




































 Agricultural Science; Vol. 2, No. 1; 2020 
ISSN 2690-5396   E-ISSN 2690-4799 

https://doi.org/10.30560/as.v2n1p128 

128                          Published by IDEAS SPREAD 
 

Potentials of Cow Dung Ash as Protectants of Cowpea Seeds against 
Callosobruchus maculatus F. (Coleoptera: Chrysomelidae) 

Mohammed Suleiman1 & Fatima M. Haruna1 
1 Department of Biology, Umaru Musa Yar’adua University, Katsina, Nigeria 
Correspondence: Mohammed Suleiman, Department of Biology, Umaru Musa Yar’adua University, Katsina, 
Nigeria. Tel: 234-806-7195-818. E-mail: mohammed.suleiman@umyu.edu.ng 
 
Received: April 1, 2020   Accepted: April 20, 2020   Online Published: April 26, 2020 
 
Abstract 
Experiments were conducted to assess the protectant ability of cow dung ash (CDA) to cowpea seeds against 
Callosobruchus maculatus F. under laboratory conditions of 50-65% R.H and 28-32oC. The CDA at different rates 
of 0.625, 1.25, 2.50, 5.00, 10.00% (w/w) incorporated with cypermethrin powder at 0.625% were applied to 20 g 
of cowpea seeds in petri dishes. None of the ashes or cypermethrin was added to the control. Ten beetles were 
introduced into each of the petri dishes and covered with muslin cloth. The experiment was arranged in a 
completely randomized design (CRD) and replicated three times. Effects of the treatments on adult mortalities as 
well as median lethal dose (LD50) of CDA against C. maculatus were determined. Adult emergence and weight 
losses in seeds treated with CDA at different doses were assessed. Findings of the study revealed that the adult 
mortality of C. maculatus among different CDA treatments differed significantly (p < 0.05) and ranged from 13.33 
to 100.00% 72 hours after treatment (HAT). Results of the study also showed that LD50 of CDA against C. 
maculatus at 24 hours after treatment (HAT) was 5.51 % (w/w). No adults emerged from the treated seeds and 
weight losses were observed in those treated with the lowest dose and the control only. Therefore, CDA could be 
utilized to protect cowpea seeds from C. maculatus infestations during storage.  
Keywords: Adult emergence, Callosobruchus maculatus, Cow dung ash, cowpea seed, mortality, protectants 
1. Introduction 
Cowpea is highly palatable, very nutritious and relatively free from anti-metabolites. Shiringani and Shimelis 
(2011) reported that dried seeds of cowpea provide cheap source of proteins in many diets in the tropics and sub-
tropics. The report further showed that cowpea seeds contain 22-33% protein, 60-66% carbohydrates, 5-6% fibre, 
3.7-4.4% ash and 1.1-3% oil. 
Callosobruchus maculatus (cowpea weevil) has been reported to be the main insect pest of cowpea (Vigna 
unguiculata) (Suleiman, 2016). Ojebode et al. (2016) reported that C. maculatus is very destructive on account of 
its short life cycle. Their damage causes losses in weight, nutritional, commercial values of cowpea as well as 
viability of stored seeds (Suleiman, 2016). Larvae feed and develop inside the seed which become unsuitable for 
human consumption and when adults emerge they leave a neat circular exit hole. Each adult consumes 
approximately 25% of the seed from which it develops (Asawalam and Anaeto, 2014). 
Synthetic insecticides may play a significant role in reducing storage losses due to insect pests. However, the 
extensive use of these chemicals poses so many problems such as insecticide resistance, health risk to consumers 
and environmental contamination (Sani and Suleiman, 2017). These problems have necessitated the replacement 
of synthetic insecticides with natural products which are eco-friendly to protect stored grains from insect 
infestations (Vanmathi et al., 2012). 
Natural products have been used for many years by small scale farmers in many parts of Africa to protect stored 
products (Suleiman and Rugumamu, 2017; Suleiman et al., 2019). Previous findings demonstrated the use of these 
materials as safe and effective protectants of stored cowpea against C. maculatus infestations (Suleiman and 
Suleiman, 2014; Asawalam and Anaeto, 2014). These products act by suffocating adult bruchids, restricting 
oviposition or even lead to the death of the insect (Sani and Suleiman, 2017).  
In order to minimize the use of chemicals there is need for a search of alternative eco-friendly methods such as 
application of natural products like cow dung ash (CDA). Therefore, this study was aimed at evaluating the efficacy 
of CDA in controlling C. maculatus infesting stored cowpea. 



