ЗВІТ З НДР 29-81 ЗА 2007 – 2009 Р 368 Journal homepage: www.fia.usv.ro/fiajournal Journal of Faculty of Food Engineering, Ştefan cel Mare University of Suceava, Romania Volume XII, Issue 4 – 2014, pag. 368 - 374 THE APPLICATION OF BASALTIC TUFFS IN THE TECHNOLOGY OF CULTIVATION THE LIVE F EED FOR FISH – PRELIMINARY STUDY Oleksii KHUDYI1, *Igor KOBASA1, Olga KUSHNIRYK1, Lidiia KHUDA1 1 Yuriy Fedkovych Chernivtsi National University, Ukraine, imk-11@hotmail.com * Corresponding author Received November 24th 2015, accepted December 18th 2015 Abstract: The research work is meant to investigate the possibility of using the zeolite-containing tuffs in the technology of live fish feed cultivation by example of Cladocera. It is shown that the use of natural and treated at the temperature of 150 °C basaltic tuffs from „Polytske-2” basalt deposit during Moina macrocopa cultivation leads to an increase of the culture density by 2 and 2.3 times, respectively. In the experiment with Simocephalus vetulus the increase of culture density has not been found, however basaltic tuffs allow to stabilize the culture growth and slow its extinction rate. Besides, it is observed the effective purification of cultural medium of dissolved nitrogen forms probably by adsorption. The use of basaltic tuffs, calcined at a temperature of 1000 °C, leads to the death of M. macrocopa and S. vetulus cultures in 6 days. This circumstance makes the use of calcined tuffs im- practical. Keywords: zeolites, Cladocera, cultivation medium, ammonium, nitrites, nitrates 1. Introduction Basaltic tuffs are natural aluminosilicates from zeolite group of minerals. Zeolites possess several unique features, including adsorption, and therefore, they are used in the technological process of fluids purifi- cation, including wastewaters [1-3]. This is especially true for systems with closed cy- cle of water use. In this regard, tuffs are widely used in the recirculation of aquacul- ture systems as components of water treatment [4, 5]. Live feed is widely used to obtain quality planting material for commercial fish farming and for decorative aquaculture ob- jects [6]. Among all the diversities of for- age organisms, an important place is taken by Cladocera crustaceans. One of the problems that occur with periodic method of zooplankton cultivation is exhaustion of culture medium caused by the accumula- tion of metabolic products in it [7, 8]. The use of substances with high adsorption properties, such as zeolites, may have ef- fects on prolonging use of culture media without their update that reduces the costs of live feed. In addition, zeolites possess the ion-exchange properties which can positively affect the availability of micro- and macro-elements for cultivating organ- isms. Given the above, the purpose of the research was to find the applicability of zeolites from «Polytske-2» basalt deposit during Cladocera cultivation. 2. Materials and methods The mineral samples received from „Poly- tske-2” basalt deposit, situated in the Rivne Region of Ukraine, were used to study the possibility of basaltic tuffs applied in the http://www.fia.usv.ro/fiajournal mailto:imk-11@hotmail.com Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XII, Issue 4 – 2014 Oleksii KHUDYI, Igor KOBASA, Olga KUSHNIRYK, Lidiia KHUDA, The application of basaltic tuffs in the technology of cultivation the live feed for fish – preliminary study, Food and Environment Safety, Volume XIV, Issue 4 – 2015, pag. 368 – 374 369 technology of cultivation of live food for fish. The determination of basaltic tuffs compo- sition was performed using classical meth- ods of chemical analysis [9], the content of Si, Al, Fe, Mg, Ca, P, S – by flame pho- tometry method, Na and K – by atomic