover, considering the fluctuations in tea prices and the rising demand for changes in land use, assessing the effectiveness of tea land utilization offers a scientific foundation for strategic planning, enhancing livelihoods, and guaranteeing the sustainability of land resources in quang binh. 2. methods data survey: secondary data on the current status of tea production was collected from the statistical office of ha giang province and the statistical office of quang binh district. primary data on the current status of tea production (yield, selling price, costs, etc.) was collected from a random survey of 60 tea-growing households in quang binh district using a prepared questionnaire. data processing: collected data is examined, cleaned, and imported into excel in accordance with each survey indicator system, including cost, workforce size, and productivity. it is then processed according to standards for assessing the efficiency of the economy, society, and environment. economic efficiency is related to gross output (go), intermediate cost (ic), added value (va), and capital efficiency (ce) [10]. gross output is the total economic value of tea production in an accounting period and a unit area (go = total products × product value); intermediate cost is the total cost used to pay for materials and services during tea production; added value is the difference between gross output and intermediate cost or the profit of a crop (va = go ic). capital efficiency is the profitability of capital (ce = va/ic). social efficiency is evaluated through indicators such as employment opportunities, working-day value [10], people's acceptance [12, 13], and security of land use rights [9]. environmental efficiency is evaluated through indicators such as increased land cover, maintenance and protection of soil, and reduction of land degradation [13]. 3. empirical results 3.1. current status of tea production in quang binh district table 1 illustrates that the tea cultivation area in quang binh district remained relatively stable during the 2020-2024 period. according to the ha giang’s provincial statistical office [11], the district's total tea-growing area in 2024 reached 33,286.7 hectares, with cultivation concentrated mainly in the communes of tien nguyen, xuan minh, and tan trinh (accounting for approximately 56% of the total tea cultivation area of the entire district). in contrast, vị thường recorded the smallest tea area, with only around 2 hectares under cultivation [14]. during this period, the area of tea under productive harvest showed a consistent upward trend, increasing from 2,701.7 hectares in 2020 to 3,145.7 hectares in 2024, an approximate growth of 16.4%. while the total area remained largely unchanged, the proportion of productive tea area to total tea area rose markedly, from 83.0% to 95.7% [11]. this indicates a significant improvement in land use efficiency and reflects the accelerated conversion of immature agriculture and food sciences research, 2025, 12(2): 110-115 112 © 2025 by the authors; licensee asian online journal publishing group plantations to commercial tea production. however, the rate of expansion began to decelerate after 2022, suggesting that the district may be approaching a relatively saturated stage regarding exploitable tea land. this trend signals a necessary strategic shift from land expansion to the enhancement of yield and product quality. although the growth in productive tea area has contributed to improved output and farmer income, it also presents challenges, requiring a focus on deeper investment, particularly in improved cultivars, intensive farming techniques, and market connectivity to secure sustainable growth. table 1. current status of tea production in quang binh district. indicators 2020 2021 2022 2023 2024 tea cultivation area (ha) 3.256,50 3.234,2 3.268,0 3.293,60 3.286,7 productive tea area (ha) 2701.7 2.861,1 2.997,5 3.085,90 3.145.7 output (tons) 11.036,5 14.897,1 16.780,5 16.972,5 17.301.2 source: ha giang’s provincial statistical office [11]. during the 2020-2024 period, tea output in quang binh district increased steadily from 11,036.5 tons in 2020 to 17,301.2 tons in 2024, representing a growth of approximately 56.8%. the most significant increase occurred between 2020 and 2021, with a rise of over 35%, followed by moderate and stable growth in subsequent years. this upward trend reflected improvements in production efficiency, primarily driven by the expansion of the productive tea area (table 1). according to survey results, quang binh's tea yield steadily increased and surpassed the provincial average. as illustrated in figure 1, the district's yield consistently rose and remained higher than the ha giang provincial average throughout most of the period. in 2020, quang binh's yield was 4.09 tons/ha, lower than the provincial average of 4.78 tons/ha. however, from 2021 to 2024, quang binh exceeded the provincial level and maintained this lead. quang binh district's tea yield in 2024 was 5.50 tons/ha, which is 17.34% higher than the province's total yield (ha giang province's yield in 2024 was 4.6 tons/ha). nevertheless, compared to other significant tea-producing regions, especially thai nguyen province, the yields of tea in quang binh district and ha giang province as a whole remain considerably lower. according to statistical office of quang binh district [14], the average tea yield in thai nguyen province in 2024 reached approximately 12.5 tons/ha [15]. in comparison, the lower yield in quang binh district is primarily attributed to insufficient intensive farming practices that do not meet technical standards, the aging condition of many tea plantations, and suboptimal planting density. figure 1. tea yield in ha giang province and quang binh district. source: ha giang’s provincial statistical office [11]. tea cultivation in quang binh district primarily focuses on industrial shan tea, which is suitable for the cool, humid climate of the highlands. according to custom, tea trees are occasionally planted on slopes or in garden plots adjacent to residential areas. it is not technically ensured that tea will be planted, pruned, fertilized, and pestcontrolled; many tea estates do not utilize organic fertilizers. the application of fertilizers is mainly empirical, lacking balance among nutrients, while pest control measures still rely on chemicals, with little attention paid to biological methods and organic farming. additionally, the harvesting and preservation of raw materials are not conducted at the right time and using proper techniques, which also affects the quality of finished tea. these limitations not only reduce the productivity and quality of tea but also negatively impact the development of sustainable agriculture. 3.2. land use efficiency for tea cultivation 3.2.1. economic efficiency economic efficiency refers to the evaluation of the outcomes achieved from an economic activity [16]. economic efficiency is a multifaceted concept that represents the beneficial outcome of an economic activity in relation to the required expenditures or effort needed for its implementation [17]. the economic efficiency of tea cultivation serves as a key indicator, reflecting not only the productive capacity of the land but also integrating various natural and socio-economic factors. it thus helps assess the suitability of natural conditions for tea planting and production. research findings indicated a marked decline in the economic efficiency of tea in quang binh district during the 2022-2024 period, with a particularly sharp drop in 2024. as shown in table 2, added value reached 47,139,524 vnd in 2022, decreased to 46,173,333 vnd in 2023, and fell significantly to 17,573,690 vnd in 2024. this decline was not primarily due to production costs, as total expenditures in 2024 (22,738,810 vnd) were even lower than in the previous two years. the primary cause was disruptions in the market for consumption, specifically the temporary suspension of tea imports by china, which is ha giang province's main export market. this disruption resulted in a agriculture and food sciences research, 2025, 12(2): 110-115 113 © 2025 by the authors; licensee asian online journal publishing group sharp decline in tea prices and limited market access, leading to significant reductions in both total gross output and added value. these changes highlighted how susceptible tea production is to market uncertainties and emphasized the urgent need for market diversification and added value techniques to lessen dependence on exports. according to field surveys conducted in the research region, many households in quang binh district neglected or abandoned their tea gardens as a result of the low income from tea cultivation. table 2. economic efficiency of tea cultivation in quang binh district. years go ic va ce 2022 70,330,000 23,190,476 47,139,524 2.03 2023 70,070,000 23,896,667 46,173,333 1.93 2024 40,312,500 22,738,810 17,573,690 0.08 the data presented in table 2 indicated that capital efficiency was expected to significantly decline from 2.03 in 2022 to 1.93 in 2023, eventually reaching a mere 0.08 in 2024. this change revealed a sharp decline in the returns on capital spent on tea, especially in 2024, indicating that these expenditures produced little value. the sharp decline in tea prices, difficulties in exporting, growing input costs, and hard-to-market goods were the main causes noted. this situation affected tea farmers' profits, but it also demonstrated the urgency of reorganizing production to increase value, reduce costs, diversify products, and enhance the efficiency of government investments in the tea sector. 3.2.2. social efficiency data in table 3 showed that the labor cost for growing one hectare of tea is between 140 and 145 workers. the cost of labor is significantly lower for mass tea cultivation than in many other tea-growing regions. research by nam [12] showed that tea cultivation in dinh hoa district, thai nguyen province attracts 450 laborers, two times higher than tea cultivation in quang binh district. another study by duc [18] in huong son district, ha tinh province, also indicated that tea cultivation attracted 348.7 laborers per hectare per year [18]. in tea farming, the process of picking tea requires the most labor. survey findings indicated that a worker can harvest 60 to 70 kg of tea. therefore, if 5 tonnes of tea are produced per hectare each year, 70 to 80 workers are needed for tea harvesting. table 3 showed that the value of labor days in tea cultivation in quang binh district decreased sharply over the period from 2022 to 2024, from 325,100.17 vnd to only 125,526.35 vnd, representing a decrease of more than 61%. the income and livelihoods of local workers were strongly impacted by this phenomenon, indicating a significant decline in the socioeconomic efficiency of tea. the purchase price of tea at the garden dropped from vnd 13,000 per kg to roughly vnd 8,000 per kg in the first months of 2024 and barely vnd 6,000 per kg by the end of 2024, mainly due to the steep drop in raw tea prices caused by a significant loss in the export market to china. this situation makes it imperative to diversify the market, enhance the value of the production chain, and promote consumption links to ensure revenue and stabilize the livelihoods of highland tea growers. table 3. social efficiency of tea cultivation in quang binh district. years number of working days working-day value local people's acceptance security of land use rights 2022 145 325,100.17 good good 2023 143 322,890.44 good good 2024 140 125,526.36 good good the evaluation results in table 3 showed that people's acceptance of tea was "good." this was a crucial sign of the social efficacy of tea development for a mountainous district like quang binh. people's positive attitudes and attachment to tea demonstrated that the crop is appropriate for local livelihood demands, growing methods, and production conditions. in regions with a large number of ethnic minorities, such as the mong, dao, and tay, tea not only helps provide steady jobs but also boosts income and reduces poverty, fostering social stability. simultaneously, the decades-long acceptance showed how satisfied and confident people are in tea as a vital crop for family economic growth. this is particularly important for sustainable rural development programs, especially in upland areas, where community and social acceptance play a key role in ensuring policy effectiveness and diffusion. the widespread acceptance of tea by the populace is intimately linked to maintaining long-term land use rights and also demonstrates societal efficiency. people are more likely to stabilize output and remain on the cultivated land rather than move or leave it when they can clearly see the socioeconomic benefits of tea planting. in reality, the degree of acceptance of tea among quang binh district residents encourages them to preserve their land use rights as a sustainable source of income while having no plans to relocate. the acceptance of tea cultivation by the people of quang binh district has led them to maintain land use rights as a sustainable livelihood asset, with no intention of transferring or selling their land. in the context of land being an important resource increasingly pressured by competition for use, this helps limit the fragmentation of agricultural land, protect the land fund for production, and establish a stable foundation for policies that support the development of highland areas. additionally, land used for growing tea is frequently linked to traditional agricultural methods and the lives of mountain people; hence, the mindset of preserving land and sustaining output persists, particularly in instances where the state still guarantees land use rights. even though the research findings indicated a temporary economic downturn, tea trees will remain a "livelihood pillar" in the area, and people do not prioritize surrendering land use rights in the current situation. according to nguyen et al. [19], thailand's land tenure security benefits from higher agricultural investment [19]. thailand's land tenure security benefits from higher agricultural investment [20, 21]. therefore, tea is not only a good choice for cultivation but also helps to ensure that people in quang binh district, ha giang province, have stable and long-term land use rights. 3.2.3. environmental efficiency reports from agricultural households indicate that growing tea helps conserve the environment while providing locals with a reliable source of income from february to november. the broad canopy and high coverage density of tea trees, which are biological traits, allow them to reduce soil erosion and runoff, particularly in steep hilly regions. shan tea varies greatly in height and canopy growth throughout a season; in the first harvest, the trees grow to a agriculture and food sciences research, 2025, 12(2): 110-115 114 © 2025 by the authors; licensee asian online journal publishing group height of roughly 80-100 cm, and in the final harvests of the season, they can reach 120-130 cm. in addition to increasing productivity, the gradual rise in biomass also enhances soil cover efficiency, helps retain moisture, and slows soil deterioration. table 4. environmental efficiency of tea cultivation in quang binh district. assessment increased land cover maintenance and protection of soil reduction of land degradation good 93.33 83.34 76.66 average 6.67 13.33 16.67 low 0.00 3.33 6.67 the results in table 4 show how important tea trees are for preventing land degradation and safeguarding soil. the maintenance and protection of soil from tea trees were graded as "good" by up to 83.34% of the assessors, "average" by 13.33%, and "low" by 3.33%. similarly, 76.66% of survey respondents gave it a "good" rating, 16.67% an "average" rating, and just 6.67% a "low" rating for the reduction of land degradation. these figures clearly reflect the positive ecological effects of tea trees, especially in the context of agricultural production in mountainous areas such as quang binh district. tea trees' broad canopy and consistent growth throughout the seasons have made a substantial contribution to preserving vegetation cover, lowering erosion, retaining moisture, and safeguarding the structure of farmed soil. 3.3. some solutions to enhance the efficiency of tea land use reasonable planning and improvement of tea-growing land: the efficiency of tea land use depends largely on the organization of production space. the tea-growing area must therefore be replanned in accordance with the slope, geography, and soil characteristics to ensure the proper planting density. replanting is required to maintain consistent coverage and maximize the amount of farmed land in tea gardens with improper planting density, such as too few trees or when many tea trees are dead or stunted. replanting is best done between february and march, planting during cold, moderate rain or after heavy rain. repairing the tea canopy twice a year, in april after the conclusion of the spring crop and again in july, is also crucial. replanting improves the ability to develop evenly among tree batches, increases soil coverage, reduces weeds, and prevents erosion in addition to stabilizing tree density according to technical criteria. furthermore, tea gardens with reasonable density also help increase the efficiency of fertilizer management, irrigation water, and care, thereby improving the productivity and quality of tea buds in a sustainable direction. applying sustainable farming techniques and developing value chain production linkage models: transitioning from traditional farming practices to sustainable techniques is an essential solution for effective and long-term land utilization. one significant way to enhance the effectiveness of tea land use is to encourage a shift in perspective from solely agricultural production to agricultural economic growth. constructing value chains emphasizes elements such as cooperative ties, product quality, consumption markets, and branding. to maximize added value on the same unit of land area, this transformation requires support from agricultural extension policies, technology transfer, and, especially, the development of cooperative economic models or connections between farmers and tea processing and consuming firms. specifically, to produce safe, high-quality tea products that are competitive in the market and generate high added value per unit area, a strong production link between the "4 houses: farmers scientists entrepreneurs managers" must be established. additionally, tea gardens require regular cleaning and weeding. young branches are susceptible to disease, so avoid pruning too early. to prevent the spread of disease, tea buds and diseased leaves should be burned and pruned as soon as symptoms appear. provide tea plants with balanced water and fertilizer applications. the optimal time and method for pruning tea are from mid-december to the end of january the following year. to ensure higher productivity, tea should also be harvested using the correct method. leave a sufficient number of buds (three to four leaves for the spring harvest and one to two leaves for the summer-autumn crop) after selecting suitable buds. it is advisable to harvest buds near the fresh leaves at the top of the main stem and branch axis. increasing the use of organic fertilizers and biological products: to improve soil quality in areas where hilly soil is susceptible to erosion and degradation, chemical fertilizers for tea plants must be replaced with organic or microbial fertilizers. organic fertilizers help improve humus content and soil structure, while biological products, particularly nitrogen-fixing or phosphorus bacteria, enhance nutrient absorption for plants. indeed, various microbes can convert insoluble phosphorus compounds into forms that plants can easily absorb and utilize [22]. substantial amounts of phosphorus are stored in biomass for plant use, and phosphorus-solubilizing bacteria in the soil are crucial for solubilizing inorganic phosphorus minerals. nitrogen-fixing microbes can employ biological nitrogen fixation to convert atmospheric molecular nitrogen (n₂) into ammonia (nh₃). plants absorb this ammonia as an accessible form of nitrogen, enhancing soil fertility and sustainability, improving plant health, and increasing productivity [23]. phosphorus-solubilizing microorganisms in the soil are essential for decomposing inorganic phosphorus minerals and storing a sizable amount of phosphorus in biomass for plant uptake. enhancing training and technology transfer: the efficiency of tea land use is closely linked to the cultivation level and awareness of the producer. tea productivity and quality will remain low even with favorable site potential if producers lack the necessary knowledge and skills to manage soil, water, fertilizer, and pests. consequently, enhancing farmers' capacity through consistent coaching and training programs must go hand in hand with increasing land use efficiency. advanced agricultural practices suitable for the region should be the main focus of training materials. these should include balanced nutrient management, fertilization based on the developmental needs of tea plants, the use of organic fertilizers, and soil cultivation methods that prevent erosion and preserve fertility. simultaneously, encouraging the implementation of integrated pest management (ipm) strategies, prioritizing biological pesticides and safe pest control methods, will help reduce adverse effects on the soil environment and safeguard the agricultural ecosystem. to increase land use efficiency in a stable and long-term manner, it is also vital to emphasize the role of agricultural extension agents, cooperatives, and businesses in facilitating connections, sharing technologies, and assisting individuals in engaging in sustainable tea production. 4. conclusion tea cultivation plays an essential role not only in generating household income but also in contributing to agriculture and food sciences research, 2025, 12(2): 110-115 115 © 2025 by the authors; licensee asian online journal publishing group broader economic, social, and environmental objectives in quang binh district. the overall area used for tea production remained relatively stable between 2020 and 2024; however, in 2024, economic efficiency fell precipitously due to disruptions in export markets and a sharp decline in tea prices, underscoring the susceptibility of local life to market swings. tea cultivation continues to be highly accepted by people, especially among ethnic minorities, due to its compatibility with traditional practices and its role in ensuring employment and land tenure stability. additionally, tea cultivation contributes to increasing land cover, maintaining and protecting the soil, and reducing land degradation. however, the declining value of daily labor wages and limited market 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[23] j. j. wewalwela, m. seneviratne, and s. a. shields-menard, role of nitrogen-fixing microorganisms for plant and soil health. in r. soni, d. c. suyal, p. bhargava, & r. goel (eds.), microbiological activity for soil and plant health management. singapore: springer, 2021. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.tifs.2020.06.009 https://doi.org/10.3390/foods13223580 https://doi.org/10.1016/0264-8377(85)90069-9 https://doi.org/10.2307/142548 https://doi.org/10.1016/j.techfore.2018.03.015 https://doi.org/10.1016/j.oneear.2021.08.015 https://doi.org/10.26459/hueunijard.v131i3c.6761 https://doi.org/10.3390/su8020178 https://doi.org/10.1016/j.euroecorev.2005.02.001 https://doi.org/10.1257/000282802762024575 54 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 1, 54-68, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i1.6819 © 2025 by the authors; licensee asian online journal publishing group on-farm evaluation of fertilizer recommendation methods: impacts on rice yield and economic benefits in subtropical soils md. enamul haque1 richard w. bell2 miaomiao cheng3 md. abdus satter4 md. baktear hossain5 md. jahiruddin6 md. mofizur rahman jahangir7 md. aminul islam8 sohela akhter9 md. monayem miah10 a. k. m. anisur rahman11 ( corresponding author) 1,2,3school of agricultural science, murdoch university, murdoch 6150, australia. 1email: enamul.haque71@gmail.com 2email: r.bell@murdoch.edu.au 3email: miaomiao.cheng@dpird.wa.gov.au 4,5bangladesh agricultural research council, farmgate, dhaka, bangladesh. 4email: a.satter1959@gmail.com 5email: baktear@gmail.com 6,7department of soil science, bangladesh agricultural university, mymensingh, bangladesh. 6email: m_jahiruddin@yahoo.com 7email: mmrjahangir@bau.edu.bd 8soil science division, bangladesh rice research institute, gazipur, bangladesh. 8email: aminbrri@gmail.com 9soil science division, bangladesh agricultural research institute, gazipur, bangladesh. 9email: sohela_akhter@yahoo.com 10agricultural economics division, bangladesh agricultural research institute, gazipur, bangladesh. 10email: monayem09@yahoo.com 11department of medicine, bangladesh agricultural university, mymensingh, bangladesh. 11email: arahman_med@bau.edu.bd abstract bangladesh publishes fertilizer recommendations but most farmers do not use these recommendations. in this study, we tested the performance of the fertilizer recommendation guide (frg), rice crop manager (rcm) and soil testing kit (stk) for determining fertilizer requirements of monsoon rice (oryza sativa l) against farmers’ fertilizer practice (ffp). rates of nitrogen, phosphorus, potassium, sulfur and zinc were determined by four methods and used on 72 farmer’s fields in five agro-ecological zones (aez) over two years. all methods produced significantly higher grain yield than ffp (4.12 t ha-1): they were 13.8%, 9.6% and 8.3% higher for stk, rcm and frg, respectively. rcm didn’t perform consistently across the locations. overall, the stk performed the best followed by frg. the superiority of stk is attributed to its assessment of current soil status while the frg recommendation is grounded on an older data set of soil analysis for each aez. the n dose was comparable among the methods while p dose was much higher for ffp, and rcm underestimated the k requirement of rice. farmers declined to adopt the stk. by contrast, the supply of frg information to the farmers by providing a simplified card with training is a simple and accessible technology. keywords: adoption, frg card, nitrogen, phosphorus, potassium, rice crop manager, soil test kit, sulfur, zinc. contents 1. introduction ...................................................................................................................................................................................... 55 2. materials and methods ................................................................................................................................................................... 55 3. results ................................................................................................................................................................................................ 61 4. discussion .......................................................................................................................................................................................... 64 5. conclusion ......................................................................................................................................................................................... 66 references .............................................................................................................................................................................................. 66 mailto:enamul.haque71@gmail.com mailto:r.bell@murdoch.edu.au mailto:miaomiao.cheng@dpird.wa.gov.au mailto:a.satter1959@gmail.com mailto:baktear@gmail.com mailto:m_jahiruddin@yahoo.com mailto:mmrjahangir@bau.edu.bd mailto:aminbrri@gmail.com mailto:sohela_akhter@yahoo.com mailto:monayem09@yahoo.com mailto:arahman_med@bau.edu.bd https://www.doi.org/10.20448/aesr.v12i1.6819 https://orcid.org/0000-0002-6551-5174 https://orcid.org/0000-0002-7756-3755 https://orcid.org/0000-0002-2979-6976 https://orcid.org/0000-0002-3636-1057 https://orcid.org/0009-0009-6936-5634 https://orcid.org/0000-0002-7944-2156 https://orcid.org/0000-0002-9501-3239 https://orcid.org/0000-0003-0110-4488 https://orcid.org/0000-0002-8387-0727 https://orcid.org/0000-0002-1054-4437 https://orcid.org/0000-0001-9660-4949 agriculture and food sciences research, 2025, 12(1): 54-68 55 © 2025 by the authors; licensee asian online journal publishing group citation | haque, m. e., bell, r. w., cheng, m., satter, m. a., hossain, m. b., jahiruddin, m., jahangir, m. m. r., islam, m. a., akhter, s., miah, m. m., & rahman, a. k. m. a. (2025). on-farm evaluation of fertilizer recommendation methods: impacts on rice yield and economic benefits in subtropical soils. agriculture and food sciences research, 12(1), 54–68. 10.20448/aesr.v12i1.6819 history: received: 7 april 2025 revised: 15 may 2025 accepted: 16 june 2025 published: 25 june 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the australian centre for international agricultural research (aciar), australia (grant number: lwr/2016/136) and the krishi gobeshona foundation (kgf), bangladesh (grant number: cn/frpp: icp-ii). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contribution of this paper to the literature: tools have been developed to assist farmers in making better fertilizer decisions, but they are rarely tested against one another and demonstrated to be more effective than farmers' current fertilizer use. we demonstrated that an expert-driven fertilizer recommendation system was accessible to smallholder farmers and achieved an 8% average yield increase. 1. introduction rice (oryza sativa l.) is the most dominant crop in asia while about 40% people of the world consume rice as a major staple food. rice crops in bangladesh occupy about 75% of the crop area and rice-based cropping system is predominant [1]. the rice soils are commonly deficient in n, p, k, s and zn. in the intensive, high yield cropping systems of south asia, especially in india and bangladesh, farmers generally apply excessive p fertilizer to rice but generally insufficient k, s or zn [2]. nitrogen applications relative to recommendations vary with farm size, cropping pattern and crop species [2]. however, improvements in n use efficiency can increase crop yields [3] since commonly less than 50% of urea-n is taken up by the crops and the rest is lost through leaching (no3), volatilization of nh3 [4] and denitrification (n2o, n2) processes [5] and thereby contaminating environment [6]. optimising the use of fertilizers continues to be a global concern since crop yield loss occurs with underuse while the risk of environmental loss increases with overuse. there are numerous fertilizer recommendation systems for crop cultivations globally. according to beneduzzi, et al. [7] a literature search identified 12 methods for recommending n, eight methods for p, and seven methods for recommending k, in addition to five computer programs to make fertilizer recommendations at varying rates. buresh, et al. [8] determined fertilizer k and p requirements for rice (oryza sativa l.) cultivation by the site-specific nutrient management (ssnm) approach using estimated target yield, nutrient balances, and yield gains from added nutrients. that approach was based on model estimates without reliance on fresh soil analysis [9]. dobermann and cassman [10] argued that effective strategies for site-specific nutrient management should be based on a quantitative understanding of the relationships between nutrient supply and crop demand. usually, soil analysis is considered as an essential step for evaluation of soil fertility and thus on the fertilizer recommendation. soil tests provide a scientific basis for fertilizer recommendations. understanding and measuring spatial variability regarding nutrient availability in the soil is crucial to defining site-specific fertilizer management strategies to increase production efficiency and sustainability of agricultural production [11]. however, soil chemical analysis is expensive and time consuming to the point of being economically unfeasible for site-specific nutrient application in low value-added crops [12]. for smallholder farms, spatial and temporal variability in nutrient status on many small fields adds to the cost of using soil testing for site specific nutrient management. there are three methods of fertilizer recommendation now available in bangladesh – frg [13] rice crop manager (rcm) and soil testing kit (stk). however, the majority of farmers do not use those methods [2] and employ their own calculation which in this study we have called farmers’ fertilizer practice (ffp). the fertilizer rates proposed may vary among recommendation methods but their impact on crop yield and profitability is not known. with this understanding, we studied performances of three fertilizer recommendation methods relative to ffp, with two objectives: (i) to determine the fertilizer (urea, tsp, mop, gypsum and zinc sulphate) requirement for transplanted aman rice, and (ii) to identify the best method of fertilizer recommendation for rice in terms of yield benefits, economic return and adoptability by smallholder farmers. 2. materials and methods 2.1. experimental site and soil the field trials were conducted in six locations representing five agro-ecological zones (aez) of the country. in each location, the experiment was set up in six sites representing six farmer plots and it was done in two years (2018 and 2019) giving a total of 72 field trials (6 locations × 6 sites × 2 years) (figure 1). all the experimental sites were on medium-high land. bangladesh has a sub-tropical humid climate and is characterized by hot and humid summer and cool dry winter. the country experiences monsoon rainfall, 80% of which occurs between june and october when transplanted aman rice is grown. general characteristics of the soils under the experiments are presented in table 1. the methods for soil analysis were followed as outlined by page, et al. [14]. https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ agriculture and food sciences research, 2025, 12(1): 54-68 56 © 2025 by the authors; licensee asian online journal publishing group figure 1. map of bangladesh showing trial locations. agriculture and food sciences research, 2025, 12(1): 54-68 57 © 2025 by the authors; licensee asian online journal publishing group table 1. general characteristics of soils in different locations and sites under experiments. parameters durgapur godagari mymensingh sadar thakurgaon sadar dacope amtali methods aez 11 26 9 1 13 13 gis texture silt loam silty clay loam silt loam sandy clay loam silty clay silty clay hydrometer ph 6.5-7.2 5.7-6.9 6.0-7.3 5.6-6.1 6.6-7.9 6.6-7.9 soil-water ratio 1:2.5 organic matter % 1.45-2.25 1.45-2.25 1.45-2.25 1.45-2.55 1.45-2.25 1.45-2.25 wet oxidation total n (%) 0.045-0.085 0.045-0.085 0.045-0.085 0.045-0.085 0.045-0.085 0.045-0.085 kjeldahl extractable p (mg /kg) 3.75-11.3 11.3-18.8 3.75-11.3 11.3-18.8 3.75-11.3 3.75-11.3 nahco3 extraction exch. k ((cmol/kg) 0.085-0.225 0.085-0.225 0.045-0.085 0.045-0.085 0.085-0.225 0.085-0.225 nh4oac extraction extractable s (mg /kg) 5.4-16.2 5.4-16.2 5.4-16.2 5.4-16.2 37.8-48.5 37.8-48.5 cacl2 extraction extractable zn ((mg /kg) 0.27-0.81 0.27-0.81 0.81-1.36 0.27-0.81 1.36-1.89 1.36-1.89 dtpa extraction note: aez 1 = old himalayan piedmont plain, 9 = old brahmaputra floodplain, 11 = high ganges river floodplain, 13 = ganges tidal floodplain, 26 = high barind tract. source: fao/undp [15]. agriculture and food sciences research, 2025, 12(1): 54-68 58 © 2025 by the authors; licensee asian online journal publishing group three types of fertilizer recommendation methods frg, rcm and stk, and ffp were tested in 72 farmers’ (36 farmers each year) plots during two consecutive aman rice seasons of 2018 and 2019. in bangladesh, soil fertility experts have developed a fertilizer recommendation guide (frg) for all 30 aezs for the farmers to select a recommended dose of fertilizers in their crops. here fertilizer recommendation for a crop or cropping pattern is given on the basis of general fertility level of an aez. the bangladesh agricultural university (bau), mymensingh has developed a low-cost rapid soil testing technology i.e. soil testing kit (stk) based on an on-site soil test, from which a fertilizer requirement for a crop is estimated. the rice crop manager (rcm) is a web-based fertilizer recommendation system developed by irri for site-specific nutrient management (ssnm) in the specific rice fields [8]. before initiating crop establishment with rcm (http://webapps.irri.org/in/od/rcm), each farmer is interviewed to collect essential data inputs. the data collected includes: the farmer’s field location, field size, rice variety, anticipated age of seedlings at transplanting, water management (irrigated or rainfed), rice yield in previous years with the same or similar variety, portion of aboveground residues from the previous crop retained in the field, and the farmer’s choice of fertilizer sources. rcm utilizes this data to calculate a field-specific fertilizer recommendation tailored to achieve a target yield set by rcm. the rcm recommendation includes rates and timings for the application of fertilizer sources selected by the farmer [16]. 2.2. treatments and design each method of fertilizer estimation is considered as a treatment. the experiment consisted of four treatments: t1 = farmer fertilizer practice (ffp), t2 = frg based fertilizer dose, t3 = rcm based fertilizer dose and t4 = stk based fertilizer dose. urea (46% n), triple superphosphate (tsp 20% p), muriate of potash (mop 50% k), gypsum (18% s) and zinc sulphate heptahydrate (21% zn) were used to supply n, p, k, s, & zn, respectively. the doses of nutrients based on the four methods including ffp are given in table 2 and their fertilizer conversions are shown in supplementary table s1. each experiment was laid out in a randomized complete block design (rcbd) with six dispersed replications, i.e. each farmer/site represented one replication. so, there were 12 plots in each location for both years; farmers/sites were different for the two years. each plot size ranged from 115 to 334 m2 (table 3). table 2. estimation of n, p, k, s and zn rates based on four different methods for five locations for use in transplanted aman rice crops (average of 2 years). location (upazila) fertilizer rate estimation methods nitrogen (kg/ha) phosphorus (kg/ha) potassium (kg/ha) sulphur (kg/ha) zinc (kg/ha) durgapur, rajshahi ffp 93.8 26.1 39.5 5.2 1.89 frg 79.9 8.5 38.0 9.9 1.61 stk 88.7 8.0 75.5 11.2 2.14 rcm 95.4 10.3 24.0 5.1 1.56 mean ± sd 89.4 ± 7.0 13.2 ± 8.6 44.3 ± 22.0 7.9 ± 3.1 1.8±0.28 godagari, rajshahi ffp 106.0 15.3 36.8 12.0 0.91 frg 88.9 10.7 62.5 8.5 0.49 stk 88.7 3.0 53.0 13.4 4.25 rcm 117.0 12.2 29.8 7.5 2.45 mean ± sd 100 ± 13.9 10.3 ± 5.2 45.5 ±14.9 10.3 ±2.8 2.03±1.71 mymensingh, sadar ffp 50.0 12.6 29.5 0.5 0.00 frg 89.1 9.1 43.0 7.7 1.28 stk 88.7 9.2 69.3 5.6 2.14 rcm 139.3 24.6 33.8 3.4 1.31 mean ± sd 91.7 ±36.6 13.9 ± 7.3 43.9 ± 17.8 4.3 ±2.8 1.18±1.71 thakurgaon, sadar ffp 99.2 18.9 48.3 14.1 3.15 frg 86.4 8.4 39.5 8.5 1.59 stk 86.2 10.1 64.8 8.7 3.17 rcm 70.2 9.7 35.8 6.3 2.38 mean ± sd 92.0 ±11.9 12.3 ± 4.8 45.1 ±12.9 7.9 ± 3.3 1.86±0.75 dacope, khulna ffp 69.1 17.3 42.3 4.2 0.00 frg 81.7 12.7 34.3 4.3 1.28 stk 88.9 11.0 44.5 6.8 2.64 rcm 73.4 9.7 24.3 8.7 0.79 mean ± sd 85.5 ± 8.8 11.8 ±3.3 47.1 ± 9.2 9.4 ± 2.2 2.57 ± 1.1 amtali, barguna ffp 84.4 20.1 12.0 3.0 0.00 frg 72.7 8.6 29.3 3.4 1.37 stk 89.1 13.0 65.5 4.5 2.15 rcm 84.4 9.0 24.0 1.2 0.88 mean ± sd 82.6 ± 7.0 12.7 ± 5.3 32.7 ± 23.0 3.0 ± 1.4 1.10±0.90 note: ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, stk=soil testing kit, rcm = rice crop manager. 2.3. crop management rice (varieties mentioned in table 3) was grown as a rainfed crop during july to november (kharif season) in each site and location. thirty-day-old rice seedlings were transplanted at three seedlings per hill with a spacing of 20 × 20 cm. urea was applied in three splits 50% urea during final land preparation, 25% urea at 30 days (tiller stage) and 25% urea at 50 days after transplanting (panicle initiation stage). the other fertilizers viz. tsp, mop, gypsum and zinc sulphate were added during final land preparation before transplanting rice seedlings. pre and post emergence herbicides were used to all plots as per requirement. insecticides ‘brifer 5 g’ and ‘cidial 5 g’ were used to control insect attack principally stem borer of rice. irrigation was required in some sites before final land preparation. the crop was harvested at maturity from four randomly selected quadrats of 10m2 from each plot to record their yields. the yields were reported at 14% grain moisture. agriculture and food sciences research, 2025, 12(1): 54-68 59 © 2025 by the authors; licensee asian online journal publishing group 2.4. statistical analysis yield data were statistically analyzed by mstat-c statistical software program (michigan state university, usa) based on a randomized complete block design (rcbd) and the mean differences were compared by duncan’s multiple range test (dmrt) [17]. correlation statistics was done to examine the relationship between the methods of fertilizer estimation (ffp, frg, rcm and stk) and pairwise treatment comparisons were done by tukey’s hsd (honest significant difference) method. the impact of nutrients or fertilizers (n, p, k, s & zn) on rice yield were evaluated by mixed-effects linear regression models [18]. 2.5. economic analysis total variable costs were calculated considering costs for land preparation, seedlings, fertilizers, irrigation, harvesting, and labor for all operations including threshing and drying. gross return was calculated by multiplying the amount of produce by its corresponding price at harvest. gross margin was calculated by subtracting variable cost from gross return. each of the treatments was evaluated based on total variable cost, gross return, gross margin and benefit-cost ratio. the labor required to complete each operation (land preparation, irrigation, and herbicide or insecticide application) in a particular treatment plot was recorded and converted to person-days /ha considering 8h as equivalent to one person-day and the daily labor wage was tk. 400 (1us$ = 85 tk.) per person per day (bangladesh government wage rate). prices of urea, tsp, mop, gypsum and zinc sulphate were 16, 22, 15, 10 and 180 tk./kg; urea, tsp and mop prices are government subsidized. agriculture and food sciences research, 2025, 12(1): 54-68 60 © 2025 by the authors; licensee asian online journal publishing group table 3. locations, rice varieties, seedling age, and transplanting and harvesting dates of crops during 2018 and 2019. year location no. of field sites gps location individual plot size (m2) rice varieties seedling age (days) seedling transplanting period harvesting date latitude longitude 2018 durgapur, rajshahi 6 24°27'27"n 24°28'8"n 88°45'23"e 88°45'54"e 264 brridhan72 24-25 29-31 jul 2018 9-17 nov 2018 godagari, rajshahi 6 24°23'46"n 24°24'05"n 44°26'27"e 44°26'57"e 238 brridhan51 30-35 11-18 jul 2018 9-17 nov 2018 sadar, mymensingh 6 24°40'53"n 24°41'14"n 90°26'6"e 90°27'0"e 213 brridhan49 30-35 5-13 aug 2018 22-27 nov 2018 sadar, thakurgaon 6 25°59'59"n 26°0'0"n 88°28'27''e 88°30'34''e 334 brridhan51 25-30 30 jul 8 aug 2018 13-19 nov 2018 dacope, khulna 6 22˚34'59"n 22˚36'30"n 89˚28'2"e 89˚30'81"e 214 br-23 30-35 3-15 sep 2018 12-24 dec 2018 amtali, barguna 6 22˚2'14"n –22˚2'26" n 90˚14'30"e90˚14'34" e 227 br-23 30-35 9-16 sep 2018 13-19 dec 2018 2019 durgapur, rajshahi 6 24˚25'36"n 24˚25'38"n 88˚44'38"e 88˚44'39"e 119 brridhan48 24-25 26-28 aug 2019 16-nov 2019 godagari, rajshahi 6 24˚23'42"n 24˚43'05"n 88˚26'27"e 88˚26'47"e 179 brridhan51 24-27 16-19 jul 2019 16-18 nov 2019 sadar, mymensingh 6 24°40'56"n 24°41'49"n 90°25'07"e 90°26'49"e 115 brridhan49 29-34 1-11 aug 2019 12-26 nov 2019 sadar, thakurgaon 6 26°01'17"n 26°03'36"n 88°28'49"e 88°30'40"e 175 brridhan51 22-35 4-28 aug 2019 17-25 nov 2019 dacope, khulna 6 22°36'20"n 22°36'38"n 89°18'36"e 89°30'9"e 187 br-23 30-40 30 aug 18 sep 2019 8-22 dec, 2019 amtali, barguna 6 22°2'16"n –22°2'30" n 90°14'32"e-90°14'51" e 131 br-23 28-30 7-19 sep 2019 9-18 dec 2019 agriculture and food sciences research, 2025, 12(1): 54-68 61 © 2025 by the authors; licensee asian online journal publishing group 3. results 3.1. variations in the estimated fertilizer rates the nutrient or fertilizer rates varied with different methods of estimation. the average n rate ranged from 83.1 to 96.6 kg/ha, with the highest estimation by rcm and the lowest by frg, while for ffp and stk rates were 83.7 and 88.4 kg n/ha, respectively (figure 2). for p, the ffp had the highest rate, 18.4 kg/ha, while stk recommended the lowest, 9.1 kg/ha. the stk estimated the highest rate of k (62.1 kg/ha) and rcm proposed the lowest (27.3 kg/ha). for s and zn also, stk estimation was the highest followed by rcm. the zn rate was <1 kg/ha for ffp. hence, the ffp rate of p was double the value of stk while in case of k the ffp rate was half of the stk rate. figure 2. nutrient rates (kg/ha) as estimated by different methods over the locations. note: ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, rcm=rice crop manager, stk=soil testing kit. rcm = rice crop manager. 3.2. grain yield the two years’ average yields of rice against each fertilizer assessment method for each location and sites are presented in figure 3. the year-wise grain yield for 2018 and 2019 are shown in table s2 (supplementary). the grain yield varied with different fertilizer recommendation methods, locations and sites. the stk method always gave the highest grain yield and ffp gave the lowest. overall, rcm and frg methods produced an identical effect on rice yield. the result trends were stk > rcm = frg > ffp. figure 3. effects of different methods of fertilizer estimation on rice yield at different locations. note: ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, rcm = rice crop manager, stk=soil testing kit agriculture and food sciences research, 2025, 12(1): 54-68 62 © 2025 by the authors; licensee asian online journal publishing group considering location variations, the grain yield recorded at durgapur (rajshahi district) varied from 3.96 to 4.58 t/ha, godagari (rajshahi) from 5.23 to 6.16 t/ha, mymensingh sadar from 5.07 to 5.52 t/ha, thakurgaon sadar from 4.32 to 4.69 t/ha, dacope (khulna) from 4.38 to 5.07 t/ha and amtali (barguna) from 4.09 to 5.57 t/ha (table 4). the mean grain yield of rice for the six locations over the treatments were 4.25, 5.68, 5.36, 4.52, 4.67, and 4.64 t/ha, respectively. it reveals that stk demonstrated the highest yield (4.69 t/ha) and ffp exhibited the lowest (4.12 t/ha). comparing the results for rcm and frg, similar results were noted for durgapur, mymensingh and thakurgaon locations, but the yield was significantly higher for rcm over frg at dacope and amtali while the reverse was true for godagari i.e. frg gave higher yield over rcm. the location average yield was found to be 5.09 t/ha for stk, 4.94 t/ha for rcm, 4.88 t/ha for frg and 4.51 t/ha for ffp; the grain yields recorded with rcm and frg were statistically identical. table 4. grain yield (t/ha) of rice in different locations as influenced by different fertilizer doses determined by various methods (results are the average of 2 years). fertilizer rate estimation methods durgapur godagari mymensingh sadar thakurgaon sadar dacope amtali methods average ffp 3.96 b 5.23 b 5.07 b 4.32 b 4.38 b 4.09 c 4.51 c frg 4.29 ab 5.83 a 5.38 b 4.51 b 4.38 b 4.91 b 4.88 b rcm 4.19 b 5.49 b 5.46 ab 4.58 ab 4.86 a 5.07 a 4.94 b stk 4.58 a 6.16 a 5.52 a 4.69 a 5.07 a 4.51 a 5.09 a f-test ** ** * * ** ** * location average 4.25 5.68 5.36 4.52 4.67 4.64 note: * indicate p < 0.05. ** indicate p<0.01. within a column, the mean values followed by the same letter are not significantly different at the 0.05 level of probability by dmrt. ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, stk=soil testing kit, rcm = rice crop manager. 