ISSN: 2456–5474 RNI No.  UPBIL/2016/68367 VOL.- VII , ISSUE- III April  - 2022
Innovation The Research Concept
A Survey On Aflatoxin Contamination In Animal Feed In Uttar Pradesh
Paper Id :  15947   Submission Date :  2022-04-12   Acceptance Date :  2022-04-19   Publication Date :  2022-04-21
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Karuna Shanker Mishra
Lecturer
Chemistry
SRM Inter College
Bisalpur, Pilibhit,Uttar Pradesh, India,
R K Agarwal
Retd Asso. Professor
Chemistry
Bareilly College,
Bareilly, U.P., India
Kamal Kishore
Associate Professor
Pharmacy
MJP Rohilkhand University
Bareilly, U.P., India
Abstract
Aflatoxins are mold-synthetized secondary metabolites that are ubiquitously present in agricultural commodities, such as maize, groundnut cake, pulse, and cereals etc which are the part of animal feed formulation. The aflatoxin contamination in different types of animal feed viz. Poultry feed, Husk of pulse, Maize, Groundnut cake, Linseed cake, Wheat bran and Rice bran, collected from the retail as well as wholesale shops, godown and local market of various cities, from Uttar Pradesh in three seasons i.e. during rainy season, before rainy season and after rainy season. Of 233 samples analysed, 65 (27.89%) were positive for aflatoxin B1 (AFB1) and the highest number of samples contaminated with aflatoxin B1 was found in poultry feed 38.93% followed by maize 35.00%, groundnut cake 34.28%, lindseed cake 31.03%, husk of pulse 29.72 %, and wheat bran 6.89%. None of rice bran samples were found affected during different seasons. After rainy season samples of groundnut cake 50.00% were the highly contaminated than other ingredients. Environment plays a significant role for the growth of fungi. Higher the relative humidity and higher temperature, higher was the contamination of aflatoxin B₁ in animal feed.
Keywords Aflatoxin, Animal feed, Carcinogens
Introduction
Aflatoxins are one of many natural occurring mycotoxins that are found in agricultural products, soil, grains, nuts, dairy products, tea, spices as well as animals and humans. Aflatoxins are specially problematic in dry and hot climates. Aflatoxins, known carcinogens, are polycyclic unsaturated organic compounds consisting of a coumarin nucleus flanked with a highly reactive bifuran ring system on one side and a six membered lactone ring on the other side. The carcinogenic properties are attributed to the presence of a lactone ring (Asao et al., 1963). Aflatoxins are the toxic metabolites of two closely related fungi namely Aspergillus flavus and Aspergillus parasiticus (Heathcote, 1984) which are distributed worldwide. (Diener et al., 1987; Ranjan et al.,1991) and may colonize and contaminate variety of feeds and food stuffs (Verma and Sinha, 1992). Chronic contamination of aflatoxin in feed lead to a gradual deteriorating of health through liver damage and immunosuppressant(Choudhary et al.,2010; Bryden et al.,2012; Choudhary et al.,2020) .Substantial literature is available regarding aflatoxin contamination in common food and feed but there are few reports about the contamination of aflatoxin in animal feed in Uttar Pradesh.
Objective of study
Substantial literature is available regarding aflatoxin contamination in common food and feed but there are few reports about the contamination of aflatoxin in animal feed in Uttar Pradesh.The main objective of the present survey is to provide data on occurrence of aflatoxins in various animal feed commodities under natural conditions used for animals in the different geographical areas of Uttar Pradesh.
Review of Literature

Chronic contamination of aflatoxin in feed lead to a gradual deteriorating of health through liver damage and immunosuppressant(Choudhary et al.,2010; Bryden et al.,2012; Choudhary et al.,2020) .Choudhary et.al. (2020) reported that the amount of aflatoxin B1 was significantly high (P<0.05%) in household cattle feeds followed by commercial cattle feeds, where the mean value of AFB1 was 5.02 ± 2.46µg/kg and 4.19 ± 1.05µg/kg, respectively. Freshly harvested cattle feeds (maize, paddy husk, paddy straw, maize straw and green grasses) were comparatively safe. Seasons had marked influence on aflatoxin B1 contamination on various types of cattle feeds. Maximum levels of aflatoxin B1 was detected during wet seasons in household cattle feeds (Mean value = 7.79 ± 2.07µg/kg) .Similar reports are also reported by various workers in India as well as in abroad.