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2. Method 
2.1 Rearing of Callosobruchus maculatus 
All experiments were conducted in Laboratory 3 of the Department of Biology, Umaru Musa Yar’adua University, 
Katsina (UMYUK), Nigeria. Adults of C. maculatus were obtained from infested cowpea sold in Katsina central 
market. Cowpea seeds used for the rearing were brought from local market in Katsina and kept in an oven at 40oC 
for 4 hours to disinfest the seeds and allowed to cool for 24 hours. Fifty pairs of adult C. maculatus were placed 
in 500 ml volume glass bottle containing 250 g of the disinfested cowpea seeds. The set up was kept on the 
laboratory shelf at room temperature. The parent weevils were removed by sieving after 7 days of their introduction. 
Seeds were kept under laboratory condition until emergence of F1 progeny. 
2.2 Preparation of CDA 
Cow dung was collected from Barhim village of Katsina, Nigeria. The cow dung was sun-dried and crushed into 
small particles. The small sized particles were placed in the muffle furnace and heated for 3 hours. The ash 
collected was allowed to cool and then stored in air-tight container under prevailing laboratory condition of 50-
65% R.H and 28-32oC. 
2.3 Assessment of Adult Mortality 
Healthy disinfested cowpea seed (20 g) were placed in Petri dishes and treated with 0.125, 0.25, 0.50, 1.00 and 
2.00 g of cow dung ash, separately. This was equivalent to 0.625, 1.25, 2.50, 5.0 and 10.0% (w/w), respectively. 
Cypermethrin (0.625%) was added to another 20 g seeds while no powder was added the control seeds. Five pairs 
of newly emerged adult beetles were introduced to each of the treated and untreated seeds in the Petri dishes. They 
were covered with nylon mesh and held in place with rubber hands. The set up was arranged in a completely 
randomized design (CRD) with three replications. This was inspected daily to remove, count and record dead 
insects for seven days. At the end of seven days, all beetles (dead and alive) were removed and percentage adult 
mortality was assessed as the number of dead beetles divided by total number of beetles introduced and multiplied 
by one hundred as indicated hereunder. Adult Mortality (% ) =  (Number of Dead Insects)/(Total Number of Insects Introduced)  x 100   
2.4 Determination of Median Lethal Dose (LD50) of CDA 
To evaluate LD50, methods of Ebadollahi and Mahboubi (2011) were employed. The number of dead weevils in 
each bottle containing sorghum grains treated with the botanical powders was counted at the end of 24 hours after 
treatment HAT). The LD50 was calculated by using probit analysis with SPSS (version 16.0) software package. 
2.5 Examination of Adult Emergence of Callosobruchus maculatus 
The remaining insects (dead and alive) were sieved from the treated cowpea seeds immediately after mortality test. 
The seeds were left undisturbed under laboratory condition for adult emergence. To calculate the adult emergence 
direct examination was performed daily from the first emergence for seven days. All emerged individuals were 
removed, counted and recorded for 7 days. 
2.6 Damage Assessment 
Both treated and untreated seeds were sieved to remove the ash and any dust. To determine percentage grain 
damage, the final weight of the grain was subtracted from the initial weight of the grain, divided by initial weight 
of the grain and then multiplied by one hundred (100) as shown in the formula below: Weight Loss (%) =  (Initial Weight (g) −  Final Weight (g))/(Initial Weight (g) )  x 100   
2.7 Data Analysis 
All data were tested for normality using KS normality test and found to be non-parametric. For this reason, 
Kruskal-Wallis test was employed to test if there was significant difference in the effects of CDA on adult mortality 
and weight losses in the treated grains. Significantly different means were separated using Dunn’s multiple 
comparison tests. 
3. Results 
3.1 Adult Mortality of Callosobruchus maculatus Exposed to CDA  
Adult mortality of C. maculatus in cowpea seeds treated with different concentration of CDA varied. The 
mortalities were directly proportional to the varying concentrations of CDA applied and ranged from 13.33 to 
53.33% among the treatments while it was 3.33% in the control at 24 hours after treatment (HAT) (Table 1). 
Kruskal-Wallis test showed the existence of significant difference (KW = 18.809; p = 0.0045) in adult mortality of 