3.3. economics of fertilizer use total variable costs include all inputs costs (fertilizers, seedlings, pesticides, irrigation, etc.) and labor costs for operations from land preparation, transplanting etc. to harvest and then processing for marketable produce. yearwise variable costs are displayed in table s3 (supplementary) and the two years’ average cost is presented in table 5 which indicates that the variable costs for stk were us$ 817 followed by 799, 797 and 792 us$ in case of rcm, frg and ffp, respectively. fertilizer rates and likely costs varied with the different fertilizer assessment methods. it is noted that generally the fertilizer doses or costs were lower for rcm than stk particularly in saline (dacope and amtali) and calcareous soils (durgapur), but the opposite was the case for the other three locations (barind, non-calcareous and piedmont soils). however, the costs varied among the methods of fertilizer estimation and obviously between the doses of fertilizer applied for different methods. agriculture and food sciences research, 2025, 12(1): 54-68 63 © 2025 by the authors; licensee asian online journal publishing group table 5. total fertilizer costs, gross margin and benefit-cost ratio of crop in different locations as influenced by different fertilizer doses determined by various methods (results are the average of 2 years). location total fertilizer costs (us$/ha) gross margin (us$/ha) benefit-cost ratio ffp frg rcm stk ffp frg rcm stk ffp frg rcm stk durgapur 93 98 86 102 587 722 672 793 1.63 1.81 1.75 1.86 godagari 112 103 107 130 734 930 830 1034 1.88 2.13 2.00 2.22 mymensingh sadar 52 81 97 77 867 885 841 909 2.07 2.05 1.94 2.05 thakurgaon sadar 117 98 65 106 739 805 828 870 2.00 2.16 2.20 2.21 dacope 77 84 78 98 531 528 678 688 1.70 1.69 1.89 1.87 amtali 55 79 52 108 363 502 569 397 1.51 1.69 1.78 1.52 note: ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, rcm = rice crop manager, stk=soil testing kit. agriculture and food sciences research, 2025, 12(1): 54-68 64 © 2025 by the authors; licensee asian online journal publishing group gross return calculated from the price of rice grain and straw against each treatment is presented for year-wise data in table s4 (supplementary). like crop yield, the gross return was relatively lower in saline zones (dacope and amtali) that was followed by piedmont plain (thakurgaon), and gross return for the other three locations (dur-gapur, godagari and mymensingh) was relatively higher. for all locations except amtali, the stk recorded the highest gross return (us$ 1,598) and ffp had the lowest return (us$ 1,434). the gross return in durgapur ranged from us$ 1,517 to 1,709, godagari us$ 1,567-1,879, mymensingh us$ 1,676-1,772, dacope us$ 1,2901,479 and in amtali the return was us$ 1,0781,294 per ha (table 5). the year-wise values of gross margin for different fertilizer recommendation tools and locations are shown in table s5 (supplementary) and 2-year average results are displayed in table 5. the gross margin followed the trend: stk > rcm = frg > fp. for locations, the gross margin followed the order: godagari > mymensingh > thakurgaon > durgapur > dacope > amtali indicating that gross margin in saline regions was relatively lower which can be attributed to lower yield. the gross margin in durgapur ranged between 587 and 793 us$, godagari between 734 and 1034 us$, mymensingh between 841 and 909 us$, thakurgaon between 739 and 870 us$, dacope between 531 and 688 us$ and in amtali it was between 363 and 569 us$ per ha. 4. discussion we tested the performance of frg, rcm, and stk in determining the fertilizer requirements of monsoon rice compared to ffp. additionally, we assessed the total fertilizer costs, gross margin, and benefit-cost ratio of the crop in different locations for various fertilizer doses determined by each method. although performance of the stk method was better than frg, the lack of a profitable business model for operators has prevented its widespread use by farmers. consequently, we recommend the distribution of pertinent frg information to farmers through the implementation of frg cards. these cards would contain information on fertilizer requirements for each crop, providing a simple and adoptable technology to enhance farmers' decision-making regarding fertilizer usage. 4.1. effects of methods of fertilizer rate estimation on rice yield the different rates of application of fertilizers produced differences in rice yield but the stk method consistently had the highest grain yield while the ffp had the lowest. for the other two methods (frg and rcm), the yield was comparable over the locations. thus, the grain yield followed the order of stk > rcm = frg > ffp (table 4). overall, the stk yield was 0.5-1.0 t/ha higher than the ffp yield. the potential yield (same as frg target yield) of rice varieties used in this study is 5.0 ± 0.5 t/ha. hence, the stk method enabled rice to reach the potential yield in all locations, the rcm in all places except durgapur, frg in all locales except durgapur and dacope, and ffp in only two locations, godagari and mymensingh. 4.2. relationship between fertilizer estimation methods in terms of rice yield all the methods of fertilizer rate assessment showed significant and positive correlation between each other indicating that yield trend for every method over the locations was similar. it is noted that stk is most correlated with the other methods and ffp is very less correlated (table 6). the highest correlation occurs between frg and stk (r=0.813) because both methods are soil analysis based; the stk value is from fresh soil sample analysis while the frg from an average of historical soil test value of a whole aez. the yield differences among the methods were assessed by ‘t’ statistics. it reveals that the yield increase over ffp for all other methods was statistically significant. specifically, stk showed significance at the 0.1% level, rcm at the 1% level, and frg at the 5% level of significance while the increases as a percentage were 12.9%, 9.6% and 8.3% higher over ffp, respectively. the average yield differences of frg versus rcm, frg versus stk, and rcm versus stk across the locations were not significant (table 6). table 6. correlation and ‘t’ statistics to determine the relationship between fertilizer estimation methods with respect to rice yield (n=72, 6 locations, 2 years). fertilizer rate estimation methods ‘r’ value ‘t’ value+ ffp vs frg 0.750*** 2.64* ffp vs rcm 0.745*** 3.08** ffp vs stk 0.780*** 4.10*** frg vs rcm 0.756*** 0.43 frg vs stk 0.813*** 1.46 rcm vs stk 0.771*** 1.05 note: *, p<0.05; **, p<0.01; ***, p<0.001. + pairwise treatment comparisons by tukey’s hsd method. 4.2. relationship between nutrient rates determined by different methods and rice yield the rate of different nutrients (n, p, k, s & zn) for rice production was determined by four methods. their indi-vidual contribution to rice yield is evaluated by mixed-effects linear regression models and a t-test that uses sat-terthwaite’s method (table 7). all the nutrients except p had positive contribution to rice yield in the decreasing order of k> zn> n> s: rates of p had a negative effect on yield. statistically, the effect of p, k and zn on rice yield was significant. noticeably the ffp-p dose was 2-3 times higher than that of other methods’ dose in all locations except mymensingh where rcm-p dose was 2 times higher than ffp-p dose. on the other hand, the stk-k dose was 2-3 times higher than the rcm-k dose in all places except godagari where it was about 10 kg/ha lower compared to rcm-k. probably the low estimation of k rates accompanied with inconsistency yield performances across the locations is a weakness of rcm method. additional research to improve the calibration of the rcm k rates may improve its utility as a recommendation tool. like rcm, a major weakness of frg method is the low valuation of k dose. the next revision of the frg may need to increase the recommendations for k as suggested by islam, et al. [2] and islam, et al. [19]. furthermore, nutrient balance studies show that negative k balance is quite large and that increases in k rate by 25 to 50% can increase crop yield [19, 20]. the ffp method agriculture and food sciences research, 2025, 12(1): 54-68 65 © 2025 by the authors; licensee asian online journal publishing group has failed to offer potential yield presumably due to use of very high dose of p, low or no application of zn and low or high rate of s application (table 2). interestingly, all the n doses were similar, with only a 10% variation among the methods. table 7. correlation and ‘t’ statistics to determine the relationship between nutrient rates with respect to rice yield (n=24, 6 locations, 4 fertilizer methods). fertilizer rate ‘r’ value ‘t’ value (n=24) (satterthwaite's method) yield vs n rate 0.253 1.49 yield vs p rate -0.380* -3.39** yield vs k rate 0.242 2.32* yield vs s rate 0.178 0.39 yield vs zn rate 0.181 2.14* note: *, p<0.05; **, p<0.01. the initial soil analysis indicated consistently low to very low levels of soil exchangeable k across all study locations [13]. a study estimated the relationship between grain yield and nutrient accumulation in dry matter of irrigated rice, especially focusing on harvest index values ≥0.4. predicted reciprocal internal efficiencies (ries) at 60–70% of yield potential corresponded to specific nutrient accumulations. for irrigated rice with a harvest index ≥0.4, the estimated accumulation per ton of grain yield included 14.6 kg of n, 2.7 kg of p, and 15.9 kg of k [8]. based on that consideration, we estimate that the uptake by rice grain could range from 66 to 74 kg n /ha, 12-14 kg p /ha and 72-81 kg k/ha. it is important to note that the k uptake for a rice crop may be considerably higher, given that the concentration of k in rice straw is generally more than four times that in the grain [21]. while a portion of the k uptake is anticipated to be supported by the soil (though the exact amount remains unknown due to the absence of a k control), irrigation water, and accretion from rainfall [19] the remaining requirement must be fulfilled through the application of k fertilizer (mop). inadequate application of k also restricts the n uptake which lowers n use efficiency and increases the leaching risk of soluble forms of n. most of the k in soil is chemically bound in insoluble forms and is slowly available for plant growth. this is especially true in soils that have been depleted due to continuous farming [22]. while a rice crop removes a large amount of k from soil for growth of straw and less through grain, some of the straw k is recycled to soils. increased crop residue retention as in conservation agriculture practice increases the k recycling to soil and decreases the magnitude of negative k balance. panaullah, et al. [23] reported highly negative k balance for rice-based cropping system in bangladesh. recently hasan, et al. [24] reported an area of 0.287-2.43 mha (out of 8.87 mha arable land) have very low to low k status. 4.4. economic benefits for different methods of fertilizer requirement assessment the fertilizer cost in stk method was highest in four locations durgapur, godagari, dacope and amtali – but not in mymensingh and thakurgaon. for the case of ffp, fertilizer cost was the lowest in durgapur, mymensingh, dacope and barguna. across all locations, average fertilizer cost follows the order: stk>frg>ffp>rcm, with an overall variation of 10-20%. so, although the cost variation from different fertilizer management strategies methods was not large, due to more effective balances of nutrients supplied, the impact on rice yield was substantial. hence, the highest bcr was for stk (1.96) and the lowest for ffp (1.8), while bcr for both rcm and frg was 1.92 (table 5). the consistent benefit from the stk method of fertilizer requirement assessment was due to a combination of reasonable fertilizer costs with higher income from output sales. improved economic performance of any practice is an important factor for farmer adoption [25]. hence based on profitability, the stk method of fertilizer requirement assessment was expected to be most attractive to farmers but other factors as discussed below mitigated against its acceptance by farmers. 4.5. adoption of fertilizer recommendation information by farmers to gain widespread adoption, a fertilizer recommendation system must effectively tackle the dual challenges of optimizing recommendations for both yield and profit, while also ensuring acceptance among farmers for various crops and cropping patterns. the endorsement of such a system by policies is crucial for its success. in bangladesh, and in many other nations, the transfer of crop technologies primarily occurs through government-funded agricultural extension agencies. despite this established pathway, , a substantial percentage (60-85%) of bangladeshi farmers do not adhere to fertilizer recommendations provided by the fertilizer recommendation guide (frg) [2]. consequently, the current extension system appears to be falling short in effectively disseminating fertilizer management technology to farmers. furthermore, while stk method demonstrated superior performance compared to both frg and rcm, practical challenges hinder its widespread implementation. operating the soil test involves continual procurement of chemicals and demands skilled personnel for handling. unfortunately, establishing a profitable business model for making the stk widely accessible to smallholder farmers remains elusive. inconsistent results of the current rcm method discourage its promotion to farmers. as discussed above, further research to improve its calibration for n, p, k, s and zn fertilizer recommendations may improve its utility and adoptability. hence, we explored the frg as an alternative, effective and adoptable method of disseminating information. to address the issue, we developed frg cards taking information from the latest fertilizer recommendation guide [13] and tested them with farmers. across the ten project hubs, 33,237 (including 1,336 women farmers) farmers received frg cards followed by a short training session (about 20 mins each) for use in cropping on 21,160 ha of arable land. an evaluation was done to assess the effectiveness of the frg card in disseminating frg information to the farmers [26]. the key findings were as follows: (i) 100% of respondents were familiar with frg card, but less than 50% with stk and rcm, (ii) about 95% of the frg card holders followed the recommendation of bangladesh agricultural research council (barc) [13] in t. aman rice due to user-friendliness, and (iii) a small percentage of farmers used soil testing kit (stk). agriculture and food sciences research, 2025, 12(1): 54-68 66 © 2025 by the authors; licensee asian online journal publishing group hence, frg card when provided with training to the farmers was accessible, while applying the frg information in their crops increased aman rice grain yield by 84% and net profit increased by 143% over ffp. the next step is to scale-out the use of the frg card by larger numbers of farmers. this will require lower cost methods of distributing the frg card and training farmers in its use including mass media and social media. 5. conclusion the study indicates that the stk (soil testing kit developed by bangladesh agricultural university, mymensingh) had the best performance with regard to rice yield and gross margin. next to it was the fertilizer recommendation guide (frg) based on agroecological zone-specific recommendation. in contrast, the rice crop manager (rcm) did not consistently yield reliable results. a notable weakness observed across all three methods was the consistent underestimation of potassium (k) requirements, except for the stk method, which directly assessed the current soil k availability. this underestimation of k needs is a common issue in subtropical asia, where rice cropping systems deplete substantial amounts of k annually, leading to an increasing demand for k fertilizer over time. although performance of the stk method was better than frg, the lack of a profitable business model for operators has prevented its widespread use by farmers. previous barriers to supplying relevant frg information to farmers can be overcome by providing necessary packaged information on fertilizer requirements for each crop to farmers in the form of a frg card. the frg card is a simple and adoptable technology for improved fertilizer decision-making by the farmers. abbreviations the following abbreviations are used in this manuscript: % percentage aez agro-ecological zones bau bangladesh agricultural university cmol centimole dmrt duncan’s multiple range test fao food and agriculture organization of united nations ffp farmer fertilizer practice frg fertilizer recommendation guide gps geo-graphical positioning system ha hectarage k potassium kg kilogram mg milligram mha million hectarage mop muriate of potash n urea numan nutrient management for diversified cropping in bangladesh p phosphorus rcbd randomized complete block design rcm rice crop manager s sulphur (gypsum) ssnm site-specific nutrient management stk soil testing kit t tonne t/ha ton per hectarage tk. taka tsp triple superphosphate undp united nations development program us$ us dollar zn zinc sulphate fertilizer references [1] m. nasim et al., "distribution of crops and cropping patterns in bangladesh," bangladesh rice journal, vol. 21, no. 2, pp. 1-55, 2017. 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[14] a. l. page, r. h. miller, and d. r. keeney, methods of soil analysis, 2nd ed. madison, wi. usa: american society of agronomy, 1982. [15] fao/undp, land resources appraisal of bangladesh for agricultural development: agroecological regions of bangladesh. rome: food and agriculture organization of the united nations, 1988. [16] s. sharma et al., "field-specific nutrient management using rice crop manager decision support tool in odisha, india," field crops research, vol. 241, p. 107578, 2019. [17] k. a. gomez and a. a. gomez, statistical procedures for agricultural research. new york: john wiley & sons, 1984. [18] d. bates, m. mächler, b. m. bolker, and s. c. walker, "fitting linear mixed-effects models using lme4," journal of statistical software, vol. 67, no. 1, pp. 1-48, 2014. [19] m. islam et al., "conservation agriculture in intensive rice cropping reverses soil potassium depletion," nutrient cycling in agroecosystems, vol. 125, no. 3, pp. 437-451, 2023. [20] m. islam et al., "conservation agriculture improves yield and potassium balance in intensive rice systems," nutrient cycling in agroecosystems, vol. 128, no. 2, pp. 233-250, 2024. [21] p. csathó, t. arendas, n. fodor, and t. németh, "evaluation of different fertilizer recommendation systems on various soils and crops in hungary," communications in soil science and plant analysis, vol. 40, no. 11-12, pp. 1689-1711, 2009. [22] c. srinivasarao et al., "soil potassium fertility and management strategies in south asian agriculture," advances in agronomy, vol. 177, pp. 51-124, 2023. [23] g. panaullah et al., "nutrient uptake and apparent balances for rice-wheat sequences. iii. potassium," journal of plant nutrition, vol. 29, no. 1, pp. 173-187, 2006. [24] m. n. hasan, m. a. bari, and m. r. lutfar, soil fertility trends in bangladesh 2010 to 2020 (srsrf project). dhaka, bangladesh: soil resource development institute, ministry of agriculture, 2020. 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[26] m. w. rahman, evaluative research survey report. dhaka, bangladesh: numan project, 2022. table s1. estimation of rates of urea, tsp, mop, gypsum and zinc sulphate fertilizers based on four different methods and locations for use in t. aman rice cultivation in 2018 and 2019. location treatment fertilizer dose (kg/ha) urea tsp mop gypsum zinc sulphate 2018 2019 2018 2019 2018 2019 2018 2019 2018 2019 durgapur, rajshahi ffr 76 341 151 110 76 82 0 58 3.3 7.5 frg 197 158 50 35 71 81 76 34 2.7 6.5 stk 197 197 40 40 151 151 62 62 0.0 12.2 rcm 180 244 44 59 39 57 19 38 1.4 7.5 godagari, rajshahi ffr 221 250 72 81 72 75 83 50 4.0 1.2 frg 199 196 57 50 100 150 50 44 2.8 0.0 stk 199 195 15 15 138 74 75 74 12.2 12.1 rcm 257 263 56 66 60 59 42 41 8.4 5.6 sadar, mymensingh ffr 97 125 57 69 38 80 0 6 0.0 0.0 frg 198 198 51 40 71 101 64 22 2.8 4.5 stk 197 197 40 52 151 126 0 62 0.0 12.2 rcm 354 265 178 68 70 65 0 38 0.0 7.5 sadar, thakurgaon ffr 214 227 71 118 71 122 0 157 7.1 10.9 frg 186 198 48 36 57 101 60 34 2.6 6.5 stk 186 197 37 64 104 155 47 50 11.6 6.5 rcm 157 155 63 34 98 45 36 34 7.1 6.5 dacope, khulna ffr 132 175 89 84 85 84 47 0 0.0 0.0 frg 165 198 51 76 36 101 25 23 2.8 4.5 stk 198 197 40 70 78 100 25 50 2.8 12.3 rcm 163 163 43 54 43 54 43 54 0.0 4.5 amtali, barguna ffr 250 125 125 76 0 0 0 0 0.0 0.0 frg 165 158 51 35 37 80 25 13 2.8 5.0 stk 198 198 70 60 151 111 0 50 0.0 12.3 rcm 250 125 62 28 36 30 0 13 0.0 5.0 note: ffr=farmer’s fertilizer rate, frg=fertilizer recommendation guide, stk=soil testing kit, rcm=rice crop manager mop=muriate of potash, tsp = triple superphosphate. table s2. grain yield (t/ha) of t. aman rice in different locations and years as influenced by different fertilizer doses determined by various methods in 2018 and 2019. fertilizer tools durgapur, rajshahi godagari, rajshahi mymensingh, sadar thakurgaon sadar dacope, khulna amtali, barguna 2018 ffp 3.76b 4.95b 4.56 3.55 3.48b 3.47b frg 3.98ab 5.51a 4.68 3.6 3.57b 3.79ab stk 4.14a 5.75a 4.59 3.66 3.99a 3.67b rcm 3.95ab 5.29ab 4.38 3.81 4.19a 4.23a f-test ** ** ns ns ** ** 2019 ffp 3.50b 4.55b 4.86b 4.3 4.48b 3.99c frg 3.81ab 5.09ab 5.1b 4.61 4.39b 5.14a stk 4.19a 5.46a 5.45a 4.87 5.24a 4.52b rcm 3.68b 4.69b 5.56a 4.52 4.65b 5.00a f-test ** ** ** ns ** ** note: ** indicate p<0.01; ns = not significant. within a column, the mean values followed by the same letter are not significantly different at the 0.05 level of prob ability by dmrt. ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, stk=soil testing kit, rcm = rice crop manager. agriculture and food sciences research, 2025, 12(1): 54-68 68 © 2025 by the authors; licensee asian online journal publishing group table s3. variable costs (usd/ha) of t. aman rice in different locations and years as influenced by different fertilizer doses determined by various methods in 2018 and 2019 fertilizer tools durgapur, rajshahi godagari, rajshahi mymensingh, sadar thakurgaon sadar dacope, khulna amtali, barguna 2018 ffp 785 833 778 710 762 786 frg 791 815 815 709 749 763 stk 795 840 810 731 761 789 rcm 767 829 891 717 748 773 2019 ffp 1074 833 840 767 756 643 frg 1001 830 869 681 780 697 stk 1,38 851 917 704 821 743 rcm 1025 835 897 661 774 676 note: ns = not significant. within a column, the mean values followed by the same letter are not significantly different at the 0.05 level of probability by dmrt. ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, stk=soil testing kit, rcm = rice crop manager. table s4. gross return (usd/ha) of t. aman rice in different locations and years as influenced by different fertilizer doses determined by various methods in 2018 and 2019 fertilizer tools durgapur, rajshahi godagari, rajshahi mymensingh, sadar thakurgaon, sadar dacope, khulna amtali, barguna 2018 ffp 1.748 2.120 1.989 1.822 1.409 1.317 frg 1.841 2.374 2.015 1.807 1.437 1.410 stk 1.897 2.545 2.001 1.902 1.601 1.384 . 1,799 2.269 1.933 1.865 1.668 1.559 2019 ffp 1.285 1.014 1.363 1.133 1.171 838 frg 1.395 1.131 1.439 1.193 1.148 1.054 stk 1.521 1.213 1.543 1.273 1.356 942 rcm 1.337 1.054 1.537 1.169 1.209 1.028 note: ns = not significant. within a column, the mean values followed by the same letter are not significantly different at the 0.05 level of probability by dmrt. ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, stk=soil testing kit, rcm = rice crop manager. table s5. gross margin (usd/ha) of t. aman rice in different locations and years as influenced by different fertilizer doses determined by various methods in 2018 and 2019 fertilizer tools durgapur, rajshahi godagari, rajshahi mymensingh sadar thakurgaon sadar dacope, khulna amtali, barguna 2018 ffp 963 1287 1211 1112 647 531 frg 1050 1559 1200 1098 688 647 stk 1102 1705 1191 1171 840 595 rcm 1032 1440 1042 1148 920 786 2019 ffp 211 181 523 366 415 195 frg 394 301 570 512 368 357 stk 483 362 626 569 535 199 rcm 312 219 640 508 435 352 note: ns = not significant. within a column, the mean values followed by the same letter are not significantly different at the 0.05 level of probability by dmrt. ffp = farmer fertilizer practice, frg=fertilizer recommendation guide, stk=soil testing kit, rcm = rice crop manager. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. 87 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 87-95, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6169 © 2024 by the authors; licensee asian online journal publishing group impact of bank of agriculture credit’s on agricultural productivity in south western nigeria rasaki, wasiu adedamola1 omotoso, abeeb bababtunde2 ( corresponding author) 1department of agribusiness, oyo state college of agriculture and technology, igboora, nigeria. email: awrasaki@oyscatech.edu.ng 2department of agricultural technology, oyo state college of agriculture and technology, igboora, nigeria. email: omotosoabeebtunde@yahoo.com abstract this study examined the impact of bank of agriculture (boa) credit facilities on agricultural productivity in the south-west region of nigeria. a multi-stage sampling technique was employed to select 225 beneficiaries and 630 non-beneficiaries farmers. descriptive statistics were implemented to investigate the socio-economic attributes of the respondents. impact of credit was analyzed using propensity score matching (psm), and the productivity of producers was analyzed using total factor productivity (tfp). the respondents' average age was 47.6 years, and 87.0% of them were male. the average psm score was 0.269, with four matching methods (nearest neighbour, radius, kernel, and stratification matching) being implemented. the maximum and minimum propensity were 0.8207 and 0.0525, respectively. both the balancing property and the region of common support (0.0503, 0.8209) were satisfied. the average treatment effect on the productivity differences of the treated (att) was approximately 0.347, and this difference was statistically significant at the 1% level. productivity of att varied by approximately 21.0%. in summary, this investigation provided compelling evidence that the boa credits have a positive impact on agricultural productivity in the south west. it was suggested that administrations at all levels and financial stakeholders should collaborate to ensure that producers have access to credit. keywords: credit, productivity, impact, radius, kernel, stratification matching. citation | adedamola, r. w., & bababtunde, o. a. (2024). impact of bank of agriculture credit’s on agricultural productivity in south western nigeria. agriculture and food sciences research, 11(2), 87–95. 10.20448/aesr.v11i2.6169 history: received: 16 october 2024 revised: 18 november 2024 accepted: 22 november 2024 published: 29 november 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the tertiary education trust fund (grant number: oyscatech/rg/2021/08). institutional review board statement: the ethical committee of the oyo state college of agriculture and technology, igboora, nigeria has granted approval for this study on 12 march 2023 (ref. no. oyscatech/et/08). transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: developed the topic, and drafted the proposal, r.w.a.; developed the questionnaire, o.a.b. both authors have read and agreed to the published version of the manuscript. contents 1. introduction ...................................................................................................................................................................................... 88 2. literature review ............................................................................................................................................................................ 89 3. methodology ..................................................................................................................................................................................... 90 4. result and discussion ..................................................................................................................................................................... 92 5. conclusions and policy recommendations ................................................................................................................................ 95 references .............................................................................................................................................................................................. 95 mailto:awrasaki@oyscatech.edu.ng mailto:omotosoabeebtunde@yahoo.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6169 https://orcid.org/0009-0005-9203-3874 agriculture and food sciences research, 2024, 11(2): 87-95 88 © 2024by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study uniquely compares the productivity impacts between beneficiaries and nonbeneficiaries of bank of agriculture credit in south western nigeria, providing a dualperspective analysis. unlike prior research, it incorporates farmers’ socio-economic and contextual variables to reveal nuanced insights into credit utilization and barriers, guiding targeted policy and program improvements. 1. introduction agricultural sector has been characterized as the most significant sector of the economy, and it has the potential to significantly contribute to the nation's future economic development, as it has in the past [1]. the study also asserted that agriculture is a critical component of the socio-economic development of most developing economies and is a significant factor in national development. agriculture was a significant source of foreign exchange revenues and it supplied the burgeoning population with sustenance and employment, as well as basic materials for the expanding industries. according to ogen [2] agriculture was the most significant sector in the 1960s in terms of its contribution to the gdp and occupational distribution. additionally, the nigerian economy could be accurately described as an agricultural economy in the first decade following independence. agriculture is a substantial contributor to nigeria's gdp, with small-scale producers playing a dominant role in this contribution, as per rahji and fakayode [29]. nevertheless, her productivity and development are impeded by their restricted access to credit facilities [3]. according to the world bank report [4] relationship between agricultural financing and economic development has been the subject of extensive theoretical and empirical research in both developed and developing countries. in order to examine the impact of agriculture on the nigerian economy, rhaji [5] implemented the ordinary least squares method. he concluded that the absence of accessible, affordable, and sufficient credit is the cause of the systemic decline in the agricultural sector's contribution to the nigerian economy. however, agricultural credit is regarded as a strategic resource that can be used to elevate the living standards of our rural, impoverished agricultural community by promoting the production of crops and animals to new heights. consequently, it is essential for the growth of the economy. credits are critical factor in the enhancement of agricultural productivity. farmers can acquire the necessary apparatus and inputs for conducting agricultural operations as a result of the timely availability of credit [6]. the quickest method of boosting agrarian productivity is through the provision of inexpensive and accessible credit [7]. credit is granted for the acquisition of seed, fertilizer, cattle, and implements, as well as for the alleviation of distress. farmers must have the necessary funds to increase the utilization and diversity of their inputs. the producers' reserves are either negligible or nonexistent, necessitating that they borrow to finance their productive endeavours. the majority of farmers, particularly those who are minor, are unable to obtain essential commodities (such as fertilizer, enhanced seed, advanced technology, and plant protection) from their sources due to a dearth of funding. therefore, agricultural credit is a critical component in providing the necessary investment to stimulate production growth [8]. also, credit facilitated the adoption of yield-enhancing technologies while the demand for inputs was enhanced by the increased flow of credit to producers [9]. consequently, credit to farmers is usually recognized as a successful strategy for increasing agricultural output [10]. the process of improving agriculture and transforming the rural economy is said to be reliant on agricultural finance. according to mahmood, et al. [11] the fastest way to increase agricultural output is to provide affordable and easy loans. the agricultural sector, according to the thesis, is more dependent on credit than any other sector of the economy because of seasonal changes in farmer returns and the financial needs involved with the shift from subsistence to commercial farming. credit allows people to improve their level of life and earn more money [11]. as a result, there is growing concern over the producers' incapacity to access credit, despite the presence of credit rules. for example, in 2000, the nigerian agricultural cooperative and rural development bank (now the bank of agriculture) set aside n5 million for each local government area in the nation as an investment revolving fund. farmers who had access to finance faced much higher mean input expenditures per hectare, independent of their socioeconomic standing. the acquisition, administration, and repayment of credit are plagued by a multitude of issues, despite its importance in agricultural production. the nigerian agricultural cooperative and rural development bank (nacrdb), founded in 1973 and formerly known as the nigerian agricultural and cooperative bank, is a government-sponsored lending organization dedicated to meeting the credit needs of agriculture. the efficacy of nacrdb has been inadequately reviewed, due mostly to a high default rate among receivers [12]. the boa, which is entirely under the control of nigeria's federal government, is the country's most significant agency for supporting agricultural and rural development. the federal ministry of finance incorporated owns 60% of the structure, while the central bank of nigeria (cbn) owns 40%. the federal ministry of agriculture supervises the bank of agriculture limited. the nigerian agricultural bank (nab) was formed in 1973 and renamed the nigerian agricultural and cooperative bank (nacb) in 1978. the nigerian agricultural cooperative and rural development bank limited (nacrdb) was founded in 2000 as a result of a merger with the people's bank of nigeria (pbn) and the acquisition of the risk assets of the family economic advancement programme (feap). in 2010, a plan was adopted to turn the bank into a long-term and efficient national agricultural and rural development finance organization. this technique resulted in a future rebranding as boa limited [13]. it is a nationally owned development bank entrusted with providing low-cost loans to smallholder and commercial farmers, as well as small and medium-sized rural businesses. it also provides microfinancing to small and medium-sized non-agricultural businesses. the goal is to ensure the sustainable provision of agricultural and rural finance services, thereby supporting the national economic development agenda, which includes food security, poverty alleviation, job creation, reducing rural-to-urban migration, reducing reliance on imported food, and increasing foreign exchange earnings. 1.1. statement of the problem according to phillip, et al. [10] the provision of financing to farmers is generally regarded as a successful technique for augmenting agricultural productivity. mahmood, et al. [11] said that credit gives farmers the ability agriculture and food sciences research, 2024, 11(2): 87-95 89 © 2024by the authors; licensee asian online journal publishing group increasing the income and enhance level of life. agricultural financing is seen vital for enhancing agriculture and transforming the rural economy. the assertion is that the agriculture industry relies heavily on credit than any other input due to the fluctuations in farmers' earnings and the finance needed for the transition from subsistence to commercial farming. despite nigeria's huge fertile land, a large percentage of the population suffers from hunger, poverty, and unemployment as a result of agricultural neglect. several agro-industries depend largely on the importation of vital raw materials for manufacture, while a considerable proportion of nigerian youths remain unemployed. various laws have been implemented to address these issues, with banks designated to play a significant role in financing via the supply of loans. nonetheless, the truth persists that banks, namely commercial banks, have not adequately addressed the issue, as seen by the limited impact on agricultural loans [14]. conversely, in instances when loans are poised for approval, some farmers or customers lack the requisite collateral and integrity demanded by banks as a safeguard against potential losses or unexpected liabilities in the event of failure. consequently, several loans are diverted to non-agricultural projects due to the tendency of certain individuals to prioritize extravagant home expenditures, so undermining the intended aim of the credit. some reputable customers, who may be shielded from this allegation, are inevitably burdened by uncontrolled elements, like change in policies, and challenges in securing official permits [15]. numerous studies indicate that the provision of accessible and inexpensive financing is the most effective method for enhancing agricultural productivity [31]. the claim is that the agriculture industry relies more heavily on the finance necessary for transitioning from subsistence to commercial farming. in response nigeria government established financial institution (boa) saddled with responsibility of providing easy and cheaper credit to farmer. but the drawback is that many farmers have collateral, while some diverted the loan instead of using in it to boost the production. given the foregoing, the study investigated impacts of agricultural credit on agricultural productivity in southwest. specific objectives are to: 1. profile the socio-economic characteristics of the boa credit facilities beneficiaries and non-beneficiaries. 2. analyze the productivity differentials between non-beneficiaries and beneficiaries of boa credit facilities in south-western nigeria. 3. analyze the effect of boa credit on farm productivity of the beneficiaries in the south-western nigeria. 2. literature review 2.1. empirical framework the demand for commodities to enhance crop production had increased as a consequence of the credit transfer to producers, according to siddiqi, et al. [9]. omotoso and omotayo [16] and omotayo, et al. [17] found that the annual household income, per capita annual household income, rice income, rice output, and rice yield of producers are significantly and positively impacted by the utilization of credit in rice cultivation. furthermore, a study conducted by akinola and oladejo [18] highlighted that direct lending mechanisms offered by boa played a pivotal role in fostering innovation among agribusinesses within the region. this was evident through investments made by farmers towards modern farming techniques after securing funds from boa-based credit programs. ayegba and ikani [19] revealed that less has been done to boost agriculture by assisting farmers through sufficient credit. on an institutional level, omotoso and omotayo [20] noted that while there were positive impacts associated with accessing boa loans for agriculture purposes including improved infrastructure and technology adoption challenges related to loan accessibility processes hindered broader participation among small-scale farmers. 2.2. theoretical framework 2.2.1. theory of impact evaluation according to the world bank [21] impact assessment assesses the intended and, preferably, unforeseen consequences of a particular intervention. impact evaluation differs from outcome monitoring, which focusses on the achievement of objectives. it seeks to determine how participants' well-being changed in the absence of the intervention. this entails counterfactual analysis, which is defined as "a comparison between the actual events and the events that would have occurred in the absence of the intervention" [22]. impact evaluations aim to address cause-and-effect enquiries. they seek to identify outcome changes that can be directly linked to a program [23]. impact evaluation assists in addressing critical enquiries essential for evidence-based policymaking: what is effective, what is ineffective, the context, the reasons, and the associated costs. there has been a growing focus on this issue in policy-making across western and developing countries [24]. this component is crucial in the evaluation toolkit and essential for global initiatives to enhance aid delivery's effectiveness and public expenditure, ultimately contributing to improved living standards [25]. since it is impossible to see program participants' outcomes if they were not beneficiaries, an impact evaluation is fundamentally an issue of missing data. in the absence of counterfactual data, the most viable alternative is to assess the outcomes of treated individuals or households against those of a non-treated comparison group. in this process, a comparison group is selected that closely resembles the treated group, ensuring that individuals receiving treatment would have experienced outcomes akin to those in the comparison group had treatment not been administered. effective impact evaluations depend on identifying an appropriate comparison group [26]. researchers employ two primary approaches to replicate the counterfactual of a treated group: (a) establishing a comparator group via a statistical design, or (b) adjusting the program's targeting strategy to eliminate pre-existing differences between the treated and non-treated groups prior to outcome comparison. the equation below illustrates the fundamental evaluation problem by comparing outcomes y between treated and nontreated individuals i: 𝑌𝑖 = 𝛼𝑋𝑖 + 𝛽𝑇𝑖 + 𝜀𝑖 (1) where t is a dummy equal to 1 for those who participate and 0 for those who do not. x is a collection of additional noted traits of the person and maybe of his or her home and local surroundings. at last, ε is an error term indicating latent traits influencing y as well. the equation shows a method often employed in impact assessments—that of agriculture and food sciences research, 2024, 11(2): 87-95 90 © 2024by the authors; licensee asian online journal publishing group gauging the direct influence of the program “t” on results y. changes in pricing within program regions might potentially be of importance as indirect impacts of the program—that is, those unrelated to participation. the problem with estimating equation (𝑌𝑖 = 𝛼𝑋𝑖 + 𝛽𝑇𝑖 + 𝜀𝑖) is that because of these elements—a deliberate program placement and self-selection within the program—treatment assignment is not usually random. programs are therefore positioned in line with the needs of the communities and people, who then self-select depending on program design and location. one might base self-selection on observable traits, unseen elements, or both. regarding unobserved components, the estimating equation's error term will include variables likewise linked with the treatment dummy t. one cannot quantify and hence explain these unseen properties in the equation above, thereby generating unobserved selection bias. that is, cov (t, ε) ≠ 0 indicates the failure of one of the main assumptions of ordinary least squares in acquiring unbiased estimates: independence of regressors from the disturbance factor λ. naturally, the correlation between t and ε biassed the other estimates in the equation, including the estimate of the program impact β. one might also depict this issue in a more conceptual context. imagine one is assessing an antipoverty initiative meant to increase family earnings, a credit intervention. 𝐷 = 𝐸(𝑌𝑖(1) | 𝑇𝑖 = 1) – 𝐸(𝑌𝑖(0) | 𝑇𝑖 = 0) (2) where d= different in productivity of credit beneficiary and non-beneficiary yi stand in for household i's per capita income. ti = 1 represents for participants the value of yi during treatment as yi (1). ti = 0 for nonparticipators allows yi to be expressed as yi (0). the average impact of the program may be shown as follows if yi (0) is utilized across nonparticipating homes as a comparative result for participant outcomes yi (1) one may also assume, less strongly, conditional exogeneity of program placement. the validity of the effect estimations is determined by the exogeneity of program targeting across treated and non-treated regions, as well as the justified assumptions on the comparability of participant and comparison groups. still, one cannot evaluate the degree of bias "b" without any strategies or presumptions [26]. 3. methodology 3.1. study area the research was carried out in southwest nigeria. southwest nigeria consists of six states: ekiti, lagos, ogun, ondo, osun, and oyo. it is designated as the south west geopolitical or geographical region of nigeria. the area mostly speaks yoruba; nevertheless, several variants occur even within a single state. the area is located between longitudes 20° 31' e and 60° 0' e, and latitudes 60° 21' n and 80° 37' n. the area's population is 28,767,752, covering a total of 77,818 km² [26]. southwest nigeria is delineated to the north by kwara and kogi states, to the east by edo and delta states, to the south by the gulf of guinea, and to the west by the republic of benin. the climate of southwest nigeria is tropical, marked by distinct wet and dry seasons. the climatic conditions vary between nigeria's two distinct seasons: the rainy season (march november) and the dry season (november february). the dry season signifies the onset of harmattan dust, as frigid, arid winds from the northern deserts infiltrate the southern regions at this time. the temperature ranges from 21°c to 34°c, and annual precipitation varies between 1,500mm and 3,000mm [13]. agriculture is the primary occupation of the population in the area. they farm food crops such as rice, yam, cassava, maize, and cowpea, while cash crops include cocoa, oil palm, kola nut, plantain, banana, cashew, citrus, and timber. animal husbandry, including fisheries, poultry, pig farming, goat, sheep, and cow raising, is evident. figure 1 is the map of nigeria shows all the regions and states. green portion is the north west region, yellow part is north west, red part is north central, blue part south east, purple part is south south region and white part is the south west region where this research was carried out. figure 1. map of nigeria. 3.2. population of the study study populations were made up of farmers (those that benefited and those that did not benefit from boa credits) in the southwest, nigeria. agriculture and food sciences research, 2024, 11(2): 87-95 91 © 2024by the authors; licensee asian online journal publishing group 3.3. sampling techniques and sample size multi-stages sampling technique was adopted. in stage one, three states were randomly selected from six states in the south-west, nigeria (which are ogun, osun and oyo states). in stage two, all the three branches of bank of agriculture in each of the state were selected and list of applicants (successful and non-successful) were collected; make total number of nine (9) branches. branches of bank of agriculture limited in each of the three states are: ogun state (abeokuta, imeko and abigi branches), osun state (osogbo, ode-omu and ile-ife branches) oyo state (igboora, ibadan and iseyin branches). stage involved systematic selected of 25 beneficiaries (successful applicants) from each of the lists that was collected from each of the nine (9) branches; make a total of 225 beneficiaries. in stage four, 70 nonbeneficiaries (non-successful applicants) were also systematically selected from the list provided by the bank; make a total of 630 non-beneficiaries. both beneficiary and non-beneficiaries were 855 respondents. 3.4. type of data and instrument used for data collection this study used primary data acquired via a well-structured questionnaire. the questionnaire was framed in such a way to attain the objectives of the research. the questionnaire contained some useful information such as access to credit, constraint faced, inputs used, output and revenue. 3.5. methods of data analysis data were analyzed using descriptive statistics, total factor productivity and propensity score matching. 3.6. model specification 3.6.1. total factor productivity (tfp) 𝑇𝐹𝑃 = 𝑙𝑜𝑔𝑌 − 𝛽1𝑙𝑜𝑔𝐾– 𝛽2𝑙𝑜𝑔𝐿 (3) where y = output. k= capital. and l= labour costs. other necessary factors like land, fertilizer, feed cost etc. 𝐸𝑥𝑝𝑙𝑖𝑐𝑖𝑡𝑙𝑦: 𝑇𝐹𝑃 = 𝑙𝑜𝑔𝑌 − 𝛽1𝑙𝑜𝑔𝑋1 − 𝛽2𝑙𝑜𝑔𝑋2 − 𝛽3𝑙𝑜𝑔𝑋3 − 𝛽4𝑙𝑜𝑔𝑋4 − 𝛽5𝑙𝑜𝑔𝑋5(8) where tfp = total factor productivity logy = natural log of output logx1-logx5 = natural logs of inputs like; land, labour, capital, chemical, feed etc β1-β5 = coefficient of parameters 3.6.2. propensity score matching (psm) since the 1990s, propensity score matching has been widely used in economics to assess the effects of a program or intervention on the economy. ravallion [27] reiterated by mapila, et al. [28] posits that propensity score matching involves the identification of two groups: those who participated in the intervention (access to credit in this study), represented by ht=1 for beneficiary farmers, and those who did not participate, represented by ht=0 for nonbeneficiary farmers. farmers receiving intervention benefits are paired with non-beneficiary farmers based on the likelihood that the latter would have benefitted from the intervention, referred to as the propensity score. propensity score matching was used to analyze impact of agricultural credit on farmers’ productivity. it was used to compare the outcome of those benefited from agricultural credit and those who did not. 𝑌𝑖 = 𝑋1 + 𝑋2 + 𝑋3 + − − −𝑋𝑛 + є (4) let y1 = beneficiary of agricultural credit. y0 = non-beneficiary. if a farmer is a recipient of agricultural credit, z, an indicator variable, equals 1 (z=1); if not, z=0. this indicates if the farmer really got treatment (credit). x represents a vector of control variables. data on program recipients indicate the average result among the treated e(y1|x, z=1). thus, att of the scheme can be estimated as: 𝐴𝑇𝑇 = 𝐸(𝑌1 − 𝑌0|, 𝑍 = 1) (5) = 𝐸(𝑌1|, 𝑍 = 1) − 𝐸𝑝|𝑧 = 1 {𝐸𝑌(𝑌0|𝑍 = 1, 𝑃)} (6) = 𝐸(𝑌1|, 𝑍 = 1) − 𝐸𝑝|𝑧 = 1 {𝐸𝑌(𝑌0|𝑍 = 0, 𝑃)} (7) where the first term of the above expression can be estimated from the treatment group and the second term from the mean outcome of the matched (on p) comparison groups. table 1, shows the summary of the objectives of the study and analytical technique(s) used for each of the objective. table 1. summary of objectives and analytical techniques. s/n objectives description analytical technique 1 describe the socio-economic characteristics of the boa credit beneficiaries and non-beneficiaries. nigeria. age, gender, marital status. descriptive statistics such as frequency, percentages and means. 