absorption spectroscopy. The studies were conducted on laboratory cultures of Cladocera crustaceans – Simo- cephalus vetulus (Muller) and Moina mac- rocopa (Straus). Cultivation of zooplank- ton was carried out in 0.5 dm³ jars using standardized by chemical composition syn- thetic medium ADaM [10], at a tempera- ture of 21±2 °C, cold light fluorescent glow 2,5–4 klux and 16 hour photoperiod. To determine the effective amounts of minerals during formation of experimental groups, the tuff was introduced into jars in amounts of 30, 40 and 50 g per 1 liter. According to the results of previous studies we found that the porosity and specific sur- face area of basaltic tuff granules depends on the temperature of heat treatment. These parameters reach the maximum val- ues in the temperature range of 150–400 °C. There is a steady trend towards a de- crease in the specific surface area and po- rosity after tuff calcination in the tempera- ture range of 500–1000 °C. It may be caused by the removal of hygroscopic and zeolite water [11]. Taking into account above stated in the present study it was used both natural and thermally activated at temperatures of 150 °C and 1000 °C tuffs, which differ in their sorption properties. Group of organisms, in cultivation medium in which the tuff has not been introduced, served as a control. The feeding of cultures was performed every 48 hours, yeast Sac- charomyces cerevisiae at a dose of 0,05 g/l were used as feed. Measurements of pH level were carried out using I-160МI Ion Meter, determination of total mineraliza- tion level in culture medium was per- formed indirectly through conductivity us- ing conductivity meter SX-650 ULab. The content of ammonium, nitrites and nitrates was determined in the culture medium. Measurings the concentration of NH4 + and NO3 - ions were performed by potentiom- etric method using I-160МI Ion Meter with ion-selective electrodes ELIS-121 NH4 + and ELIS-121-121NO3 -, concentration of NO2 - was determined by photocolorimetric methods [12]. 3. Results and discussion The mineralogical composition of basaltic tuffs, obtained from „Polytske-2” basalt deposit which was used in the study, in- cludes: zeolites (35–40%), montmoril- lonites (30–40%), feldspars (10–15%), sil- icas (4–5%), hematites (3–5%) [13]. The statistically average results of the chemical composition analysis of the tuff are ex- pressed in mass percents of oxides. The summary of the results shows that the basaltic tuff is an aluminum silicate with a mass ratio Si/Al = 4,7÷5,9, containing Iron in amounts of 68–74 g/kg. Except ele- ments, whose compounds in terms of ox- ides were presented in table 1, the investi- gated tuffs also contain trace elements (Mn, Zn, Cu, Co, Ni) in amounts of 0,71– 0,08 g/kg. Toxic elements (As, Pb, Hg, Cr) were not found. Table 1 The chemical composition of basaltic tuff from basalt deposit „Polytske-2” in terms of oxides, the mass fraction expressed in % Oxides Mass frac- tion, % Oxides Mass frac- tion, % SiO2 67.44 CaO 0.46 ТіО2 1.75 Na2O 0.94 Al2O3 12.82 K2O 1.06 Fe2O3 10.14 P2O5 0.12 МnО 0.09 SO3 0.11 MgO 5.02 Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XII, Issue 4 – 2014 Oleksii KHUDYI, Igor KOBASA, Olga KUSHNIRYK, Lidiia KHUDA, The application of basaltic tuffs in the technology of cultivation the live feed for fish – preliminary study, Food and Environment Safety, Volume XIV, Issue 4 – 2015, pag. 368 – 374 370 The results of the studies showed that the use of basaltic tuffs from „Polytske-2” basalt deposit enhances the growth intensi- ty of both zooplankton cultures. A positive effect from tuffs using was manifested at 7–8 days of cultivation in experiment with M. macrocopa. The maximal development of Moina culture was