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C. maculatus among various concentrations of CDA, cypermethrin and control. Adult mortalities at 48 and 72 
HAT were observed to follow similar trend to that of 24 HAT where they were directly proportional to increase in 
concentration (Table 1). The treatments had significantly different (p < 0.05) effect on adult mortality of the insects 
at 24 and 72 HAT, respectively The mortalities also increased with increase in exposure periods in the order: 24 < 
48 < 72 HAT. 
 
Table 1. Mean Adult Mortality of Callosobruchus maculatus in Cowpea Seeds Treated with Different Doses of 
CDA 

Dose (%) 
Mean Adult Mortality of C. maculatus (% ± S. E.) 

Exposure Period (Hours) 
24 48 72 

0.625 13.3 ± 3.33d 46.6 ± 6.67d 83.33 ± 3.33b 

1.25 23.33 ± 3.33c 66.67 ± 3.33c 86.67 ± 3.33b 

2.5 26.67 ± 3.33c 76.67 ± 3.33b 90.00 ± 0.00b 

5.0 36.67 ± 3.33b 90.00 ± 0.00a 93.33 ± 3.33a 

10.0 53.33 ± 3.33a 96.67 ± 3.33a 100.00 ± 0.00a 

Cypermethrin (0.625%) 66.67 ± 3.33a 93.33 ± 3.33a 96.67 ± 3.33a 

Control (0.00%) 3.33 ± 3.33e 3.33 ± 3.33e 6.67 ± 3.33c 

Mean values with different superscripts on the same column are significantly different at p < 0.05 
 
3.2 Median Lethal Dose (LD50) of CDA against Callosobruchus maculatus  
Median lethal dose (LD50) of CDA against C. maculatus at 24 hours after treatment was 8.50% (w/w) with 
confidence limits (Cl) of 5.51 – 23.91% (Table 2). The Chi-square (X2) goodness of fit showed that the difference 
was not significant (p > 0.05) for LD50 of CDA. 
3.3 Adult Emergence of Callosobruchus maculatus in Treated Cowpea Seeds 
No adult emergence of C. maculatus was observed from all the treated samples. However, the mean number of 
emerged adult beetles from untreated seeds was recorded as 16.67. 
 
Table 2. LD50 of CDA against Adult Callosobruchus maculatus in Cowpea Seeds 

LD50 (%) 
95% Confidence Limits 

Slope ± S. E. X2 p 
Lower Limit Upper Limit 

8.50 5.51 23.93 2.59 ± 0.05 1.501 0.826 
 
3.4 Weight Losses in Cowpea Seeds Treated with CDA 
The only weight loss observed among the treated cowpea seeds was in those treated with the lowest concentration 
of CDA (0.625%) while the untreated seeds lost 11.33% of their weight to C. maculatus (Figure 1). Kruskal Wallis 
test showed that there was significant difference (KW = 19.857; p = 0.0029) in weight losses of cowpea seeds 
treated with cow dung ash at different doses 
 



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Figure 1. Percentage Weight Losses in Cowpea Seeds Treated with Varying Concentrations of CDA and Infested 