2 analyze the productivity differentials between the beneficiaries and nonbeneficiaries of boa credits in southwest nigeria. the diverse ways in which farmer improve their productivity. the factors to be considered are production factors: inputs and output. total factor productivity (tfp) and t-test. 3 analyze the effect of boa credit on farm productivity of the beneficiaries in the south-western nigeria. different ways in which farmers make a living and build their worlds, e.g. improve productivity, increased income, improved welfare etc. propensity score matching (psm). agriculture and food sciences research, 2024, 11(2): 87-95 92 © 2024by the authors; licensee asian online journal publishing group 4. result and discussion sex: table 2 results showed that majority (87.1 percent) of respondents were male while only 12.9 percent were female. it also indicated that 91.1 percent of credit beneficiaries were male while female beneficiaries were just 8.9 percent, 85.7 percent of non-beneficiaries were male while 14.3 percent were female. this shows that male dominated agricultural production in the study area and that male benefited more from bank of agriculture credit than female. age: it was revealed that 8 percent of beneficiaries have age below 30 years while 10.2 percent of nonbeneficiaries belong to this age bracket and the pooled response was 9.6 percent. only 8 percent and 9.7 percent of beneficiaries and non-beneficiaries were above 60 years respectively. the mean age were 48.5 years, 47.6 years and 47.8 years respectively for the beneficiaries, non-beneficiaries and pooled data. the average age of beneficiaries is slightly higher than that of non-beneficiaries. this is in tandem with findings of nosiru [29] in his work where the average age for beneficiaries was 45.48 years while that of non-beneficiaries was 43.54years. marital status: it was revealed that about 86 percent of beneficiaries and 85 percent of non-beneficiaries were married. percentage of singles beneficiaries and non-beneficiaries were 4.9 percent and 9.7 percent respectively while 9.3 percent of beneficiaries and 5.8 percent of non-beneficiaries were either divorced, widowed or separated. this implies that married respondents had access to credit more than others. this conform with olagunju [30] in her studywhere 78.30 percent of the respondents were married/engaged. education qualification: results in table 2 showed that 14.2 percent of beneficiaries had no formal education. the percentage of beneficiaries and non-beneficiaries that had education up to tertiary level were 15.6 and 6.7 percent respectively. this implies that majority of beneficiaries (85.8 percent) and non-beneficiaries (89.7 percent) had formal education. this result contradicted the finding of tilahun [31] in his study where majority (53.6 percent) of credit unconstrained and (56.2 percent) credit constrained had no formal education. farm size: revealed that majority of beneficiaries (72.0%) and non-beneficiaries (81.4%) had less or 5 hectares of land. the average farm size owned by beneficiaries was 5.9 hectares while that of non-beneficiaries was 3.7 hectares. this implies that on average beneficiaries had more land area than non-beneficiaries. this result contradicted that of kiplimo, et al. [32] where mean farm size of farmers that accessed credit was 2.6 hectares and those that did not access credit was 2.9 hectares. contact with extension agents: it was further revealed that 59.1 percent of beneficiaries and 30.2 percent of non-beneficiaries had contact with extension agents. beneficiaries and non-beneficiaries that did not have contact with extension agents were 40.9 percent and 69.8 percent respectively. this indicates that beneficiaries engaged with extension agents more frequently than non-beneficiaries. results contradicted the findings of kiplimo, et al. [32] which indicated only 21.5 percent of beneficiaries had contact with extension agents. type of labour: the table also showed that 16.9 percent of beneficiaries and 45.6 percent of non-beneficiaries used family labour. beneficiaries and non-beneficiaries that used hired labour were 48.9 and 21.2 percent respectively. percentage of beneficiaries and non-beneficiaries that used both family and hired labour were 34.2 percent and 33.2 percent respectively. this implies that beneficiaries used more of hired labours than family labours. this result negated the findings of takane [33] in his study “labour use in smallholder agriculture in malawi” where family labour accounted for 88% of total labour used. table 2. socioeconomic distribution of respondents. socio-economic characteristics beneficiaries frequency % (n=225) non-beneficiaries frequency % (n=630) pooled frequency % (n=855) sex male 205 91.1 540 85.7 745 87.1 female 20 8.9 90 14.3 110 12.9 age (years) ≤ 30 18 8.0 64 10.2 82 9.6 31-40 36 16.0 103 16.4 139 16.3 41-50 83 36.9 223 35.3 306 35.8 51-60 70 31.1 179 28.4 249 29.1 above 60 18 8.0 61 9.7 79 9.2 mean 48.5 47.6 47.8 marital status married 193 85.9 533 84.6 726 84.9 single 11 4.9 61 9.7 72 8.4 divorced 11 4.9 7 1.1 18 2.1 widow/widower 5 2.2 23 3.7 28 3.3 separated 5 2.2 6 1.0 11 1.3 education qualification non-formal 32 14.2 65 10.3 97 11.4 primary 85 37.8 289 45.9 374 43.7 secondary 73 32.4 234 37.1 307 35.9 tertiary 35 15.6 42 6.7 77 9.0 farm size (ha) ≤ 5 162 72.0 513 81.4 675 78.9 6-10 34 15.1 89 14.1 123 14.4 above 10 29 12.9 28 4.5 57 6.7 mean 5.9 3.7 4.3 source of land inheritance 109 48.5 386 61.3 495 57.9 gift 41 18.2 107 18.0 148 17.3 rent/lease 30 13.3 70 11.1 100 11.7 purchase 45 20.0 67 10.6 112 13.1 contact with extension agents agriculture and food sciences research, 2024, 11(2): 87-95 93 © 2024by the authors; licensee asian online journal publishing group socio-economic characteristics beneficiaries frequency % (n=225) non-beneficiaries frequency % (n=630) pooled frequency % (n=855) yes 133 59.1 190 30.2 323 37.8 no 92 40.9 440 69.8 532 62.2 type of labour family labour 38 16.9 287 45.6 325 38.0 hired labour 110 48.9 134 21.2 244 28.5 family & hired labour 77 34.2 209 33.2 286 33.5 4.1. productivity differentials among the respondents table 3 showed the mean productivity of beneficiaries compared to non-beneficiaries. mean productivity of beneficiaries and non-beneficiaries was 1.64 and 1.29, respectively. the mean difference in productivity was 0.35, which was significant at the 1%. this indicates a significant difference in productivity between beneficiaries and nonbeneficiaries. table 3. t-test analysis (productivity by credit). categories obs. mean std. err. std. dev. [95% conf. interval] non-beneficiaries 630 1.29 0.007 0.17 1.28 1.31 beneficiaries 225 1.64 0.031 0.46 1.58 1.70 combined 855 1.38 0.011 0.32 1.36 1.40 difference -0.35 0.022 -0.39 -0.30 diff = mean (0) – mean (1) t = -16.1 4.2. descriptive statistics of propensity score and region of common support results in table 4 showed that the total was 833 farmers and average propensity score was 0.269. maximum and minimum propensity was 0.8207 and 0.0525 respectively. this implies that the average probability to access credit from boa by respondents was about 27 percent. the region of common support (0.0503, 0.8209), as presented in table 4, indicates the point at which treatment (beneficiaries) and control (non-beneficiaries) exhibit similar propensity scores, thereby satisfying the balancing property. minimum propensity score is 0.0503, while maximum propensity score is 0.8207. table 4. descriptive statistics of propensity score and region of common support. intervention obs. mean std. dev. min. max. credit (propensity score) 833 0.27 0.17 0.05 0.82 credit (common support) 833 0.05 0.82 4.3. distribution of treated and controls across blocks result in table 5 showed that optimum number of blocks was 6 which was determined by the programme. beneficiaries and non-beneficiaries that were matched at lowest p-score (0.05) were 7 and 133 respectively while only 1 farmer each for beneficiaries and non-beneficiaries were matched at highest p-score. this implies that very few respondents were matched at level of high propensity score. table 5. distribution of treated and controls across blocks. access to boa credit block of p score non-beneficiaries beneficiaries total 0.05 133 7 140 0.10 167 38 205 0.20 224 84 308 0.40 74 66 140 0.60 9 29 38 0.80 1 1 2 total 608 225 833 4.4. predictive performance of probit the summary of predictive performance of the boa credit was presented in table 6. it showed a very high performance (92.70 percent) in non-treatment case and about 26 percent performance in the treatment case. the overall performance was about 75 percent. this implies the boa credit performed well in agricultural productivity in study area. table 6. predictive performance of probit (in %). performance indicator credit overall correct classification rate 75.09 correct non-treatment classification rate 92.70 correct treatment classification rate 25.78 4.5. average treatment effect on the treated (att) productivity differences result table 7 presents att regarding productivity differences. att was determined by calculating the difference in contract productivities between the treated group (beneficiaries) and the control group (non-beneficiaries) with comparable propensity scores. the methods employed for att productivity comparisons included nearest neighbour, radius, kernel, and stratification matching. table 7 presents the total number of treated contracts, the corresponding number of control contracts utilized for matching, att productivity differences, the att productivity differences expressed as a percentage of average productivities, and the t-statistics for the productivity comparisons. radius, kernel, and stratification matching utilize all control contracts, whereas nearest neighbour agriculture and food sciences research, 2024, 11(2): 87-95 94 © 2024by the authors; licensee asian online journal publishing group matching employs only a subset of these contracts (151 out of 608) that exhibit the closest propensity scores to the treated contracts. each treated contract was paired with one or more control contracts. nearest neighbour matching involved pairing each treated contract with one or more control contracts that exhibited the closest propensity score. all 225 beneficiary farmers were matched with 151 non-beneficiary farmers. att for productivity differences is 0.347, which is significant at the 1%. the att percent productivity difference was 25.04%. this result implies that boa credit increases the agricultural productivity by 25.0 percent in the study area. radius matching matched each treated contract with one or more control contracts that has the similar radius. it matched all 225 beneficiaries with 608 non-beneficiaries farmers. it used more control than nearest neighbour. att productivity for radius matching was 0.351 which was significant at 1% level of significance and att percent productivity difference was 25.33%. this result implies that boa credit increases the agricultural productivity by 25.33 percent in the study area. kernel matching paired each treated contract with a weighted average of all control contracts, assigning weights that are inversely proportionate to the distance between the propensity scores of the treated and control contracts. it correlated all 225 beneficiaries with 608 non-beneficiary farms. it used the same quantity of controls as radius matching. the average treatment effect (att) productivity for kernel matching was 0.358, significant at the 1% level, with an att percent productivity difference of 25.84%. this result implies that boa credit increases the agricultural productivity by 25.84 percent in the study area. stratification matching, matched each treated contract with one or more control contracts that have the similar strata. it matched all 225 treatments with 608 controls farmers. att productivity for stratification matching was 0.360 which is significant at 1% and att percent productivity difference was 25.98%. this result implies that boa credit increases the agricultural productivity by 25.84 percent in southwest. table 7. att productivities differences. matching method number of treated number of control att productivity differences att percent productivity differences standard error t statistic nearest neighbour 1.641(225) 1.294(151) 0.347 21.04% 0.036 9.66*** radius 1.641(225) 1.290(608) 0.351 21.39% 0.031 11.18*** kernel 1.641(225) 1.283(608) 0.358 25.84% stratification 1.641(225) 1.281(608) 0.360 25.98% note: values in parentheses are numbers of observations. *** represents 1% level of significance. 4.6. bootstrapped standard errors table 8 presented the result of bootstrapped standard error. the essence of bootstrapping was to adjust the standard errors and therefore, correct the bias in the estimate. the result showed very little or no difference in the standard errors of the estimate and that of bootstrapped standard errors. this implies that there were little or no bias in the estimate of the four matching methods used. the t-statistics for all the four matching methods used were significant at 1% level of significance. table 8 presents compelling evidence that the boa credit positively influences agricultural output in the research region. table 8. bootstrapped standard errors of the matching methods. matching method number of treated number of control att productivity differences standard error t-statistic nearest neighbour 1.641(225) 1.294(151) 0.347 0.038 9.15*** radius 1.641(225) 1.290(608) 0.351 0.034 10.24*** kernel 1.641(225) 1.283(608) 0.358 0.027 13.35*** stratification 1.641(225) 1.281(608) 0.360 0.034 10.56*** note: values in parentheses are numbers of observations. *** represents 1%. 4.7. result of the t-test table 9 presented the results of the hypotheses. results revealed that, there were significant differences between boa credit beneficiary farmers and non-beneficiary farmers with respect to sex, years of schooling, household size, main occupation, contact with extension agents, membership of organizations and farm size. household size was significant at 10%. sex, years of schooling were significant at 5% while main occupation, contact with extension agent, membership of organizations and farm size were significant at 1%; hence, null hypotheses were rejected. there were no significant difference between the beneficiaries and non-beneficiaries with respect to age, marital status and farming years of experience; hence, null hypotheses were accepted. table 9. t-test estimates of the variables (difference between means). variables mean diff std. error t-value p-value decision treated control age 48.480 47.630 0.847 0.858 0.99 0.324 accept ho sex 0.910 0.850 0.054 0.026 2.08 0.038** reject ho married 0.857 0.846 0.011 0.028 0.42 0.673 accept ho years of schooling 7.836 7.108 0.728 0.360 2.02 0.044** reject ho household size 7.862 7.405 0.457 0.259 1.76 0.078* reject ho main occupation 0.738 0.529 0.209 0.038 5.56 0.000*** reject ho farming experience 18.711 18.051 0.660 0.899 0.73 0.463 accept ho extension contact 0.591 0.301 0.290 0.036 7.96 0.000*** reject ho member of organization 0.449 0.230 0.288 0.034 6.36 0.000*** reject ho farm size 5.861 3.700 2.161 0.433 4.99 0.000*** reject ho note: ***, ** and * represent 1%, 5% and 10%. agriculture and food sciences research, 2024, 11(2): 87-95 95 © 2024by the authors; licensee asian online journal publishing group 5. conclusions and policy recommendations subsequent conclusions were derived from the results of this investigation: the study area was dominated by masculine agricultural production, and males derived greater benefits from bank of agriculture credit than females. both beneficiaries and non-beneficiaries were married and in their prime years. it was also determined that there was a substantial disparity in productivity between beneficiaries and non-beneficiaries, and that these disparities were genuine. the boa credit performed exceptionally well in the study area. from the nearest neighbour, radius, kernel, and stratification matching results, it was determined that agricultural productivity in southwest increases approximately by 25% as a consequence of boa credit. the policy implications of the findings were to enhance and maintain access to agricultural credit, as it was determined that agricultural productivity was positively influenced by agriculture. consequently, educational stakeholders and governments at all levels (federal, state, and local governments) should collaborate to enhance farmers' access to formal education. farmers should not be apprehensive about utilizing credit in their farming operations, as research has demonstrated that credits has beneficial impacts on agricultural productivity. references [1] d. a. okolo, "regional study on agricultural support, nigeria’s case," evolution, vol. 3, no. 3, pp. 1-97, 2004. 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[33] t. takane, "labour use in smallholder agriculture in malawi: six village case studies," african study monographs, vol. 29, no. 4, pp. 183-200, 2008. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.jclepro.2023.137487 https://doi.org/10.1007/s10668-024-04681-8 https://doi.org/10.1007/s10668-024-04681-8 https://doi.org/10.3390/agriculture12030363 https://doi.org/10.1007/s10584-024-03775-6 14 © 2023 by the author; licensee asian online journal publishing group agriculture and food sciences research vol. 10, no. 2, 14-27, 2023 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v10i2.5195 © 2023 by the author; licensee asian online journal publishing group review on deep fat-fried food products safety and its implication for consumer healthiness yadesa abeshu holeta agricultural research center, eiar, ethiopia. email: abeshuy@gmail.com abstract edible oil is the most widely consumed food in worldwide. however, high-temperature cooking of fat results harmful compounds. this extended cooking times caused chemical and physical reactions at high temperatures. additionally, heat-labile chemical compositions such as phenolic compounds, fatty acids, and essential amino acids gradually changed. high cooking temperatures have a real impact on the flavor quality and stability of the oil. but, applying antioxidants and inhibitors slows down the oxidation of frying oil at lower temperatures. the primary goal of this review process was to create awareness among the public and regulatory bodies about the extent to which oil based food processed at high temperatures for long time degrades food quality and endangers consumers. to address the problem of deep fat frying, the various earlier research works were well reviewed. the results of this study were thoroughly examined and concluded that cooking fat for a long period of time reduced its quality and increased the risk of non-communicable diseases. thus, to lower the amount of free radicals in oils, frying them for a short period of time and using antioxidants. also, it is critical to implement food quality control policies and educate community to safeguard the health of consumers. keywords: chemical reactions, chronic disease, deep frying, degradation, edible oil, food, inhibitors, physical reactions. citation | abeshu, y. (2023). review on deep fat-fried food products safety and its implication for consumer healthiness. agriculture and food sciences research, 10(2), 14–27. 10.20448/aesr.v10i2.5195 history: received: 6 october 2023 revised: 17 november 2023 accepted: 24 november 2023 published: 4 december 2023 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the author confirms that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the author declares that there are no conflicts of interests regarding the publication of this paper. contents 1. introduction ...................................................................................................................................................................................... 15 2. changes throughout deep-fat cookery .................................................................................................................................... 15 3. causes of repeated fat cooking .................................................................................................................................................. 18 4. deeply fried oil and foods quality ............................................................................................................................................ 19 5. high temperature oil cooking factors ................................................................................................................... 20 6. foods nutrient changes during deep-fat frying .................................................................................................................... 21 7. deep-fat frying consumption unhealthiness .......................................................................................................................... 23 8. ways to solve the harmful effects of deep fat cooking ....................................................................................................... 24 9. conclusion ......................................................................................................................................................................................... 25 10. conflict of interest declaration ................................................................................................................................................. 25 references .............................................................................................................................................................................................. 25 mailto:abeshuy@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v10i2.5195 https://orcid.org/0000-0002-4648-2370 agriculture and food sciences research, 2023, 10(2): 14-27 15 © 2023 by the author; licensee asian online journal publishing group contribution of this paper to the literature the review paper advances scientific awareness and encourages further research to address the issue of the negative effects of deep fat cooking on food quality and consumer health. once more, it offers information on food quality control regulations and how consumers can avoid consuming excessive amounts of fat-fried food. 1. introduction commercial deep-fat frying has been estimated to be worth £45 billion in the united states and at least twice this amount for the rest of the world [1]. although oilseeds make up the bulk of ethiopia's crop production, the country relies primarily on foreign products to meet its growing demand for edible oil. according to the merchandise wholesale import and trade enterprise [2], the annual consumption of edible oil in ethiopia is approximately 530,000 tons per year. cooking can be defined as the process of submerging food in hot oil to a temperature of 150 to 190 degrees celsius while maintaining contact between the food, oil, and air. cooking oil enhances the texture and flavor of food by serving as a medium for heat transmission. food surface area, cooking duration, food wetness, breading or bashing techniques, and frying oil all affect how much oil is absorbed by food. most of the time during cooking, the absorbed oil tends to collect on the surface of cooked food and seeps inside while chilling [2]. foods that are fried below at a lower temperature or for a shorter amount of time than is ideal, but, have a white or light brown color. foods that are under-fried lack interesting deep-fat cooked flavors, vibrant colors, and crisp textures. foods that are overfried at temperatures higher and cooking times longer than ideal have surfaces that are hardened and blackened, and their texture is greasy due to the excessive absorption of oil [3]. frying produces desirable or undesirable flavors, and changes flavor stability and quality, color, texture, unsaturated fatty acid, foaming, color, viscosity, density, heat, free fatty acid content, volume, polar materials, and composite materials increase. in particular, fatty foods such as potato chips, cookies, and other street food fast foods are made from a variety of plant and animal ingredients and are highly exposed to frying. cooking temperature and time, frying oil, inhibitor, and product type influence oil hydrolysis, oxidation, and polymerization during frying [4]. long-term consumption of processed foods, frying oil, cooked oil, and unconsumed oil can significantly impair a person's antioxidant defense network and cause diseases such as hypertension, diabetes, obesity, and fallopian tube inflammation [5]. the purpose of the research is to review the effects of edible frying on oil quality and health issues and to provide integrated data on oil/fat frying changes to customers and food processors. as a result of raising awareness through a review of this seminar, it became clear that the intensive and repeated frying of fats and oils has an impact on health and deteriorates the quality of products. in ethiopia, food processing is local and industrial, causing health problems as there is no required method of control or safety awareness to deal with the problem. 2. changes throughout deep-fat cookery 2.1. changes of physical characteristics frying is a process of cooking and drying in hot oil which is the process of heat and mass transfer. when heat is transferred from the oil to the food, the moisture in the food turns into gas, and the oil is absorbed by the food. in addition to the thermal and physical properties of food and oil, food shape and size, and oil temperature, several factors affect heat and mass transfer. table 1 exhibits a series of changes in several physical parameters of oil/fat during frying and the reasons for these changes [6]. table 1. changes in some physical parameters of oils/fat during deep-frying. physical parameter changes during frying deep caused by uv increases conjugated fatty acids stored density increases polymerized triacylglycerol insulator constant decreases polar-oxidized parts color millard reaction no conduction increases polar compounds physical action surface tension decreases polar compounds smolder point decreases volatile oxidized breakdown cpds. heat increases polar compounds viscosity increases polymerized triacylglycerol source: choe and min [6]. according to table 1, deep-frying either increased or decreased the physical characteristics of oil such as density, color, conduction, surface tension, heat, and viscosity. because certain chemicals changed into new forms and some volatilized. the oil's or fat's quality is altered to have an unwanted flavor or odor. to maintain food quality and the health of consumers, it is crucial to cook oils or fat at the ideal temperature and time. 2.2. chemical changes characteristics cooking oils not only transfer heat to cooked foods, but they can also add unwanted off-flavors to fried foods due to damaged oil [7]. during frying, many harmful chemical processes (hydrolysis, oxidation, polymerization, etc.) occur, causing the oil to break down and form volatile products and non-volatile monomers and compounds. in addition, these chemical reactions cross linked by hydrolysis, oxidation, and polymerization, free fatty acids, low molecular weight alcohols, aldehydes, ketones, acids, lactones and hydrocarbons, diglycerides and monoglycerides, cyclic and epoxy compounds, rumored trans isomers, triacylglycerol monomers and dimers, oligomers, etc. [8]. likely table 2 exhibits some chemical changes occurred during deep fat frying at high temperature for long time and shows the parameters prone to be changed due causes of oxidation and polymerization. agriculture and food sciences research, 2023, 10(2): 14-27 16 © 2023 by the author; licensee asian online journal publishing group table 2. changes in some chemical parameters because of fat frying. chemical parameter changes during deepfrying caused by anisidine value rises secondary oxidation iodine value declines formation of oxidized fat products peroxide value rises /declines primary oxidation products petrol ether rises oxidized polymerization products polar compounds rises oxidized and polymerized degradation of products polymerized triacylglycerol rises oxidized and polymerized triacylglycerol acid value rises formation of products oxidation using free carboxyl groups table 2 lists the chemical parameters that changed when oil or fat was repeatedly and repeatedly deep-fried at a high temperature. oil deep-frying causes a rise in parameters such as the anisidine value, peroxide value, petrol ether, polar compounds, and polymerized triacylglycerol, while a fall in iodine value is caused by the reaction of free fatty acids with oxygen. however, depending on the type of fat or oil, peroxide value can occasionally rise or fall. these are brought on by the oxidation of oil and the polymerization of compounds when cooking or frying in an environment where air is present. 2.2.1. oil hydrolysis cooking food in hot oil immediately generates steam, which evaporates in the form of bubbles and gradually reduces as the food is fried. water, steam, and elements cause chemical reactions in cooking oils and foods. water, a weak nucleophile, attacks the organic compound bonds of triacylglycerols, producing diand monoacylglycerols, glycerol, and free fatty acids. the content of free fatty acids in frying oil increases with the amount of food being fried [9]. thermochemical reactions mainly occur in the oil region and not at the water-oil interface. chemical reactions are more pronounced in oils containing short chains and unsaturated fatty acids than in oils containing long chains and saturated fatty acids. this is because short chain and unsaturated fatty acids are more soluble in water than long chain and saturated fatty acids. water from food does access short-chain fats and oils and cause chemical reactions. large amounts of water transform oil rapidly [10]. water hydrolyzes oil faster than steam. heavy contact between the oil and food components increases the hydrolysis of the oil. monoand diacylglycerols are initially expanded in vegetable oil when cooking potato chips between 155°c and 195°c. frequently replacing the cooking oil with modern oil will slow down the chemical reaction of the frying oil. the hydroxides and alkaline substitutes used to wash poultry promote oil hydrolysis. frying time did not affect oil hydrolysis [4]. free fatty acids and their compounds produce off-flavors, making the oil unsuitable for frying. diacylglycerols and monoacylglycerols, glycerol and free fatty acids promote strong hydrolysis reactions of oils [11]. glycerol evaporates at 150 °c, and the glycerin left in the oil promotes the formation of free carboxylic acids through chemical reactions [4]. the free fatty acid content of edible oil is 0.05% to 0.08%. the basic reactions that occur during oil hydrolysis are shown in figure 1. the content of free fatty acids in frying oil increases with the amount of food being fried (figure 1). free fatty acid values are used to determine the quality of frying oil. so, the figure 1 illustrates how the triglycerides are decomposed to glycerol and free fatty acids as the oil cooked at high temperature in steam. then the oils hydrolyzed to free fatty acids which are prone to oxidation and become piousness to human. figure 1. chemical reactions in oil produce free fatty acids. 2.2.2. oil oxidization when oil fried, elements react with oil [12]. the chemical process of thermal oxidation is essentially the same as the mechanism of autoxidation. although thermal oxidation rates are faster than autoxidation, specific and detailed scientific data and comparisons of oxidation rates between thermal and autoxidation do not seem to be available. as shown in figure 1, the mechanism of thermal oxidation includes initiation, progression, and termination of the reaction. oil in its best non-radical state does not react with di-radical elements in its triplet state thanks to the spin barrier. normal oxygen in the air is a radical compound. radical oil is required for the oxidation of radical elements. the oil must be in a highly radical state so that it can react with radical oxygen for oil oxidation reactions. the chemical elements that have the weakest bond with the oil's carbon are removed and become radicals. according to rumors [12], the energy required to break the carbon-hydrogen bond at the 11th carbon atom of linoleic acid is 50 kcal/mol. as shown in figure 2, the double bond between carbon 9 and carbon 12 reduces the carbon-hydrogen bond of carbon 11 by withdrawing electrons. the carbon-hydrogen bond at carbon 8 or 11 is α to the covalent bond in oleic acid and is 75 kcal/mol. the carbon-hydrogen bond on a saturated carbon without double bonds is approximately 100 kcal/mol. differences in the strength of the bonds between chemical elements and carbon in fatty acids represent differences in the oxidation rates of stearic, oleic, linoleic, and linoleic acids during thermal or autoxidation. the weakest carbon-hydrogen bond in linoleic acid is that carbon-11 is removed first, thus removing the hydrogen on carbon-11 and creating a radical on carbon-11. agriculture and food sciences research, 2023, 10(2): 14-27 17 © 2023 by the author; licensee asian online journal publishing group figure 2. the initiation, propagation and termination of oil thermal oxidation. a chain reaction between free alkyl radicals and peroxyl radicals promotes thermal oxidation of oil. the oxygenoxygen bond strength of r-o-o-h is approximately 44 kcal/mol. this is a relatively weak chemical bond. hydroperoxides usually seem to become unstable during the frying process. hydroperoxides are degraded into alkoxy radicals and group radicals by peroxide-bound hemolysis [13]. the hydro peroxide is decomposed to supply oxyand hydroxy radicals. known mono-oxygenated product love 9-keto-10, 12and 13keto-9-, 11-octadecadienoate and hydroxyl group derivatives such as 9hydroxy-10, 12and 13-hydroxy-9and 11-octadecadienoate from hydro peroxides of thermally change alkyl group linoleate at a 150◦c. the alkoxy radical reacts with other alkoxy radicals or is decomposed to form non radical products. the formation of non-radical volatile and non-volatilizable compounds at the tip of oxidization is named the termination step. most volatiles are aloof from cookery oil by steam throughout deep fat frying [13]. as stated, adding water to the frying system reduces volatile compounds in the oil [14]. the number of volatile compounds found in oils varies widely and depends on the type of oil, food, and frying conditions. volatile compounds greatly affect the taste quality of frying oils and cooked foods. the rate of aerobic decomposition reactions increases as the concentration of elements and free radicals increases [15]. refined oils may contain less than 1 ppm of alkaline substances, such as atomic number 11 or atomic number 19 fatty acid salts. modern cooking oils should contain less than 10 ppm of alkaline substances [3]. 2.2.3. oil polymerization labile compounds are very important to the flavor properties of edible oils and frying products, but volatile components are present in the total degradation products of frying oils at concentrations of 1:500,000. the main decomposition products of frying oil are non-volatile polar compounds and triacylglycerol dimers and polymers. the number of cyclic compounds is relatively small compared to non-volatile polar compounds, dimers, and polymers [16]. dimers and polymers are large molecules with molecular weights between 692 and 1600 daltons that are formed by a mixture of -c-c, -c-o-c, and -c-o-o-c bonds. dehydroxydimer, ketodehydrodimer, monohydrodimer, dehydrodimer of linoleic acid, and dehydrodimer of oleic acid are dimers that occur in vegetable oils upon cooking at 195 °c [17]. dimers and polymers have hydroperoxy, epoxy, hydroxyl, and carbonyl groups, as well as -c-o-c and -c-o-o-c bonds. dimers or polymers can be either acyclic or cyclic, depending on the reaction method and the form of fatty acids present in the oil [18]. dimerization and chemical action during frying are radical reactions. the group radical is ideally formed on the group carbon at the alpha position of the double bond. dimers are formed by reaction of allyl radicals with c-c bonds. the formation of acyclic polymers from monounsaturated fatty acids during heating is shown in figure 3. triacylglycerols react with atomic number 8 to form alkyl radical hydroperoxides (rooh) or dialkyl peroxides (roor). these are immediately converted to alkoxy and peroxy radicals by ro-oh or roo-r cleavage. alkoxy radicals extract chemical elements from oil molecules to provide group compounds or mix with various alkyl radicals to produce oxydimers [18]. agriculture and food sciences research, 2023, 10(2): 14-27 18 © 2023 by the author; licensee asian online journal publishing group figure 3. acyclic compound formation from monounsaturated fatty acid throughout deep-fat cooking. peroxy radicals mix with alkyl radicals to form peroxy dimers. the formation of dimers and polymers depends on the type of oil, frying temperature, and frying range, with the amount of polymer increasing as the number of frying steps and frying temperature increases. oils rich in polyunsaturated fatty acids polymerize much more easily during frying than oils rich in oleic acid [19]. cyclic polymers are prepared between or among triacylglycerols by radical reactions, namely diels-alder reactions. the formation of cyclic compounds in edible oils depends on the degree of unsaturation and frying temperature. the formation of cyclic monomers and polymers accrued because the quantity of omega-6 fatty acid increased [18]. the formation of cyclic monomers was negligible till the linolenic acid content exceeds 20%. cyclic compounds don`t seem to be shaped to a big extent until the oil temperature reaches 200oc to 300oc. vegetable oil formed a dimer of antidepressants and a cyclic dimer of linoleic acid, similar to the cyclic monomers in fried foods. the polymers formed during frying are rich in oxygen. polymers further accelerate oil degradation, increase oil viscosity [20], reduce heat transfer, form foam during frying, and cause undesirable discoloration of foods. polymers also cause high oil absorption into foods. the polymer is a highly conjugated diene, and when the oil or metal comes into contact with the atomic number 8 in the air, it forms a brown, resinous residue on the edges of the fryer. resinous residues are typically formed when oil does not release water but retains water while absorbing air [3]. 3. causes of repeated fat cooking even today, the cuisine of the day is consistent with society's preferred methods of preparing more delicious meals. there is a high demand for ready-to-eat foods in developing countries. cooking forms compounds with rich flavors, attractive colors, skins, and textures that improve the sensory quality of foods and are highly appreciated by consumers. cooking fat is the main component of these fried foods [21]. therefore, oil prices are the most important economic factor. therefore, to ensure economy, vegetable oil is usually repeatedly heated. in order to reduce the cost of frying and save a lot of money, several household users and most food processors in ethiopia repeatedly use the same vegetable oil for frying until the oil completely turns an undesirable color. i am. the oil is repeatedly reused and is discarded and replaced with fresh oil only when it becomes foamy, very viscous, emits dangerous odors, and darkens in color [22]. often, they are not replaced at all. rather, fresh oil is different from already heated thick and viscous oil [23]. 3.1. oil recooking alterations eating limited amounts of fried foods does not cause any degree of health problems in conventional people. the problem begins when you heat and use the same oil over and over again. when frying food at temperatures between 170°c and 200°c, the oil used undergoes various changes. 1. hydrolysis – the water from the food being fried evaporates and the triglycerides (tg) in the cooking oil are hydrolyzed to glycerin. free fatty acids (ffa), monoglycerides (mg), diglycerides (dg). 2. oxidation – lipid molecules in frying oil undergo primary oxidation to form unstable lipid species called “hydroperoxides.” this is cleaved to produce secondary oxidation products containing non-volatile and volatile compounds [24]. some of these byproducts polymerize (tertiary oxidation), increasing the viscosity of the oil and causing surface browning and darkening of the oil. thermal polymerization at high temperatures during the cooking process, high agriculture and food sciences research, 2023, 10(2): 14-27 19 © 2023 by the author; licensee asian online journal publishing group molecular weight cyclic carboxylic acid monomers (fa) and tg dimers and oligomers are formed [24]. fried foods may absorb some aerobic products such as hydroperoxides and aldehydes produced by this method of increasing the amount of oil [25]. table 3. peroxide value (pov) for ethyl radical linolenate throughout heating at completely different temperature. 70 oc 180oc 250oc cooking time (h) pov (meq/kg) heating time (h) pov (meq/kg) heating time (h) pov (meq/kg) 6 1777 5 237 3 44 24 1058 10 251 5 77 45 505 20 119 10 198 69 283 30 80 20 67 source: nawar [26]. table 3 displays the precise chemical change parameter for variations in peroxide value at various heating times. oils that are cooked or reheated can become oxidized due to the reaction of oxygen with free fatty acids. when oil is cooked repeatedly at different temperatures, it completely changes its peroxide value. as a result, the table provided a clear explanation of the pov changes in meq/kg for ethyl radical lenolenate during heating and cooking. 4. deeply fried oil and foods quality food flavors that are created by high temperature oil cooking include fruity, grassy, burned, nutty, and gloomy. however, the frying temperature has no effect on the flavor of the oil; it relies on the oil and quantity of frying. when frying potatoes at 160°c, 180°c, and 200°c, the oil's flavor quality was superior than that of vegetable or colza oil. according to wu and chen [14] the main volatile chemicals in vegetable oil at 200°c were reported to be 2-heptenal, 2-octenal, 1-octen-3-ol, 2, 4-heptadienal, and 2, 4-decadienal. at the ideal concentration of atomic number 8, a typical intriguing deep-fried flavor is produced. high oxygen levels result in an odd flavor, whereas low oxygen levels yield a weak and bad flavor [27]. flavor compounds in fried foods are volatile compounds from polyunsaturated fatty acids and are dienals, alkenes, lactones, hydrocarbons and many cyclic compounds [27]. 4-hydroxy-2-nonenoic acid and its lactone, 4-hydroxy-3nonenoic acid and its lactone, trans, trans-2, 4-decadienal, trans, trans-2, 4-nonadienal, trans, trans-2, 4-octadienal, trans-2-heptenal, trans-2-octenal, trans-7-octenal, nonenlactone and trienals are attractive flavor compounds found in cooking oil and are made by oxidation of linolenic and it is polyunsaturated fatty acids. polyunsaturated fatty acids are responsible for the desirable flavor of frying. completely different oils give different flavors when frying due to the difference in the quality and quantity of fatty acids in the frying oil. butanal, pentanal, hexanal, heptane, pentanol, 2-hexenal, heptanal, 1-octen-5-ol, 2-pentylfuran and 2-decenal give unpleasant results when frying [28]. carbonyl compounds formed during deepfat frying will reply with amino acids, amines, and proteins and turn out fascinating and nutty pyrazines. a number of unpredictable composites shaped in deepfat cuisine, 1, 4dioxane, benzene, toluene, and hexylbenzene, do not contribute to desirable flavor and are noxious compounds [28]. figure 4. cyclic compound formation from polyunsaturated fatty acid by diels-alder reaction throughout deep-fat frying. figure 4 illustrates the formation of cyclic compounds from polyunsaturated fatty acids through chemical reactions during deep fat frying at high temperature for a longer period of processing. polyunsaturated fatty acids are liquid oils which can be easily oxidized as exposed to high temperature and oxygen. when the polyunsaturated oils treated at high temperatures free radicals occurred and immediately reacted with oxygen, as a result the oil become agriculture and food sciences research, 2023, 10(2): 14-27 20 © 2023 by the author; licensee asian online journal publishing group oxidized and deteriorate. finally, the rest chemical structures become cyclic compounds by reacting each other instead of migrated hydrogen. 5. high temperature oil cooking factors the ratio of oil, frying time and temperature, type of heating, frying oil composition, initial oil, composition of food to be fried, type of fryer, antioxidants and oxygen content material have an effect at the deterioration of oil throughout deep-fats cooking. the consequences of frying factors on the usual of frying oil are normally rumored in any other case or oppositely, due to the utilization of diverse analytical ways for high-quality determination and different experimental situations [29]. 5.1. fresh oil replecement a high quantitative relation of clean oil to total oil presents higher frying oil exceptional. frequent replenishment of clean oil decreases the formation of polar compounds, diacylglycerols, and loose fatty acids will boom the frying existence and first-rate of oils [30]. again, rumored that refilling with recent oil improved the usual of cooking oil completely 1/30 frying. also [31] mentioned that common turnover of oil prompted quite a few aerophilous response than hydrolytic reaction in the course of deep-fat frying of potatoes. a suggested every day turnover is 15% to twenty-five% of the capability of the pullet and therefore the high turnover will decrease the usage of antifoaming agent akin to silicones. 5.2. cooking conditions cooking time will increase the contents of loose fatty acids, polar compounds consisting of triacylglycerol dimers and exchange triacylglycerols, dimers and polymers. the primary 20 cooking’s boom the formation of polar compounds unexpectedly. there has been no essential growth of polar compounds 1/30 frying [18]. excessive frying temperature quickens thermal oxidization and chemical action of oils. vegetable oil showed three.09% and 1. sixty-eight% of conjugated dienes and trans-acids, respectively, after 70-m frying of potato chips at 170oc. but soybean oil that performed equal cooking at 190oc showed 4.39% and 2.60% of the several values. excessive frying temperature shrunken polymers with peroxide linkage and amassed the polymers with ether linkage or carbon to carbon linkage [32]. the intermittent heating and cooling of oils causes’ higher deterioration of oils than continuous heating because of the atomic number eight solubility growth within the oil as soon as the oil cools down from the frying temperature [29]. the 25% of polyunsaturated fatty acid of the flower oil became destroyed in the intermittent frying, while totally the five% become destroyed in non-stop cooking. 5.3. quality of cooking oil unfastened fatty acids boom the thermal oxidization of oils and their unsaturation in place of chain length lightemitting diode to vital effects on thermo oxidative degeneration; the addition of 0.53 mmol of tridecanoic, palmitic and oleic acids to virgin oil confirmed 15.zero, 14.3 and 10.1 h of induction amount with a rancimat (metrohm) [11]. chemical technique and genetic change are two of the methods to decrease the unsaturated fatty acids of cooking oil. chemical procedures will grow the frying balance of oil. however, hydrogenation produces trans-carboxylic acid or aluminous taste, and it does not in addition improve the usual of oil with low omega-6 fatty acid. modified vegetable oil with zero.1% linoleinic acid had a variety of hydrolytic degradation, but lower p-anisidine values and compound formation, than the soybean oil with 2. 3% linoleinic acid [18]. genetically modified high oleic vegetable oil improved frying stability over conventional corn oil. consequently, low linoleinic acid oil through genetic amendment changed into advised to be a likely diverse to alter cooking oil. mixing of many oils can change the carboxylic acid compositions of oils and might decrease the oxidization of oils at some point of deep-fats frying. the esterified glyceryl ester doesn't display any pro-oxidant activity in the oil oxidation during deep-fat frying. loose fatty acids in frying oil improved thermal oxidation of the oil [33]. the remedy of shortening with bleaching clay, charcoal, chelate, or mgo advanced the oil exceptional for fries. every day addition of ascorbyl palmitate to current oil shrunken loose carboxylic acid formation, however gathered stuff consistent and coloration modifications. oils with beneath 0.05% unfastened fatty acids and 1.0 milliequivalent peroxides in 1 kilogram of oil are fascinating for deep-fats cooking [18]. table 4. common indicators for deep-fat frying fats and oils. parameter levels in unused fat and oil free fatty acids 0.05–0.08% peroxide value 1.0 meq/kg oil smoke point 200 ◦c moisture 0.10% flavor and odor bland source: christopoulou and perkins [17]. before any physical or chemical alterations take place, the fresh oil parameters value is displayed in table 4. these parameters vary according to the temperature and frying time as the oil deep-fries. however, when deep fat frying, it was thought that using mixed types of oil would be more stable than using monotype oil. the table indicates that the parameters that are most likely to alter while frying are free fatty acids, peroxide values, smoke point, moisture, flavor, and odor. 5.4. food compositions moist conditions in meals create steam insurance over the pullet and decrease contact with air [34]. fantastic deal of moisture in foods will increase the oil reaction during deep-fat cookery. lots of the moisture contents in meals, the maximum the hydrolysis of oils. emulsifier from frying foods triggered foam formation at the initial stage of deepagriculture and food sciences research, 2023, 10(2): 14-27 21 © 2023 by the author; licensee asian online journal publishing group fat frying. phosphatidylcholine attenuate the oxidization of animal oil at 180oc for 3hr. starch increases the degradation of oil and amino acids guard the oil from degradation at some point of deep-fats cookery [35]. 