Methodology
The various important villages, markets, godowns and poultry farms from Uttar Pradesh in three seasons i.e. during rainy season, before rainy season and after rainy season were surveyed for sample collection of commonly used feed stuffs for the domestic animals viz. poultry feed, husk of pulse, maize, groundnut cake, lindseed cake, wheat bran and rice bran. Each sample comprised 250g representative of 100 kg animal feed lots. Each sample were collected in dried sterilized polythene bags and were subsequently taken to the laboratory for further analysis. Precautions were taken during sampling to avoid erroneous result of uneven distribution. After monitoring the moisture content sample were dried at 60°C to remove moisture and samples were stored in deep freezer to check further fungal growth. Each dried sample was ground and mixed to obtain 50g sub sample for aflatoxin analysis. Chemical extraction of aflatoxin in animal feed samples was done in methanol: water (60:40 v/v) (Thomas et al., 1975). The residue was finally extracted with chloroform. Of 2ml of finally eluted chloroform extract 50 ml was spotted on silica gel thin layer chromatographic (TLC) plates prepared in laboratory, which was subsequently developed in toulene, isoamyl alcohol and methanol (90:32:2 v/v), solvent system for separation of different aflatoxin. (Reddy et al., 1970). The Triflouroacetic Acid was used for chemical confirmation of aflatoxin. The concentration of aflatoxin was quantitated spectrophotometrically (Nabney and Nesbitt, 1965). Test of significance is done by the method of Snedecor and Cochran (1967).
Statistics Used in the Study

Table: Aflatoxin Contamination in Animal Feed in Uttar Pradesh

Animal Feed 

No. of Sample  

analysed

No. of positive  

sample for aflatoxin  (%) 

Range of aflatoxin B1 concentration (µg/kg)

Before Rainy Season

Poultry feed 

16 

6(37.50%) 

Traces to 1050

Husk of pulse 

14 

4(28.57%) 

Traces to 560

Maize 

15 

6(40.00%) 

Traces to 500

Ground nut cake 

12 

3(25.00%) 

Traces to 280

Lind seed cake 

12 

6(50.00%) 

Traces to 105

Wheat bran 

09 

1(11.00%) 

Traces to 310

Rice bran 

05 

-(0.00%) 

-

Total 

83 

26(31.32%) 

TRACES TO 1050



During Rainy Season

Poultry feed 

16 

7(43.75%) 

Traces to 850

Husk of pulse 

14 

3(21.42%) 

Traces to 900

Maize 

11 

3(27.27%) 

Traces to 450

Ground nut cake 

15 

4(26.66%) 

Traces to 778

Lind seed cake 

09 

2(22.22%) 

Traces to 200

Wheat bran 

10 

- (0.00%) 

-

Rice bran 

06 

- (0.00%) 

-

Total 

81 

19(23.45%) 

Traces to 900

After Rainy Season

Poultry feed 

15 

5(33.33%) 

Traces to 500

Husk of pulse 

09 

4(44.44%) 

Traces to 400

Maize 

14 

5(35.71%) 

Traces to 200

Ground nut cake 

08 

4(50.00%) 

Traces to 300

Lind seed cake 

08 

1(12.50%) 

Traces to 150

Wheat bran 

10 

1(10.00%) 

Traces to 100

Rice bran 

05 

- (0.00%) 

Traces to 100

Total 

69 

20(28.98%) 

TRACES TO 500

Grand total 

233 

65(27.89%) 

TRACES TO 1050

Result and Discussion

The results of the present survey, the aflatoxin contamination in animal feed in Uttar  Pradesh in three different seasons are given in table. The aflatoxin contamination in  different types of animal feed viz. poultry feed, husk of pulse, maize, groundnut cake,  lindseed cake, wheat bran and rice bran, collected from the retail as well as wholesale  shops, godown, local market, villages and poultry forms, from Uttar Pradesh in three  seasons i.e. during rainy season, before rainy season and after rainy season. The total  233 samples were analysed, 65 (27.89%) samples were positive for aflatoxin B(AFB1)  and the highest number of samples contaminated with aflatoxin B1 was found in  poultry feed 38.93% followed by maize 35.00%, groundnut cake 34.28%, ,lindseed  cake 31.03% , husk of pulse 29.72 %, and wheat bran 6.89%. None of rice bran  samples were found affected during different seasons. After rainy season samples of  groundnut cake 50.00% were the highly contaminated than other ingredients. Highest  contamination was found in linseed cake, poultry feed and groundnut cake before rainy season, during rainy season and after rainy season respectively. Environment  plays a significant role for the growth of fungi. Higher the relative humidity and higher  the temperature, higher was the contamination of aflatoxin B in animal feed. Clean, dry and insect free storage conditions are critical to prevent aflatoxin growth as noted  by the USAID desk review. Making storage options inexpensive and accessible is of  permanent importance for reducing the growth of the aflatoxins in various  commodities. 

A toxigenic strain of aspergillus flavus and aspergillus parasiticus, a suitable  base and congenial climatic conditions are the three major factors which contribute  to aflatoxin contamination in food and feed substrate under natural condition.  