by C. maculatus 
 
4. Discussion 
4.1 Effect of CDA on Adult Mortality of Callosobruchus maculatus 
Findings of this study have revealed that CDA was effective in causing high adult mortality of C. maculatus. This 
is in line with the findings of Arya and Tiwari (2013) who reported that cow dung ash applied at 2% to wheat 
grains resulted in 90.80% adult mortality of Rhyzopertha dominica Fab. Similar findings were reported by 
Narayana et al. (2019) that 2% CDA applied to maize grains caused 93.89% adult mortality of R. dominica at 24 
HAT. It was also observed that the higher doses of CDA were almost as effective as the chemical insecticide 
(Cypermethrin) in killing adults of C. maculatus. The killing effect of CDA could be as a result of their small-sized 
particles that might have blocked the insect spiracle leading to impaired respiration and eventual death as observed 
by Suleiman and Suleiman (2014).  
4.2 Effect of CDA on Adult Emergence of Callosobruchus maculatus  
Results from this study indicated that the CDA was effective in reducing adult emergence of C. maculatus this 
implied that cow dung ash has a great protectant property against adult emergence of the insect. These findings are 
similar to earlier findings that only 2.75 adults of Rhyzopertha dominica emerged in grains treated with CDA (Arya 
and Tiwari, 2013). Similar findings were reported by Adebayo and Ibikunle (2014) that 1 g of CDA applied to 
cowpea seeds reduce adult emergence of C. maculatus from 2.80 in the control to 2.08 in the treated seeds.. 
Furthermore, results of this study showed that the higher doses of cow dung ash applied were as effective as 
Cypermethrin in reducing adult C. maculatus.  
4.3 The Role of CDA in Reducing Weight Losses of Cowpea Seeds Infested by Callosobruchus maculatus 
Finding of this study indicated that the high dose applied did not allow C. maculatus to cause any damage in 
cowpea seeds. This is line with the findings of Narayana (2019) which showed that only 0.19% of weight loss was 
recorded in maize grains treated with 2% CDA and infested by R dominica. Similarly, Tesema et al. (2015) reported 
that 2% of CDA gave a complete protection to chickpea against C. chinensis. The CDA might have inhibited the 
larval penetration into the and thus showed reduced levels of seed infestation and weight losses. 
Acknowledgments 
The authors are grateful to Dr. Babangida Adulkarim of the Department of Biology, UMYUK for remarkable 
guidance during data analyses.   
References 
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This is an open-access article distributed under the terms and conditions of the Creative Commons Attribution 
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    /HRV (Za stvaranje Adobe PDF dokumenata najpogodnijih za visokokvalitetni ispis prije tiskanja koristite ove postavke.  Stvoreni PDF dokumenti mogu se otvoriti Acrobat i Adobe Reader 5.0 i kasnijim verzijama.)
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    /NLD (Gebruik deze instellingen om Adobe PDF-documenten te maken die zijn geoptimaliseerd voor prepress-afdrukken van hoge kwaliteit. De gemaakte PDF-documenten kunnen worden geopend met Acrobat en Adobe Reader 5.0 en hoger.)
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    /UKR <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>
    /ENU (Use these settings to create Adobe PDF documents best suited for high-quality prepress printing.  Created PDF documents can be opened with Acrobat and Adobe Reader 5.0 and later.)
  >>
  /Namespace [
    (Adobe)
    (Common)
    (1.0)
  ]
  /OtherNamespaces [
    <<
      /AsReaderSpreads false
      /CropImagesToFrames true
      /ErrorControl /WarnAndContinue
      /FlattenerIgnoreSpreadOverrides false
      /IncludeGuidesGrids false
      /IncludeNonPrinting false
      /IncludeSlug false
      /Namespace [
        (Adobe)
        (InDesign)
        (4.0)
      ]
      /OmitPlacedBitmaps false
      /OmitPlacedEPS false
      /OmitPlacedPDF false
      /SimulateOverprint /Legacy
    >>
    <<
      /AddBleedMarks false
      /AddColorBars false
      /AddCropMarks false
      /AddPageInfo false
      /AddRegMarks false
      /ConvertColors /ConvertToCMYK
      /DestinationProfileName ()
      /DestinationProfileSelector /DocumentCMYK
      /Downsample16BitImages true
      /FlattenerPreset <<
        /PresetSelector /MediumResolution
      >>
      /FormElements false
      /GenerateStructure false
      /IncludeBookmarks false
      /IncludeHyperlinks false
      /IncludeInteractive false
      /IncludeLayers false
      /IncludeProfiles false
      /MultimediaHandling /UseObjectSettings
      /Namespace [
        (Adobe)
        (CreativeSuite)
        (2.0)
      ]
      /PDFXOutputIntentProfileSelector /DocumentCMYK
      /PreserveEditing true
      /UntaggedCMYKHandling /LeaveUntagged
      /UntaggedRGBHandling /UseDocumentProfile
      /UseDocumentBleed false
    >>
  ]
>> setdistillerparams
<<
  /HWResolution [2400 2400]
  /PageSize [612.000 792.000]
>> setpagedevice