5.5. types of fryers transition metals just like iron, that are present in meat, had been gathered in the oil in the course of frying and this inflated the fees of oxidization and thermal degradation of oil. even and fast heat transfer to the oil can save you hot spots and the scorch of oil. polymerized fats deposited at the fryer causes gum formation, the formation of froth, color darkening and in addition deterioration of frying oil. a small floor-to-volume ratio of fryer for a minimum touch of oil with air is suggested for deep-fat frying. negishi, et al. [36] reported that oil oxidation become slowed down by means of editing a fryer to have a ratio of oil intensity (d) to grease vicinity (a) with d/a1/2 = 0.93. copper or iron fryer hastens the oxidation of frying oil. 5.6. antioxidants the evident gift or added antioxidants in oils and foods have an impact on oil exceptional during deep-fat frying. tocopherols, butylated hydroxyanisole (bha), butylated hydroxytoluene (bht), propyl gallate (pg), and tertbutylhydroquinone (tbhq) sluggish down the oxidation of oil at room temperature. but they end up less effective at frying temperatures because of losses through volatilization or decomposition [37]. the addition of spinach powder at five%, 15%, or 25% in flour dough attenuates the formation of polar compounds in oil [38]. the addition of crimson ginseng extracts to the flour dough at 1% and 3% accelerated the formation of loose fatty acids, conjugated dienoic acids, and aldehydes in oil all through the deep-fat cookery of the dough at 160oc. beef nuggets battered with glandless oilseed flour attenuate unfastened fatty acids, conjugated diene compounds, and thiobarbituric acid-reactive substances. the addition of carrot powder to the dough at 10%, 20%, or 30% increased the aerophilic balance of oil at p < 0 xss=removed> zero.05 [39]. propane and carbonyls fashioned throughout deep-fats frying aren't the maximum precursors of amide [40]. many nutrients are touchy to higher temperatures and oxidization, however high temperatures are reached solely in floor layers of deep-fried food, wherever their loss is surely terribly excessive. total losses depend mostly on internal temperature that every so often varies among seventy and 900c. throughout this variety, diet retention depends alternatively extra at the inner temperature than on the temperature of the cookery oil [41]. tocopherol is misplaced at the aspect of the oxidation of unsaturated fatty acids throughout heating. cookery oil is absorbed by means of the meals at some point of cooking and additionally the absorbed amount relies upon on the usual of the cooking oil, which impacts the net intake of tocopherol [42]. a stimulating resistance has been incontestable for tocopherol homologues for the duration of home cookery simulation with virgin olive oil, sunflower-seed oil or vegetable shortening oil for 8 sequential frying operations. vegetable frying oils are a remarkable deliver of diet e. all vegetable oils used for frying comprise nutrition e quantity of between 15 and fortynine mg/a hundred g. deep-fried ingredients, because of oil uptake are enriched with respectable amounts of the vitamin. as an instance, a touch of a hundred g gives up to 50% of the recommended dietary allowance (rda) of tocopherol [43]. tocopherol (tocopherols) from the frying oil participates in radical reactions. table 5. effects of (rosemary and sage) extract antioxidants on the quality of refined, bleached, and deodorized palm olein during frying of potato chips. characteristics heating time (d) control rosemary (0.4%) sage (0.4%) anisidine value 0 0.96 0.95 0.96 2 36.0 30.6 29.3 4 51.4 42.1 42.0 6 62.0 50.2 50.8 0 0.05 0.05 0.05 free fatty acid (%) 2 0.19 0.15 0.16 4 0.42 0.26 0.25 6 0.59 0.42 0.44 0 0.01 0.01 0.01 polymer content (%) 2 1.00 0.73 0.70 4 1.55 1.30 1.35 6 2.65 1.82 1.90 0 0.29 0.29 0.29 c18:2/c16:0 ratio 2 0.26 0.27 0.28 4 0.21 0.25 0.24 6 0.17 0.20 0.21 table 5 shows how antioxidant sources like sage and rosemary affect refined, bleached, and deodorized palm oil when it's being fried. with an increase in oil frying time, the values of the characteristics exhibit rising trends. however, as more antioxidants are added to the oil during frying, the values tend to drop. for this reason, adding antioxidants like sage and rosemary to the oil helps to stabilize it a little bit when deep-frying it. 6. foods nutrient changes during deep-fat frying the fee of nutrient decomposition in the course of deep-fats frying is contingent upon the frying length and the sort of oil applied. the decomposition price of γ-tocopherol for the duration of the deep-fat frying of potatoes in an aggregate of soybean and rapeseed oil at a hundred and eighty ◦c became located to be the maximum speedy, followed by means of δand α-tocopherol. lipid oxidation effects within the formation of aldehydes, epoxides, hydroxyketones, and dicarboxylic compounds, which react with amines, amino acids, and proteins in fried meals [44]. millard reaction happens during food frying, leading to nutrient loss and browning. the degree of browning is frequently associated with the losses of lysine, histidine, and methionine. the reaction among epoxyalkenals and agriculture and food sciences research, 2023, 10(2): 14-27 22 © 2023 by the author; licensee asian online journal publishing group proteins generates polypyrrolic polymers and volatile heterocyclic compounds [45]. carbonyl compounds produced throughout lipid oxidation react with amino acids, mainly asparagines, resulting inside the formation of acrylamide and a lower in the dietary cost and safety of meals. acrolein, shaped from oil, reacts with asparagines, generating acrylamide throughout deep-fat frying. acrylamide formation requires a heating temperature above a hundred ◦c and increases because the temperature rises [1]. the styles of oil and addition of silicone had no influence at the acrylamide concentration in ingredients. acrolein and carbonyls formed for the duration of deep-fat frying aren't the primary precursors of acrylamide [40]. 6.1. vitamins various nutrients are liable to degradation from excessive temperatures and oxidation, with the surface layers of fried meals being the number one region for such losses. but, the whole loss of nutrients is more often than not depending on the internal temperature, which typically tiers among 70 and 900c. on this variety, the retention of vitamins is more heavily motivated by using the inner temperature than the temperature of the frying oil [41]. vitamin e is especially vulnerable to degradation due to the oxidation of unsaturated fatty acids for the duration of heating. the quantity of frying oil absorbed by using the food at some point of cooking is depending on the great of the cooking oil, which could affect the internet intake of vitamin e. but, tocopherol homologues have validated tremendous resistance at some stage in home frying simulations with virgin olive oil, sunflower oil, or vegetable shortening oil for up to 8 successive frying operations. depending on the oil kind, as much as 50% of nutrition e can be retained after four to 5 consecutive frying sessions [42]. vitamin e is particularly vulnerable to degradation due to the oxidation of unsaturated fatty acids during heating. the amount of frying oil absorbed by the food during cooking is dependent on the quality of the cooking oil, which can impact the net intake of vitamin e. however, tocopherol homologues have demonstrated remarkable resistance during domestic frying simulations with virgin olive oil, sunflower oil, or vegetable shortening oil for up to eight successive frying operations. depending on the oil type, up to 50% of vitamin e can be retained after four to five consecutive frying sessions [42]. even as vitamin e from frying oil can take part in free radical reactions, it could also lower their fee, therefore having dietary and health advantages [41]. apparently, no widespread exchange within the vitamin e content of french fries was found in the course of 4 days of commercial frying, because the growth in fat intake of the fries compensated for the diet e reduction due to frying the oil. additionally, studies have shown a boom in overall diet e content material in chook nuggets and breaded shrimp after frying in soybean and corn oils [46]. earlier than frying, the vitamin e content material became 4.6 mg/a hundred g in chicken nuggets and 0.6 mg/a hundred g in breaded shrimp, which multiplied to four.9 mg/100 g and five.1 mg/100 g, respectively, after frying. table 6. retention of tocopherol in a mixture of soybean and rapeseed oils during frying of potatoes at 180 ◦c tocopherol retention (%) number of frying α γ δ 0 100 100 100 4 86.2 82.1 91.0 8 85.9 74.0 87.1 12 83.1 67.6 80.3 16 75.9 59.3 74.3 20 71.9 49.6 67.8 24 64.5 41.6 63.7 28 60.9 33.5 51.5 source: hidalgo and zamora [45]. table 6 illustrates the retention of tocopherol at frying frequencies ranging from 0 to 28 minutes. we all know that high temperatures during cooking and frying cause vitamins to be lost. thus, research on the amount of tocopherol vitamin retained during deep fat frying is summarized in the table. it makes clear that the concentration of vitamin drops as frying time increases. thus, extended oil-frying times result in the loss of vital nutrients, especially tocopherol and other vitamins. plant source food incorporates active carotenes, along with vitamin a, which can be lost for the duration of the frying process. however, if the frying technique is short, losses of beta-carotene, the most critical consultant of this organization of pro-nutrients, can be stored low. in evaluation, deep fried greens enjoy losses of beta-carotene which are two times as excessive as the ones in shallow-fried foods, with some beta-carotene probably migrating into the frying oil. studies have proven that in the frying of cabbage, 29% of beta-carotene turned into destroyed, and common losses of vitamin-a interest were discovered to be 14% for boiling, 24% for frying, 29% for fermenting, 44% for sundrying, and 60% for solar-drying observed through boiling. even as a few carotenoids are misplaced in cooking water, losses all through frying are more due to leaching into frying oil. in terms of other vitamins, research has shown that vitamins b1, b2, b6, and c are better retained in frying than in boiling, steaming, or stewing. thiamin, and vital vitamin of the b group, stories decrease losses for the duration of frying than when food is prepared the use of other strategies. the largest lack of thiamin happens during boiling, followed by steaming, parching, and frying, which can be attributed to the water-soluble nature of the diet being leached out into the water. riboflavin, any other crucial vitamin b, is often poor in human diets and is better retained when frying bird meat than thiamin is when frying darkish meat. niacin is exceptionally stable, but nevertheless reports losses all through frying pork muscle, beef, and cook meat. vitamin c retention in green greens is higher whilst stir-frying than when cooking with numerous waters or the usage of a microwave. agriculture and food sciences research, 2023, 10(2): 14-27 23 © 2023 by the author; licensee asian online journal publishing group table 7. vitamin c retention of broccoli and green beans cooked using four methods. cooking method vitamin c % retention in broccoli vitamin c % retention in green beans stir-frying 76.6 57.5 microwave 56.8 58.9 much water 44.8 59.6 little water 72.2 76.0 source: fillion and henry [47]. table 7 provides an overview of the vitamin c retention for various frying methods and meal types. when broccoli is stir-fried, the vitamin c retention is improved. however, the nature of the product makes stir-frying green beans less suitable. cooking green beans with minimal water likely results in the best retention; this also applies to broccoli. therefore, the length of time that vitamin c remains stable in various food products varies depending on the method of cooking and amount of water used. 6.2. mineral components because mineral additives dissolve in water, they undergo significant changes during cooking, especially when boiling. on the other hand, since mineral additives are best soluble in small amounts of frying oil, cooking has little effect on them. however, the weight of fried foods is reduced because of water loss. although the majority of mineral components are non-volatile, it is anticipated that their content material on moist weight will increase. even so, the frying oil absorbs simultaneously, resulting in a boom in the weight of the fried fabric. a small decrease in mineral content may be found if the metallic content is expressed on a dry weight basis. deep-fried foods lose a significant amount of minerals, ranging from 1% in potatoes to 26% in beef. mineral losses from deep-frying range from 2 to 8% in breaded meat and fish; which is significantly less than in deep-fried meat and fish fillets without coating. this is because the minerals dissolved in the meat gravy are absorbed by the breadcrumbs [41, 48]. 6.3. mineral metabolism the effects of frying and non-frying consumption of olive, sunflower, and palm oil on the bioavailability of calcium, phosphorus, and magnesium in growing rats were investigated by the researchers [49]. the results demonstrated that the type of oil consumed had no effect on magnesium's bioavailability. however, the body's ability to retain magnesium remained unaffected by any of the three frying oils; instead, they all increased the amount of magnesium absorbed and urined. calcium absorption efficiency was higher in animals fed both kinds of sunflower oil—unused and used for frying—but there was no appreciable difference in calcium retention, urine, serum, or carcass calcium. the consumption of frying oils did not significantly change the bioavailability of calcium. 6.4. protein fry food does not change the digestibility of protein when no other ingredients are added. however, the addition of substances that reduce—such as the flour-based meatballs and fishballs—can result in a slight but discernible decrease in the protein's digestibility. according to bognar [48] research, cooked foods can retain anywhere from 90% of their protein when meat is boiled to 96%–100% when meat, fish, and potatoes are deep-fried. the study looked at how different cooking techniques affected the amount of protein. these findings lend credence to the theory that frying has no impact on protein digestibility. table 8. protein digestibility coefficient of raw and fried foods state hake beef pork swordfish meat balls fish balls raw 0.92 0.93 0.92 0.94 0.90 0.92 fried 0.91 0.93 0.92 0.96 0.88 0.89 source: fillion and henry [47]. the protein digestibility coefficients for foods containing raw and fried protein sources are explained in table 8 of the research report. the findings show that, aside from some typical structural changes, very few appreciable differences exist between the raw and fried products during oil deep-frying. this indicates that protein digestibility is the foods most stable during cooking and frying. some slight alterations are noted, though, as a result of the products containing a combination of meat and flour. 6.5. carbohydrate studies on the impact of frying on the carbohydrate content of potatoes, potato products, and breaded meat and fish have been carried out. the findings indicated that, depending on the kind of food, carbohydrate retention varied from 95% to 100%, indicating that the frying process had no discernible impact. deep-frying considerably raised the resistant starch content while cooking frozen french fries had no effect on the starch composition when compared to raw samples. amylose-lipid complex formation might have a role in this. after being fried, potatoes' digestible starch content decreased, but their fiber content increased. according to fillion and henry [47] fiber is essential for preventing diseases like diabetes, heart disease, and colon cancer. 7. deep-fat frying consumption unhealthiness tasty food is made edible through deep-frying. foods fit for human consumption are deep-fried using copious amounts of oil. it is not thought that increasing oil consumption is good for human health. fried foods are generally regarded as safe, even though deep-frying generates some unquestionably toxic products (such as polar compounds or polymers) [50]. when cooking oil is used frequently and turns toxic for ingestion by humans, it works best. cooking oil's plant antioxidant content decreases and hazardous reactive oxygen species are produced when oil is heated repeatedly, hastening the oxidative breakdown of lipids. it is anticipated that eating meals that contain heated oil for an extended period of time will negatively affect the antioxidant defense system, which is a major contributor to the development of diseases like vasculitis, diabetes, hypertension, and obesity [51]. agriculture and food sciences research, 2023, 10(2): 14-27 24 © 2023 by the author; licensee asian online journal publishing group foods originating from plants fully absorb food originating from animals. fried veggies, whole eggplants, tomatoes, onions, mushrooms, and pineapple pulp are better sources of fat absorption for them. large amounts of fat are consumed by vital people collectively. cooking meats like beef, pork, lamb, or sausages reduces their fat content. cooking fats and oils enter our diets through absorption from hard foods. depending on the food and consequently the type of frying medium, the percentage of oil absorbed in food varies from 4% to 14% of the total weight. the absorption of cooking oil is influenced by its quality. the surface tension between the potatoes' surface and the frying oil is high when using fresh oil. frequent frying causes an increase in oil polarization and interfacial tension decreases. so, oil consumption increases with regular cooking of potatoes [52]. heating causes thermal and aerophilous breakdown, which produces changed, polymerized compounds with more polarity. this consequently modifies the properties of deep-fried food and dietary fat in the organic process. the world health organization (who) recommends limiting one's consumption of sugar, salt, and saturated and transfats (hydrogenated fats). snacks, processed foods, and drinks frequently contain these ingredients [53]. 7.1. cardiovascular disease (cvd) according to mozaffarian, et al. [54], cvd is the leading cause of death globally and is expected to be linked to over 23.6 million deaths by 2030. as early as the 1970s, it was proposed that people who consume a diet high in fat, primarily from olive oil, would have lower rates of coronary heart disease (chd). however, eating deep-fried food is also linked to an increased risk of cardiovascular disease. on the other hand, there is limited and inconsistent evidence supporting the link between consumption of fried foods and the risk of coronary artery disease (cad) food [55]. these null results contradict the positive associations reported in alternative studies. it is possible that mediterranean diets emphasizing fruits and vegetables, meat and whole grain coffee intake [56], and/or the types of oils used for coaching in spain (particularly vegetable and flower oil) could contribute to the distinction determined by research. it has been demonstrated that using extra virgin olive oil when frying some foods decreases the depth of macromolecule oxidation [57]. a study conducted in costa rica using a case-control sample of 485 individuals who had survived a major acute myocardial infarct and 508 controls found no correlation between nonlethal acute myocardial infarct and an increase in the frequency of deep-fried food consumption (from 4.57 to 9.75 servings/day; p < 0.05) [58]. the most important oils used for instruction on this population are palm oil, vegetable oil derivatives, and oil. in a case-control study conducted in india in 2003, involving 199 matched controls and 160 five patients with coronary cardiovascular disease, the patients with coronary heart disease reported consuming more deep-fried and shallow-fried food than the controls did. 7.2. heart failure according to a study, eating deep-fried fish has also been linked to lower glide, a lower ejection fraction, and a higher resistance to the trendy tube-shaped shape in older adults. belin, et al. [55] found that consuming more than one serving of fried fish per week at baseline was linked to a 48 % increased risk of heart failure (hf) and heart rate (hr) is 1.48 with 9.5 % confidence interval: 1.19–1.84. additionally, compared to subjects who reported consuming fried food, there was a positive and significant correlation between the consumption of fried meals and the incidence of heart failure in a prospective cohort study [55]. 7.3. hypertension changes in unhealthy cooking habits from boiling to cooking have been found to be beneficial for dominant pressure according to unit location and fat in the general population. when it comes to cooked food intake and elevated blood pressure, there may be limited and inconsistent treatment evidence at the same time. a cross-sectional study conducted in spain found that eating more fried food was linked to a higher prevalence of hypertension [59]. the solar (seguimiento universidad de navarra) mediterranean cohort study found that a baseline common intake of fried foods was linked to the following risk of high blood pressure (adjusted hazards ratios = 1.18 (95 percent confidence interval: 1.03–1.36) and 1.21 (95 percent confidence interval: 1.04–1.41) for those who consumed excessive amounts of cooked meals twice or more per week, respectively [59]. 7.4. type two diabetic disease (t2d) the risk of t2d has been directly linked to the consumption of potatoes, red meat, and opportunity processed meats [60]. restaurant cooked meals and dietary consumption had certainly been linked to t2d. information from the nurses' fitness assessment and the medical professionals' adherence to: further investigation revealed a strong correlation between the frequency of fried food consumption and, in turn, the risk of type 2 diabetes (t2d), with adjusted rrs (95 percent confidence intervals) for those who consumed fried foods [61]. 7.5. obesity according to several studies, eating deep-fried food is positively correlated with obesity, a larger waist circumference, or weight gain in pregnant women [62]. additionally, two large potential studies have suggested a link between a higher incidence of incident obesity/weight problems and the consumption of fried foods. according to tiwari, et al. [63] the prospective study revealed a strong correlation between the consumption of fried food and both general and vital obesity (adjusted odds ratios for general obesity in the highest quintile versus the lowest quintile of fried meal consumption: 1.26 (ninety five% ci: 1.09–1.forty five, p for fashion four times/week). 8. ways to solve the harmful effects of deep fat cooking since the oil will be made safe for human consumption, there will be a reduction in the harm that comes from using heated oil continuously in a number of ways. vegetable oil can be made safe by using natural antioxidants as adsorbents to delay the production of products that cause oil deterioration. numerous studies have made it clear that antioxidants such as propylgallate (pg), tert-butyl hydroquinone (tbhq), butylated hydroxylanisole (bha), agriculture and food sciences research, 2023, 10(2): 14-27 25 © 2023 by the author; licensee asian online journal publishing group propylated hydroxyl toluene (bht), and tocopherols reduce the reaction of oil at temperature. nevertheless, due to losses from volatilization or breakdown, their effectiveness decreased at cooking temperature [37]. it has been observed that adding different absorbents during frying, such as sugarcane bagasse, rosemary, and antioxidants from turmeric extract, reduces the detrimental effects of the deterioration products. the addition of curcumin, a naturally occurring inhibitor found in turmeric (olantof zingiberaceae) family, is said to lessen the negative effects of products that cause oil deterioration. when turmeric extract was added to the cooked food at a concentration of 0.03%, the amount of trans-carboxylic acid decreased. this relates to curcumin, which inhibits the rate of auto-oxidation by converting lipoid radicals into a highly stable form, which typically inhibits the fat reaction [64]. antioxidant supplements added to vegetable oil that is frequently used can slow down the rate at which the oil oxidizes during cooking and improve the way that cooked food is perceived by the senses. the findings of this study are corroborated by the claim made by abriana and johannes [65] that the addition of an inhibitor to vegetable oil verifies the integrity of the reaction during cooking and that curcumin is a useful antioxidant due to its ability to scavenge radicals by donating a h atom from synthetic resin as its active group. other than this, a number of adsorbents like ash, magnesol xl, and sugarcane bagasse can also be used to reduce the formation of the decay product produced by continuous frying [66]. 9. conclusion edible oils are widely consumed and popular food worldwide but the high temperature cooking of oils causes deterioration of food and health problems. this oil long time and high temperature cooking oils problems occurred because of the chemical and physical changes upon cooking. reactions in oils increase the number of free fatty acids, mono and diacylglycerols and glycerols. these compounds with free radical hydrogen react with oxygen in the air and causes great health problems on the consumers. but using fresh oil, temperature of cooking, time of cooking, food materials, fryer and antioxidants have an effect on the quality and safety of oil during cooking. also using artificial antioxidants like turmeric and natural antioxidants like tocopherols, bha, bht, pg, and tbhq can decrease oil oxidations, even if they diminished normally at cooking temperature. but using lignin compounds instead of using antioxidants are more effective to stabilize oil during cooking. therefore, the consumption of fried fat/oil at high temperature for long time becomes dangerous for human causing chronic diseases like cvd diseases, heart failure, hypertension, polygenic disorder and fatness. so, the review warns us to take care with oil cooking and consumption so as not expose to chronic diseases. 10. conflict of interest declaration the authors declare that there are no potential competing financial interests or personal relationships that could have appeared to influence the work reported in this review paper. the authors mentioned in this paper are the only owner of the research paper work both in finance and technical works. references [1] f. pedreschi, p. moyano, k. kaack, and k. granby, "color changes and acrylamide formation in fried potato slices," food research international, vol. 38, no. 1, pp. 1-9, 2005. https://doi.org/10.1016/j.foodres.2004.07.002 [2] r. g. moreira, x. sun, and y. chen, "factors affecting oil uptake in tortilla chips in deep-fat frying," journal of food engineering, vol. 31, no. 4, pp. 485-498, 1997. https://doi.org/10.1016/s0260-8774(96)00088-x [3] r. moreira, m. castell-perez, and m. barrufet, fried product processing and characteristics. in: deep-fat frying: fundamentals and applications. gaithersburg, md: chapman & hall food science book, 1999, pp. 11–31. 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[66] r. wannahari and m. f. m. nordin, "reduction of peroxide value in used palm cooking oil using bagasse adsorbent," american international journal of contemporary research, vol. 2, no. 1, pp. 185-191, 2012. https://doi.org/10.3844/ajeassp.2012.59.62 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.3945/ajcn.2009.28682 https://doi.org/10.1016/s0162-0908(09)79539-x https://doi.org/10.1007/s11746-008-1335-6 https://doi.org/10.1007/s11746-008-1335-6 https://doi.org/10.3844/ajeassp.2012.59.62 228 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 228-235, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6260 © 2024 by the authors; licensee asian online journal publishing group a review of the role of lectins in animal gametes and embryo development touba nadri1,2 andres gambini3,4 ( corresponding author) 1department of animal science, faculty of agriculture, university college of agriculture and natural resources, university of tehran, karaj, iran. 2department of animal science, college of agriculture and natural resources, urmia university, urmia, iran. 1,2email: t.nadri7070@yahoo.com 3school of agriculture and food sustainability, the university of queensland, 4343, gatton, queensland, australia. 4school of veterinary sciences, the university of queensland, 4343, gatton, queensland, australia. 3,4email: a.gambini@uq.edu.au abstract lectins are carbohydrate binding proteins found in most of the plants and in some animals. sperm glycocalyx modifications are known to occur during capacitation and the acrosome reaction. these changes are very important for gamete recognition and fertilization in mammals but are not fully understood. moreover, studies on lectin treatment of oocytes have yielded inconsistent effects on fertilization rates, suggesting a complex interplay of mechanisms. beyond fertilization, the mitogenic properties of lectins are associated with germinal vesicle breakdown and cumulus cell expansion, indicating their involvement in oocyte maturation. lectins have been also implicated in modulating cell adhesion, cell surface remodeling, and signaling pathways during early embryonic cleavage stages, influencing successful preimplantation embryo development. this article discusses the roof lectins in sperm, oocyte and embryo biology. keywords: embryogenesis, lectin, oocyte, reproductive, sperm, spermatozoa. citation | nadri, t., & gambini, a. (2024). a review of the role of lectins in animal gametes and embryo development. agriculture and food sciences research, 11(2), 228–235. 10.20448/aesr.v11i2.6260 history: received: 17 october 2024 revised: 19 november 2024 accepted: 3 december 2024 published: 27 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: drafting, writing and revision, t.n. and a.g. both authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 229 2. conclusions ..................................................................................................................................................................................... 232 references ............................................................................................................................................................................................ 232 mailto:t.nadri7070@yahoo.com mailto:a.gambini@uq.edu.au https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6260 https://orcid.org/0000-0003-0186-1120 https://orcid.org/0000-0002-3652-2068 agriculture and food sciences research, 2024, 11(2): 228-235 229 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature in the past years, a review study on the effect of lectins on animal reproduction had not been conducted. the purpose of this study was to review the studies that have been conducted on the effect of lectins on the reproductive system in one article. this study reviews the effect of lectins on the male and female reproductive systems, which has not been done so far. 1. introduction lectins are carbohydrate binding proteins present in most of the plants and in some animals [1]. the term 'lectins' is derived from the latin 'legree', meaning to pick or select. unlike antigens, lectins do not stimulate the immune system; however, they have the innate ability to bind in a manner analogous to antibodies [1]. these lectins selectively bind to carbohydrate moieties of glycoproteins that are present on the surface of most animal cells. structurally, these lectins have a diverse class of proteins, which have the ability to bind carbohydrates with considerable specificity [2]. numerous studies have explored the in vivo and in vitro effects of lectins. in vitro, they have been shown to affect lymphocyte mitogenesis, exhibiting both stimulating and inhibitory effects, particularly in lymphocytes from the gastrointestinal tract (git). they possess the ability to aggregate immunoglobulin’s, to trigger the alternative complement pathway, to inhibit fungal growth and also to induce histamine release from basophilic and mast cells. lectins are relatively resistant to both heat (withstanding over 30 min at 70°c) and digestion. moreover, some of the lectins are highly resistant to gastric acid and proteolytic enzymes [3]. 1.1. structure of lectins according to goldstein, et al. [4] lectins primarily consist of carbohydrate-binding proteins or glycoproteins of nonimmune origin, which bind cells, precipitate glycoconjugates, or sometimes both. the specific ability of lectins to bind with the cell surface mainly depends on the presence of monosaccharides or simple oligosaccharides, which, when present, inhibit the lectin-associated reaction [4]. in some cases, carbohydrate-associated enzymes with multiple combining sites may agglutinate or precipitate glycoconjugates and thus may be classified as lectins [5]. some lectins are structurally differentiated such as: 1. erythrine c lectins. 2. concanavalin c lectins. 3. ulex europaeus lectins. 4. c type lectins. the binding of lectins is reversible and noncovalent, involving either simple or complex carbohydrate conjugates, whether free in solution or on cell surfaces. surfaces containing glycoconjugates act solely as lectin receptors. the specificity of lectins generally relies on the hapten inhibition test, in which various sugars are tested for their ability to inhibit hemagglutination of erythrocytes. all lectin molecules possess two or more carbohydrate-binding sites, essential for their capacity to agglutinate cells or to interact with complex carbohydrates. some lectins can accommodate up to five or more sugar residues at their combining sites. multibranched oligosaccharides typically exhibit stronger lectin binding reactivities than linear ones due to cooperative binding effects of lectin and carbohydrate complexes. some lectins have dual or multiple affinities for different disaccharides. interactions between these molecules are hydrophobic, and electrostatic forces are rarely involved. lectins are purified from crude aqueous buffer or saline extracts of various tissues using standard protein chemistry methods, including ammonium sulfate or ethanol precipitation, and affinity chromatography [6]. 1.2. biological role the biological role of lectins is speculative. lectins may be involved in sugar transport or carbohydrate storage. some lectins may be associated with the binding of symbiotic rhizobia to form root nodules. due to their crucial role in adhesion and agglutination, lectins have been considered important in both symbiotic and pathogenic interactions between some microorganisms and hosts. microbial lectins may play an important role in adhesion to surfaces colonized by the microorganisms. for example, tomato lectins bind to mucosal cells and resist denaturation by acids and proteolytic enzymes [7]. there are several studies which have established lectins as specific agglutinins for blood groups of erythrocytes. due to their multifaceted biological properties, lectins were later developed by cell biologists as probes to investigate cell surface structures and functions. some lectins have been used to fractionate animal cells, including b and t lymphocytes, and to demonstrate changes in cell surface architecture following virus or parasite infection [8]. lectins have been used as carriers for the delivery of chemotherapeutic agents. they are significant reagents for investigating cell surface receptors in bacteria, protozoa, and higher organisms. the interactions of plant lectins with microorganisms have been applied for typing bacteria, fungi, and protozoa. lectins serve an important role in the characterization of bacterial cell components and in the identification of bacteriophage receptors. many bacteria contain surface-associated lectins that enable these microorganisms to adhere to surfaces. one of the major advantages of applying lectins in microbiology is that cellular or surface receptor sites can be partially characterized by hapten inhibition [9]. 1.3. advantages of lectins • stability. • activity in small concentration. • commercial availability of many lectins. • ability to probe subtle surface structural differences between various isolates [10]. 1.4. disadvantages of lectins in humans, lectins have been reported to cause damage, including mass food poisoning from uncooked kidney beans and it also causes hemolytic anemia and jaundice from mexican fava beans. lectins may cause acute gastrointestinal symptoms including nausea and vomiting. they bind to the luminal surface of absorptive agriculture and food sciences research, 2024, 11(2): 228-235 230 © 2024 by the authors; licensee asian online journal publishing group erythrocytes in the small intestines. this will cause severe damages to the microvilli of the intestine, thus disrupting the digestion and absorption [11, 12]. lectins can also promote the growth of harmful bacteria in the gut. lectins also disrupt proteins and carbohydrate malabsorption. in protein malabsorption, gut lectins bind to erythrocytes, causes inflammation which blocks the production of enterokinase, a protein enzyme. in case of carbohydrate malabsorption, it reduces intestinal glucose uptake by 50%. wheat germ agglutinin and other lectins can even bind to insulin receptors on cells, disrupting glucose metabolism. grains will have high content of lectins, which may cause inflammatory bowel and celiac diseases in humans [11, 12]. 1.5. the role of lectin on primary sperm attachment sperm glycocalyx modifications, known to occur during capacitation and the acrosome reaction, are critical for gamete recognition and fertilization in mammals but are not fully understood [13]. during the process of spermegg interactions, at least three separate events involving cell surface recognition occur: (1) the interaction between sperm and egg components (e.g., egg jelly or glycoprotein coat) before the acrosome reaction; (2) the attachment of sperm to the egg envelope surface after the acrosome reaction; and (3) the fusion of sperm and egg plasma membranes [14]. secondary attachment examinations have been conducted across various species of invertebrates, including the adhesion of either the sperm acrosome process or acrosome granule contents to the egg vitelline layer or chorion [15-18]. in sea urchins, the acrosome reaction is induced when spermatozoa initially contact the egg jelly coat, triggered by a sulfated fucose component [19]. the investigation by rosati and de santis [20] focused on the attachment process and revealed that binding sites for concanavalin a (con a) and asparagus pea lectin (apl) are present on both the egg chorion and sperm surfaces [20]. moreover, competition experiments involving monosaccharides have suggested that fucose may be implicated in gamete attachment [21, 22]. lectins, proteins with high saccharide-binding specificities, have become popular membrane probes [22]. they have been employed in studies to identify the presence of sugars on gamete surfaces. it has been demonstrated that both spermatozoa and the egg chorion of ciona have binding sites for fluorescein-conjugated con a and apl [20]. despite a decrease in fertilization following lectin treatment of eggs, the outcomes were inconsistent. huang, et al. [23] found that fitc-conjugated and gold-labeled wga attached to the chorion of the ascidian, phallusia mammillata, and hindered fertilization huang, et al. [23]. in limulus, fitcapl and fitc-con a adhered to the envelope surface of the egg; however, only apl reduced sperm attachment. there was no indication of fitc-gpl binding except on the egg cortex, and gpl did not impede sperm attachment. 1.6. lectin receptors changes during spermatozoon epidydimal maturation and capacitation sperm glycocalyx composition in mammals has been extensively studied by various authors bearer and friend [24]; schröter, et al. [25]; tecle and gagneux [26] and töpfer-petersen [27]. similarly, studies suggest that the distribution of lectin receptors changes during spermatozoon epididymal maturation, capacitation, and the acrosome reaction in mammals [22, 28, 29]. according to other researchers, lectins may prove useful in selecting spermatozoon subpopulations with the highest fertilizing capacity [30-32] highlighting the importance of understanding how the glycocalyx varies in other biologically or commercially valuable species. the knowledge of molecular changes undergone by spermatozoa during in vitro capacitation could facilitate the selection of spermatozoa subpopulations with the highest fertilizing potential for use in artificial insemination processes. this would also promote genetic exchange and increase genetic variability, helping to maintain group stability, avoid animal transport, or reduce the number of breeding males needed in a single facility [13]. sperm receptors comprised of sugars are also present on egg membranes of brown algae, fucus serratus [33] and the ascidian, ciona intestinalis [21]. isolated fucus egg membranes have shown to compete with whole eggs for spermatozoa, thereby lowering the percentage of eggs fertilized [33]. the lewis-a antigen, a type of carbohydrate antigen that belongs to the lewis blood group system, was found attached to viable sperm with intact acrosomes. however, when sperm were fixed, having their acrosomes removed through freezing and thawing, no binding was observed. furthermore, enzymatic removal of fucose from the epithelium led to a significant decrease in binding [34]. in the case of hamster sperm, it appears that a lectin on the sperm can identify a carbohydrate structure containing sialic acid. colloidal gold-labeled fetuin (a biochemical reagent used in microscopy and binding assays to visualize interactions involving carbohydrate-binding molecules) was observed binding to the heads of fresh epididymal hamster sperm, but did not bind to the apical regions of the heads of sperm incubated under capacitating conditions until they were hyperactivated [35]. the hyperactivated or capacitated hamster sperm failed to adhere to the epithelium upon introduction into the oviduct [36]. this suggests that a lectin present on the sperm surface undergoes a loss or modification during capacitation, enabling the sperm to detach from the epithelium. the glycosylated chains present on the membrane play a crucial role in gamete recognition, adhesion, and fertilization in mammals. changes in the plasma membrane during in vitro capacitation and the acrosome reaction (ar), such as the removal or appearance of specific proteins or glycoproteins, have been investigated using lectins that selectively bind to carbohydrate residues [13]. different lectins, each tagged with distinct fluorochromes, have been utilized to investigate the expression, distribution, and alteration of membrane glycoconjugates. studies involving flow cytometry and electron microscopy in human sperm have demonstrated alterations in lectin binding patterns throughout the capacitation process and the ar [37]. for instance, in humans, concanavalin a (con-a) binds to the anterior head region of capacitated sperm, but following the induction of the ar, this lectin binds to the equatorial region and the internal acrosome membrane. notably, con-a exhibited weak binding to the head membrane and midpiece of fresh boar spermatozoa. an elevation in mannose residues was noted in the acrosomal region of capacitated sperm, with the labeling becoming especially pronounced at the acrosomal border following the acrosome reaction. consequently, these findings illuminate the glycosylation patterns of boar spermatozoa [13]. alterations in the distribution of lectin binding sites during capacitation and the acrosome reaction have been documented in various mammalian species, although there are conflicting findings. while lee and ahuja [38] reported no significant changes in distribution patterns in mouse sperm, other researchers have observed alterations in the distribution of plasma membrane glycoproteins during capacitation in boar spermatozoa [39]. additionally, vázquez, et al. [40] identified a notable decrease in lectin binding (con-a) in capacitated and acrosome-reacted boar agriculture and food sciences research, 2024, 11(2): 228-235 231 © 2024 by the authors; licensee asian online journal publishing group spermatozoa [40]. furthermore, the distribution of n-acetylglucosamine, sialic acid, mannose, and fucose residues in the boar sperm glycocalyx undergoes changes during capacitation and the acrosome reaction [13]. 1.8. the role of lectin receptors sites in the zona pellucida of follicular and ovulated oocytes the mammalian oocyte is surrounded by a transparent, extracellular coat called the zona pellucida (zp) [41]. this extracellular matrix plays a crucial role during fertilization [42-44]. the zp restricts interspecies fertilization, is involved in the induction of the acrosome reaction in bound sperm, and assists in establishing the block to polyspermy after fertilization. in a wide variety of species studied, the zp is composed of only a few glycoproteins. it has been suggested that carbohydrate determinants of the zp glycoproteins are crucial in sperm-egg interaction [45, 46]. the most studied species in this respect is the mouse, where it has been shown that the zp consists of three glycoproteins, namely zp1, zp2, and zp3, each of which has a unique polypeptide chain and oligosaccharides [47]. it was demonstrated that sperm-egg interaction in the mouse is controlled by o-linked oligosaccharides of the zp glycoprotein, serving as sperm binding sites and the polypeptide chain as an inducer for the acrosome reaction [48, 49]. in that respect, specific sugar residues of zp o-linked oligosaccharides, e.g., α-galactosyl (gal) residues, were shown to play a role in sperm binding activity [50]. it may be assumed that the ability of specific sugar residues to function as sperm receptors largely depends on their spatial distribution and availability on the zp surface before and after fertilization, and on their species-specific biochemical nature. therefore, there is an increasing demand to expand our ability to localize and characterize specific sugar residues in the zp at the ultrastructural level and to correlate their distribution with maturational changes and fertilization. lectins, sugar-binding proteins or glycoproteins of non-immune origin, which agglutinate cells and/or precipitate glycoconjugates having saccharides of appropriate complementarity [4] have been used extensively in numerous studies to inhibit sperm-egg adhesion [51-55]. lectins have previously been used for the characterization of zp at the electron microscopic (em) levels [56-58]. it was shown by fitc-labeled lectins that mouse zp contains receptors to several lectins, particularly those that bind to gal, n-acetylgalactosaminyl (galnac), and nacetylglucosaminyl (glcnac) residues [58]. em analysis of lectin binding sites in the zp of several mammalian species [57] revealed a high density of specific lectin receptors, including rca-i and wga, at the outer regions of the zp, whereas con a receptors were sparsely distributed throughout the entire thickness of the zp. shalgi, et al. [59] demonstrated that in ovulated oocytes, most sugar residues were homogeneously distributed throughout the entire depth of the zp or located mainly in its inner portion, whereas in follicular oocytes, at all developmental stages, the sugar residues were mainly located at the outer regions of the zp, adjacent to the cumulus cells [59]. the results clearly reflect a change in both the content and distribution of sugar residues in the zp following ovulation, which may have significance in fertilization. previous studies have indicated that after hormonal stimulation for ovulation, the zona pellucida exhibits a progressive enhancement in its permeability to sperm [59-62]. other studies reported that new antigens originating from the oviduct epithelium are found on the zp after ovulation [56, 63-67]. 