achieved both in ex- perimental and control groups on the 14th day, but the density of cultures that were grown using tuffs was 2 times higher (fig- ure 1 a). The culture of S. vetulus is characterized by much lower growth rates with achieving of maximum density on the 4th day, the introduction of natural basaltic tuff in the experimental concentrations (30, 40 and 50 g/l) does not affect the density of S. vetulus culture (figure 1 b). a b c d Figure 1. The dynamics of M. macrocopa (a, c) and S. vetulus (b, d) density using different concentrations of natural unmodified (a, b) and thermally (150 °С) calcined (c, d) basaltic tuffs However, it was found that the use of ba- saltic tuff allows stabilizing the develop- ment of culture and slowing its extinction rate. It is known that the temperature of basaltic tuffs calcination affects their elemental composition and adsorption properties. Re- spectively, the heat treatment can influence the biological objects that are grown in the presence of basaltic tuffs [14]. In accord- ance with the research results, it was estab- lished that thermally activated tuffs differ- 0 400 800 1200 1600 2000 2400 2800 3200 3600 0 2 4 6 8 10 12 14 16 18 in d . / l days 30g/l 40g/l 50g/l К 0 20 40 60 80 100 120 140 160 180 0 2 4 6 8 10 12 14 16 in d . / l days 30g/l 40g/l 50g/l К 0 400 800 1200 1600 2000 2400 2800 3200 3600 0 2 4 6 8 10 12 14 16 18 in d . / l days 30g/l 40g/l 50g/l К 0 20 40 60 80 100 120 140 160 180 0 2 4 6 8 10 12 14 16 in d . / l days 30g/l 40g/l 50g/l К Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XII, Issue 4 – 2014 Oleksii KHUDYI, Igor KOBASA, Olga KUSHNIRYK, Lidiia KHUDA, The application of basaltic tuffs in the technology of cultivation the live feed for fish – preliminary study, Food and Environment Safety, Volume XIV, Issue 4 – 2015, pag. 368 – 374 371 ently affect density and dynamics of both zooplankton species. The introduction of tuffs, thermally treated at 150 °C, into jars for M. macrocopa cul- tivation is accompanied by increase of cul- ture density by 2.3 times as compared to the control in contrast to the experiment with natural tuffs, where the difference was 2 times (figure 1 c). The use of tuffs treated at high temperature (150 °C) was accompanied by no changes in S. vetulus culture density, while the stabilization of the culture and slowdown of its extinction have been seen like in the case where natu- ral tuffs were used (figure 1d). The nega- tive impact on the culture in both cases was not observed. Due to this fact, the use of zeolite-containing tuffs to increase the mentioned species density is not appropri- ate. A negative effect of tuffs, calcined at the temperature of 1000 °C, on the growth of both studied cultures (M. macrocopa and S. vetulus) was observed in the experiment (figure 2). a b Figure 2. The density of M. macrocopa (a) and S. vetulus (b) cultures using thermally calcined (1000 °С) basaltic tuffs During the first 6 days of the experiment the complete extinction of two zooplank- ton species has been seen, that is associat- ed with significant changes in the structure and functions of basaltic tuffs. Changes of pH level during M. macrocopa cultivating were observed in the cultivation medium due to the accumulation of meta- bolic products of living organisms (figure 3). Figure 3. The pH dynamics during M. macrocopa cultivation with use of different concentrations of natu- ral unmodified basaltic tuffs 0 10 20 30 40 50 60 0 2 4 6 in d . / l days 30g/l 40g/l 50g/l 0 10 20 30 40 50 60 0 2 4 6 in d . / l days 30g/l 40g/l 50g/l 7.1 7.6 8.1 8.6 0 2 4 6 8 10 12 14 16 18 days p H 30g/l 40g/l 50g/l К Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XII, Issue 4 – 2014 Oleksii KHUDYI, Igor KOBASA, Olga