Diener and Davis (1969), showed that moisture and relative humidity of the  surroundings are the important factor for the growth of fungus which leads to  aflatoxin production in various animal feeds. The sorting, cleaning, drying, packaging  and hygienic conditions in stores and transport are also the factors of fungal growth.  The food grains are normally harvested at higher moisture content leads to increase  risk of mould growth and alfatoxin production Singh et al. (1997) analysed 67 dairy  cattle feed samples and reported 65% positively for aflatoxin ranging from 50-400  mcg/kg. Choudhary et.al. (2020) reported that the amount of aflatoxin Bwas  significantly high (P<0.05%) in household cattle feeds followed by commercial cattle  feeds, where the mean value of AFB1 was 5.02 ± 2.46µg/kg and 4.19 ± 1.05µg/kg,  respectively. Freshly harvested cattle feeds (maize, paddy husk, paddy straw, maize  straw and green grasses) were comparatively safe. Seasons had marked influence on  aflatoxin B1 contamination on various types of cattle feeds. Maximum levels of  aflatoxin B1 was detected during wet seasons in household cattle feeds (Mean value  = 7.79 ± 2.07µg/kg) .Similar reports are also reported by various workers in India as  well as in abroad.The percentage of aflatoxin contamination in the present study may  be due to association of toxigenic strain of aspergillus flavus and aspergillus 

parasiticus along with the favourable climatic conditions prevailing in these areas,  poor harvesting, storage as well as marketing facilities greatly stimulate the  contamination of aflatoxin in food and feed stuffs. Although the present study did not  survey the incidence of disease related to this problem but it can certainly be  presumed that the contaminated samples may cause some physiological  abnormalities in genital, digestive and respiratory tracts through different  mechanisms. This will depend on both the concentration of aflatoxin in feed and its  daily consumption.

Conclusion
Environment plays a significant role for the growth of fungi. Higher the relative humidity and higher temperature, higher was the contamination of aflatoxin B₁ in animal feed. Hence it is essential to have rigorous monitoring programme for aflatoxin, so that better feed management can be done for livestock. Monitoring and preventive programmes should be one of the most effective strategy to decrease the risk of exposure to both human and animals.
References
1. Asao, T., Buchi, G., Abdel, K.M.M., Chang, S.B., Wick, E.L. and Wogan, G.N. (1963), Aflatoxin and G. J. Am. Chem. Soc., 85: 1706-1707. 2. Bryden, W.L. (2002), Mycotoxin contamination of the feed chain: Implications for animal productivity and feed security. Animal Feed Science and Technology, 173(1-2): 134-158. 3. Choudhary, A., and Kumari, P. (2010), Management of mycotoxin contamination in preharvest and post harvest crops: present status and future prospects. Journal of Phytology, 2(7): 37-52. 4. Choudhary, A.K., Tuda, Smita, Kumari, P., Ranjan, A., (2020), Present status, prevalence and seasonal variations of aflaxtoxin in cattle feed, Bihar, India. Indian Journal of Science and Technology, 13(17): 1738-1745. 5. Diener, U.L., Cole, R.J. Sanders TH., Payne, G.A., Lee, L.S. and Klich, M.A. (1987), Epidemiology of aflatoxin formation by Aspergillus flavus. Ann. Rev. Phytopathol., 25: 249-270. 6. Diener, U.L. and Davis, N.D. (1969), In Aflatoxin (Ed. L.A. - Goldblott) Academic press New York, pp. 13-54. 7. Heathcote J.G. (1984), Aflatoxins and related toxins in mycotoxin production, isolation, separation and purification (Ed. Betina, V.A. Elsvier Sci. Pub., pp. 89. 8. Nabney, J. and Nesbitt, B.F. (1965), A spectrophotometric method for determination the aflatoxin, analyst, 90: 155-160. 9. Ranjan, K., Sinha, A. (1991), Occurrence of mycotoxigenic fungi and mycotoxins in animal feed from Bihar, India. Journal of the Science of Food and Agriculture, 56(1): 39-47. 10.Reddy, T.V. Vishwanathan, L. and Venkitasubramanian, T.A. (1970), Thin layer chromatography. Anal. Biochem., 38: 568-571. 11.Singh, P.P., Jand, S.K., Baxi, K.K. and Singh, N. (1997), Occurrence of aflatoxins in dairy cattle feed in relation to climatic condition. Indian Vet. Med. J., 21: 121- 123. 12.Snedecor, G.W. and Cochran, W.G. (1967), Statistical methods (The Iowa State University Press, Ames, I.A. VI Education, 91-119). 13.Thomas, F., Eppley, R.M. and Trucksess, M.W. (1975), Rapid screening method for aflatoxins and Zearalenone in corn. J. Assoc. Off. Anal. Chem., 58:114-116. 14.Verma, C.S. and Sinha, D.P. (1992), Aflatoxin problem in Pulse in Vihar. Bio Journal Patna, 4(1-2): 63-69.