1.9. the effect of lectin on the physiology of oocyte maturation the developmental potential of oocytes matured in vitro is often reduced compared to those matured in vivo [68]. to enhance the bubaline oocyte maturation rate, it is essential to modify a suitable oocyte maturation medium with various supplements. in vitro matured oocytes are commonly used for the production of ivf or cloned calves. complete maturation of oocytes is critical for the developmental competence of embryos [69]. although in vitro maturation is achievable, it is associated with a loss of developmental competence, which is linked to the absence of specific proteins in oocytes cultured to metaphase ii in vitro [70]. the production of embryos in the laboratory involves three major steps: oocyte maturation, fertilization, and embryo culture. sirard, et al. [71] characterized five levels of oocyte developmental competence, including the ability to resume meiosis, to cleave upon fertilization, to develop into blastocysts, to induce pregnancy, and to generate healthy offspring. cytoplasmic changes accompanying oocyte growth include mrna transcription, protein translation, and post-translational modifications [72, 73]. oocytes are complex cells containing many organelles and molecules, each of which must be in the appropriate state for the egg to be competent to sustain subsequent embryo development [74]. despite at least 80% of bovine oocytes collected from antral follicles undergoing spontaneous nuclear maturation in culture [75] gonadotropins are often added to maturation media to induce cytoplasmic maturation, cumulus expansion, and to improve embryonic development. follicle-stimulating hormone (fsh) induces expansion of cumulus oocyte complexes in vitro [76] enhancing bovine fertilization and regulating the cleavage rate, growth, and reproductive processes of the mammalian body [77]. luteinizing hormone (lh) controls the length and sequence of the female menstrual cycle, including ovulation and ovarian production of both estrogen and progesterone [78]. additionally, it has been reported that serum is required for hormonally induced cumulus expansion of cocs, although the percentage may be as low as 0.01–5% [76]. in most cases, supraphysiological hormone concentrations are added to in vitro maturation (ivm) media, and it is not clear if these high concentrations are strictly required [79]. recently, recombinant gonadotropins have become commercially available; these are very pure sources of hormone that can allow the individual roles of fsh and lh to be investigated without the hormone cross-contamination of pituitary, serum, or urinary preparations. general mitogens, specifically lectins, can stimulate the maturation of isolated mouse cumulus oocyte complexes (cocs) [80]. lectins are carbohydrate-binding proteins or glycoproteins of nonimmune origin that have profound effects on a variety of cultured cells, including agglutination and mitogenesis [8]. they serve many different biological functions in animals, from the regulation of cell adhesion to glycoprotein synthesis and the control of protein levels in the blood. the mitogenicity of lectins is strongly associated with the stimulation of germinal vesicle breakdown (gvb) as well as cumulus expansion. funsho fagbohun and downs [81] reported that the dosage of lectin concentration and maturation rate of the cocs are directly proportional to each other. studies demonstrated that lectin doses enhance both oocyte maturation and the regulation of gene expression in oocytes. the processes of meiotic maturation and the acquisition of developmental competence determine the ability of the oocyte to undergo successful fertilization, cleavage, and embryonic development. these important steps depend upon various factors that lead to proper nuclear and cytoplasmic maturation [82]. oocyte meiotic maturation is a complex process involving germinal vesicle breakdown, chromosome condensation and segregation, formation of the metaphase plate, completion of meiosis i, extrusion of the first polar body (pb), and arrest at metaphase ii [83]. it agriculture and food sciences research, 2024, 11(2): 228-235 232 © 2024 by the authors; licensee asian online journal publishing group is well known that oocyte developmental potential is a reflection of proper cytoplasmic maturation. even though most bovine oocytes resume meiosis and progress to metaphase ii following ivm [84] cytoplasmic in vitro maturation is generally compromised, leading to lower rates of development compared to in vivo matured oocytes. lectins have been used as probes for determining the presence and physiological role of carbohydrates on cell surfaces [57, 85, 86]. binding of lectins to the plasma membrane and investing layers of mouse, hamster, rat, amphibian, ascidian, and sea urchin eggs has been documented. treatment of oocytes or sperm of some species with lectins has been shown to negatively affect fertilization, implicating oligosaccharides as important participants in gamete-gamete interaction [87-90]. furthermore, treatment of mature oocytes with lectins has been reported to stimulate activation, as manifested in the cortical granule reaction [88, 91, 92]. of particular interest to the present study is the finding reported many years ago by shida, et al. [93] that the lectin phytohemagglutinin stimulated gvb in starfish oocytes. although this are needs further research, variety of important physiological processes in germ cells could potentially be mediated by surface glycoconjugates [93]. in line with this, it will be important to identify the glycoconjugates and receptors responsible for the actions of lectins on the cumulus cell-enclosed oocyte and to determine if they are related or identical to receptors for other stimulatory ligands such as growth factors and gonadotropins. the primary objective of these studies is to comprehend the mechanisms through which the oocytecumulus cell complex is prompted to initiate maturation. while lectins cannot be considered physiological inducers of cumulus expansion and oocyte maturation, they nevertheless can serve as useful tools in elucidating the mechanism(s) involved. 1.10. the effect of lectin on the physiology of embryogenesis by influencing cell adhesion, guiding cell-cell communication, modulating intracellular signals, and affecting morphogenetic movements, lectins contribute in shaping the physiological and morphological landscape of the developing embryo. francoise poirier, et al. [94] provided the first evidence that soluble lectins may play a role in mammalian development as early as implantation. the significance of soluble lectins during early embryogenesis has been demonstrated in two other species as well [94]. in xenopus embryos, a soluble lectin with a relative molecular mass of 43 × 10^3 is first detected in fertilized eggs and later secreted into the extracellular matrix during gastrulation [95, 96]. in contrast, in chick embryos, two similarly sized lactose-binding proteins (one being l14) are detected in some migrating cells during gastrulation [97-99]. these findings suggest that interactions between lectins and associated glycoconjugates may play a crucial role in the dynamic changes of the extracellular environment during embryogenesis in these species. francoise poirier, et al. [94] found that the expression of l14 lectin is tightly regulated during several stages of embryogenesis. the striking regulation of its synthesis at the time of implantation strongly suggests that this carbohydrate-binding protein may play a key role in the attachment of the hatched blastocyst to the uterine epithelium [94]. as another example, wang, et al. [100] demonstrated the differential impacts of lectins (wheat germ agglutinin (wga), soybean agglutinin (sba), and peanut agglutinin (pna)) from three commonly used plant protein sources (wheat, soybean, and peanut meals) on embryonic development and liver cells of zebrafish. supported by cell signaling and metabolic analyses, their study showed that both wga and sba caused apoptosis, while pna stimulated cell proliferation in contrast. this study cautions against the general consideration of plant lectins as toxic without discrimination. therefore, further characterization of the distinct functions of individual lectins could be valuable for both nutrition and other potential applications [100]. 2. conclusions throughout this exploration of lectin's roles in reproductive processes reproductive processes involving the sperm, egg, and embryo, significant insights have emerged regarding their function in mammalian fertilization and oocyte maturation. modifications to the sperm glycocalyx, which occur during capacitation and the acrosome reaction, are pivotal for gamete recognition and successful fertilization, yet the intricacies of these modifications are not fully elucidated. this article has highlighted the potential of lectins to serve as a proxy in assessing the fertilizing capacity of sperm, reflecting their binding affinity to specific glycosylated regions on the sperm surface. moreover, lectins have proven instrumental in inducing oocyte maturation and cumulus expansion within mouse oocytecumulus cell complexes. this induction points to lectins' ability to stimulate specific signal transduction or metabolic pathways that are crucial for both mitogenesis and the comprehensive maturation of the oocyte-cumulus cell complex. such findings underscore the dual functionality of lectins, suggesting that they can significantly enhance our understanding of, and capabilities in, reproductive biology. by leveraging lectins to manipulate and study reproductive processes, we gain a deeper understanding of cellular interactions and developmental mechanisms essential for successful fertilization and embryo development. this comprehensive approach not only clarifies the biological functions of lectins but also highlights their potential therapeutic applications in addressing fertility issues and optimizing in vitro fertilization techniques. abbreviations map: mitogen activated protein; git: gastro intestine tract; con a: concanavalin a; apl: asparagus pea lectin; zp: zona pellucida; gal: a-galactosyl; em: electron microscopic; galnac: gal, n-acetylgalactosaminyl; glcnac: nacetylglucosaminyl; fsh: follicle stimulating hormone; lh: luteinizing hormone; ivm: in vitro maturation; cocs: cumulus oocyte complexes; gvb: germinal vesicle breakdown; map: mitogen activated protein kinase; wga: wheat germ agglutinin; sba: soybean agglutinin; pna: peanut agglutinin; ar: acrosome reaction; mrna: messenger rna; fitc: fluorescein isothiocyanate; rca-i: ricinus communis agglutinin i; pha-m: phytohemagglutinin m. references [1] s. s. komath, m. kavitha, and m. j. swamy, "beyond carbohydrate binding: new directions in plant lectin research," organic & biomolecular chemistry, vol. 4, no. 6, pp. 973-988, 2006. https://doi.org/10.1039/b515446d [2] j. rini, "lectin structure," annual review of biophysics and biomolecular structure, vol. 24, pp. 551-577, 1995. https://doi.org/10.1039/b515446d agriculture and food sciences research, 2024, 11(2): 228-235 233 © 2024 by the authors; licensee asian online journal publishing group [3] j. rocca, "lectins as next generation mucoadhesives for specific tarheting of the gastrointestinal tract," gastroinstestinal target drug develop, vol. 4, pp. 12-19, 2004. 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[100] k. wang et al., "differential apoptotic and mitogenic effects of lectins in zebrafish," frontiers in endocrinology, vol. 10, p. 356, 2019. https://doi.org/10.3389/fendo.2019.00356 asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1002/(sici)1098-2795(199802)49:2%3c119::aid-mrd3%3e3.3.co;2-m https://doi.org/10.1016/s0074-7696(08)60939-0 https://doi.org/10.1002/mrd.1120170406 https://doi.org/10.1038/257321a0 https://doi.org/10.1016/0014-4827(74)90356-5 https://doi.org/10.1530/jrf.0.0470299 https://doi.org/10.1016/s0021-9258(17)40653-3 https://doi.org/10.1016/0014-4827(72)90081-x https://doi.org/10.1016/0014-4827(72)90081-x https://doi.org/10.1242/dev.115.1.143 https://doi.org/10.1016/0012-1606(88)90071-1 https://doi.org/10.1083/jcb.97.6.1875 https://doi.org/10.1242/dev.107.4.909 https://doi.org/10.3389/fendo.2019.00356 100 © 2025 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 2, 100-109, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i2.7312 © 2025 by the authors; licensee asian online journal publishing group effects of additives and ensiling period on groundnut shell silage and in sacco rumen degradability characteristics abba ibrahim abubakar1 abubakar masud2 muktar umar muktar3 bilkisu ahmad4 tukur sani tajo5 sani ibrahim6 mahmud suraj abdullahi7 ( corresponding author) 1,2,3,4department of animal health and production technology, federal polytechnic, kabo, kano state, nigeria. 1email: abbamusa35@yahoo.com 2email: masudabk79@gmail.com 3email: muktarumarmuktar91@gmail.com 4email: ummanabba122@gmail.com 5department of science laboratory technology, federal polytechnic, kabo, kano, state, nigeria. 5email: sanitajo88@gmail.com 6department of animal health technology, audu bako college of agriculture dambatta, kano state, nigeria. 6email: ibrahimsanirogo@gmail.com 7department of animal science, bayero university kano, nigeria. 7email: mahmudsuraj70@gmail.com abstract this study evaluated the effects of additives and ensiling periods on the chemical composition and in sacco rumen degradability of groundnut shell (gns). in phase one, a 3 × 5 factorial crd was used with three ensiling periods (3rd, 5th, and 7th weeks) and five additives (control, urea, yeast, molasses, and naoh). in phase two, in sacco degradability was determined using a 3 × 3 switchover design. chemical composition was analyzed following aoac procedures, and data were subjected to anova. ph decreased with longer ensiling periods, reaching desirable levels (3.8– 5.0). additives, period, and their interactions significantly (p<0.05) influenced proximate constituents and fibre fractions but not dry matter (dm) degradability. urea and yeast increased crude protein (8.97% and 7.15%, respectively), while dry matter (dm) and crude fibre decreased after ensiling with additives. crude protein rose and crude fibre declined with longer ensiling. the highest crude protein (9.58%) was from urea-treated gns at 7 weeks. after 48 hours of incubation, dm disappearance peaked at 7 weeks for all additives, with urea and yeast yielding the highest potential degradability (100%). it is concluded that ureaor yeast-treated gns ensiled for 7 weeks improves nutrient quality and degradability, presenting a practical feed option for farm animals. keywords: additives, ensiling, degradability, groundnut shell, silage, in sacco, rumen. citation | abubakar, a. i., masud, a., muktar, m. u., ahmad, b., tajo, t. s., ibrahim, s., & abdullahi, m. s. (2025). effects of additives and ensiling period on groundnut shell silage and in sacco rumen degradability characteristics. agriculture and food sciences research, 12(2), 100–109. 10.20448/aesr.v12i2.7312 history: received: 28 march 2025 revised: 8 august 2025 accepted: 19 august 2025 published: 26 august 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 101 2. materials and methods ................................................................................................................................................................. 101 3. results and discussion ................................................................................................................................................................. 102 4. conclusion ....................................................................................................................................................................................... 108 references ............................................................................................................................................................................................ 108 mailto:abbamusa35@yahoo.com mailto:masudabk79@gmail.com mailto:muktarumarmuktar91@gmail.com mailto:ummanabba122@gmail.com mailto:sanitajo88@gmail.com mailto:ibrahimsanirogo@gmail.com mailto:mahmudsuraj70@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i2.7312 https://orcid.org/0000-0001-9763-4795 https://orcid.org/0009-0009-4312-1156 https://orcid.org/0009-0008-6397-3877 https://orcid.org/0009-0004-5235-6348 https://orcid.org/0009-0001-2792-863x https://orcid.org/0009-0001-0514-4172 https://orcid.org/0009-0000-6132-4680 agriculture and food sciences research, 2025, 12(2): 100-109 101 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this work provides novel evidence that ureaor yeast-treated groundnut shells, when ensiled for seven weeks, significantly improve crude protein content and rumen degradability. it establishes an efficient, low-cost method or technique for upgrading an underutilized agroindustrial by-product into a high-quality ruminant feed. 1. introduction processing crops to obtain desired products typically generates large amounts of residues as by-products. in ruminant nutrition, many of these residues are valuable because they supply nutrients to animals, especially during periods of feed scarcity [1]. one such crop residue is the groundnut shell (gns), which is produced in large quantities in many communities. an average of 1018 kg/ha of groundnut shell (gns) is produced annually in nigeria [2]. with the exception of a small quantity used as fuel or mulching material, most are discarded as waste. it is common practice in nigeria to either burn or leave them on the farm to decompose. burning gns contributes to global warming; therefore, there is a need to convert it into a useful feed resource [3]. however, one of the disadvantages of crop residues as feed is generally low palatability, low degradability, and low nutrient content [4]. improved management of crop residues enables their efficient utilization as valuable feed resources. one important way of achieving protein self-sufficiency for the growing nigerian population is through enhanced utilization of crop residues [5]. improving the digestibility of poor-quality forages using physical, biological, or chemical methods contributes greatly to this goal [6]. physical, biological, and chemical treatments disrupt the bonds between cellulose, hemicellulose, and lignin in plant cell walls. this disruption increases the surface area available for microbial attachment, thereby improving digestibility [7, 8]. the in sacco technique of feed evaluation was developed to enable the routine assessment of different feeds. rumen degradability is commonly determined using this method, which involves incubating feed samples in nylon bags placed in the rumen. the in sacco technique is a robust tool widely applied in ruminant nutrition studies [9]. it is particularly valuable for describing the degradability characteristics of proteins and other feed fractions in forages, as well as in rumen simulation studies [10]. the in-situ method is the most frequently used approach for estimating the degradability of dry matter (dm), organic matter (om), protein, fiber, and other nutrients [11]. the rate and extent of dm fermentation in the rumen are critical factors influencing nutrient availability to ruminants [12]. 2. materials and methods 2.1. study area the experiment was conducted at the laboratory and research farm of the department of animal science, bayero university, kano. the facilities are located at the new site campus of the university, about 13 km from kano metropolis, in ungogo local government area of kano state. kano state lies between longitude 9°30′ and 12°30′ east, and latitude 8°42′ and 9°30′ north, within the semi-arid region of northern nigeria [13]. the state occupies a total land area of 24,400 km² [14] with mean annual rainfall ranging from 600 to 1000 mm [15]. the climate is characterized by 4–8 months of dry season, with average maximum and minimum temperatures of 33.0°c and 15.2°c, respectively. 2.2. experimental design the experiment was laid out in a 3 x 5 factorial arrangement in a completely randomized design, with five (5) different additives (control, molasses, sodium hydroxide, urea, and yeast) and three ensiling periods (3rd, 5th, and 7th week). in sacco degradability was laid out in a 3 x 3 switch-over design. 2.3. ensiling procedure before ensiling, 31.5 kg of groundnut shells were ground into smaller pieces of about 1 cm using a medium crusher with a 2 mm size. five combinations of groundnut shell, treated with molasses, urea, laboratory yeast, sodium hydroxide, and untreated groundnut shell were used. forty-five bottles were employed, each filled with either treated or untreated groundnut shell, compressed using a wooden stick to remove air and tightly closed. grease was applied at the brim to ensure an airtight condition after filling with silage materials and compression. each bottle was replicated three times and divided into three ensilage periods of 3rd, 5th, and 7th weeks, stored under shade. at the end of each ensilage period, samples were sun-dried and prepared for chemical analysis and degradability studies. an untreated (control) groundnut shell was prepared by adding 3000 ml of distilled water to 1 kg of groundnut shell and ensiling it. molasses at 5% of groundnut shells was diluted in 3000 ml of water (50 ml of molasses was diluted in 3 liters of water to treat 1 kg of gns) and mixed with groundnut shells. good silages have been reported when molasses is applied at 3-5% [16]. urea at a level of 2% of groundnut shell was dissolved in 3000 ml of distilled water (20 g of urea was dissolved in 3 liters of water to treat 1 kg of gns) and mixed with groundnut shell as described in the procedure roy and rangnekar [17]. yeast at 1% of groundnut shell was dissolved in 3000 ml of distilled water (10 g of yeast was dissolved in 3 liters of water to treat 1 kg of gns) and mixed with groundnut shell as described in the procedure of gattass et al. [18]. sodium hydroxide at 1% of groundnut shell was dissolved in 3000 ml of distilled water (10 g of naoh was dissolved in 3 liters of water to treat 1 kg of gns) and thoroughly mixed with groundnut shell. 2.4. quality determination of ensiled groundnut shell ensiled materials were opened at the 3rd, 5th, and 7th weeks after ensiling. at opening, ph, colour, and aroma were determined, and quality characteristics were assessed according to babayemi et al. [19]. immediately after agriculture and food sciences research, 2025, 12(2): 100-109 102 © 2025 by the authors; licensee asian online journal publishing group opening, a laboratory thermometer was inserted to determine the temperature. the ph was measured by adding 100 ml of distilled water to 25 g of each treatment in a beaker, and a ph meter was used to determine the ph. colour and aroma were assessed by descriptive statistics. the conditions were scored for aroma and colour by three independent panelists on a scale of 1-4 (table 1). table 1. description of colour and aroma rating used as indices of silage quality. rating colour aroma 1 dark or deep brown putrid or rancid 2 light brown pleasant 3 pale yellow sweet 4 yellowish green very sweet source: muhammad, et al. [20]. 2.5. in sacco degradability study degradability study of groundnut shell was carried out using three fistulated kano brown bucks with an average age of 48 months, according to the nylon bag technique described by ørskov et al. [21]. the animals were given a ten-day period for adaptation to the new feed and housing conditions prior to the suspension of bags. samples were shade-dried and ground through a 2 mm sieve before rumen incubation. duplicate samples of about 5 g each were placed in nylon bags (bag size 80 mm x 140 mm; pore size 45 μm) and suspended in the rumen of three fistulated bucks for 8, 12, 18, 24, 36, and 48 hours as outlined by ørskov and mcdonald [22]. after removal from the rumen, the bags were immediately dipped into cold water to stop microbial activity, then washed under running water to remove rumen contents from the outside of the bags. they were subsequently rinsed for 30 minutes to ensure thorough cleaning. zero-hour samples were prepared by washing the bags containing test samples for 30 minutes without rumen incubation. the residues remaining in the bags were ovendried at 60°c for 48 hours, following the procedure of nocek [23]. the bags were then cooled, weighed, and the data obtained were used to calculate the percentage dry matter loss. the results from the in sacco study were fitted to the model of ørskov and mcdonald [22] (p = a + b (1 – e^(-ct))) to determine the degradation characteristics of the incubated samples. where, p = potential degradability after time ‘t’. a = water soluble fraction (zero hour). b = insoluble but degradable fraction after time ‘t’. c = rate of degradation of the slowly degradable fraction b. t = incubation length, i.e. 8, 12, 18, 24, 36, and 48 hrs. e = exponential. 2.6. statistical analysis descriptive statistics were used to assess silage quality. for in sacco data, analysis of variance (anova) was performed using the general linear model procedure of spss (spss version 21.0). differences among means were separated using lsd at a 95% probability level. 2.7. chemical composition of groundnut shell samples ensiled (gns) were analyzed for proximate composition (cp, cf, ee, dm, nitrogen-free extract and ash) according to procedures of aoac (association of official analytical chemists) [24]. neutral detergent fibre (ndf) and acid detergent fibre (adf) were determined according to procedures outlined by van soest and robertson [25]. 3. results and discussion 3.1. results 3.1.1. effects of additives on groundnut shell silage characteristics the result of additives on gns characteristics is shown in table 2. the ph values ranged from 5.63 – 3.87 and there was a significant (p<0.05) difference among the treatments. the silage made with naoh had a higher ph value (5.63), while the lowest value was recorded in yeast treatments. the silage treated with additives had ph values less than 6.0. table 2. effects of additives on groundnut shell silage characteristics. parameter treatment ph colour aroma untreated gns 4.67b light brown pleasant urea+gns 4.55b pale yellow sweet yeast+gns 3.87c pale yellow sweet molasses+gns 4.55b pale yellow sweet naoh+gns 5.63a light brown pleasant sem 0.043 note: means with different superscripts (a, b, c) within the same column are significantly different (p<0.05). 3.1.2. effect of ensiling period on groundnut shell silage characteristics the results of the ensiling period on gns silage characteristics are presented in table 3. the ph values of silages were significantly affected (p<0.05) by the ensiling period. the ph values ranged from 4.39 at 7 weeks, 4.58 at 5 weeks, and 4.98 at 3 weeks. in terms of colour and aroma, all silage produced at different periods was light brown with a pleasant aroma at 3 weeks, and pale yellow with a sweet aroma at 5 and 7 weeks, respectively. agriculture and food sciences research, 2025, 12(2): 100-109 103 © 2025 by the authors; licensee asian online journal publishing group table 3. effects of ensiling period on groundnut shell silage characteristics. parameter period ph colour aroma week 3 4.98a light brown pleasant week 5 4.58b pale yellow sweet week 7 4.39c pale yellow sweet sem 0.033 note: means with different superscripts (a, b, c) within the same column are significantly different (p<0.05). 3.1.3. interaction effect of additives and ensiling period on groundnut shell silage characteristics results of additives and the ensiling period on gns silage characteristics are shown in table 4. the result revealed that urea and yeast gns treatment had a pale yellow colour with a sweet aroma. control and yeast treatment had a combination of both pale yellow and light brown colours with a sweet and pleasant aroma in all the periods of ensiling. meanwhile, sodium hydroxide had a light brown colour with a pleasant aroma and a ph range of 3.62–5.21 across all treatments and periods of ensiling. table 4. interaction effects between additives and ensiling period of groundnut shell (gns) silage characteristic. parameters treatments period ph colour aroma untreated gns 3 4.66 light brown pleasant 5 4.73 light brown pleasant 7 4.62 pale yellow sweet urea+gns 3 4.59 pale yellow sweet 5 4.55 pale yellow sweet 7 4.50 pale yellow sweet yeast+gns 3 4.16 pale yellow sweet 5 3.38 pale yellow sweet 7 3.62 pale yellow sweet molasses+gns 3 4.72 light brown pleasant 5 4.72 pale yellow sweet 7 4.20 pale yellow sweet naoh+gns 3 5.21 light brown pleasant 5 5.10 light brown pleasant 7 5.03 light brown pleasant sem 0.075 t x p * note: t x p = treatment x period interaction. 3.1.4. effect of additives on chemical composition of groundnut shell silage the results of additives on the chemical composition of gns silage are shown in table 5. the result revealed that there was a significant (p<0.05) difference among all the parameters evaluated. percent dry matter (dm) of silage made from gns (control) has the highest value of 92.60%, while the lowest value of 90.77% was recorded from the silage made from gns (yeast). treatment (urea) had the highest crude protein (cp) content at 8.97%, and the lowest cp at 4.33% was recorded in the control, followed by naoh. crude fibre (cf) was higher in the control at 51.42%, while the lowest value of 33.71% was recorded from silage made from gns (urea). the treatment with yeast and molasses had the highest value of nfe, and the lowest value was recorded in the control. the lowest value of acid detergent fibre (adf) was recorded in the urea treatment, while the highest adf was in the control. the lowest neutral detergent fibre (ndf) was recorded in the urea treatment, and the highest ndf was in the control. table 5. effects of additives on chemical composition (%) of groundnut shell silage. parameters treatment dm ash cp cf ee nfe adf ndf untreated gns 92.60a 4.78b 4.33c 51.42a 0.99d 38.45d 57.71a 75.04a urea+ gns 90.89b 5.45a 8.97a 33.71d 3.41c 39.36c 41.25d 65.19d yeast+ gns 90.77bc 4.59c 7.15b 39.49c 4.88a 43.90a 52.09b 71.39b molases+gns 90.86c 3.91d 7.01b 46.88b 3.71b 42.58b 46.19 b 72.29b naoh+gns 90.81bc 3.91d 7.00b 47.88b 3.64b 38.47d 43.23b 67.24c sem 0.032 0.37 0.210 0.157 0.063 0.204 0.427 0.431 note: means with different superscripts (a, b, c, d) within the same column are significantly different (p<0.05). dm: dry matter, cp: crude protein, cf: crude fibre, ee: ether extract, nfe: nitrogen free extract, adf: acid detergent fibre, ndf: neutral detergent fibre. 3.1.5. effect of ensiling period on chemical composition of groundnut shell silage the effects of the ensiling period on chemical compositions of gns silage are shown in table 6. the parameters evaluated are dm, ash, cp, cf, ee, nfe, adf, and ndf, and they were all significant (p<0.05) at all the ensiling periods. the dm percentage was highest at the 3rd week of the ensiling period (91.35%) compared to the 5th and 7th weeks, respectively. ash content was highest at the 7th week (5.26%) and lowest at the 3rd week (3.84%). crude protein (cp) content was highest at the 7th week (7.35%), while the lowest cp content was observed at the 3rd week (6.50%). crude fiber (cf) content was highest at the 3rd week (47.36%) and lowest at the 7th week of the ensiling period (40.26%). agriculture and food sciences research, 2025, 12(2): 100-109 104 © 2025 by the authors; licensee asian online journal publishing group table 6. effects of ensiling period on chemical composition (%) of gns-treated silage. parameter period dm ash cp cf ee nfe adf ndf week 3 91.35a 3.84a 6.50b 47.36a 2.78c 37.84b 72.06a 67.81a week 5 91.24a 4.52b 6.82b 43.41b 3.03b 41.95a 70.88b 74.80b week 7 91.06b 5.26c 7.35a 40.26c 4.17a 41.87a 68.06b 71.46c sem 0.025 0.028 0.163 0.122 0.49 0.158 0.334 1.040 note: means with different superscripts (a, b, c) within the same column are significantly different (p<0.05). dm: dry matter, cp: crude protein, cf: crude fibre, ee: ether extract, nfe: nitrogen-free extract, adf: acid detergent fibre, ndf: neutral detergent fibre. 3.1.6. interaction between additives and ensiling period on chemical composition of groundnut shell silage the effect of interaction between additives and the ensiling period on the chemical composition of gns silage is shown in table 7. additives and the ensiling period significantly affect the chemical composition of silage. the percentage of dry matter (dm) in silage made without additives (control) at the 3rd week of the ensiling period was highest at 92.74%, while at the 7th week, yeast-treated silage had the lowest dm content at 90.57%. the percentage of ash shows an increasing trend as the ensiling period progresses, with the highest value recorded at the control at week 7 and the lowest at molasses at week 3. the crude protein (cp) content was highest in urea-treated silage at the 7th week, reaching 9.58%, and lowest in the control at 3.50% during the same period. the lowest crude fiber (cf) interaction was observed in urea at week 7, at 29.14%. the highest ether extract was found in yeast-treated silage at week 7, at 5.67%, while the lowest was in the control at week 3, at 0.45%. the nitrogen-free extract (nfe) was highest in molasses at week 7, at 45.94%, and lowest in the control at week 7, at 36.28%. these findings highlight the influence of different additives and the duration of ensiling on the chemical composition of silage, which is crucial for optimizing feed quality and nutritional value. the interaction between additives and ensiling period on the chemical composition of gns was significantly (p<0.05) different for all parameters evaluated. 7. interaction effects between additives and ensiling period on chemical composition (%) of groundnut shell silage. parameters treatment period dm ash cp cf ee nfe adf ndf untreated gns 3 92.74 3.72 3.50 54.00 0.45 37.63 59.42 75.19 5 92.56 4.33 4.20 51.19 0.62 40.29 57.93 76.40 7 92.52 6.34 5.29 49.05 1.89 36.28 55.77 73.54 urea +gns 3 90.98 4.61 8.44 39.37 2.28 37.42 59.71 69.14 5 90.94 5.61 8.88 32.66 3.64 40.21 32.41 64.27 7 90.75 6.13 9.58 29.14 4.31 41.58 31.60 62.18 yeast +gns 3 90.81 4.18 7.24 43.26 4.18 42.03 57.23 76.39 5 90.92 4.43 7.38 38.15 4.78 45.29 45.37 73.64 7 90.57 5.15 8.02 37.08 5.67 44.30 53.66 68.35 molasses + gns 3 90.96 3.34 6.34 50.07 3.15 36.92 47.17 72.66 5 90.92 4.12 6.72 47.55 3.49 44.48 46.09 73.81 7 90.69 4.33 7.07 43.04 4.48 45.94 45.32 69.88 naoh + gns 3 90.93 4.21 7.18 47.55 3.95 38.00 44.41 67.78 5 90.92 4.32 7.08 43.04 4.48 41.68 41.93 65.51 7 90.76 4.46 7.28 42.21 5.24 38.03 53.63 68.08 sem 0.055 0.633 0.364 0.273 0.109 0.354 0.740 0.747 t x p * * * * * * * * note: t= treatment x period interaction, 3= weeks 3, 5= weeks 5, 7= weeks 7. 3.1.7. dry matter (dm) disappearance (%) of untreated groundnut shell the results of dry matter disappearance of untreated gns at different ensiling period is presented in figure 1. the untreated gns disappearance curve revealed that the 7th week silage had the greatest disappearance values during 0-48 hours of incubation, followed by the 5th week, and finally, the least was the 3rd week silage. figure 1. dry matter disappearance (%) of untreated groundnut shell. agriculture and food sciences research, 2025, 12(2): 100-109 105 © 2025 by the authors; licensee asian online journal publishing group 3.1.8. dry matter (dm) disappearance (%) of urea treated groundnut shell the results of dry matter disappearance of urea treated gns are shown in figure 2. the urea dry matter curve revealed that gns treated with urea and ensiled at 7th week produced slightly higher dm disappearance values after a 48-hour incubation period, followed by the 5th week, and the least was at the 3rd week ensiling period. figure 2. dry matter disappearance (%) of groundnut shell treated with urea. 3.1.9. dry matter disappearances (%) of groundnut shell treated with yeast figure 3 the analysis revealed that gns treated with yeast at the 7th week exhibited the highest dry matter (dm) disappearance, followed by the 5th week, while the lowest was recorded at the 3rd week ensiling period after a 48-hour incubation period. figure 3. dry matter disappearance (%) of groundnut shell treated with yeast. 3.1.10. dry matter disappearance (dm) of groundnut shell treated with molasses the results of dry matter disappearance of molasses treated gns at different ensiling period are shown in figure 4. the dry matter disappearance curve revealed that molasses-treated gns at 7th weeks produced the greatest dm disappearance after 48 hours, followed by 5th weeks, and the least was at three weeks after the 48hour incubation period. agriculture and food sciences research, 2025, 12(2): 100-109 106 © 2025 by the authors; licensee asian online journal publishing group figure 4. dry matter disappearance (%) of groundnut shell treated with molasses. 3.1.11. dry matter disappearance (dm) of groundnut shell treated with naoh the results of dry matter disappearance of naoh treated gns at different ensiling period are shown in figure 5. the dry matter disappearance curve revealed that naoh-treated gns at 7th weeks produced the greatest dm disappearance after 48 hours, followed by 5th weeks, and the least was at three weeks after the 48-hour incubation period. figure 5. dry matter disappearance (%) of groundnut shell treated with naoh. 3.1.12. effect of additives on dry matter degradability of groundnut shell silage the results of the rumen degradation characteristics of treated and untreated gns silage are shown in table 8. groundnut shell shows no significant difference (p>0.05) between treatments. the results indicate that yeast has a higher quickly soluble fraction ‘a’ (45.55), while urea has a higher ‘b’ slowly degradable fraction (328.79). the degradation rate constant ‘c’ was higher in the control (0.048) and lower in yeast and molasses (0.002). the highest potential degradability (pd) was recorded in yeast (100%), and the lowest was in naoh (88.61%). the highest effective degradability (ed2, ed5, and ed8) was recorded in molasses (67.50%, 53.17%, and 48.81%, respectively), and the lowest was obtained in the control (54.07%, 44.78%, and 41.58%, respectively). table 8. effect of additives on dry matter degradation characteristics of groundnut shell silage. parameters treatments a b c a b pd ed2 ed5 ed8 untreated gns 29.27 189.98 0.048 28.77 66.24 95.03 54.07 47.54 41.58 urea + gns 31.33 328.79 0.041 32.43 66.70 99.14 59.88 44.78 46.07 yeast + gns 45.55 271.32 0.041 30.18 69.81 100.00 57.74 49.58 46.73 molasses+ gns 23.58 286.82 0.002 35.42 54.61 90.67 67.50 53.17 48.81 naoh + gns 9.52 295.19 0.002 30.92 57.71 88.61 59.70 47.64 44.24 sem 15.76 123.17 0.028 1.379 4.453 4.267 1.99 3.05 3.22 note: means with different superscripts within the same column are significantly different (p<0.05). aquickly soluble fraction; bslowly degradable fraction; cdegradation rate constant (fraction/hour); awashing lost, brumen degradability fraction, pdpotential degradability; edeffective degradability (%). agriculture and food sciences research, 2025, 12(2): 100-109 107 © 2025 by the authors; licensee asian online journal publishing group 3.1.13. effect of ensiling period on dry matter (dm) degradation characteristics of groundnut shell silage the results of the effects of the ensiling period on dm degradation characteristics are presented in table 9. the results revealed that no significant difference was observed for all the parameters evaluated. from the results obtained, period 7 had a higher value of the ‘a’ quickly soluble fraction; the ‘b’ slowly degradable fraction was higher at period 5, while the ‘c’ degradation rate constant was higher at period 3. the potential degradability (pd) obtained at the 7th week was higher (99.15%) compared to the 3rd and 5th weeks (92.25% and 92.67%, respectively). table 9. effect of ensiling period on dry matter (dm) degradation characteristics of groundnut shell silage. period parameters a b c a b pd ed2 ed5 ed8 week 3 6.81 230.46 0.054 33.53 58.55 92.25 56.64 44.77 41.04 week 5 43.68 400.94 0.003 29.13 63.32 92.67 60.87 50.17 46.96 week 7 33.18 311.85 0.026 32.00 67.18 99.15 61.83 48.72 44.87 sem 12.204 95.413 0.022 1.068 3.449 3.305 1.546 2.360 2.498 note: means with different superscripts within the same column are significantly different (p<0.05). aquickly soluble fraction; bslowly degradable fraction; cdegradation rate constant (fraction/hour); awashing lost, brumen degradability fraction, pdpotential degradability; edeffective degradability (%); 3.1.14. interaction effect between additives and ensiling period on dry matter degradation characteristics of groundnut shell silage table 10 presents the interaction effects between additives and ensiling period on the dry matter (dm) degradation characteristics of groundnut shell silage. the results show that neither additives nor the ensiling period significantly affected parameters a, b, or c across treatments. however, ureaand yeast-treated silages at 7 weeks generally exhibited higher potential degradability (pd = 100%) compared to other treatments, while molasses-treated silage showed the highest effective degradability (ed2, ed5, and ed8). these findings suggest that treatment type and storage duration influence degradability patterns, with urea and yeast improving potential degradability and molasses improving effective degradability. table 10. interaction effect between additives and ensiling period on dry matter degradation characteristics of groundnut shell silage treatment period a b c a b pd ed2 ed5 ed8 untreated gns 3 30.00 348.59 0.001 26.37 73.63 100 54.08 40.03 36.37 5 36.67 113.04 0.006 32.77 52.33 85.10 53.17 43.86 41.20 7 21.15 108.31 0.125 27.23 72.77 100 54.97 49.53 47.17 urea + gns 3 50.19 164.41 0.141 22.07 66.67 100 51.53 43.73 40.97 5 39.39 323.71 0.002 29.33 70.47 100 63.07 49.33 45.70 7 39.36 397.48 0.001 27.10 72.90 100 64.50 49.87 46.07 yeast +gns 3 35.31 229.84 0.003 38.43 61.57 100 63.33 47.43 43.07 5 31.27 357.04 0.002 22.07 75.34 100 54.87 41.13 37.46 7 28.00 399.46 0.002 36.80 63.20 100 61.47 45.06 39.70 molasses + gns 3 10.23 162.67 0.119 34.53 49.83 85.33 59.23 51.63 47.93 5 41.16 244.76 0.003 36.87 48.73 86.33 70.50 53.93 49.50 7 39.80 453.04 0.002 34.87 65.27 100 72.76 53.97 49.00 naoh + gns 3 29.15 246.81 0.006 34.37 41.63 75.93 55.03 41.00 36.87 5 69.93 966.19 0.005 24.40 69.73 94.13 62.73 62.60 60.93 7 37.58 200.95 0.002 34.00 61.77 95.76 55.47 45.17 42.40 sem 27.28 213.35 0.05 2.39 7.71 7.39 3.46 5.28 5.58 note: means with different superscripts within the same column are significantly different (p<0.05). aquickly soluble fraction; bslowly degradable fraction; cdegradation rate constant (fraction/hour); awashing lost, brumen degradability fraction, pdpotential degradability; edeffective degradability (%). 3.2. discussion ph is widely used as a simple indicator of silage quality, with well-fermented silages typically exhibiting low ph values. kung and shaver [26] reported that tropical grass and legume silages generally have ph values ranging between 4.3 and 4.7. in this study, the ph of groundnut shell (gns) silage decreased with increasing ensiling duration, reflecting progressive fermentation. sodium hydroxide treated silage maintained a relatively higher ph (5.21), which agrees with henderson et al. [27]. in contrast, urea-, yeast-, and molasses-treated silages had lower ph values (3.62–5.21), indicating effective fermentation and good preservation quality. the colour and aroma of gns silage further supported these findings. all silages appeared pale yellow to light brown, with pleasant or sweet odours. according to kung and shaver [26] a sweet aroma is an indicator of wellfermented silage, while oduguwa et al. [28] emphasized that silage resembling the original forage colour and having a pleasant smell reflects desirable fermentation. these observations suggest that additives contributed positively to the physical quality of gns silage. proximate composition results revealed slight variations across treatments, which may be attributed to differences in additives, crop variety, cultivation methods, and environmental factors, as previously noted by larbi, et al. [29]. untreated silages recorded higher dry matter (dm) content than treated silages, possibly due to reduced fermentation losses [30]. urea and yeast treatments resulted in significantly higher crude protein (cp) contents, consistent with earlier studies demonstrating that ammoniation enhances protein levels in crop residues [31, 32]. in contrast, crude fibre contents decreased with urea, yeast, molasses, and naoh treatments, supporting the view that these additives disrupt lignocellulosic bonds, thereby improving digestibility [33]. the fibre fractions (adf and ndf) of gns silage also declined with additive treatment and longer ensiling periods. this observation is in agreement with tadesse et al. [34], who reported reductions in fibre fractions of ensiled crop residues following chemical and biological treatment. such reductions suggest improved availability of nutrients for microbial fermentation in the rumen. agriculture and food sciences research, 2025, 12(2): 100-109 108 © 2025 by the authors; licensee asian online journal publishing group the in sacco degradability trial demonstrated that both the ensiling period and additives influenced the degradability characteristics of gns. dry matter (dm) disappearance rates increased with longer incubation times, with the highest degradability observed after 48 hours of rumen incubation. ureaand yeast-treated silages ensiled for 7 weeks showed markedly higher potential degradability (100%), indicating that these treatments improved rumen breakdown of the silage. molasses-treated silages, however, recorded the highest effective degradability across different passage rates, suggesting that molasses enhanced the readily fermentable fraction of the silage. these findings are consistent with reports by ørskov and mcdonald [22] who highlighted the usefulness of the in sacco method in estimating degradability parameters. similar improvements in degradability following urea treatment have also been reported in crop residues such as maize stover and wheat straw [35]. yeast supplementation has likewise been associated with increased microbial activity and improved fibre utilization [36]. the present results confirm that biological and chemical additives can significantly enhance the degradability of groundnut shell, a by-product that is otherwise poorly utilized in animal feeding. overall, the study demonstrates that both the ensiling period and additive type influence the fermentation quality, proximate composition, and degradability characteristics of gns silage. urea and yeast were most effective in improving crude protein and potential degradability, particularly after 7 weeks of ensiling, while molasses improved effective degradability. these findings suggest that groundnut shell, when properly treated and ensiled, can serve as a valuable feed resource for ruminant production. 4. conclusion this study demonstrated that both additive type and ensiling duration significantly influence the fermentation quality, proximate composition, and degradability of groundnut shell silage. urea and yeast treatments produced the highest crude protein contents and markedly improved potential degradability, particularly after seven weeks of ensiling. molasses treatment enhanced effective degradability, while sodium hydroxide maintained higher ph values. overall, the results confirm that groundnut shell, a widely available agricultural by-product, can be transformed into a valuable ruminant feed resource when properly ensiled with suitable additives. 4.1. recommendations based on the results and drawbacks of this research work, the following recommendations are made: silage made from urea or yeast ensiled at the 7th week period is recommended for feeding farm animals. further investigation needs to be conducted using feeding trials with groundnut shell ensiled with urea or yeast. references [1] a. zarah, b. bello, i. mohammed, and f. abbator, "effect of agro-industrial by-products and crop residues based concentrate on the performance of balami lambs," journal of agriculture, vol. 2, pp. 313-315, 2013. 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[36] k. phesatcha and m. wanapat, "study on ruminal degradability of local feeds using the nylon bag technique," khon kaen agricultural journal, vol. 40, no. supplement 2, pp. 123–127, 2012. asian online journal publishing group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. any queries should be directed to the corresponding author of the article. https://doi.org/10.1016/j.anifeedsci.2006.05.021 https://doi.org/10.1017/s0021859698005590 170 © 2024 by the authors; licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 170-174, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6238 © 2024 by the authors; licensee asian online journal publishing group performance of juvenile pangasianodon hypophthalmus cultivated in a recirculating aquaculture system with tanks of different colors túlio, pacheco boaventura1 fábio, aremil costa santos2 pedro, paulo cortezzi pedras3 ronald kennedy luz4 ( corresponding author) 1,2,3,4aquaculture laboratory, department of animal science, school of veterinary medicine, federal university of minas gerais – ufmg, avenida antônio carlos, 6627, cep 31.270-901, belo horizonte, mg, brazil. 1email: tuliopb1@hotmail.com 2email: fabioaremil@gmail.com 3email: pedrocortezzi@hotmail.com 4email: luzrk@yahoo.com abstract the tanks used in rass can be manufactured in any color. ras tank color can have a great influence on fish, with direct impacts on food capture and, consequently, performance and survival. this study investigated the influence of different tank colors on the performance and coloration of pangasianodon hypophthalmus. juvenile were cultivated in tanks of different colors (white, blue, and black) for 60 days.one hundred and forty-four juvenile p. hypophthalmus, weighing 2.18±0.5 g and measuring 6.76±0.17 cm in total length, were distributed among three rass. each ras was equipped with a 200-liter rectangular filter, with mechanical and biological filter, a heating and water pumping system and four 30-liter tanks covered with colored adhesive (white, blue or black) that were filled with 28 liters of water. therefore, each ras was considered a color treatment (white, blue or black) with four replicates (tanks), which had a density of 0.43 juveniles per liter (12 juveniles in each replicate). juveniles cultivated in darker tanks exhibited darker coloration. juveniles cultivated in blue tanks showed higher final weight (fw), total length (tl), and weight gain (wg) and lower feed conversion ratio (fcr). juveniles cultivated in white tanks had lower tl and higher fcr. in conclusion, p. hypophthalmus juveniles performed better when cultivated in blue tanks. keywords: aquaculture, catfish, fish, panga. citation | boaventura, t. p., santos, f. a. c., pedras, p. p. c., & luz, r. k. (2024). performance of juvenile pangasianodon hypophthalmus cultivated in a recirculating aquaculture system with tanks of different colors. agriculture and food sciences research, 11(2), 170–174. 10.20448/aesr.v11i2.6238 history: received: 15 november 2024 revised: 3 december 2024 accepted: 17 december 2024 published: 23 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by conselho nacional de desenvolvimento científico e tecnológico (grant number: cnpq-brasil – 150480/2023-7 and 402952/2021-9, 308547/2018-7), fundação de amparo à pesquisa do estado de minas gerais (grant number: fapemig-brasil – bpd00406-22 and apq-01531-21), and coordenação de aperfeiçoamento de pessoal de nível superior (grant number: capes-brasil – finance code 001). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction ............................................................................................................................................................................................................ 171 2. materials and methods ......................................................................................................................................................................................... 171 3. results ...................................................................................................................................................................................................................... 171 4. discussions .............................................................................................................................................................................................................. 172 5. conclusion ............................................................................................................................................................................................................... 173 references .................................................................................................................................................................................................................... 173 mailto:tuliopb1@hotmail.com mailto:fabioaremil@gmail.com mailto:pedrocortezzi@hotmail.com mailto:luzrk@yahoo.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6238 https://orcid.org/0000-0002-6143-5417 https://orcid.org/0000-0001-5682-8040 https://orcid.org/0000-0001-8456-8880 https://orcid.org/0000-0002-1021-5772 agriculture and food sciences research, 2024, 11(2): 170-174 171 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study evaluates the effect of tank color in a recirculating aquaculture system (ras) on the performance of juvenile pangasianodon hypophthalmus over 60 days. blue tanks yielded better growth (weight, length, gain) and feed conversion compared to white tanks, which had the poorest results. 1. introduction pangasianodon hypophthalmus, popularly known as panga, is a freshwater fish native to the mekong river in asia. characteristics, such as resistance, rapid weight gain and tolerance to high stocking densities, make the species appealing for aquaculture [1]. the sustainability of p. hypophthalmus production has been questioned in recent years, mainly due to disease outbreaks [2] and water pollution from untreated effluents [2, 3]. implementation of the recirculating aquaculture system (ras) for p. hypophthalmus cultivation is one way to increase production while mitigating the effects of environmental impacts [2]. the tanks used in rass can be manufactured in any color, although the majority sold are blue, green or black [4, 5]. ras tank color can have a great influence on fish, with direct impacts on food capture and, consequently, performance and survival [4-7]; however, the influence of tank color is specific to each fish species, with some performing best in light-colored tanks and others in darker-colored tanks [5]. it was recently reported that juvenile p. hypophthalmus, cultured for 20 days, perform better in brighter (green and white) tanks [8]. however, no study has evaluated the influence of tank color on juvenile p. hypophthalmus when cultured for longer period. 2. materials and methods this studywas approved by committee for ethics in animal use (ceua/ufmg 106/2021) and aimed to evaluate the influence of tank color on the performance and survival of juvenile p. hypophthalmus. this estud was conducted at the laboratorio de aquacultura (laqua) of the universidade federal de minas gerais (ufmg) for a period of 60 days. one hundred and forty-four juvenile p. hypophthalmus, weighing 2.18±0.5 g and measuring 6.76±0.17 cm in total length, were distributed among three rass. each ras was equipped with a 200-liter rectangular filter, with mechanical and biological filter, a heating and water pumping system and four 30-liter tanks covered with colored adhesive (white, blue or black) that were filled with 28 liters of water. therefore, each ras was considered a color treatment (white, blue or black) with four replicates (tanks), which had a density of 0.43 juveniles per liter (12 juveniles in each replicate). the tank colors were analyzed using the cie l*, a*, b* coordinate system through a calorimeter application created by research lab tools, são paulo, brazil (accessible via google play) [9]. the values measured included l* (luminosity: white = 100 to black = 0), a* (red = +60 to green = -60 ), and b* ( yellow = +60 to blue = -60) [4, 5] (table 1). table 1. mean chromaticity parameters (mean ± standard deviation) for the tested tank colors (l: lightness; a: redness when positive, grey when zero, and greenness when negative; b: yellowness when positive, grey when zero, and blueness when negative). tank colors l* a* b write 78.30±1.38 2.37±0.23 2.60±0.15 blue 64.81±1.64 -12.03±1.07 -45.06±3.90 black 18.72±0.96 2.18±0.23 -1.70±0.03 the animals were cultured for a period of 60 days, during which they were fed until apparent satiety twice a day (08:00 and 16:00), with 2–3 mm pellets of extruded commercial diet containing 38% crude protein. the food provided was previously weighed in individual containers. thirty minutes after feeding, feces were removed using a siphon, and any remaining food was gathered, dried, and weighed to determine the amount consumed. water quality parameters were assessed every three days. temperature and ph were measured with a portable combo ph meter from hanna, dissolved oxygen (do) was determined with a digital oximeter from the same brand, and ammonia was analyzed using a labcontest colorimetric kit. in the experiment final (60 days after cultivation), all fish were carefully wrapped in a damp cloth for weighing and measuring to determine their final weight (fw) and total length (tl). these biometric data were used to calculate feed consumption metrics, such as weight gain (wg) [((final weight initial weight) / initial weight) × 100] and feed conversion ratio (fc) (feed consumption / weight gain). for photographic documentation, three juveniles from each tank (n = 12 per treatment) were euthanized using a eugenol overdose (285 mg/l)[10]. survival rates were evaluated by counting the remaining individuals. data were subjected to the shapiro-wilk normality test and levene's test for homoscedasticity, followed by analysis of variance (anova) with tukey's post hoc test at 5% probability. infostat software was used for all data analyses. 3. results there were no differences in water quality parameters among rass (p>0.05) (table 2). juvenile p. hypophthalmus cultivated in white tanks had lighter skin color and those cultivated in black tanks had darker skin color (figure 1 a). note: the values of l*, a* and b. are data obtained by the calorimeter application. where: l: lightness; a: redness when positive, grey when zero, and greenness when negative; b: yellowness when positive, grey when zero, and blueness when negative. agriculture and food sciences research, 2024, 11(2): 170-174 172 © 2024 by the authors; licensee asian online journal publishing group table 2. water parameters obtained after 60 days of the cultivation of juveniles of pangasianodon hypophthalmus in tanks of different colors. tank colors temperature (°c) ph toxic ammonia (mg/l) dissolved oxygen (mg/l) write 27.2±1.3a 6.97±0.21a 0.010±0.003a 8.02±1.12a blue 27.1±1.1a 7.01±0.19a 0.012±0.001a 7.98 ±1.03a black 27.2±0.9a 6.98±0.21a 0.012±0.003a 8.12±0.98a figure 1. photograph of pangasianodon hypophthalmus after 60 days of cultivation in tanks of different colors. white (a), blue (b) and black (c). the background of the photographs was removed using photoshop software. survival did not differ significantly among treatments (p>0.05) (table 3). no dead individuals were found inside tanks and reductions in survival were due to animals jumping out of the tanks, which occurred during the night. table 3. performance parameters (mean ± standard deviation) obtained after 60 days of the cultivation of juveniles of pangasianodon hypophthalmus in tanks of different colors. tank color fw (g) fl (cm) wg (%) fc sur (%) white 16.6±0.8b 12.9 ±0.2b 636.0±99.47b 0.92±0.07a 96.4±7.1a blue 20.3±1.2a 14.2±0.2a 824.62±30.8a 0.66±0.08b 97.6±4.1a black 19.2±0.6 ab 13.9±0.1a 736.0±30.5ab 0.72±0.04b 96.45±4.2a juveniles cultivated in the blue tanks had significantly higher fw, fl and wg and lower fc than juveniles cultivated in white tanks (p<0.05) (table 3). juveniles cultivated in black and blue tanks did not differ significantly for fl and fc. juveniles cultivated in black tanks had fw and wg values intermediate to those cultivated in blue and white tanks. juveniles cultivated in white tanks had the lowest fl and highest fc (p<0.05). 4. discussions during the experiment, water temperature remained within the optimal range indicated for the cultivation of p. hypophthalmus [11]. the remaining parameters (ph, oxygen and toxic ammonia) remained within acceptable levels for fish cultivation in general [12, 13]. the difference in color observed for juveniles cultivated in thanks of different colors may be related to a change in skin pigmentation for camouflage in the environment, which is common to several species of fish [5] and explains the darker colors for fish cultivated in black tanks. there were no significant differences in survival rate among the juveniles cultured in tanks of different colors. sebesta, et al. [14] studied the cultivation of coregonus peled larvae in tanks of different colors and found that those cultivated in black tanks had the highest survival rate. takeshita and soyano [15] studied the effects of green, white, red and black tanks during juvenile epinephelus coioides cultivation and found that those cultivated in black tanks had a higher mortality rate, which was attributed to a greater incidence of cannibalism. monk, et al. [16] studied the cultivation of gadus morhua larvae in white and black tanks and found no significant difference in survival rate. nawang, et al. [8] also found no differences in survival rate for juvenile p. hypophthalmus cultured in green, black note: the letters inserted in the table indicate the statistical difference in the results. since all treatments received the same letter (a), it indicates that there is no difference in the results. results with distinct letters represent a significant difference (p<0.05) according to anova followed by tukey’s test at 5%. note: the letters (a and b) entered in the table indicate the statistical difference in the results. results with different letters indicate a significant difference (p<0.05) as determined by anova followed by tukey's test at 5%. lowercase letters denote a significant difference (p<0.05) between treatments. final weight (fw) in g, final length (fl) in cm, weight gain (wg) in %, feed conversion (fc), and survival rate (sur) in %. agriculture and food sciences research, 2024, 11(2): 170-174 173 © 2024 by the authors; licensee asian online journal publishing group and white tanks. therefore, the survival rate of fish in tanks of different colors can vary depending on the species and its cultivation phase. the influence of tank color on animal performance is specific to each species and may depend on dietary characteristics and vision capacity. boaventura, et al. [4] evaluated the influence of different tank colors (white, blue, green and black) on the cultivation of juvenile colossoma macropomum and found that those cultivated in lighter colored tanks performed better than those cultivated in darker tanks. these authors hypothesized that this difference may have been due to a greater ease in locating and capturing food. furthermore, these authors also reported that juveniles cultivated in darker tanks (black) had higher hemoglobin and lower triglyceride levels in the blood, which was attributed to greater movement, mainly to locate and capture food. in contrast, studying the influence of different tank colors on the catfish lophiosirulus alexandri, costa, et al. [17] found no significant differences in performance and survival, which was attributed to the ability of the species to locate food through smell, taste buds and free neuromasts. still, okomoda, et al. [18] studied the influence of tank color on juvenile clarias gariepinus and found the highest performance for those cultivated in darker tanks (black), which was attributed to their preference for darker environments and possession of barbels to locate food in the absence light. ferosekhan, et al. [19] reported that pangasius pangasius larvae had their best performance when cultivated in darker colored (black) tanks. in contrast, nawang, et al. [8] found juvenile p. hypophthalmus to have their best performance when cultivated in green and white tanks for a period of 20 days. the juveniles of the present study were cultivated for 60 days, so the difference found in relation to the data for larvae presented by ferosekhan, et al. [19] may have been due to the adaptation of juveniles to the colors. juveniles of several fish species were reported to have their best performance when cultivated in blue tanks, such as: nile tilapia, oreochromis niloticus [20]; beluga, huso huso [21]; european catfish. silurus glanis [22] and the betta fish betta splendens saekhow, et al. [23]. opiyo, et al. 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licensee asian online journal publishing group agriculture and food sciences research vol. 12, no. 2, 123-130, 2025 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v12i2.7621 © 2025 by the authors; licensee asian online journal publishing group cryopreservation and its effects in coffee-tree (coffea spp.) culture yanelis castilla valdés1 maría esther gonzález vega2 ( corresponding author) 1,2department of plant genetics and breeding, national institute of agricultural sciences, mayabeque, cuba. 1email: yanelis@inca.edu.cu 2email: esther@inca.edu.cu abstract coffee is one of the most commercialized beverages worldwide. it is obtained from the fruits of the coffee plant (coffea spp), a genus that is difficult to conserve because its seeds tolerate a certain amount of desiccation but become sensitive to the low temperatures used in conventional germplasm banks. hence, the importance of using other methods, such as cryopreservation, which allows for storage over extended periods. during this process, tissues are subjected to different treatments and methods that can cause cryogenic damage and stress. to determine their occurrence and rule out the appearance of genetic variability, various analyses are performed, ranging from morphological to epigenetic. the present bibliographic review was conducted to identify the main research on cryopreservation and its effects on coffee cultivation. the analyses applied to evaluate cryogenic damage were studied at the morphological, histological, physiological, and biochemical levels. the effects detected represent tissue responses to stress caused by this long-term preservation technique. molecular studies have not been reported so far; thus, it is necessary to continue deepening these analyses to achieve their complementarity, in order to corroborate the genetic stability of the coffee plants recovered from cryopreservation. keywords: conservation, cryodamage, genetic stability, in vitro culture, ros, stress, vitrification. citation | valdés, y. c., & vega, m. e. g. (2025). cryopreservation and its effects in coffee-tree (coffea spp.) culture. agriculture and food sciences research, 12(2), 123–130. 10.20448/aesr.v12i2.7621 history: received: 15 september 2025 revised: 20 october 2025 accepted: 27 october 2025 published: 31 october 2025 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support. institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: both authors contributed equally to the conception and design of the study. both authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 124 2. cryopreservation ............................................................................................................................................................................ 124 3. conclusions ..................................................................................................................................................................................... 129 references ............................................................................................................................................................................................ 129 mailto:yanelis@inca.edu.cu mailto:esther@inca.edu.cu https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i2.7621 https://orcid.org/0000-0003-1859-9210 https://orcid.org/0000-0001-5841-8272 agriculture and food sciences research, 2025, 12(2): 123-130 124 © 2025 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study consolidates, for the first time in the literature, research conducted on the effects of cryopreservation on coffee cultivation. it provides new insights into the methods used to detect genetic stability after long-term preservation of this genus and enhances understanding of the causes of cryogenic damage. 1. introduction coffee represents one of the tropical agricultural products of higher importance, contributing to the economy of about 80 coffee grain producer countries. its cultivation currently faces numerous challenges, with climate change being one of the most relevant, as it influences biotic and abiotic factors that affect its phytogenetic resources [1, 2]. although 130 species of coffea spp. are currently known [3], world production is based mainly on two species: coffea arabica l. and c. canephora pierre [4]. in view of their characteristics during conventional storage, coffee seeds have been classified as intermediate, since they tolerate a certain level of desiccation but do not survive total desiccation or the combined effects of desiccation and low temperatures [5]. for this reason, other ex situ conservation methods are applied to achieve their preservation for longer periods of time, such as cryopreservation. cryopreservation is a process in which cells or tissues are preserved by freezing at ultra-low temperatures, below zero degrees, such as -196°c (boiling point of liquid nitrogen). at these low temperatures, metabolic processes are slowed down [6], including biochemical reactions that can cause aging and cell death, so that material maintained under these conditions can theoretically be stored for prolonged periods [7]. this is the method of choice to ensure long-term storage of germplasm of unorthodox vegetatively propagated seeded plant species or important cell lines [8]. the different cryopreservation methods for long-term preservation of germplasm in vitro comprise numerous stages, ranging from tissue culture, pre-growth or dehydration, cryoprotection, freezing, rewarming, material recovery, and regeneration. consequently, tissues are exposed to physical, chemical, and physiological stresses, which can result in cryodamage. successful cryopreservation is often evaluated by the survival of plant tissues and their ability to regenerate whole plants [9]. several cryogenic studies have been conducted to preserve coffee plant genotypes using various methods over extended periods. however, many of these studies are limited to reporting only the germination results or the viability of cryopreserved explants, without further analysis or long-term assessment [10-13], while others refer to morphological, histological, biochemical, or physiological aspects of the seeds, embryos, or seedlings recovered after the application of this technique [14-26]. the present bibliographic review was conducted to identify the main research conducted on cryopreservation and its effects on coffee cultivation. 2. cryopreservation cryopreservation, or the storage of biological material at the temperature of liquid nitrogen (-196 °c), is an alternative to ex situ germplasm conservation, which makes it possible to maintain plant genetic resources in the long term in a safe and cost-effective manner [27]. liquid nitrogen is “relatively inexpensive, chemically inert, readily accessible, and independent of electricity” [28]. at this temperature, metabolic processes are slowed down so that plant material can be stored for very long periods of time [5]. in addition, cryopreservation eliminates the need for regular renewal of the collection, which reduces the risk of genetic erosion caused by pests, diseases, climatic conditions, contamination, and genetic variations [29]. it therefore represents a safe and cost-effective option for the long-term preservation of all categories of species that are difficult to preserve by other methods [5]. among the main cryopreservation methods are: the conventional protocols or classical method (slow freezing), encapsulation-dehydration, pre-culture-desiccation and desiccation, vitrification, and derived protocols (dropletvitrification, encapsulation-vitrification, and cryo-plate) [30], and the more recently developed cryo-mesh [31]). classical method: the classical method is based on slow freezing of the explants (0.1 to 2 °c min-1) usually down to -40 °c, and then their rapid immersion in liquid nitrogen. dehydration of the samples is achieved during the temperature decrease, although solutions with cryoprotective properties or a physical dehydration step can also be used previously. disadvantages of this method include the complexity of the operation, the use of equipment that controls the cooling rate (programmable freezers), and the formation of ice crystals in the extracellular solutions, which can cause loss of water from the cells in the so-called freeze-dehydration process [32]. encapsulation-dehydration: this technique has the advantage of being relatively simple, since it facilitates handling a large number of explants enclosed in calcium alginate capsules at the same time and uses only sucrose as an osmotic agent. it replaces the use of expensive and sophisticated programmable freezing equipment with rapid immersion in liquid nitrogen. return to normal culture temperature is usually accomplished by exposing the encapsulated material to flowing air in a laminar flow hood, instead of using a 40 °c water bath [30]. vitrification method: vitrification protocols have the common characteristic that the critical step to achieve the survival of the cryopreserved material is the dehydration stage, not the freezing stage, as occurs in classical protocols. in these procedures, the removal at positive temperatures of most or all of the freezable water from the cells is promoted through osmotic dehydration using highly concentrated solutions [32]. among these are the socalled plant vitrification solutions or pvs. they are composed of penetrating and non-penetrating cryoprotective agents used to increase osmolarity in plant tissues and reduce the free water content of the cells [33]. to avoid phase transitions at ultra-low temperatures, without excessively reducing the water content of the cells, vitrification, or solidification of a liquid forming an amorphous or glassy structure, must be achieved [6, 34]. thus, in the amorphous solid state, the molecules associate without acquiring the organized structure characteristic of crystallization [35]. highly concentrated solutions sufficiently dehydrate the cells and cause the solute molecules to hinder the organization of water molecules that can form ice crystals. the ultra-fast cooling rate prevents sufficient time for ice crystals to form before all molecules are immobilized by the temperature of the liquid nitrogen. these two conditions allow the formation of the glassy state [34]. agriculture and food sciences research, 2025, 12(2): 123-130 125 © 2025 by the authors; licensee asian online journal publishing group the most commonly used plant vitrification solutions are: pvs 2, composed of 30% glycerol +, 15% ethylene glycol +, 15% dimethyl sulfoxide +, and 0.4 m sucrose [36], and pvs 3, composed of 50% glycerol + 50% sucrose [37]. • droplet-vitrification. it is derived from the vitrification method and differs from it in the ultra-fast cooling and heating speeds achieved to treat the samples. instead of using cryovials, the tissues are transferred to a reduced volume of a drop of the vitrification solution, placed on a small sheet of aluminum foil, in which they are directly immersed in liquid nitrogen [38]. • for rewarming, the foil with the samples is immersed directly in liquid culture medium supplemented with 1.2 m sucrose [39] at room temperature. the excellent thermal conductivity of the aluminum foil, combined with the small volume of cryoprotective solution in contact with the tissues, allows both cooling and heating to proceed at a very high rate [40]. • encapsulation-vitrification. in this method, the tissues are encapsulated in calcium alginate, similar to the procedure used in the encapsulation-dehydration technique. the loading and dehydration treatments with the pvs solution are performed as described for the vitrification method. the capsules are placed in cryovials with the vitrification solution and immersed in liquid nitrogen until heated in a water bath [30]. • cryo-plate. in the cryo-plate method, the explants are also encapsulated, but they are immobilized in a thin layer of calcium alginate that gels on the surface of an aluminum plate [32]. this method has two variants: v cryo-plate and d cryo-plate. in the case of the v cryo-plate, the explants are subjected to the vitrification solution for dehydration, and then the same steps of the vitrification procedure are followed. in the d cryoplate, dehydration is performed physically (with silica gel or in the laminar flow cabinet) and proceeds with the steps of the encapsulation-dehydration technique [41]. an advantage shared by the encapsulation-vitrification and cryo-plate methods is that they allow the handling of numerous explants immobilized in calcium alginate, unlike the vitrification method, which involves the direct handling of each tissue. it is also possible to shorten the total duration of the cryopreservation protocol compared with that required for encapsulation-dehydration [42]. • cryo-mesh. it is a recently developed method [31]. in general, it resembles the v cryo-plate protocol, with the difference that the samples are placed in a stainless steel mesh that also allows high cooling and reheating rates. vitrification-based procedures are appropriate for complex organs such as apical meristems and embryos, which contain a wide variety of cell types with specific requirements. moreover, being operationally simpler than classical methods, they have the potential for wider applicability, as they demand only minor modifications for different cell types [43]. desiccation method: equally important is the desiccation method, which is simple and very appropriate for cryopreserving seeds, and which, combined with a previous pre-culture in medium supplemented with sugars, results in the so-called pre-culture-drying protocol and can be effective for successfully cryopreserving zygotic and somatic embryos [44]. for seed cryopreservation, the pre-culture phase is omitted, which implies the use of a more simplified and shorter protocol. desiccation can be achieved in different ways: by exposing the explants to the air current of a laminar flow cabinet; by placing them in hermetically sealed containers with silica gel; or by achieving controlled atmospheres with saturated saline solutions [30]. 2.1. differential scanning calorimetry the most widely used technique, so far, to identify and quantify crystallization and glass transition events is differential scanning calorimetry (dsc or differential scanning calorimetry) [45], which is a very valuable tool to optimize liquid nitrogen immersion conditions and to develop low-temperature protection strategies [46]. the fundamental principle of dsc is based on applying regulated temperature changes to the sample under study, while measuring the heat flow and temperature associated with its thermal transitions. the equipment detects temperature differences between a reference and the sample, and recalculates the differential heat flux [47]. this data is of great interest because it allows obtaining valuable information on endothermic, exothermic events, or changes in the heat capacity of the plant material. it can be used to determine the glass transition, ice nucleation temperature, melting, boiling, and crystallization time, which are important characteristics in cryopreservation processes. it is considered a very advantageous method and provides a relatively fast, real-time measurement of the thermal characteristics during the dehydration of the evaluated sample [47]. 2.2. cryodamage cryopreservation techniques for long-term preservation of germplasm in vitro comprise numerous steps, ranging from tissue culture, pre-growth or dehydration, cryoprotection, immersion in liquid nitrogen, rewarming, material recovery, and regeneration, all of which expose tissues to physical, chemical, and physiological stresses. successful cryopreservation is often evaluated by the survival of plant tissues and their ability to regenerate whole plants [8]. when water is removed from the cells of dehydration-sensitive organisms, two main factors can explain the damage caused, according to some authors [48]: damage resulting from mechanical stresses that disrupt structural organelles at high moisture contents (greater than -5 mpa), and damage to macromolecular structures following more extreme desiccation. consistent with these approaches, recent research evidence indicates damage to cellular metabolic activity at an intermediate level of humidity. these cells can continue to respire but cannot eliminate the toxic metabolic products that accumulate [48]. this metabolic disorder that occurs in partially dehydrated cells can also cause the death of cells that remain partially hydrated for a certain period of time, which is the basis of the accelerated aging test [49]. generally, seeds lose viability when they are kept at a moisture content where respiration is possible (-15 to -5 mpa), but repair processes are not efficient [50]. this may contribute to damage to the moist stored seeds or sensitive seeds that are slowly dried, as they are kept at a critical moisture level for a long period of time [48]. agriculture and food sciences research, 2025, 12(2): 123-130 126 © 2025 by the authors; licensee asian online journal publishing group when biological material is exposed to decreased temperature, all kinds of processes can be affected; chemical reactions are delayed or stopped (thus metabolism as well), and membrane stability is affected. the formation of extracellular ice crystals causes tissue rupture and cell disconnection. during slow freezing, effects related to dehydration occur as the first extracellular ice crystals are formed, resulting in a concentrated solution surrounding the cells [6]. since intracellular and extracellular fluids are in osmotic equilibrium, water leaves the cells, leading to freeze dehydration and, consequently, irreversible lethal plasmolysis. the formation of intracellular ice crystals during exposure to subzero temperatures causes penetration of the membrane and loss of its semipermeability, ultimately resulting in cell death [6]. 2.2.1. oxidative stress oxidative stress refers to the state of imbalance between oxidation and antioxidation caused by the massive production of reactive oxygen species in cells under extreme conditions, such as low temperatures [51]. management of oxidative stress is vital for the successful application of cryopreservation to plant tissues. freezing damage induces the production of free radicals, mainly reactive oxygen species or ros, which attack the lipid fraction of membranes [52]. the steps prior to storage in liquid nitrogen (conditioning and dehydration) can induce an excess of ros, such as hydroxyl radical, superoxide, and hydrogen peroxide, the accumulation of which causes negative effects on plant tissues. some of these effects may include lipid peroxidation, protein oxidation, and dna (deoxyribonucleic acid) damage. in some cases, programmed cell death (pcd) can be induced, which ultimately kills the cells after cryopreservation [53]. enzymes that remove reactive oxygen species, such as catalase, superoxide dismutase, and peroxidase, among others, can mitigate or reverse the damage caused by oxidative stress, making biochemical studies important to elucidate the effect of cell damage caused by cryopreservation [54, 55]. in this regard, some results have been obtained in embryos of barley (hordeum vulgare l.) [56] and the palm livistona chinensis (jacq.) r.br. ex mart. [57] and in castor bean (ricinus communis l.) seeds [58]. 2.3. studies for the determination of stability and genetic variability in cryopreservation of plant species in vitro tissue culture is an integral part of cryopreservation protocols; it is currently used as a basis for the establishment and maintenance of plant species and during the post-culture process for the recovery of cryopreserved tissues [53]. unlike animal cells, plant cells exhibit the phenomenon of cell totipotency, which is the ability of a cell to regenerate a whole plant [59]. when differentiated and fully organized plant tissues are introduced into in vitro culture and induced into the cell division phase by growth hormones, this can result in genetic changes, a phenomenon known as somaclonal variation [60]. somaclonal variation can manifest as modifications in plant morphology, chromosome number, accumulation of genetic mutations, gene expression levels in rna (ribonucleic acid), protein profiles, and molecular changes in dna sequences [8]. considering also the damage that can occur in dna as a consequence of oxidative stress caused by cryopreservation [61] before using this technique routinely for the long-term maintenance of plant genetic resources, it is necessary to verify that the genetic stability of the cryopreserved material is not altered [62]. different techniques can be applied for this purpose: phenotypic variation analysis (morphological and biometric studies), histological-cytological analysis (mechanisms, techniques, and cytological studies), biochemical analysis (metabolite, enzyme, or protein analysis), molecular genetic analysis (genome structure, dna hybridization, pcrpolymerase chain reaction, rapdrandom amplified polymorphic dna, ssrsimple sequence repeats, aflpamplified fragment length polymorphisms), epigenetics (chromatin analysis, dna methylation studies, dna hybridization, and methylation-specific pcr) [8]. 2.4. cryopreservation studies in coffee cultivation 2.4.1. encapsulation-dehydration method among the first approaches to the subject of the effects of cryopreservation in coffee cultivation is a study on the histo-cytology of apices of the species c. racemosa and c. sessiliflora, subjected to the encapsulation-dehydration method. a survival rate of 38% was obtained for c. sessiliflora, whose apices required a pre-growth of 3 to 10 days in murashige and skoog [63] liquid medium with 0.75 m sucrose, and 4.5 hours of dehydration in a laminar flow cabinet. likewise, c. racemosa apices demanded a progressive increase in sucrose concentration from 0.5 to 1 m, and a dehydration period of 6 hours, to reach 27% survival. after the dehydration phase, the cells of some apices were severely damaged. in these cells, membrane rupture, retracted cytoplasm, non-visible organelles and nucleoli, osmophilic granules towards the periphery of the plasmalemma, and the presence of plasmolysis were observed. however, cytological analysis 21 days after cryopreservation of the plant material showed that most of the meristematic cells recovered from immersion in liquid nitrogen. the apex was able to form leaf primordia, with little difference between the survival of dehydrated apices and those that were dehydrated and cryopreserved [14]. 2.4.2. desiccation method/classical method some studies differ in the convenience of applying a slow desiccation treatment (saturated salt solutions) or a rapid desiccation (silica gel) to coffee seeds. according to some authors [64], balancing the moisture content under the relative humidity obtained with saturated salt solutions allows reaching the optimum moisture content for cryopreservation with little loss of viability, in a simple and reproducible manner. however, a minimum of 14 days is required for the seeds to reach equilibrium moisture content. other researchers report that desiccation in silica gel is more beneficial, since, as it occurs more quickly, there is not enough time for the accumulation of damage that occurs during desiccation, and the tissues are at moisture contents in which, if there are degradation processes, these occur in a shorter time and before the damage can accumulate to high levels [18]. in the coffee tree culture, the desiccation method (rapid cooling by direct immersion in liquid nitrogen) was frequently compared with the classical method (slow cooling with the use of a programmable freezer) to evaluate the effect caused by the different cooling speeds on the survival of cryopreserved explants. such is the case of a study led by researchers from ird (institut de recherche pour le développement), from france [15]. they agriculture and food sciences research, 2025, 12(2): 123-130 127 © 2025 by the authors; licensee asian online journal publishing group cryopreserved seeds of nine coffee species that differ in their sensitivity to desiccation, with the use of saturated saline solutions, to achieve slow dehydration until the moisture content reached equilibrium, depending on the type of salt. the species studied were classified into three groups, taking into account the germination obtained in relation to the cooling rate. from later studies, the french research team suggested two strategies for routine cryopreservation in coffee germplasm banks [65]. both methods coincide in the stage of drying the seeds using saturated saline solutions. however, they differ in that strategy 1 involves slow cooling at -1 °c min-1 to -50 °c before immersion in liquid nitrogen, osmo-conditioning of the seeds, and germination under greenhouse conditions. this strategy has the advantage of not requiring in vitro tissue culture, but it necessitates a programmable freezer, which is not available in all laboratories. the second strategy involves direct immersion in liquid nitrogen and inoculation of the embryos in vitro, with acclimatization of the vitro plants before transfer to the greenhouse. this approach yields higher survival percentages and does not require a programmable freezer, although it takes more time due to the in vitro culture stage, which carries an additional risk of contamination. nevertheless, it was the alternative chosen by catie (centro agronómico tropical de investigación y enseñanza), from costa rica, to establish a coffee seed cryobank in 2002. brazilian researchers from the federal university of lavras, embrapa brazil, and other institutions have conducted extensive studies on the cryopreservation of different coffee genotypes by comparing the drying and cooling rates of the samples and analyzing their physiological, biochemical, and histological indicators. in a study conducted in 2017 on coffea arabica (cultivars arara, catiguá, catuaí amarelo, and mundo novo), seeds were subjected to rapid desiccation using silica gel and slow desiccation using a saturated nacl solution, followed by immersion in liquid nitrogen for 24 hours. in general, rapid drying with silica gel to 20% moisture content resulted in higher percentages of normal seedlings, expanded cotyledonary leaves, and dry mass, as well as improved physiological indicators. the seeds of these cultivars showed different levels of tolerance to cryopreservation, with catuaí amarelo being the most tolerant and arara the most sensitive, regardless of the desiccation rate [18]. the activity of catalase, peroxidase, and esterase enzymes increased after desiccation and exposure to ultra-low temperatures. seeds at their initial moisture content showed no or very low activity compared to seeds subjected to desiccation, indicating that these enzymes are activated when tissues undergo the stress of water loss and ultra-low temperatures. stress causes an increase in free radicals in cellular metabolism, which requires the expression and action of these enzymes, which induce defense mechanisms in seeds and protect them against cryogenic damage [18]. additional research determined the effect of moisture content, different slow cooling rates, and final temperature on cryopreservation of catuaí amarelo iac 62 seeds. only seeds dried in silica gel up to a moisture content of 20% germinated, regardless of the cooling rate and final temperature. in general, when the temperature was lowered to -40 °c, the highest values of radicle protrusion, normal seedlings, strong normal seedlings, seedlings with expanded cotyledons, root dry mass, and shoot dry mass were obtained, with significant statistical differences compared to the rest of the treatments. the results of the tetrazolium test indicated that embryos extracted from seeds stored in liquid nitrogen were less sensitive to cryopreservation than whole seeds, and embryos showed high viability when subjected to rates of -3 or -5 °c min-1 up to the temperature of -60 °c, before declining to -196 °c [19]. however, a more in-depth study on this same cultivar determined that desiccation in saturated saline solutions also favored the quality of the recovered seedlings, although a moisture content of 17% was the main factor for seed preservation, regardless of the type of cryopreservation protocol used. under these conditions, they maintained better physiological quality (radicle protrusion, normal seedlings, strong normal seedlings, cotyledonary leaves, root and aerial part dry matter, embryo viability) and better preserved cellular structures. slow cooling before storage in liquid nitrogen did not provide advantages compared to direct immersion. in general, for catalases, esterases, and peroxidases, activity was higher in seeds that were dehydrated to 17% moisture content. the better physiological quality of the plants recovered from these treatments indicated that the higher activity of these enzymes favored overcoming desiccation stress. the pattern of cellular structure observed in all the seeds analyzed indicated that the effects of cryopreservation are less drastic in embryo cells than in endosperm cells, which are more sensitive to stresses caused by dehydration, pre-freezing, and rewarming [24]. more recent research has corroborated part of these results [26]. when comparing the cryopreservation of seeds (cv. catuaí amarelo iac 62), with storage in a cold chamber (at 10 °c and 45% relative humidity) after drying in a stationary dryer, it was determined that dehydration in silica gel up to 17% of the moisture content, and preservation in liquid nitrogen, was the most effective method. seedlings from seeds dried in silica gel and cryopreserved were vigorous and showed vegetative development (number of pairs of leaves, average height of the seedling, average diameter of the cap) similar to that of seedlings produced from seeds with 32% humidity and stored for six months in cold storage. besides, cryopreservation only requires the liquid nitrogen supply for maintenance, which allows the reduction of storage space and the long-term preservation of germplasm [22]. the use of an osmo-conditioning treatment is essential to promote the proportion of seeds that develop into normal seedlings after exposure to liquid nitrogen. its effect is associated with a reduction of membrane imbibition damage [65]. this was corroborated in zygotic embryos of catuai amarelo iac 144 that were successfully cryopreserved using dehydration in silica gel for 60 min (23% moisture content), followed by osmotic rehydration using solutions with decreasing sucrose concentrations (from 1 m to 0.10 m), after rewarming. according to the ttc test, the embryos showed maximum viability (75%) and vigor (26%) with this treatment, among all those evaluated. the percentages of germination and normal seedlings reached 90% and 75%, respectively. during acclimatization, 100% survival of seedlings from cryopreserved zygotic embryos was obtained. for the morphological variables evaluated during this stage, statistical differences were observed in shoot length and root length between seedlings originating from dehydrated and cryopreserved embryos and those from fresh embryos, as well as from dehydrated and non-cryopreserved embryos. this could be due to osmotic rehydration, which is considered beneficial in seed germination and plant vigor of different crops under stress conditions. however, there were no phenotypic differences between treatments in leaf shape, color, or organ formation [17]. in cryopreservation of c. canephora seeds, the effectiveness of three slow cooling rates was also compared with direct agriculture and food sciences research, 2025, 12(2): 123-130 128 © 2025 by the authors; licensee asian online journal publishing group immersion in liquid nitrogen. desiccation to 0.25 g g-1 of water content did not affect viability, and although both cooling rates (fast or slow) were somewhat harmful to seeds, seeds responded better to direct immersion in liquid nitrogen because higher percentages of germination, normal seedlings, and viability were obtained. catalase and esterase enzymes proved to be good biochemical markers for cryopreserved seeds, and their activity was higher in those of higher physiological quality. on the other hand, for superoxide dismutase and peroxidase, the effect of the different cryopreservation treatments was insignificant, since no differences were observed between the electrophoretic activities, so in this case, they were not considered effective biochemical markers in the cryopreservation of seeds of this species [20]. a more exhaustive study on c. canephora compared two desiccation rates to determine the optimal physiological conditions of seeds after cryopreservation. for dried, non-cryopreserved seeds, a significant effect of desiccation speed was observed for all the variables studied, except for embryo viability: seeds dried quickly (in silica gel) exhibited higher physiological quality than those dried slowly (in saturated nacl salt solution). no significant differences were observed between different moisture contents (0.20, 0.25, and 0.28 g g-1). conversely, for dried and cryopreserved seeds, a moisture content of 0.25 g g-1, achieved by rapid drying, was most effective in obtaining the highest germination percentage (43%) and the best physiological characteristics. the activity of enzymes such as catalase, esterase, glutamic oxaloacetic transaminase, and polyphenol oxidase served as indicators of seed quality following cryopreservation [21]. a recent investigation evaluated the effect of moisture content and packaging types (mesh bags, falcon tubes, and trilaminated aluminum foil envelopes) on the cryopreservation of c. racemosa and c. liberica var dewevrei seeds. it was determined that there was an interaction between the moisture content and the type of seed packaging. desiccation of seeds to 20% and packaging in trifoliate aluminum foil envelopes was the most effective protocol among those studied, with survival rates of 79% for c. racemosa and 8% for c. liberica. in general, for c. racemosa, glutamic oxaloacetic transaminase, peroxidase, and esterase isoenzymes showed lower activity when seeds were cryopreserved at 20% or 18% moisture content and packed in mesh bags. in addition to the oxidative stress damage caused by the drying process, this type of packaging tended to increase stress due to the direct contact of liquid nitrogen with the endosperm. the opposite was observed with aluminum foil packaging, which was more effective for seed storage in terms of survival and physiological quality [25]. 