KUSHNIRYK, Lidiia KHUDA, The application of basaltic tuffs in the technology of cultivation the live feed for fish – preliminary study, Food and Environment Safety, Volume XIV, Issue 4 – 2015, pag. 368 – 374 372 Instead, significant increase in pH level in the experiment with S. vetulus was not ob- served, being associated with minor zoo- plankton density. The influence of changes on pH, using different concentrations of natural form of zeolite-containing tuff, has not been observed, and does not differ greatly from the control experiment. As mentioned above, tuff calcined at a temperature of 1000 °C inhibited the de- velopment of both investigated cultures. However, despite the degradation of cul- ture, the trend of pH level raise is ob- served, which is obviously associated with the accumulation of food residues, decom- position of dead organisms and ion- exchange processes between solution and tuffs. The dynamics of total mineralization val- ues during the period of zooplankton culti- vation is caused by the accumulation of waste products in the medium, food resi- dues and dead organisms. The marked in- fluence of changing on total mineralization when using thermally activated (150 °C) and natural form of basaltic tuffs at inves- tigated concentrations was not found. Indi- cators of total mineralization in experi- mental groups did not differ from the con- trol (figure 4). a b c d Figure 4. The change of total mineralization values of medium using natural form and thermally calcined tuffs: M. macrocopa (a) and S. vetulus (b) with natural tuffs; M. macrocopa (c) and S. vetulus (d) with thermally calcined at 150 °C tuffs The impact of basaltic tuffs on the dynam- ics of metabolites accumulation in the cul- tivation medium, including ammonium ion and the products of its oxidation, nitrates and nitrites, attracts a particular attention. The increased density of the culture is ac- companied by increased concentrations of soluble forms of nitrogen. The research results show that the use of the basaltic tuffs provides the slowdown accumulation of NH4 + ions and products of 750 790 830 870 910 950 990 1030 2 4 6 8 10 12 14 16 18 m g /l days 30g/l 40g/l 50g/l 750 790 830 870 910 950 990 1030 2 4 6 8 10 12 m g /l days 30g/l 40g/l 50g/l 750 790 830 870 910 950 990 1030 2 4 6 8 10 12 14 16 18 m g /l days 30g/l 40g/l 50g/l К 750 790 830 870 910 950 990 1030 2 4 6 8 10 12 m g /l days 30g/l 40g/l 50g/l К Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XII, Issue 4 – 2014 Oleksii KHUDYI, Igor KOBASA, Olga KUSHNIRYK, Lidiia KHUDA, The application of basaltic tuffs in the technology of cultivation the live feed for fish – preliminary study, Food and Environment Safety, Volume XIV, Issue 4 – 2015, pag. 368 – 374 373 its oxidation NO2 - and NO3 -, apparently due to adsorption processes (figure 5). Accordingly, the increase of concentration of the zeolite-containing tuff from 30 to 50 g/l improves the efficiency of cleaning the liquid phase of cultivation medium due to increasing of adsorption area. a b c d e f Figure 5. The dynamics in content of nitrogen soluble forms in the medium for M. macrocopa (a, c, e) and S. vetulus (b, d, f) cultivation using different concentrations of basaltic tuffs 4. Conclusion 0 0,05 0,1 0,15 0,2 0,25 0 2 4 6 8 10 N H 4 + , m m o l / l days 30g/l 40g/l 50g/l К 0 0,05 0,1 0,15 0,2 0,25 0 2 4 6 8 10 N H 4 + , m m o l / l days 30g/l 40g/l 50g/l К 0 0,05 0,1 0,15 0,2 0,25 0,3 0,35 0,4 0,45 0 2 4 6 8 10 N O 2 - , m m o l / l days 30g/l 40g/l 50g/l К 0 0,05 0,1 0,15 0,2 0,25 0,3 0,35 0,4 0,45 0 2 4 6 8 10 N O 2 - , m m o l / l days 30g/l 40g/l 50g/l К 0 0,1 0,2 0,3 0,4 0,5 0,6 0,7 0,8 0,9 1 0 2 4 6 8 10 N O 3 - , m m o l / l days 30g/l 40g/l 50g/l К 0 0,1 0,2 0,3 0,4 0,5 0,6 0,7 