2.4.3. vitrification methods although vitrification-based methods are considered very successful due to their ease, reproducibility, and applicability to a wide range of species and explants [66], they are still little studied in coffee plants. among the first investigations referring to the vitrification method in coffee plants were those carried out by castilla, et al. [67] in c. arabica and c. canephora using vitrification solutions pvs 2 and pvs 3, which demonstrated the possibility of using this method successfully in both species. in c. arabica cv. catuaí amarelo iac 144, the vitrification method was used to develop a cryopreservation protocol for zygotic embryos, with dehydration in pvs 2 for different periods of time between 10 and 250 min and at two temperatures (0 and 25 °c). the selected treatment was dehydration in pvs 2 for 100 min at 0 °c. under these conditions, the solution was not toxic, and 87% of normal seedling formation was achieved. in the histological analysis, it was determined that the cryopreserved embryos showed cells with characteristics very similar to those of the control treatment. however, in the protoderm and primary meristem, an increase in the number of plasmolyzed cells was observed. in this case, the plasmolysis detected was due to an excess in water efflux caused by the osmotic action of the cryoprotective solution. rewarming directly in the recovery solution at 25 °c for 15 min favored the percentages of germination and formation of normal seedlings, although it did not differ statistically from the values obtained in embryos that were rewarmed for 1 min in a water bath [16]. for zygotic embryos of c. arabica, two cryopreservation methods based on vitrification were developed. in the encapsulation-vitrification method, embryos were encapsulated in alginate and dried in the laminar flow cabinet for 30 or 60 minutes prior to immersion in liquid nitrogen. in the droplet-vitrification method, dehydration in pvs 3 was evaluated for different periods of time (40, 50, 60 minutes). for the encapsulation-vitrification method, the maximum germination obtained was 83%, while the embryos cryopreserved by the droplet-vitrification method reached 100% germination, regardless of the time they remained in pvs 3. in the temporary immersion system, the seedlings reached a higher multiplication rate (4.2 shoots per explant) compared to the same culture medium in a semi-solid state (1.3 shoots per explant). the results of the root initiation experiments showed that 90% of the shoots developed roots in the liquid medium, and all shoots showed two to three primary roots with an average length of 5.5 cm at 6 weeks. rooted seedlings transplanted to the greenhouse showed a survival rate of 95% after one month of growth under ex vitro conditions. dsc analysis confirmed the absence of phase transitions in zygotic embryos cryopreserved by the droplet-vitrification method, which reached 100% germination [23]. in general, the literature contains few reports and events that highlight genetic modifications related to cryopreservation, and if such modifications occur, the exact mechanisms and the nature of genetic instability have not been elucidated, considering the different stages involved in the process (in vitro culture-cryopreservationregeneration) [68]. thus, cryopreservation is considered a cost-effective alternative to ex situ germplasm conservation, which ensures the genetic stability of germplasm over time [34]. considering that the most recent research on the storage of coffee seeds under conventional conditions reaffirms the increasing loss of longevity and the impossibility of obtaining normal plants after a period of two years [69], in our opinion, it confirms the need to employ cryopreservation as an alternative for the ex situ conservation of coffee germplasm. in fact, it is considered advisable to test other methods, such as cryo-plate and cryo-mesh in order to achieve new results, adapted to the conditions of different laboratories. based on all the benefits of this alternative for the preservation of plant genetic resources, the global plant cryopreservation initiative, jointly conceived by cgiar (consultative group on international agricultural research) and the global crop diversity trust, has emerged in recent years. the initiative aims to create a sustainable system for conserving clonal crop collections and non-orthodox seeds to safeguard genetic diversity for future generations. due to its global importance, the coffee plant is considered to be one of the five crops vital to the livelihoods of developing countries and is therefore included among the priority species of this project [70]. agriculture and food sciences research, 2025, 12(2): 123-130 129 © 2025 by the authors; 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(2024). properties of boiled or steamed procambarus clarkii girard and procambarus zonangulus crawfish tail meat during refrigerated and frozen storage. agriculture and food sciences research, 11(2), 156–169. 10.20448/aesr.v11i2.6232 history: received: 13 november 2024 revised: 1 december 2024 accepted: 9 december 2024 published: 20 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this study received no specific financial support institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: conceptualization, j.s., d.b., l.l.; methodology, j.s., d.b. l.l., m.j.; software, j.s., l.l.; validation, m.j., l.l.; formal analysis, j.s.; resources, k.m., m.j., l.l.; data curation, j.s.; writing—original draft preparation, j.s., l.l.; writing—review and editing, k.m, l.l.; supervision, k.w., d.b., m.j., l.l.; project administration, l.l.; funding acquisition, l.l. all authors have read and agreed to the published version of the manuscript. acknowledgement: the louisiana state university agricultural center lsu aquaculture research station and tony’s seafood market and deli provided the crawfish for this study. contents 1. introduction .................................................................................................................................................................................... 157 2. materials and methods ................................................................................................................................................................. 157 3. results and discussion ................................................................................................................................................................. 160 5. conclusions ..................................................................................................................................................................................... 167 references ............................................................................................................................................................................................ 167 mailto:jshack2@gmail.com mailto:kmcmillin@agcenter.lsu.edu mailto:entropyrd@gmail.com mailto:mjanes@nwacc.edu mailto:lucinalampila@gmail.com https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6232 https://orcid.org/0000-0001-9249-6071 https://orcid.org/0000-0002-1644-8841 https://orcid.org/0000-0002-0797-6060 agriculture and food sciences research, 2024, 11(2): 156-169 157 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this research compares two different methods of heating two species of crawfish for consumption and the subsequent properties of the crawfish during refrigerated and frozen storage. it provides data on the composition, texture, color, and microorganism growth during 11 days of refrigerated and 6 months of frozen storage of boiled and steamed crawfish tail meat. 1. introduction crawfish/crayfish have emerged as a highly desirable and valued culinary delicacy because of their distinctive sensory, textural and flavor characteristics [1, 2]. use of the term “crawfish” in 1817 predates the term “crayfish” used in the huxley 1880 textbook [3] and so “crawfish” will refer to the crustacean in this manuscript. of the many species of crawfish around the world, only two species, procambarus clarkii, the red swamp crawfish, and procambarus zonangulus, the white river crawfish, are commercially important for harvest in louisiana, usa [4-6]. the white river crawfish is less tolerant of poor water conditions and warmer temperatures than the red swamp crawfish [5]. consumers and processors prefer the red swamp crawfish to the white river crawfish for appearance and flavor [4, 7]. crawfish may be purchased live or boiled, if for immediate consumption, depending upon the preference of consumers and restauranteurs [8] and more recently as peeled tail meat, fresh or frozen, of cooked crawfish. crawfish tail meat possess a high protein content [approximately 20%] and eight essential amino acids for human consumption [9]. crawfish, like other seafood, are a perishable product with postmortem changes occurring rapidly that cause off-odors and flavors even before spoilage miget [10] and zeng, et al. [11]. davis [5] suggested that crawfish tail meat with adhering hepatopancreas can be frozen, but typically becomes rancid within 30 days of storage. a mushy texture may occur after relatively short periods of iced storage, but deterioration of texture has also been observed in crawfish through extended frozen storage [12]. freezing rate, frozen storage conditions and thawing rate influence frozen food properties [13, 14]. frozen seafood can experience color changes [15] protein denaturation [16] lipid oxidation [17] and ice sublimation and recrystallization [1, 18] which may result in drip loss, dehydration, toughening, off-flavor, and lipid oxidation [18, 19]. drum/belt mechanically separated mince of boiled crawfish had 22% less lipid oxidation with cryogenic liquid nitrogen freezing than with air blast freezing after 180 d of frozen storage [20]. the two most prevalent commercial thermal treatments for cooking of crawfish are steaming and boiling because of their low cost and large scale efficiency [2]. raw crawfish meat may have only faint fishy odors while the desirable flavor of cooked crayfish meat is mainly from complex reactions of flavor precursors, intermediates and their interaction products [21]. crawfish cooking can produce lipid oxidation degradation, the maillard reaction and the interaction between the two reactions, resulting in volatile flavor compounds, such as aldehydes, alcohols, ketones and other compounds, that provide a unique flavor [21]. microorganisms are also a major concern in the spoilage and safety of crawfish [11, 22] as farmed or wild caught crawfish are apt to come in contact with harmful organisms in their habitats [10]. an added microbiological concern with crawfish tail meat is microbial contamination by hand-peeling from the shells if proper sanitation practices are not practiced [23]. an increase in apc to levels exceeding 106 colony forming units(cfu)/g is generally a sign of a long period of time at refrigerated temperatures or temperature abuse before freezing [24]. aerobic plate count (apc) and e. coli/coliform limits have been suggested by the international commission on microbiological specifications for foods [24]. the limits are appropriate for both refrigerated and frozen cooked crawfish. any sample having an apc count exceeding log 7 cfu/g should result in the product being discarded. for e. coli/colifoms, any sample exceeding 500 cfu/g should result in product being discarded. procambarus clarkii crawfish harvested from rice fields had higher aroma and texture acceptability scores from a consumer panel than those from moist-soil wetlands, but there were no differences in flavor, overall acceptability, composition, or fatty acid contents [25]. the total amounts of protein, fat, and ash increased in tail meat after both microwave and frying cooking while water content decreased, especially in the fried samples [26]. steaming and boiling significantly influenced the fatty acids and free amino acids and were highly correlated with flavor and key flavor compounds in pulverized freshwater crawfish (procambarus clarkia) tail meat [2]. color, texture, flavor components, and microorganism growth were measured through 11 days of refrigerated and 6 months of frozen storage of boiled and steamed crawfish tail meat from procambarus clarkia and procambarus zonangulus species. 2. materials and methods 2.1. procurement and initial preparation live freshwater crawfish consisting of both red swamp crawfish (procambarus clarkia), and white river crawfish (procambarus zonangulus) were obtained from the louisiana state university agcenter aquaculture research station in baton rouge, louisiana, usa and from tony’s seafood market and deli of baton rouge, louisiana, usa. the live crawfish were transported to the food science building in baton rouge, louisiana in mesh sacks varying in weight from 12 to 18 kg and stored two days at 4°c until processed. prior to cooking the crawfish were removed from the mesh sacks and placed in 1.5m x 0.6m x 0.6m metal containers. the crawfish from the different locations were mixed and washed with municipal tap water to remove any mud and debris. dead crawfish were separated from the live crawfish and discarded. the crawfish were then kept moist by regularly spraying with municipal tap water until they were processed. two lots of 28.8 kg of crawfish destined for boiling were removed from the metal lugs and placed in a stainless steel basket. temperatures were obtained with an t-type thermocouple and data logger (omega om-daqpro5300, omega engineering inc., stamford, ct). the crawfish were placed in approximately 75.7 l of unseasoned municipal tap water that had been brought to a boil at atmospheric pressure in a 150 l jacketed steam kettle (b.h. hubbert & son, inc., baltimore, md) with university-generated 206.85 cm hg food-grade steam as the steam source. the temperature of the water in the kettle was measured at periodic (~30 second) intervals during the cook process (comark c28 k-type thermocouple, comark instruments, norfolk, england). crawfish were lowered into the steam agriculture and food sciences research, 2024, 11(2): 156-169 158 © 2024 by the authors; licensee asian online journal publishing group kettle after the water had been brought to a boil, completely submerged in the water, and continuously stirred with a plastic paddle to ensure that the crawfish were evenly heated. it took seven minutes for the water to return to a boil. the crawfish were then boiled for exactly three minutes. the cook time of ten minutes (as used by the d&t crawfish company in abbeville, louisiana) satisfied the adequate cook time of seven minutes or more [4] that would ensure the deactivation of proteolytic enzymes. at the end of the three-minute boil, steam was immediately turned off and several crawfish were retrieved to measure internal temperature using the comark c28 k-type thermocouple inserted into the thickest part of the tail. the internal temperature of the crawfish from the boiling trials ranged from 86.7 to 91.1°c. once the steam was turned off, crawfish were submerged in a sanitized plastic lug (1.2m x 1.2m x 0.9m) containing ice-water to rapidly cool the crawfish and prevent continued cooking. after five minutes of cooling crawfish temperatures were measured using the comark c28 k-type thermocouple before pouring into plastic lined, waxed fish boxes (0.6m x 0.3m x 0.3m) for cooling at 5°c for 4 hr prior to being separated into treatments of 3.5°c refrigerated storage and frozen storage at -18°c. a commercial batch vegetable blancher was used to steam crawfish. eight trials of approximately 7.3 kg of live crawfish were conducted because of the limited blancher capacity. live crawfish (~7.3kg) were placed in stainless steel trays and inserted into the chamber of the blancher. thermocouples (omega om-daqpro-5300 type-t) connected to a data logger (omega engineering inc., stamford, ct) were placed in the tails of two crawfish per trial. initial temperatures (uncooked) and final temperature of the crawfish during the cook process were recorded. the crawfish were steamed in the blancher using food-grade steam at 206.86 cmhg to the same internal temperature (90 to 92.2°c) reached during the boiling process. the crawfish were immediately removed from their trays and placed in an ice bath similar for five minutes of cooling with periodic stirring. crawfish were placed in plastic containers [0.6m x 0.3m x 0.3m] and stored in a cooler at 5°c for 4 hr prior to allotment of refrigerated storage at 4°c for 11 d and frozen storage at -18°c for 6 mo. at each month of frozen storage, crawfish were removed from the boxes, placed in shallow trays and thawed at 4°c for analyses. deactivation of proteolytic enzymes by the heating treatments was determined by a gelatin test [4]. following the 4-hour cooling period at 5°c, five g of hepatopancreas from each treatment including raw crawfish hepatopancreas were obtained by peeling the tails of 5-10 crawfish of each treatment. the hepatopancreas was then minced using a metal spatula and set aside at 5°c until used. then, in triplicate for each treatment, 0.2 g of mince was placed into labeled 22 mm tubes (pyrex®, corning corporation, tewksbury, ma). concurrently, blanks in triplicate, which contained no hepatopancreas, were prepared. five ml of cooled 12% unflavored gelatin (knox®, kraft foods group, inc., northfield, il) in water were then added to each tube and the contents homogenized using a vortex-genie® 2 mixer (scientific industries inc., bohemia, ny). the samples were then allowed to incubate for 1h at room temperature followed by holding at 3°c for 23 h. after the 23 hour refrigerated storage, the samples were removed and analyzed subjectively to determine the presence of a firm gel. a loose gel or no gel formation at all indicated that proteolytic enzymes present in the hepatopancreas had not been deactivated whereas firm gels revealed that the enzymes had been deactivated by an adequate cooking process. 2.2. microbiological analysis the microbiological analyses conducted on the crawfish included the quantification of the aerobic plate count (apc) and e.coli/coliforms of refrigerated samples on days 0, 1, 3, 5, 7, 9, and 11 and on frozen samples at months 0, 1, 2, 3, 4, 5, and 6. crawfish tail meat with adhering hepatopancreas was removed by hand peeling and placed in 17.78cm x 19.05cm reclosable storage bags (qwik seal®, reynolds®, lake forest, illinois). twenty-five gram samples [in duplicate for each treatment] were prepared and placed in 1.56kg bags (whirl-pak®, nasco, fort atkinson, wi) along with 25.0 ml of phosphate buffered saline (pbs), which was composed of 0.24% sodium phosphate monobasic [sigma-aldrich corporation, st. louis, mo], 0.28% sodium phosphate dibasic (sigma-aldrich corporation, st. louis, mo), and 0.85% sodium chloride (amresco llc, solon, oh) in distilled water. the 25 g of tail meat and 25 ml of pbs were homogenized for 60 seconds (easymix blender, aes chemumex, bruz cedex, france). from each treatment, serial dilutions were prepared and samples plated on both 3m™ e.coli/coliform and aerobic count petrifilms™ (3m company, st. paul, mn), then incubated at 35° ± 1°c. e.coli and coliforms were counted and recorded after 24 ± 3 hours as colony forming units and apc was counted and recorded after 48 ± 3 h (official methods 991.14 and 998.08 for e.coli/coliforms and official methods 990.12 for aerobic plate count, [aoac international, rockville, md], 2005). 2.3. color analysis hunter color scale values (l*, a*, and b*) were measured on ten randomly selected tails with hepatopancreas at the widest point at the back of the peeled tails for both boiled and steamed crawfish [calibrated cm-508d spectrophotometer, konica minolta sensing americas inc., ramsey, nj]. measurements on refrigerated samples were conducted on days 1, 3, 5, 7, 9, and 11 of refrigeration (4°c) and on frozen samples on months 1, 2, 3, 4, 5, and 6 of frozen storage (-18°c). 2.4. texture analysis texture analysis was conducted using a ta-xt plus texture analyzer (texture technologies corporation, scarsdale, ny) with a 5-blade kramer shear attachment and a 30 kg load cell. texture of peeled tails with adhering hepatopancreas from both boiled and steamed treatments was measured on days 0, 1, 3, 5, 7, 9, and 11 for refrigerated samples and after 0, 1, 2, 3, 4, 5, and 6 months of frozen storage. peak shear force (n) and work of shearing (n/s) were measured on 100 grams of peeled tail meat from each treatment in triplicate. one hundred grams of sample were placed randomly in the kramer cell, which filled the cell to roughly 40-50% capacity. the blade was set at 45 mm (~10mm above the sample) and the blade speed was set at 3 mm per second. following the texture determination, the samples were fully homogenized (oster® osterizer 14 speed all metal drive blender, jarden consumer solutions, providence, ri) and used for subsequent proximate and tbars analyses. agriculture and food sciences research, 2024, 11(2): 156-169 159 © 2024 by the authors; licensee asian online journal publishing group 2.5. proximate analysis proximate analysis (moisture, ash, protein, and fat) was conducted on refrigerated crawfish tail meat with adhering hepatopancreas on days 0, 1, 3, 5, 7, 9, and 11 and on frozen crawfish tail meat with adhering hepatopancreas at months 0, 1, 2, 3, 4, 5, and 6 for both boiled and steamed samples. moisture analysis was conducted in triplicate for each cooking treatment. three grams of homogenized tail meat were placed in ceramic crucibles, weighed, and then placed in an oven (model 20 gc lab oven, quincy lab inc., chicago, il) at 100°c for 24 hours. samples were removed from the oven and placed in a desiccator to cool before weighing. the percent moisture of tail meat with adhering hepatopancreas was determined by the calculation: % moisture = ((wet weight – dry weight)/wet weight)×100. ash measurements were conducted on the samples after moisture determination. the samples in triplicate were heated to 550°c for 18-24 hours (type 6000 furnace, thermolyne inc., dubuque, ia), removed from the furnace and placed in a desiccator to cool before weighing. the percent ash was determined using the calculation: %ash = (weight after ashing – tare weight of crucible)/(dry sample weight after oven drying – tare weight of crucible) × 100. lipid extraction was performed according to a modified method of bligh and dyer [27] described by woyewoda, et al. [28]. total lipid quantification was performed in duplicate for each treatment. following lipid quantification on a wet weight basis, fatty acid analysis was conducted on the lipid. peeled crawfish tail meat with adhering hepatopancreas were homogenized (oster® osterizer 14 speed all metal drive blender with a rosin blade, jarden consumer solutions, providence, ri). a measured amount of homogenized tail meat (approximately 50.0 grams) was added to a commercial blender (waring® model 51bl31, waring®, stanford, ct) along with 100.0 ml of high performance liquid chromatography (hplc) grade anhydrous methyl alcohol (avantor™ performance materials inc., center valley, pa) and 50.0 ml of hplc grade chloroform (mallinckrodt baker inc., phillipsburg, nj) and blended for exactly two minutes. an additional 50.0 ml of chloroform was added to the blender and the mixture was blended for an additional 30.0 seconds. the mixture was then filtered through a buchner funnel containing a #1 filter paper (whatman,™ ge healthcare uk limited, buckinghamshire, uk). aspiration was used to expedite the filtering process. the filtered product contained lipid, chloroform, and methanol. fifty ml of distilled water was added and the mixture was then stirred vigorously. the mixture was transferred to a 250-ml separatory funnel and then allowed to rest at 5°c overnight. the following day, the chloroform-lipid layer was then filtered from the separatory funnel through a glass funnel containing #4 filter paper inside #1 filter paper (whatman™, ge healthcare uk limited, buckinghamshire, uk) that was filled with american chemical society (acs) grade anhydrous sodium sulfate (thermofisher scientific, waltham, ma) into a pre-weighed 250 ml round bottom boiling flask (corning corporation, tewksbury, ma). the chloroform was then removed from the round bottom flask using a rotoevaporator (buchi rotovapor r114, buchi corporation, new castle, de). to ensure that all solvent was removed and only crude lipid remained, ultrahigh pure (uhp) nitrogen (air liquide corporation, paris, france) was sprayed into the flask for 10-15 minutes or until no odor or appearance of solvent remained. the crude lipid remaining was weighed and the percent fat was determined by the calculation: % fat = ((weight of flask containing lipid – weight of empty flask)/ weight of sample used) × 100. protein analysis was conducted on homogenized tail meat with adhering hepatopancreas from each cooking method. the samples were dried in the same manner as samples dried for moisture analysis. the dried samples were then blended to a fine powder using a mortar and pestle and coffee grinder (custom grind™, hamilton beach brands inc., south pines, nc). approximately 1.5 grams of finely ground sample were placed in 15 ml clear, screw top vials (supelco®, bellefonte, pa) and transported to the soil testing & plant analysis laboratory on the lsu campus for combustion and total nitrogen determination. total nitrogen determination was conducted on 0.25 gram samples in triplicate using a leco® truspec micro chns analyzer (leco corporation, st. joseph, mi). the results were reported as percent total nitrogen in the sample. the total percent protein was determined by multiplying the percent total nitrogen by the appropriate conversion factor of 6.25 and converting from a dry weight basis to a wet weight basis. 2.6. ph analysis the ph of crawfish tail meat with hepatopancreas was measured in triplicate for each treatment using a ph meter (sms115, milwaukee instruments inc., rocky mount, nc) that had been calibrated at ph values of 4,7, and 10. ten grams of homogenized crawfish tail meat were blended with 90 ml of distilled/de-ionized water for one minute using a commercial blender (waring® model 51bl31, waring, stanford, ct). the samples were then transferred to 400 ml beakers (pyrex, ®corning corporation, tewksbury, ma) and the ph was measured and recorded. 2.7. tbars analysis thiobarbituric acid reactive substances (tbars) analyses were conducted on boiled and steamed crawfish tail meat refrigerated for 0, 1, 3, 5, 7, 9 and 11 days and on frozen samples at months 0, 1, 2, 3, 4, 5, and 6 using a modified method of vyncke [29] by lemon [30]. solutions were prepared the day prior to analysis. the extraction solution consisted of 7.5% trichloroacetic acid (tca) (thermofisher scientific, waltham, ma), 0.1% propyl gallate (sigmaaldrich corporation, st. louis, mo), and 0.1% ethylene diaminetetraacetic acid (edta) (sigma-aldrich corporation, st. louis, mo) in 92.3% deionized water. the thiobarbituric acid (tba) solution consisted of 2.883 g/l (0.02m) of tba (sigma-aldrich corporation, st. louis, mo) in deionized water. the standard solution for standard curve determination was prepared by dissolving 0.22 grams of 1,1,3,3-tetraethoxypropane (tep) (sigma-aldrich corporation, st. louis, mo) in one l of water. the working solution to actually obtain the standard curve was formed by diluting the standard solution 100 fold. fifteen grams of tissue from each treatment [in triplicate] were blended with 30 ml of extraction solution for 30 seconds using a waring® blender. the samples were then filtered through a whatman #1 filter paper into a 100 ml pyrex® beaker. five ml of the clear filtrate in the beaker was added to five ml of tba reagent in pyrex® (120 x 10 mm) test tubes with screw caps. the test tubes were then heated in boiling water in 1000 ml beakers (pyrex®, corning corporation, tewksbury, ma) on a hot plate (corning pc-420d, corning corporation, tewksbury, ma). test tubes containing only five ml of water and five ml of tba agriculture and food sciences research, 2024, 11(2): 156-169 160 © 2024 by the authors; licensee asian online journal publishing group reagent were added to beakers as blanks. after boiling for exactly 40 minutes, the tubes were removed from the beaker and cooled in running tap water. using a transfer pipette, each sample was transferred to a cuvette and their optical density was measured at 530nm against the blanks of water and tba reagent (thermo spectronic genesys™ 2 spectrophotometer, thermofisher scientific, waltham, ma). the tbars values were calculated from the standard curve obtained from the tep working solution and the values were reported in mg malonaldehyde (mda) equivalent/100 grams of tissue. 2.8. mineral analysis mineral analyses were conducted on the ashed samples remaining from the proximate analysis of boiled and steamed crawfish tail meat that had been refrigerated for days 0,1,3,5,7,9, and 11 and frozen samples on months 1,2,3,4,5, and 6. ten ml of 10% nitric acid (avantor™ performance materials inc., center valley, pa) solution in distilled water were added to each crucible for ten minutes to solubilize the ash. the solubilized ash samples were drawn into a sterile 10 ml tip syringe (luer-lok™, becton dickinson & company, franklin lakes, nj). a 0.2μm, 25-mm surfactant free cellulose acetate membrane, acrylic housing, syringe filter was placed on the syringe (nalgene™, thermofisher scientific, waltham, ma) and the sample was filtered into labeled 15 ml clear, screw top vials (supelco®, bellefonte, pa). two of the three samples were then transported to the lsu agcenter’s w.a. callegari environmental center central research station in baton rouge, louisiana for analysis in duplicate via inductively coupled plasma optical emission spectroscopy (icp-oes, varian vista-mpx ccd simultaneous icpoes unit, varian medical systems inc., palo alto, ca). the results for minerals present in the samples were reported in parts per million [ppm] of crawfish tail meat with adhering hepatopancreas on a wet weight basis. 2.9. fatty acid analysis fatty acid profiles were obtained from samples of lipid collected during proximate analysis (2.5). in duplicate, 1.3-2.0 grams of crude lipid was solubilized in exactly ten ml of hplc grade hexane (honeywell, morristown, nj) and transferred to a 15 ml clear, screw top vial (supelco®, bellefonte, pa). samples were then placed in the freezer (-18°c) until they were transported to the lsu agcenter’s w.a. callegari environmental center central research station in baton rouge, louisiana for analysis. fatty acid analysis was conducted using a gas chromatograph with an ion trap mass spectrometer (varian 450 and varian 240, respectively, varian inc., palo alto, ca) with a 75 m, 0.18 mm diameter, 0.14 μm film thickness, silica capillary column (sp2560, supelco®, bellefonte, pa) with hydrogen as the carrier gas at a flow rate of 40 cm per second. 2.10. statistical analysis results were expressed as least squares means (ls-means) ± standard deviation. the experimental design used was a 2x7 factorial design for both refrigerated and frozen studies. statistical analysis was performed using analysis of variance (anova). separation of means and difference between control and treatments were determined by the generalized linear model (glm) procedure with a t comparison for least squares means (sas institute, version 9.3). statistical significance was set at p < 0.05. 3. results and discussion 3.1. cooking, cooling, and gelatin test crawfish were submerged in an ice bath immediately after cooking to retard the cook process and to cool the crawfish within recommended guidelines [31] which slowed the rate of spoilage reactions that include bacterial and autolytic enzyme activity and reduced the rate at which bacteria multiply [32]. an average internal temperature of 87.6° c was obtained [5 crawfish per each of 2 replications] for the boiled crawfish, which killed pathogen listeria monocytogenes [31]. the average chilled temperature of the crawfish after the ice bath for the two trials was 30.11°c. after four hours of cooling in refrigeration, the temperature of all the crawfish was reduced to an average of 3.5°c. the average internal temperature of the cooked tails in the eight replications of steaming was 86.9°c. after the ice bath, the average internal temperature of the steamed samples was 29.6°c. the slightly lower internal temperature of the boiled samples might be due to the smaller batch size of ~7.26 kg versus the boiled batch sizes which were ~ 27.22 kg. once the steamed samples had been boxed and allowed to cool under refrigeration for four hours, the average internal temperature was 5.0°, similar to that of the boiled samples. boiled and steamed crawfish samples contained cooked hepatopancreas and formed stable, firm gels after cooking, which indicates that the cook time was sufficient to deactivate the proteolytic enzymes in the hepatopancreas [33]. the heat-labile proteolytic enzymes present in the hepatopancreas of crawfish promote the development of mushiness in the tail meat [34]. it is important to limit texture deterioration before freezing of crawfish tail meat as godber, et al. [12] found extended frozen storage would cause poor texture. 3.2. tail meat yield the edible tail meat yields for the boiled and the steamed crawfish through refrigerated and frozen storage are in table 1. it was expected that the edible yield would decrease during storage, especially frozen storage from freezethaw transitions and thawing due to purge. slow freezing causes large ice crystal formation that damages cell walls and promotes water loss upon thawing [1, 35]. since crawfish are generally hand peeled, there can be variations in the yield obtained from peeling depending on the technique and experience of the peeler. the average abdominal [tail] meat yield for cooked crawfish is about 15% of the live weight of the crawfish [8]. the yield also depends on the sexual maturity of the crawfish, with immature crawfish having a higher yield because they have smaller claws and thinner shells [8]. this is typical of crawfish early in the season, when yields can be as high as 22-23%. later in the season, when the crawfish have matured and have larger claws and thicker shells, the yield can be as low as 10 to 11% [8]. the yields obtained in this study averaged around 18%. this was higher than expected because these crawfish were harvested later in the season and their shells were quite thick. a reason why the yields may have been higher than expected is that the agriculture and food sciences research, 2024, 11(2): 156-169 161 © 2024 by the authors; licensee asian online journal publishing group crawfish were very carefully peeled. also, the hepatopancreas was left attached as well as the vein/intestine that runs along the dorsal side of the abdomen/tail. typically, this is removed during commercial peeling, but was left on in this research because the goal was to mimic how louisiana consumers typically ingest boiled crawfish. cooking losses of 31.17% and 18.33% from fried and microwave cooking, respectively, were reported [28]. table 1. percent (%) edible yield of peeled crawfish tail meat with adhering hepatopancreas during refrigerated (3.5°c) storage or after frozen (-18°c) storage and thawing. refrigerated day 0 day 1 day 3 day 5 day 7 day 9 day 11 avg. % yield boiled 19.14 18.41 20.16 18.09 18.02 19.46 17.43 18.67 ± 0.95 steamed 18.58 18.63 18.60 17.84 17.63 19.19 15.80 18.03 ± 1.11 frozen mo. 0 mo. 1 mo. 2 mo. 3 mo. 4 mo. 5 mo. 6 avg. % yield boiled 19.14 17.69 17.36 19.16 17.28 18.13 17.52 18.04 ± 0.81 steamed 18.58 17.08 17.98 18.71 16.84 18.92 19.45 18.22 ± 0.96 3.3. proximate analysis results the proximate analyses ls-means for the boiled and steamed refrigerated and frozen crawfish tail meat with the attached hepatopancreas are in table 2. the proximate analyses from the usda [36] and sidwell [37] are also shown for comparison. it was not specified whether or not the tail meat samples included hepatopancreas from in the usda [36] or sidwell [37] studies. table 2. means for % moisture, ash, protein, and fat of crawfish tail meat during refrigerated (3.5°c) and frozen (-18°c) storage. proximate values from references 26, 36, and 37 are shown for comparison. proximate composition boiled steamed usda [36] boiled sidwell [37] boiled sidwell [37] raw range aboutaleb, et al. [26] raw % moisture (3.5°c) 79.29 79.40 % moisture (-18°c) 77.91 77.86 80.8 75.0 (72.1 83.4) 78.61 % ash (3.5°c) 1.26 1.31 % ash (-18°c) 1.22 1.32 1.07 1.5 (0.7 3.60 1.33 % protein (3.5°c) 15.89 15.30 % protein (-18°c) 16.69 16.30 17.5 16.3 (11.9 24.10 18.46 % fat (3.5°c) 2.73 2.96 1.58 % fat (-18°c) 3.00 3.22 1.3 0.8 (0.5 2.5) total (3.5°c/-18°c) 99.17 ⁄ 98.82 98.97 ⁄ 98.70 100.67 98.9 99.98 moisture, ash, and protein in crawfish boiled for 10 m were 80%, 1.2%, and 17%, respectively, [25] while the moisture for crawfish boiled for 5 m was 86% [20]. lower moisture (38.2 and 58.2%) and higher protein (35.8 and 35.2), fat (18.9 and 2.73%), and ash (6.35 and 3.25%) were in cooked crawfish by frying and microwaving [26]. the moisture content of the peeled crawfish tail meat with adhering hepatopancreas through refrigerated and frozen storage from boiled and steamed treatments did not vary greatly. there was little change in the moisture content for the boiled samples through 0 to 11 d refrigerated storage (table 3). the moisture content for the steamed samples varied a bit more than the boiled samples throughout the 11-day storage. the differences in moisture could be from natural variation, which corresponds to previously reported large range of moistures (72.1 83.4%) [37]. table 3. percent (%) moisture of peeled crawfish tail meat with adhering hepatopancreas stored during refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 78.23d ± 0.08 78.77cd ± 0.12 78.93cd ± 0.05 80.33ab ± 2.04 80.19ab ± 0.08 78.46c ± 0.10 80.11ab ± 0.14 steamed 79.00cd ± 0.17 79.30abcd ± 0.02 78.22d ± 0.05 79.84abc ± 0.36 79.77abc ± 0.02 79.27bcd ± 0.15 80.38a ± 0.11 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcd) are not different (p < 0.05). a greater difference in moisture content with cooking treatments was observed between frozen samples (which were thawed prior to moisture determination) than for samples in refrigerated storage (table 4). ice crystal formation, protein denaturation, and increases in salt concentration during frozen storage can decrease the moisture content of the thawed product [38, 39]. it was thought that ice crystal formation may have been a factor because slow freezing rates cause larger ice crystal formation, which would result in cellular disturbance and rupturing [1, 40] and increased water loss or drip-loss from the crawfish upon thawing. there was a 13 to 20% drip loss during freeze-thaw cycles of crawfish previously boiled for 15 m [41]. there was a significant difference between the boiled and steamed samples at all time intervals except for months four and five. table 4. percent (%) moisture of peeled crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 month 6 boiled 78.23bc ± 0.08 77.05j ± 0.13 78.36bc ± 0.04 78.21cd ± 0.07 77.90efg ± 0.06 77.65h ± 0.17 77.95ef ± 0.10 steamed 79.00a ± 0.17 77.72gh ± 0.06 77.34i ± 0.14 78.42b ± 0.11 78.02de ± 0.26 77.79fgh ± 0.12 76.71k ± 0.04 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdefghijk) are not different (p<0.05). the ash contents obtained in this study did not exhibit significant differences between the boiled and steamed crawfish samples in refrigerated storage, except on d 9 (table 5). as sidwell [37] reported, the range for percent ash in an unspecified species of raw crawfish was 0.7-3.6% which indicates quite a large range of natural variation. agriculture and food sciences research, 2024, 11(2): 156-169 162 © 2024 by the authors; licensee asian online journal publishing group another study had similar results, an ash content of 1.5% [41]. the values for percent ash content of the refrigerated samples (boiled and steamed) are shown in table 5. table 5. percent (%) ash of peeled crawfish tail meat with adhering hepatopancreas stored during refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 1.30bcd ± 0.10 1.56a ± 0.16 1.15e ± 0.03 1.25bcde ± 0.08 1.21bcde ± 0.03 1.17de ± 0.06 1.19cde ± 0.08 steamed 1.25bcde ± 0.04 1.56a ± 0.04 1.23bcde ± 0.12 1.31bc ± 0.02 1.28bcde ± 0.03 1.33b ± 0.01 1.19cde ± 0.08 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcde) are not different (p<0.05). the percent ash contents of the boiled and steamed samples after frozen storage and thawing were slightly higher, on average, than the samples in refrigerated storage (table 6). this might indicate more leaching of minerals in the boiled samples compared to that of the steamed samples. blanching, or boiling, of foods has a tendency to cause leaching of vitamins and minerals; steaming has the benefit of less leaching than blanching/boiling [42]. table 6. percent (%) ash of peeled crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 month 6 boiled 1.29abc ± 0.10 1.01d ± 0.39 1.31abc ± 0.04 1.15cd ± 0.02 1.21bc ± 0.02 1.22bc ± 0.04 1.33abc ± 0.00 steamed 1.25abc ± 0.04 1.40ab ± 0.05 1.22bc ± 0.08 1.21bc ± 0.07 1.40ab ± 0.07 1.34abc ± 0.03 1.44a ± 0.02 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcd) are not different (p<0.05). the protein contents of boiled samples were significantly higher than for steamed crawfish on day of refrigerated storage, except d 7 (table 7). although significant, the actual differences were small. for both the boiled and steamed samples, there was an overall decrease in percent protein from d 0 to 11. this may be due to increased microbial counts and the hydrolysis and consumption of free amino acids and other soluble non-nitrogenous substance in the crawfish that serve as nutrients for microbial growth [11]. table 7. percent (%) protein of peeled crawfish tail meat with adhering hepatopancreas stored during refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 16.32c ± 0.04 16.46b ± 0.06 16.93a ± 0.04 15.06g ± 0.00 14.95h ± 0.07 16.23d ± 0.04 15.27f ± 0.05 steamed 16.15c ± 0.03 15.34f ± 0.06 16.02e ± 0.06 14.56i ± 0.10 15.07g ± 0.04 15.05g ± 0.02 14.91h ± 0.10 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdefgh) are not different (p<0.05). the protein contents of the boiled samples during frozen storage were higher (p<0.05) than steamed samples at every month except month six. there was a difference between the protein content of the frozen samples and the refrigerated samples. the frozen values were larger by almost one % (table 8). these values also fall within the range (11.9 24.1%) for percent protein in raw samples [37]. table 8. percent (%) protein of peeled crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 month 6 boiled 16.32ef ± 0.04 17.31a ± 0.06 16.52d ± 0.03 16.71c ± 0.03 16.21fg ± 0.19 17.26a ± 0.02 16.51d ± 0.04 steamed 16.15gh ± 0.03 16.02hi ± 0.05 16.32de ± 0.06 16.44de ± 0.16 15.90i ± 0.04 16.37e ± 0.05 16.90b ± 0.01 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdefgh) are not different (p<0.05). there were some differences in the protein content between boiled and steamed crawfish during refrigerated and frozen storage. the differences could simply arise from natural variation in the samples that were selected. the protein content of frozen samples tended to be higher than that of the refrigerated samples. the percent fat values obtained in this study (2.12 to 3.76%) were higher than literature values (table 9). this may be due to the inclusion of the hepatopancreas (fat) with the tail meat in the present study. in louisiana, crawfish are traditionally consumed with the fat [7] and so the hepatopancreas was kept attached to the tail meat to mimic the way that crawfish are commonly consumed in louisiana. most studies delineating the fat content of crawfish tail meat do not include the hepatopancreas. the hepatopancreas is roughly 30% fat by weight [43]. therefore, the fat content of crawfish with hepatopancreas would be expected to be higher than crawfish tail meat without hepatopancreas. table 9. percent (%) average fat of peeled crawfish tail meat with adhering hepatopancreas during refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 2.99cde ± 0.04 2.91de ± 0.04 2.58f ± 0.08 2.27g ± 0.13 2.24g ± 0.06 3.27bc ± 0.07 2.88de ± 0.11 steamed 2.76def ± 0.16 2.75ef ± 0.27 3.04cd ± 0.03 2.12g ± 0.11 3.41a ± 0.27 2.90de ± 0.08 3.76a ± 0.06 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdefg) are not different (p<0.05). the average fat content of the frozen crawfish was higher for both the boiled and steamed treatments than for the same treatments with refrigerated storage (table 10). this might be explained by the reduced moisture in the frozen samples compared to the refrigerated samples. the variation is much more pronounced for the steamed samples than for the boiled samples. agriculture and food sciences research, 2024, 11(2): 156-169 163 © 2024 by the authors; licensee asian online journal publishing group table 10. percent (%) fat of peeled crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 month 6 boiled 2.99def ± 0.04 3.10de ± 0.0 2.72h ± 0.08 3.13cde ± 0.10 3.00def ± 0.08 2.95efg ± 0.01 3.15cde ± 0.04 steamed 2.76gh ± 0.16 3.07def ± 0.04 3.34bc ± 0.06 3.87a ± 0.10 2.86fgh ± 0.03 3.21bcd ± 0.26 3.42b ± 0.11 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdefgh) are not different (p<0.05). 3.4. microbiological results there were no e. coli/coliforms present in any of the water samples or any of the boiled or steamed crawfish at any time. there was no presence of e. coli, but there was an average coliform count of 295 colony forming units (cfu)/g in raw crawfish. this suggests that the cook procedure for both boiling and steaming was adequate to kill these organisms. an apc count of log 5.7 cfu/g is suggested by icmsf [24] as the upper limit of acceptability and, in this study, taken as the upper limit for shelf life determination. as seen in table 11, the boiled and steamed samples in refrigerated storage did not exceed the limit of log 5.7 cfu/g until after day three. both the boiled and steamed samples exceeded the chosen limit by day 5, the data point immediately after day 3, so the acceptable shelf life of refrigerated crawfish, either boiled or steamed, was taken as three days. it is not clear why the difference in apc occurred. further study with analyses conducted on all days may provide better insight. total bacteria and psychrophilic bacteria counts were 4.6 and 4.7 and 3.3 and 3.5 log cfu/g for fried and microwaved crawfish in refrigerated storage [26]. table 11. aerobic plate counts (log10 cfu/g) for peeled crawfish tail meat with adhering hepatopancreas during refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 3.03c ± 2.87 3.10c ± 2.87 3.03c ± 2.91 6.10c ± 5.36 6.12c ± 5.05 8.10b ± 7.08 8.01b ± 6.94 steamed 2.57c ± 2.53 2.78c ± 1.97 5.46c ± 4.89 6.22c ± 5.08 tnc* 8.62a ± 7.53 8.65a ± 8.00 note: ls-means ± sd of 4 measurements at each time period. ls-mean values with the same letter (abc) are not different (p<0.05). * tnc = too numerous to count. the apc counts were constant during frozen storage and below 5.7 log cfu/g at all data points as shown in table 12 which would be expected during frozen storage at -18°c. this suggests that the shelf life, based upon apc counts, of crawfish in frozen storage is at least six months. this contrasted with results of decreased ca 2 + -atpase activity, salt soluble protein content, total sulfhydryl and reactive sulfhydryl content and increased hardness at week 4 to suggest that the recommended shelf-life of frozen crayfish should be 1 month {1]. table 12. aerobic plate counts (log10 cfu/g) for peeled crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 month 6 boiled 3.02a ± 2.87 2.27d ± 2.18 2.48d ± 1.98 2.90abc ± 2.19 2.69bcd ± 2.00 2.78bcd ± 2.00 2.79bcd ± 2.77 steamed 2.57cd ± 2.53 2.70bcd ±2.02 2.69bcd ±1.83 2.95ab ± 2.56 2.56cd ± 1.70 2.92ab ± 2.41 3.04a ± 2.41 note: ls-means ± sd of 4 measurements at each time period. ls-mean values with the same letter (abcd) are not different (p<0.05). 3.5. tbars results tbar values increased during refrigerated storage for both boiled and steamed samples but the level of tbars measured at the end of the refrigerated storage was lower than anticipated (table 13). it was anticipated that the values would be higher than previous studies because the crawfish in the current study had the hepatopancreas and vein, increasing the amount of fat that might be oxidized and result in higher tbars values. surprisingly, the tbars values obtained in this study were lower than literature values. an average tbars value of 2.24 mg mda/kg after 7 days [44] and 0.113 after 9 days of refrigerated storage [26] were obtained on crawfish tail meat. table 13. tbars (mg mda/kg) for peeled crawfish tail meat during refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 0.19fg ± 0.02 0.15h ± 0.01 0.24e ± 0.00 0.33cd ± 0.03 0.34c ± 0.01 0.42b ± 0.04 0.45b ± 0.02 steamed 0.16gh ± 0.01 0.14h ± 0.02h 0.22ef ± 0.00 0.29d ± 0.03 0.30d ± 0.04 0.32cd ± 0.01 0.53a ± 0.03 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdefgh) are not different (p<0.05). during refrigerated storage, tbars values of the boiled and steamed samples gradually increased from d 0 to 11. it was unexpected that tbars on d 0 were more than on d 1. one possible explanation is that the samples on d 0 remained at room temperature slightly longer before tbars analyses than on the other days of the study. tbars values, as expected, increased with refrigerated storage with boiled samples having slightly higher values than the steamed samples on all days except d 11. tbars of 1.5 mg mda/kg was the point at which humans can detect any off flavors and 3.0 mg mda/kg of tbars was the level at which crawfish are considered rancid and not desirable for consumption [44]. using these criteria, the fat in the crawfish tail meat and hepatopancreas was not rancid at any time during the course of the study. however, it was noticed that the crawfish in the last 5 to 6 days of refrigeration had an odor that was unpleasant. the tbars values were similar to those of abou-taleb, et al. [26] but cremades, et al. [45] identified much higher levels of tbars of crawfish in refrigerated storage. agriculture and food sciences research, 2024, 11(2): 156-169 164 © 2024 by the authors; licensee asian online journal publishing group table 14. tbars (mg mda/kg) for peeled crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 boiled 0.19f ± 0.02 0.22def ± 0.02 0.16g ± 0.02 0.21def ± 0.01 0.23cd ± 0.01 0.2cde ± 0.01 steamed 0.16g ± 0.01 0.25bc ± 0.00 0.20ef ± 0.00 0.22cde ± 0.01 0.19f ± 0.01 0.26b ± 0.04 after frozen storage (table 14), the tbars values for both boiled and steamed samples, although having statistically significant differences, were essentially stable in contrast to the tbars values of the refrigerated samples. [46] observed an average tbars value of 4.00 mg mda/kg over the course of 10 months of frozen storage, with a maximum value at month ten of 5.60 mg mda/kg in crawfish tails containing the hepatopancreas. tbars in crawfish tail meat greatly increased after 30 d of frozen storage [20] and a linear increase in tbars was found with repeated freeze-thaw cycles [41]. 