0,8 0,9 1 0 2 4 6 8 10 N O 3 - , m m o l / l days 30g/l 40g/l 50g/l К Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefan cel Mare University - Suceava Volume XII, Issue 4 – 2014 Oleksii KHUDYI, Igor KOBASA, Olga KUSHNIRYK, Lidiia KHUDA, The application of basaltic tuffs in the technology of cultivation the live feed for fish – preliminary study, Food and Environment Safety, Volume XIV, Issue 4 – 2015, pag. 368 – 374 374 We can remark that the use of natural and thermally activated at a temperature of 150 °C basaltic tuffs has a positive impact on the development dynamics of zooplankton cultures. Maximal density of M. macro- copa cultures in the experiments with natu- ral and activated tuffs was higher than in control group by 2 and 2.3 times respec- tively. The application of tuffs during S. vetulus cultivation did not cause an in- crease in the density of culture, but slowed the rate of extinction. The use of basaltic tuffs, calcined at a tem- perature of 1000 °C, leads to the death of M. macrocopa and S. vetulus cultures in 6 days. Basalt tuffs provide the effective cleaning of cultivation medium from nitro- gen soluble forms. The maximal effect is observed when using 50 g of tuffs for 1 liter of cultivation medium. 5. References [1]. KALLO D., Applications of natural zeolites in water and wastewater treatment, Rewiews in Mineralogy and Geochemistry, 45 (1): 519-550, (2001). [2]. TARASEVYCH YU.I., KLIMOVA G.M., Ob- taining of modified sorbents and their applica- tion for water purification from heavy metals, Journal of Water Chemistry and Technology, 28 (2): 107-116, (2006). (In Russian). [3]. PETRUS R., MALYOVANYY M., VAR- CHOL J., ODNORIG Z., PETRUSHKA I., LESKIV G., Technologies of wastewater treat- ment using natural dispersed sorbents, Chemical Industry of Ukraine, 2: 20-22, (2003). (In Ukrainian). [4]. VATIN N.I., CHECHEVICHKIN A.V., CHECHEVICHKIN V.N., Sorption treatment of industrial highly concentrated waters from the ammonium ion using natural zeolites. St. Petersburg: 36, (2007) (In Russian). [5]. SIRAKOV I., VELICHKOVA K., STOYANOVA S., DINEV D., STAYKOV Y., Application of natural zeolites and macrophytes for water treatment in recirculation aquaculture systems, Bulgarian Journal of Agricultural Sci- ence, 21 (1): 147-153, (2015). [6]. DAS P., MANDAL S.C., BHAGABATI S.K., AKHTARM.S., SINGHS.K. Important live food organisms and their role in aquaculture. in: Frontiers in Aquaculture. (Ed.) M. Sukham, Narendra Publishing House: 69-86, (2012). [7]. HEDSTROM A., Ion exchange of ammonium in zeolites: a literature review, J. Environ. Eng., 127: 673-681, (2001). [8]. PABALAN R.T., BERTETTI F.P., Cation- exchange properties of natural zeolites, Rewiews in Mineralogy and Geochemistry, 45 (1): 453- 518, (2001). [9]. TSIMBALYUK V.V., VOLOSHCHUK A.G., KOBASA I.M., Effect of heat treatment on sorption properties of basaltic tuff, Ukrainian Chemistry Journal, 75 (12): 85-90, (2009) (In Ukrainian). [10]. KLUTTGEN B., DULMER U., ENGELS M., RATRE H.T., ADaM, an artificial freshwa- ter for the culture of zooplankton, Water Res., 28 (3): 743-746, (1994). [11]. KOBASA I., MIKULA J., WOLOSCHUK A, TSYMBALIUK W., Influence of thermal treatment on the physical, mechanical and sorp- tion characteritics of the basalt tufa, Technical Transactions. Mechanics, 5-M (15): 3-9, (2011). [12]. LINNIK P. Determination of the nitrite ni- trogen concentration in water. in: Methods of hydroecological researches of surface water. Logos. Kyiv: 242-244, (2004). (In Ukrainian). [13]. GOLIAR N., Tuffs: the use in economic sec- tors. Rivne: 30, (In Ukrainian). [14]. MELNYCHUK V. Zeolite-smectite tuff of Rivne Region: the biological aspects of applica- tion. Volynski oberegy. Rivne: 184, (2005). (In Ukrainian).