3.6. texture texture analyses were conducted on crawfish tail meat with adhering hepatopancreas over the course of refrigerated and frozen storage. table 15 shows the peak shear force for boiled and steamed peak shear force values for crawfish during 11 days of refrigerated storage. table 15. peak shear force (kg) for peeled crawfish tail meat with adhering hepatopancreas after refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 231.54abcd ± 8.63 240.66abc ± 20.89 262.43a ± 28.24 218.59bcde ± 11.77 230.95bcd ± 5.28 248.99ab ± 33.73 228.10bcd ± 8.73 steamed 224.57cd ± 30.30 208.39de ± 14.51 217.41cde ± 19.22 217.81bcde ± 14.71 236.14abcd ± 7.75 220.65bcd ± 15.98 188.19e ± 14.12 the peak shear force (kg) for the boiled samples in refrigerated storage gradually increased from day zero to day three even though the values were not different (p<0.05); there was greater variability from day three to eleven. the trend was somewhat similar for steamed samples, but the final value for steamed samples was lower than for boiled crawfish on d 11. the frozen shear force values (table 16) are statistically similar but the shear force value on day eleven was the lowest which follows previous results [47] that proteolytic enzyme activity and an increased bacterial presence through the refrigerated storage caused the deterioration in the texture of the crawfish. the results indicated that toughening due to moisture loss or mushy or deteriorative texture due to residual activity of native proteinases in the hepatopancreas or tail that had not been deactivated by the thermal treatment (cooking) process did not substantially influence texture. however, d 7 variations in texture may also be due to the inherent nature of crawfish and the kramer analysis. the uniformity of the sample and direction of the muscle fibers plays a role in the outcome of the analysis [48]. that the crawfish were placed in the kramer cell at random and did not completely cover the bottom of the cell in the present study may have had an impact. the weight of crawfish added to the cell was kept constant for all samples, however the number of crawfish changed since the weights of individual crawfish tails varied. table 16. peak shear force (kg) for peeled thawed crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 month 6 boiled 231.54f ± 8.63 277.23abc ± 24.12 266.15abcde ± 19.81 266.05abcde ± 16.97 291.06a ± 26.08 272.72abcd ± 3.63 282.23ab ± 18.93 steamed 224.57f ± 30.30 236.14e ± 18.63 242.42def ± 7.75 247.52cdef ± 9.71 252.62bcdef ± 27.75 265.17abcde ± 16.18 285.86a ± 8.34 the peak shear forces (kg) for the thawed previously frozen crawfish tail meat with attached hepatopancreas were more consistent than for the refrigerated samples, with the boiled samples generally tougher than the steamed samples. however, at month six, the steamed sample had a higher peak shear force than the boiled sample. the hardness, springiness, and chewiness of crawfish varied among freeze-thaw cycles [41]. the work of shearing was chosen as a measure of toughness perceived when biting into a food product by measuring the force needed to shear the sample as a product of time. the results of the work of shearing mimic the values of peak shear force, which might be expected considering the measurements were taken simultaneously. values for work of shearing of boiled and frozen crawfish tails through refrigerated storage are in table 17 and through frozen storage are in table 18. the hardness, resilience, cohesiveness, and chewiness changed with the times of steaming and boiling of crawfish tail meat [2]. table 17. total shear work (n/s) for peeled crawfish tail meat with adhering hepatopancreas after refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 1059.31bcde ± 86.00 1081.67abcd ± 152.40 1241.42a ± 52.56 1045.29bcde ± 133.57 1045.49bcde ± 57.07 1092.56abc ± 35.01 1011.07bcdef ± 89.34 steamed 1105.11ab ± 147.59 907.31ef ± 15.59 994.69bcdef ± 110.62 923.59def ± 118.56 1105.80ab ± 70.51 933.20cdef ± 86.00 849.26f ± 101.99 note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letters (abcdef) are not different (p<0.05). note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcde) are not different (p<0.05). note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdef) are not different (p<0.05). note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdef) are not different (p<0.05). agriculture and food sciences research, 2024, 11(2): 156-169 165 © 2024 by the authors; licensee asian online journal publishing group table 18. total shear work (n/s) for peeled thawed crawfish tail meat with adhering hepatopancreas after frozen storage (-18°c) and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 month 6 boiled 1059.31cd ± 86.00b 1157.48bc ± 138.57 1130.41bc ± 115.72 1073.44bcd ± 117.29 1347.63a ± 36.38 1226.71ab ± 33.05 1101.48bc ± 76.3 steamed 1105.11bc ± 147.59 832.19e ± 43.64 1008.71cde ± 33.34 895.94de ± 52.86 1170.52abc ± 181.33 1182.68abc ± 200.45 1138.75bc ± 54.43 3.7. ph results there was no general trend for the ph in the boiled crawfish during refrigerated storage, presented in table 19. for the steamed samples, the initial ph declined during storage to a ph value of 7.77 on day 11. the variable ph for boiled and steamed samples through the refrigerated storage can be attributed to changes in production of basic compounds such as ammonia and other biogenic amines [49, 50]. the ph of fried and microwaved crawfish meat were 5.75 and 5.80, respectively, for refrigerated samples [26]. there were also significant decreases from month zero to month six (which had the lowest ph values) for both boiled and steamed crawfish tail meat in frozen storage (table 20). reported ph values were 7.66 to 7.79 during 180 d frozen storage [20] and from 7.35 to 7.85 during repeated freeze-thaw cycles [41]. table 19. ph of peeled crawfish tail meat with adhering hepatopancreas during refrigerated (3.5°c) storage. treatment day 0 day 1 day 3 day 5 day 7 day 9 day 11 boiled 8.10ef ± 0.00 8.20cd ± 0.00 8.10ef ± 0.00 8.17de ± 0.06 8.40a ± 0.00 8.27bc ± 0.06 8.03f ± 0.06 steamed 8.30b ± 0.00 8.20cd ± 0.00 8.10ef ± 0.00 8.13de ± 0.06 8.17de ± 0.06 8.03f ± 0.06 7.77g ± 0.06 table 20. ph of peeled thawed crawfish tail meat with adhering hepatopancreas after frozen storage (-18°c) and thawing. treatment month 0 month 1 month 2 month 3 month 4 month 5 month 6 boiled 8.10de ± 0.00 8.20bc ± 0.00 8.27ab ± 0.06 8.23abc ± 0.06 8.20bc ± 0.00 7.87fg ± 0.06 7.87g ± 0.06 steamed 8.30a ± 0.00 8.03e ± 0.0 8.20bc ± 0.00 8.17cd ± 0.06 8.20bc ± 0.00 7.93f ± 0.06 7.83g ± 0.06 3.8. color results the results of the color analyses for lightness (l*), green/red (a*), and blue/yellow (b*) showed only slight differences throughout refrigerated and frozen storage. for boiled samples during refrigerated storage, the l* values were similar except for the difference (p<0.05) in l* between d 7 and 11 (table 21). table 21. l* of peeled crawfish tail meat with adhering hepatopancreas during refrigerated (3.5°c) storage. treatment day 1 day 3 day 5 day 7 day 9 day 11 boiled 53.30bc ± 6.26 54.06abc ± 3.45 53.23bc ± 6.39 50.61c ± 6.23 54.73abc ± 4.28 57.84ab ± 6.70 steamed 53.65bc ± 5.57 52.91bc ± 5.85 51.34c ± 3.85 55.10abc ± 3.91 56.06abc ± 7.20 59.33a ± 10.64 for the steamed samples, lightness values slowly decreased from day one to day five, then increased to day eleven. there were no differences (p<0.05) between the two treatments at each individual day. table 22. l* of peeled thawed crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 1 month 2 month 3 month 4 month 5 month 6 boiled 52.32ab ± 4.28 54.45ab ± 5.76 53.74ab ± 6.66 56.60a ± 7.43 48.63b ± 5.91 48.58b ± 6.92 steamed 52.12ab ± 9.18 53.11ab ± 4.45 52.46ab ± 6.58 52.84ab ± 7.29 49.86b ± 6.72 54.23ab ± 7.71 the l* values of the boiled samples through frozen storage were all similar except that month 4 was higher (p<0.05) than months 5 and 6 and the boiled and steamed frozen crawfish l* values were not different (p<0.05) from each other at any month, as shown in table 22. these l* values through refrigerated storage were slightly higher than the samples in frozen storage. l* values showed a general decrease with increased frozen storage time in the study by bonilla, et al. [20]. the a* values or green/red scale of color showed little differences (p<0.05) in table 23 for boiled crawfish samples throughout refrigerated storage with day 3 having the highest (p<0.05) a* value. values for the steamed samples fluctuated and on day 11 the steamed samples had much smaller values a* values than the boiled samples. note: ls-means ± sd of 3 measurements at each time period. abcdels-mean values with the same letter (abcde) are not different (p<0.05). note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcdefg) are not different (p<0.05). note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abcedfg) are not different (p<0.05). note: ls-means ± sd of 3 measurements at each time period. ls-mean values with the same letter (abc) are not different (p<0.05). note: ls-means ± sd of 10 measurements at each time period. ls-mean values with the same letter (ab) are not different (p<0.05). agriculture and food sciences research, 2024, 11(2): 156-169 166 © 2024 by the authors; licensee asian online journal publishing group table 23. a* of peeled crawfish tail meat with adhering hepatopancreas in refrigerated (3.5°c) storage. treatment day 1 day 3 day 5 day 7 day 9 day 11 boiled 32.02bcd ± 6.39 35.52ab ± 5.73 32.58bcd ± 4.63 31.65bcd ± 3.74 29.35cd ± 3.63 31.06bcd ± 5.77 steamed 34.18abc ± 8.04 38.23a ± 6.94 31.47bcd ± 5.27 27.35de ± 4.03 29.35de ± 3.63 22.91e ± 10.45 the a* values for the boiled and steamed frozen samples were similar at each month except month 6, where the a* values of the boiled samples were larger than the steamed samples (table 24). table 24. a* of peeled thawed crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 1 month 2 month 3 month 4 month 5 month 6 boiled 25.80bc ± 4.08 26.58bc ± 5.04 27.24bc ± 7.27 24.55bc ± 8.27 30.15ab ± 6.23 32.93a ± 5.77 steamed 23.82c ± 8.18 30.12ab ± 3.48 27.21bc ± 6.21 26.99bc ± 4.83 28.93abc ± 7.80 25.39c ± 7.40 the b* values [blue/yellow color scale] fluctuated during the eleven days of refrigerated storage (table 25). the b* value of boiled samples increased (p<0.05) from day one to day three, then decreased from d 3 to 7 when the lowest value of 22.91 occurred. the b* values then steadily increased to d 11. steamed crawfish tail meat also had variable b* values in refrigerated storage, with a high value of 40.24 on day three and values that decreased from day five through day 9. it is not understood what caused the fluctuation in the b* values during the refrigerated storage. table 25. b* of peeled crawfish tail meat with adhering hepatopancreas in refrigerated (3.5°c) storage. treatment day 1 day 3 day 5 day 7 day 9 day 11 boiled 32.48c ± 4.66 37.78ab ± 5.99 30.78cd ± 4.88 22.91f ± 3.40 26.29def ± 4.63 32.35c ± 4.30 steamed 34.68bc± 6.31 40.24a ± 5.35 30.10cde ± 4.10 26.77def ± 4.98 25.49ef ± 3.30 27.60de ± 8.46 during frozen storage, the b* values were lower than the b* values during refrigerated storage (table 26) which corresponds to the differences in the l* and a* values with type of storage. the steamed and boiled samples were the same throughout the six months of storage at every month. cryogenically frozen crawfish showed minimal changes in b* values while blast frozen crawfish had variation in b* values during 180 d frozen storage [20]. table 26. b* of peeled crawfish tail meat with adhering hepatopancreas after frozen (-18°c) storage and thawing. treatment month 1 month 2 month 3 month 4 month 5 month 6 boiled 22.57ab ± 5.37 24.30ab ± 5.04 24.24ab ± 4.73 25.60a ± 3.86 26.12a ± 5.56 25.59a ± 5.13 steamed 20.94b ± 7.84 25.03ab ± 2.22 22.50ab ± 2.22 25.82a ± 2.13 24.86ab ± 4.47 25.11a ± 3.78 the l*, a*, and b* values all decreased during frozen storage in comparison to their refrigerated samples. l*, a*, and b* values varied greatly during consecutive freeze-thaw cycles of crawfish during frozen storage [13]. 3.9. mineral results mineral analyses shown in table 27 were conducted to determine the presence and quantity of 29 different minerals and metals that might have caused differences in crawfish properties throughout the refrigerated and frozen storage. the amount of arsenic is pertinent as many crawfish are farmed in rice ponds, as in this study, and levels of inorganic and organic arsenic are of concern in rice and rice products [51, 52]. the arsenic levels observed in the crawfish in the present study are higher than that which is approved by the environmental protection agency (epa) for allowable limits in drinking water. the limit for arsenic in drinking water is 10 parts per billion (ppb) or 0.01 parts per million (ppm). the mineral values varied greatly from those of frozen crawfish in the bonilla, et al. [20] possibly due to the inclusion of the hepatopancreas and veins in the samples analyzed in the current study. table 27. selected minerals (ppm) in tail meat with adhering hepatopancreas. mineral boiled avg. ± sd (ppm) boiled range (ppm) steamed avg. ± sd (ppm) steamed range (ppm) other sources (ppm) al 9.67 ± 2.98 4.72 – 13.78 8.54 ± 3.68 4.01 – 16.83 _ mg 86.08 ± 4.86 78.02 90.36 81.76 ± 4.73 70.97 – 87.64 330 [35] mn 1.04 ± 0.24 0.41 – 1.29 1.06 ± 0.24 0.36 – 1.29 4.2-7.28 [36] k 576.32 ± 46.15 641.46 – 494.79 647.13 ± 64.06 535.89 – 738.37 5,000 [36] na 341.98 ± 21.80 300.21 – 364.26 383.51 ± 39.82 325.93 – 431.24 1,820 [36] si 15.77 ± 2.91 11.18 – 21.30 15.71 ± 3.75 10.22 – 21.58 _ b 4.27 ± 0.32 3.73 – 4.77 4.43 ± 0.28 3.84 – 4.85 _ note: *ls-means ± sd of 10 measurements at each time period. ls-mean values with the same letter (abcde) are not different (p<0.05). note: ls-means ± sd of 10 measurements at each time period. ls-mean values with the same letter (abc) are no different (p<0.05). note: ls-means ± sd of 10 measurements at each time period. ls-mean values with the same letter (abcdef) are not different (p<0.05). note: ls-means ± sd of 10 measurements at each time period. ls-mean values with the same letters (ab) are not different (p<0.05). agriculture and food sciences research, 2024, 11(2): 156-169 167 © 2024 by the authors; licensee asian online journal publishing group mineral boiled avg. ± sd (ppm) boiled range (ppm) steamed avg. ± sd (ppm) steamed range (ppm) other sources (ppm) cu 2.60 ± 0.81 1.48 – 4.47 2.40 ± 0.92 0.73 – 3.98 7-11.21 [36] fe 14.55 ± 2.68 10.32 – 18.61 12.80 ± 2.57 6.53 – 16.22 9-373 [36] ca 416.16 ± 68.71 333.74 – 545.91 371.20 ± 71.10 297.09 – 515.43 650-2,700 [36] p 435.74 ± 51.60 346.44 – 506.66 426.01 ± 64.05 308.32 – 524.12 1,010-1,920 [36] zn 5.92 ± 0.72 4.08 – 6.91 5.62 ± 0.90 3.68 – 6.76 16.38 [36] as 0.08 ± 0.05 0.01 – 0.15 0.08 ± 0.04 0.01 – 0.13 0.02 [36] 3.10. fatty acid results table 28 gives fatty acids and their levels (%) observed in boiled and steamed crawfish samples through 11 days of refrigerated storage. table 29 represents the fatty acid values (%) of boiled and steamed crawfish through six months of frozen storage. this analysis was conducted to add to the body of knowledge regarding crawfish and the types and levels of fatty acids in crawfish containing hepatopancreas fat. the fatty acid contents of steamed crawfish were determined to be 25.77, 30.86, and 43.36 per cent for sfa, mufa, and pufa, respectively [2]. sfa varied from 24.37 to 27.00 while mufa increased from 29.82 to 33.31 and pufa decreased from 45.42 to 39.68 with increased steaming time for crawfish, but sfa, mufa, and pufa were variable among boiling times with similar ranges in values [2]. fatty acid composition varied between commercially harvested crawfish and those from moistsoil wetlands [25]. table 28. percentages of fatty acid groups in boiled crawfish fat in refrigerated (3.5°c) storage. fatty acid group day 0 day 1 day 3 day 5 day 7 day 9 day 11 % sfa boiled 38.33 33.51 36.20 72.86 63.00 42.78 26.05 % sfa steamed 41.20 24.93 58.39 56.81 56.94 27.55 16.18 % mufa boiled 23.34 25.32 24.43 8.47 7.90 18.16 29.67 % mufa steamed 24.69 26.29 9.60 9.80 13.71 41.23 37.95 % pufa boiled 38.33 41.47 40.36 18.67 29.20 39.07 44.28 % pufa steamed 34.10 48.78 32.01 33.39 19.35 31.22 45.86 note: saturated fatty acids [sfa], mono-unsaturated fatty acids [mufa], and polyunsaturated fatty acids [pufa]. table 29. percentages of fatty acid groups in boiled crawfish fat in frozen (-18°c) storage. fatty acid group month 1 month 2 month 3 month 4 month 5 month 6 % sfa boiled 41.04 44.58 36.50 45.83 42.21 36.17 % sfa steamed 32.95 38.24 33.32 37.15 29.67 35.50 % mufa boiled 16.01 22.73 26.51 27.05 29.00 26.19 % mufa steamed 27.87 24.98 28.32 29.76 30.39 27.12 % pufa boiled 42.95 32.65 36.99 27.11 28.78 37.64 % pufa steamed 39.18 36.79 38.36 33.09 39.94 37.38 note: saturated fatty acids [sfa], mono-unsaturated fatty acids [mufa], and polyunsaturated fatty acids [pufa]. 5. conclusions the quality and shelf life of whole cooked crawfish through refrigerated and frozen storage was similar in steamed and boiled crawfish tails with adhering hepatopancreas. these observations suggest that commercial crawfish processors would not benefit by investing in commercial steamers and steaming their crawfish, but they should continue the traditional boiling processes for crawfish. a further study with analyses conducted on each day of storage would allow for a more precise determination of the exact shelf life of the crawfish regarding aerobic counts. with either method of cooking, the refrigerated shelf life is very short for whole cooked crawfish in most marketing channels. further research on sensory properties would be advantageous to determine consumer acceptability on the refrigerated and frozen crawfish to compare with the analytical results observed in this study. also, sensory analysis may reveal a difference between boiling and steaming from consumer perspectives even though analytical results in this study do not suggest there would be major differences. references [1] l. shi et al., "effects of freezing temperature and frozen storage on the biochemical and physical properties of procambarus clarkii," international journal of refrigeration, vol. 91, pp. 223-229, 2018. https://doi.org/10.1016/j.ijrefrig.2018.04.027 [2] j. li et al., "exploration on the quality changes and flavour characteristics of freshwater crayfish (procambarus clarkia) during steaming and boiling," lwt, vol. 190, p. 115582, 2023. https://doi.org/10.1016/j.lwt.2023.115582 [3] j. walls, crawfishes of louisiana. baton rouge, la, usa: louisiana state university press, 2009. 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licensee asian online journal publishing group agriculture and food sciences research vol. 11, no. 2, 195-202, 2024 issn(e) 2411-6653/ issn(p) 2518-0193 doi: 10.20448/aesr.v11i2.6252 © 2024 by the authors; licensee asian online journal publishing group genomic selection strategies in crop science for sustainable agriculture in korea sang hoon han1 yu-na kim2 soo-kwon park3 jung jae lee4 ( corresponding author) 1,4jj company 54, hyeonchon 1-ro, pyeongtaek-si, gyeonggi-do, republic of korea. 1email: aslksang@gmail.com 4email: jungjae.ansc@gmail.com 2,3crop foundation research division, national institute of crop science, rural development administration, wanju-gun, republic of korea. 2email: kyn7622@korea.kr 3email: sookwonpark@korea.kr abstract genomic selection (gs) is an effective approach to address the growing need for increasing and improving food production and quality, respectively, under challenging environmental conditions. in korea, gs significantly advanced major crops such as rice and wheat by improving critical agronomic traits. however, minor crops such as mung bean and sesame still require development because of limited resources and breeding infrastructure. these crops, though less prioritized, are vital for their nutritional and functional properties and contribute significantly to agricultural diversity and rural livelihoods. addressing the research gap in minor crops is essential to prevent the loss of genetic diversity and to enhance their resilience to environmental changes. unlike traditional selection methods, gs enhances the accuracy of complex trait selection, accelerates breeding cycles, and boosts genetic gain. it leverages genome-wide markers to predict genomic estimated breeding values (gebvs) with high precision, enabling faster and more effective breeding strategies, especially for traits controlled by multiple genes. genotyping techniques, such as genotyping by sequencing and whole genome resequencing, are integral to gs, each offering specific benefits and limitations. multispecies single nucleotide polymorphism arrays present a cost-effective solution for improving gs in minor crops by capturing the genetic diversity across species and enabling cross-species data integration. expanding the gs in minor crops is essential for preserving genetic diversity, improving adaptability, and supporting sustainable agriculture and food security in south korea. by addressing these challenges, gs can transform breeding programs, ensuring a sustainable and resilient agricultural system for the future. keywords: breeding, crops, genomic selection, genotyping by sequencing, single nucleotide polymorphism platform, sustainable agriculture. citation | han, s. h., kim, y.-n., park, s.-k., & lee, j. j. (2024). genomic selection strategies in crop science for sustainable agriculture in korea. agriculture and food sciences research, 11(2), 195–202. 10.20448/aesr.v11i2.6252 history: received: 15 november 2024 revised: 18 december 2024 accepted: 20 december 2024 published: 26 december 2024 licensed: this work is licensed under a creative commons attribution 4.0 license publisher: asian online journal publishing group funding: this research is supported by the cooperative research program for agriculture science and technology development, rural development administration, republic of korea (grant number: rs-2023-00232329). institutional review board statement: not applicable. transparency: the authors confirm that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. this study followed all ethical practices during writing. competing interests: the authors declare that they have no competing interests. authors’ contributions: all authors contributed equally to the conception and design of the study. all authors have read and agreed to the published version of the manuscript. contents 1. introduction .................................................................................................................................................................................... 196 2. advantages of gs ........................................................................................................................................................................... 196 3. genotyping techniques for gs in crop science ..................................................................................................................... 197 4. limitation of application in korea ............................................................................................................................................ 198 5. future challenges .......................................................................................................................................................................... 199 6. conclusions ..................................................................................................................................................................................... 200 references ............................................................................................................................................................................................ 200 mailto:aslksang@gmail.com mailto:jungjae.ansc@gmail.com mailto:kyn7622@korea.kr mailto:sookwonpark@korea.kr https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v11i2.6252 https://orcid.org/0009-0008-5219-0754 https://orcid.org/0009-0003-5323-6738 https://orcid.org/0000-0002-4941-3848 https://orcid.org/0000-0002-6145-8862 agriculture and food sciences research, 2024, 11(2): 195-202 196 © 2024 by the authors; licensee asian online journal publishing group contribution of this paper to the literature this study uniquely focuses on applying genomic selection (gs) to under-researched minor crops in korea, including mung bean and sesame. by proposing multispecies single nucleotide polymorphism (snp) arrays as a cost-effective tool, it bridges the gap between major and minor crop research, emphasizing genetic diversity preservation and adaptability. 1. introduction although more food is required for the rapidly growing human population, food quality needs to be improved, particularly with increased nutrient content [1]. the ability to increase or even sustain crop yield and quality in the face of dynamic environmental and biotic threats is particularly challenging in the face of rapid global environmental changes [2]. in response to challenges such as climate change and mechanization, the gradual shrinkage of cultivation areas led to an increased reliance on imports. concurrently, a demand exists for highquality varieties, including those rich in functional compounds, adaptable to climate change, resistant to pests and diseases, and suitable for mechanization and processing. modern crop breeding must prioritize the development of crop varieties with agronomic traits to address the dual demands of increased food production and improved nutritional quality. particularly in korea, both major and minor crops are essential not only for food security but also for nutritional quality and economic stability. major crops, such as rice, wheat, and maize, have seen considerable advancements in breeding, focusing on yield, disease resistance, and environmental adaptability [3]. contrastingly, minor crops, which are valuable for their nutritional and functional properties, have lagged in breeding improvements and require significant genetic advancements to meet their full potential. genomic selection (gs) is a transformative tool for genetic improvement that enables breeders to predict the genetic value of crops based on genome-wide markers [4]. unlike traditional phenotypic selection, which depends on observable traits that may not be consistently expressed under various environmental conditions, gs allows for the accumulation of minor genetic effects across the genome essential for complex traits, such as yield, disease resistance, and drought tolerance [5, 6]. many countries extensively adopted gs for major crops, benefiting from faster breeding cycles and greater selection accuracy, particularly for traits controlled by multiple genes [4]. in korea, gs have begun to influence major crops, including rice and wheat, showing promise in improving agronomic traits [7]. however, for minor crops, the adoption of gs faced limitations due to economic challenges, lack of accessible high-throughput genotyping resources, and less-established breeding infrastructure compared to those of staple crops. consequently, minor crops receive limited breeding resources despite their cultural and nutritional significance, raising concerns regarding the possible loss of valuable genetic diversity. applying gs to minor crops can safeguard their unique genetic traits and enhance yield stability and environmental adaptability, making them better suited to the climate challenges and agricultural shifts anticipated in korea. in this study, we examined the potential of gs in addressing breeding challenges for both major and minor crops in korea. while gs demonstrated substantial gains in yield, resilience, and disease resistance in major crops such as rice and wheat, minor crops remain underserved due to limited resources and infrastructure. we discuss the critical need to expand gs applications to minor crops to ensure the preservation of genetic diversity and enhance their adaptability to climate and agricultural demands. furthermore, we highlight that the methodologies explored in this study can be applied to regions with agricultural challenges similar to korea, thereby providing solutions to enhance food security and economic stability in these contexts. 2. advantages of gs conventional plant breeding methods historically relied on phenotypic selection where observable traits such as plant height, disease resistance, and grain yield are used to select individuals for breeding programs. although phenotypic selection proved successful for simple traits, it struggles with complex traits due to their polygenic nature and environmental sensitivity. one of the major challenges is the gxe interactions (genotype-byenvironment), in which the same genotype may be expressed differently under varying environmental conditions [8]. hence, phenotypic selection is less effective for complex traits controlled by multiple genes and significantly influenced by environmental factors [9]. to overcome these challenges, marker-assisted selection (mas), which utilizes deoxyribo nucleic acid (dna) markers linked to specific traits to guide breeding decisions, was developed. mas showed particular success for simple mendelian traits such as disease resistance or other qualitative traits involving one or a few major effect loci [10]. however, complex traits like grain yield or drought resistance influenced by numerous small-effect genes scattered across the genome, mas proved to be less effective due to its dependence on a limited number of markers [11]. gs represents a major leap forward by enabling the prediction of breeding values based on genome-wide markers without the need to identify individual trait-specific markers [12]. gs uses statistical models estimating the contribution of each single nucleotide polymorphism (snp) across the genome to the overall genetic merit of an individual, thereby capturing major and minor allele effects. this allows for the prediction of genomic estimated breeding value (gebv) an individual with high accuracy, even in the absence of phenotypic data. a key advantage of gs over mas is its ability to capture the cumulative effects of numerous small effect genes across the genome often missed by mas [13]. this comprehensive genomic information enables more accurate selection and improves genetic gains over time. one of the main benefits of gs is that it reduces breeding cycle time. traditional breeding methods rely on the phenotypic evaluations of multiple generations, whereas gs allows for selection based on genetic information alone [4]. additionally, gs is particularly advantageous for crops with long generation intervals or those grown in environments where phenotyping is difficult or expensive, such as minor crops [14]. in these crops, gs can potentially accelerate genetic improvement, even in the absence of extensive phenotypic or genomic resources [15, 16]. agriculture and food sciences research, 2024, 11(2): 195-202 197 © 2024 by the authors; licensee asian online journal publishing group in contrast to traditional breeding methods, gs allows breeders to maximize genetic gain per unit of time by focusing on genomic predictions rather than on phenotypic performance alone [15, 17]. as gs evolves with advances in sequencing technologies and reduced genotyping costs, it is set to revolutionize breeding programs for both the animal and crop industries by improving the selection of traits difficult to measure or have low heritability [18, 19]. table 1 illustrates the characteristics of breeding methods from phenotypic selection to genomic selection. table 1. advancements in selection strategies in crop breeding. method features advantages limitations phenotypic selection relies on observable traits and phenotype measurements. simple time-consuming low-cost long breeding cycles widely applicable inefficient for complex traits markerassisted selection uses specific dna markers linked to traits of interest. high accuracy for simple traits limited for small-effect loci specific dna markers ineffective for complex traits effective for major genes requires known marker-trait association genomic selection utilizes genome-wide markers to predict breeding values. genome-wide coverage high costs with high density platforms accurate for complex traits requires computational resources shortens breeding cycles data-intensive process 3. genotyping techniques for gs in crop science 3.1. genotyping by sequencing (gbs) the gbs has been widely adopted in modern crop breeding, particularly for species with complex genomes or limited genomic resources. this innovative technique, which was developed as a cost-effective, high-throughput alternative to traditional genotyping methods, allows for a more efficient exploration of genetic diversity [20, 21]. gbs operates by digesting genomic dna with restriction enzymes to simplify the genome and simultaneously sequence multiple dna fragments, effectively capturing a wide range of genetic variations across the genome [22]. another key feature of gbs is its flexibility in handling diverse species and populations, including biparental, multi-parental, and natural populations. this adaptability allows researchers to conduct genome-wide association studies (gwas), quantitative trait locus (qtl) mapping, and gs in a wide range of species [22-25]. furthermore, gbs can generate dense marker sets crucial for capturing recombination events and improving the accuracy of genomic predictions in breeding programs [20]. however, despite its cost effectiveness and utility, gbs has some limitations. it focuses only on specific parts of the genome, and certain regions may not be represented or completely missed. additionally, imputing missing genotypes can be challenging, particularly in species with high heterozygosity or complex structural variation [20]. to mitigate these issues, gbs data require stringent quality control and thorough bioinformatic processing to ensure reliable downstream analyses [26]. as sequencing technologies improve, gbs has advanced with better bioinformatics pipelines, such as trait analysis by association, evolution and linkage (tassel)-gbs, which increase the efficiency of snp discovery and genotype calling [27]. these innovations, combined with a reduction in sequencing costs, make gbs an indispensable tool for accelerating genetic gain in crop breeding programs, particularly for minor crops such as mung bean and adzukibean, which are critical for global food security and agricultural sustainability. 3.2. whole genome resequencing (wgr) the wgr emerged as one of the most powerful tools for unraveling genetic diversity and identifying genetic variation across the entire genome. unlike targeted sequencing approaches, wgr enable researchers to capture the complete genome sequence of an individual, thereby providing the most comprehensive dataset for genetic studies [28]. wgr was instrumental in mapping the genetic architecture of complex traits, enabling the identification of snps, structural variants, insertion-deletions, and other genomic alterations across both coding and noncoding regions [29]. the ability to detect both common and rare variants of wgr is particularly beneficial for crops with complex genomes or limited reference data [30]. a high depth of coverage of entire genomes can reveal subtle genetic differences that may contribute to important agronomic traits such as disease resistance [31]. for instance, cook, et al. [32] reported resistance to soybean cyst nematodes (scn), and maron, et al. [33] revealed that a higher copy number of the mate1 gene is linked to superior ai tolerance in maize. this high resolution makes wgr particularly useful for gs and gwas because it offers specific genomic markers for agronomic traits across the entire genome [34]. however, despite its high performance, wgr remains more expensive and data-intensive than those by reduced-representation sequencing (rrs) methods such as gbs and restriction site-associated (rad) seq [35]. wgr provides much more comprehensive genome coverage than that by rrs but also comes at a higher cost in both sequencing and data processing. rrs methods selectively sequence portions of the genome, which makes them more cost-effective and faster to analyze [35]. a major challenge with wgr is managing the depth of coverage necessary for accurate variant calling, particularly when distinguishing rare variants or completing structural variations. for example, in species with large repetitive genomes, aligning sequenced reads to a reference genome can be computationally intensive and prone to errors if coverage is insufficient [36]. despite these challenges, as sequencing costs gradually decreased and computational analyses have developed, wgr has become accessible, especially for crops where detailed genomic information is required for selection [31]. 3.3. snp array with the development of next-generation sequencing (ngs) technology, large-scale sequencing enabled the discovery of genetic variants. the identification of large numbers of snps in crops was accompanied by the agriculture and food sciences research, 2024, 11(2): 195-202 198 © 2024 by the authors; licensee asian online journal publishing group development of high-throughput genotyping tools, such as snp arrays, which allow for the simultaneous genotyping of thousands to millions of molecular markers [37]. currently, more than 40 k snps array tools that play a crucial role in accelerating the rate of genetic gains in crop breeding have been developed [38-40]. the major crops include wheat (35, 90, and 820 k) [41-43] rice (44 and 50 k) [44, 45] maize (50 and 600 k) [2, 46] and cotton (35 and 63 k) [47]. such high-throughput and easy to genotyping snp arrays can cost-effectively and quickly proceed with gwas, quantitative trait locus (qtl) mapping, and gs [48-50]. in addition to their cost-effectiveness and high-throughput capacity, snp arrays offer several advantages for crop-breeding programs. they provide excellent reproducibility and consistency across different platforms and studies hiraoka, et al. [51] and boichard, et al. [52] which is critical for large-scale breeding initiatives where data comparability is key. furthermore, allowing precise and accurate genotyping with a high call rate and low error rate, it is suitable for crops with complex genomes [37]. accordingly, snp arrays for particular species were developed and can be utilized diversely across different populations, studies, and purposes. the conflict with the prominent functions of snp arrays and the development of novel snp arrays associated with agronomic traits can be restricted by the reference population or cost [31]. although snp arrays are effective for detecting known variants, they are designed based on reference populations, which can limit their ability to detect novel or rare alleles in other populations [53]. if the genetic diversity of a reference population is not representative, snp arrays may miss important variants in different breeding lines or crops. consequently, the development of a novel snp array can be timeand cost-intensive due to the identification of new genetic variants with agronomic traits in other reference populations and validation tests to determine whether they are useful [54]. table 2 presents the advantages and limitations of various genotyping methods. table 2. advantages and limitations for each various genotyping methods. genotyping method advantages limitations gbs cost-effective and high-throughput limited genome coverage generates dense marker sets across the genome imputation needed suitable without full reference relies on restriction enzyme sites snp array high consistency limited to pre-selected snps low error rates may miss novel or rare variants fast and high-throughput genotyping whole-genome sequencing (wgs) complete genome data high cost detects novel variants data-intensive storage needed includes non-coding regions wgr full variant detection high cost detailed genetic mapping complex analysis requirements best with reference genome 4. limitation of application in korea as described in the ‘advantages of the gs’, it has significant advantages over traditional mas or phenotypic selection, primarily because it enables breeders to capture small genetic effects involved in complex traits or quantitative traits [14]. gs is now widely adopted in crop breeding programs globally for major crops, such as rice, wheat, and maize. cui, et al. [55] reported the gs for ten agronomic traits, whereas huang, et al. [56] focused on disease resistance in rice. in maize and wheat, many researchers are working to maximize breeding efficiency through gs by following breeding strategies of the international maize and wheat improvement center (cimmyt) [18, 57-59]. in addition to rice, wheat, and maize, other major crops such as soybean and cotton have also seen extensive applications of gs in breeding programs. for example, in soybean, researchers developed highdensity snp arrays, such as soy50k and soysnp arrays, to facilitate the identification of key traits such as seed composition, flooding tolerance, and disease resistance. advancements in soybean genomics accelerated breeding for improved yield and stress resilience [60, 61]. similarly, in cotton, genomic resources were employed to improve fiber quality [62]. in korea, studies increasingly focused on the application of gs in various major crops. kang, et al. [7] conducted a study using the korean wheat (k-wheat) core collection to optimize the gs for key agricultural traits. similarly, kim, et al. [63] developed a customized 580 k chip array to enhance gs and gwas in rice. kang, et al. [7] and kim, et al. [63] emphasized the importance of tailoring gs tools to local crop varieties and specific traits of interest. in wheat, kang et al. focused on optimizing gs for traits related to adaptability and yield, whereas kim et al. addressed key productivity and quality traits in rice. these efforts led to significant advancements in the use of high-throughput genotyping platforms to improve the precision and efficiency of breeding programs for major crops in korea, ultimately contributing to the development of more resilient and high-yield varieties adapted to local agricultural conditions [7, 63]. genetic diversity analyses and gs were actively pursued for other major crops in korea. for example, cho, et al. [64] conducted soybean research focusing on identifying key adaptive traits using whole-genome sequencing data. they analyzed soybean accessions from various countries, including korea, china, japan, and the united states, and found that korean accessions revealed significant genetic diversity. minor crops, similar to those of major crops, have significantly high nutritional value and require gs to improve their productivity and adaptability. however, in korea, research on these crops faces challenges due to their relatively low domestic demand, which makes funding and research initiatives difficult to prioritize. despite their importance in enhancing agricultural diversity and nutrition, research in korea traditionally focused on major crops such as rice, wheat, and soybean. ha, et al. [65] conducted a detailed genomic study on mung bean, providing essential genetic resources by identifying quantitative trait loci (qtls) associated with important traits, such as drought tolerance and pod maturity. however, even with these foundational genetic resources, gs has not yet been widely applied in mung bean breeding programs. similarly, kim, et al. [66] reviewed the current genomic resources available for mung agriculture and food sciences research, 2024, 11(2): 195-202 199 © 2024 by the authors; licensee asian online journal publishing group bean, emphasizing the need for further gs development to optimize breeding for traits such as disease resistance and seed quality [66]. similarly, sesame, another important minor crop, has seen advances in genetic studies, particularly in the identification of key loci associated with complex traits, such as disease resistance. asekova, et al. [67] combined qtl mapping and gwas to identify candidate genes related to phytophthora blight resistance in sesame. they identified snp markers on chromosome 10 associated with resistance to phytophthora nicotianae. these findings highlighted the potential use of genetic markers to improve disease resistance in sesame through breeding strategies. however, despite the discovery of numerous qtls and genetic variants related to various agronomic traits, the application of gs has not progressed in minor crops. the primary challenges are limited domestic demand and high costs associated with gs implementation, which have significantly hindered the development of comprehensive gs programs for these crops [14, 68]. although the genetic groundwork is in place, including significant progress in marker discovery and the mapping of complex traits, the practical implementation of gs in korea remains a challenge [14]. 5. future challenges to overcome the current challenges of gs for minor crops in korea, cost-effective genotyping methods such as gbs or custom snp chip arrays offer a viable solution. gbs is a cost-effective, high-throughput method for generating genome-wide markers, making it a popular tool for gs, particularly in species with limited genomic resources [20]. customized snp arrays also offer a targeted approach, allowing breeders to focus on known genetic variants relevant to specific traits [69]. however, even cost-effective methods face limitations when applied to minor crops because the scale of research and available resources remain insufficient to justify the high cost of customized chip development. a promising approach to address the various challenges in minor crops, where gs cannot play a critical role, the multi-species snp array is proposed, where a single chip can be used across multiple species, significantly lowering the costs per species. for example, silva‐junior, et al. [70] developed a flexible multi-species 60 k snp chip for eucalyptus, demonstrating its utility across 12 species and its potential to extend multi-species genotyping for practical breeding applications in closely related species. similarly, a multispecies snp array was designed for colossoma macropomum (tambaqui) and piaractus mesopotamicus (pacu), providing an efficient genomic tool for both species, thereby reducing costs and enhancing genetic research in aquaculture [71]. furthermore, multispecies snp arrays were developed for applications in both plant and aquaculture, covering species such as rubus spp. (raspberry and blackberry), leptospermum scoparium (mānuka), chrysophrys auratus (australasian snapper), and pseudocaranx georgianus (silver trevally). these arrays facilitate comparative genetic studies across species, enabling researchers to optimize genetic resources for use in breeding programs and conservation and addressing important economic and ecological goals in both fields [72]. table 3 presents the criteria used for snp selection in the development of multi-species snp arrays for each species, detailing sequencing methods, call rates, minor allele frequencies (maf), and final snp counts, along with their references. table 3. summary of criterions of snps selection for multi-species snp array. species sequencing method call rate maf final snps reference eucalyptus spp. wgr >97.2% >0.01 60,904 silva‐junior, et al. [70] rubus spp. gbs, wgs 96.4 >0.05 12,723 montanari, et al. [73] manuka wgr 9,002 snapper illumina novaseq 98.3 18,489 trevally wgs 20,234 tambaqui rad-seq >97% >0.01 23,739 mastrochiricofilho, et al. [71] pacu 23,768 multispecies snp arrays offer several advantages beyond cost-effectiveness for gs in minor crops. first, they enable researchers to capture a broader range of genetic diversity by including polymorphisms conserved across multiple species, allowing for more comprehensive assessments of genetic variation, and aiding in the study of population structure and evolutionary dynamics. this diversity also strengthens genomic prediction models, especially when resources for unique species-specific arrays are limited, making multispecies snp arrays ideal for minor or underfunded crops [73, 74]. furthermore, multispecies snp arrays support data integration across species, facilitating meta-analyses and improving the statistical power of genetic predictions. this approach is particularly beneficial for breeding programs because it enables researchers to perform cross-species comparisons more effectively, thereby enhancing the understanding of complex traits influenced by environmental interactions [75]. additionally, by integrating genomic data across species, multi-species arrays can significantly increase the accuracy and power of predictions for agronomic traits such as disease resistance and yield, as pooling diverse datasets strengthens the reliability of genetic predictions and accelerates breeding cycles [72]. conclusively, multi-species snp arrays present an optimal approach for implementing gs in minor crops and provide a cost-effective and robust solution. this approach allows the application of a single snp chip across multiple species, thereby significantly reducing the expenses associated with developing species-specific genotyping platforms. for minor crops, which often face funding and resource constraints, this method is especially practical, as it not only reduces costs, but also captures a wider range of genetic diversity. such diversity is instrumental in improving the accuracy and statistical power of genetic predictions, enabling a more effective selection of complex traits across diverse environmental conditions. figure 1 illustrates the process of snp selection for multi-species snp arrays, while figure 2 presents the overall flowchart for the development and application of these arrays in genomic selection. agriculture and food sciences research, 2024, 11(2): 195-202 200 © 2024 by the authors; licensee asian online journal publishing group figure 1. process of snps selection for multi-species snp array. figure 2. overall flowchart of development and application of multi-species snp array. 6. conclusions summarily, multispecies snp arrays provide an effective and economical approach for advancing gs in minor crops, enhancing genetic diversity capture, and enabling the robust selection of complex traits, thereby supporting sustainable agricultural productivity and adaptability across diverse environments. references [1] m. a. augustin et al., "role of food processing in food and nutrition security," trends in food science & technology, vol. 56, pp. 115125, 2016. 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