APPLICATION OF ENZYME-LINKED IMMUNOSORBENT ASSAY FOR AFLATOXIN CONTENT EVALUATION OF DIFFERENT POWDERED MILK AVALABLE IN ANAMBRA STATE, NIGERIA

Dr. Enuneku Patrick Omuku; ORJI, L.C.; ONUIGBO, A.U.; OZUKWE, A.E

doi:10.5281/zenodo.8119362

Dr. Enuneku Patrick Omuku Department of Chemistry, Nnamdi Azikiwe University
ORJI, L.C. Department of Chemistry, Nnamdi Azikiwe University
ONUIGBO, A.U. Department of Chemistry, Nnamdi Azikiwe University
OZUKWE, A.E Department of Chemistry, Nnamdi Azikiwe University

DOI: https://doi.org/10.5281/zenodo.8119362

Keywords: Aflatoxin, Immunoassay, Enzyme, Crude Protein, Centrifugation, Test kit

Abstract

The application of enzyme-link immunosorbent assay for aflatoxin contamination of 20 different powdered milk samples consumed within Anambra State, Nigeria was carried out primarily to ascertain their health implications. The samples were subjected to Ridascreen ELISA competitive enzyme immunoassay for the quantitative determination of aflatoxins in the milk. All the reagents required for the enzyme immunoassay including statndards were contained in the test kit used. Preparatory samples stages were grinding, extraction, filtration/centrifugation followed by acid mixture digestion process with tablet of selenium catalyst. The proximate analyses of the samples were done via AOAC standard procedures. The results revealed relatively low moisture contents with a range of 1.64 to 4.72%, with sample T recording highest value of 4.72% . The highest value of ash content was observed for sample D ( 6.57%) while sample R had lowest value of 0.85%. The lowest amount of crude fat was recorded by sample F (2.60%) as against high value of 21.33% for sample B. Crude protein showed least amount in samples A (20.19%) with the highest amount found in sample H (23.67%). The quantification of aflatoxins in the 20 powdered milk samples using ELIZA method revealed relatively low concentration of aflatoxins in the samples and the trend observed had a range of 0.36ppm (sample B) to 0.81ppm (sample F). The average aflatoxin concentration was estimated to be 0.61ppm. Samples C, E and F had mean level of aflatoxin recorded as 0.71ppm while the mean amount implicated for samples A, B, G and K was 0.51ppm. Conclusively, the results of the proximate analysis of the milk samples revealed appreciable occurrence of crude protein and relatively low amount of aflatoxins. The level of aflatoxins in all the samples was found to occur below National Regulatory threshold value of 4ppm, hence all the samples analyzed can be adjudged to be safe for consumption with little precautionary measures for hygiene and preservation.

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References

1.D.R., Milićević,, M., Skrinjar, T. Baltic, Real and perceived risks for mycotoxin contamination in foods and feeds: challenges for food safety control, Toxins, 2 (2010), pp. 572-592
2.Food and Agriculture Organization of the United Nations, Final report. FAO/WHO regional conference on food safety for Africa, Harare, Zimbabwe, 3 to 6 October, Food and Agriculture Organization of the United Nations, Rome. (2005). Review on biological degradation of mycotoxins, Anim. Nutr, 2 (2016), pp. 127-133
3.M.E., Flores-Flores,, E., Lizarraga, A., Lopez de Cerain, E. Gonzalez-Penas, Presence of mycotoxins in animal milk: a review, Food Control, 53 (2015), pp. 163-176
4.Azziz-Baumgartner, Eduardo, et al. "Mycotoxins and Child Health: A Systematic Review." International Journal of Hygiene and Environmental Health, vol. 220, no. 5, 2017, pp. 711-725. doi: 10.1016/j.ijheh.2017.02.011
5 Azziz-Baumgartner, Eduardo, et al. "Mycotoxins and Child Health: The Need for Health Risk Assessment." International Journal of Environmental Research and Public Health, vol. 13, no. 3, 2016, pp. 277. doi: 10.3390/ijerph13030277
6. Bennett, Joan W. "Mycotoxins in Infants and Children." Pediatric Clinics of North America, vol. 64, no. 2, 2017, pp. 367-382. doi: 10.1016/j.pcl.2016.11.0
7. Osman, Kamal N., and Stephen W. Page. "Mycotoxin-Related Neurological and Developmental Abnormalities in Infants and Children." Pediatric Neurology Briefs, vol. 31, no. 5, 2017, pp. 1-2. doi: 10.15844/pedneurbriefs-31-5-5
8. Augustin M.A, and Craven h. encyclopedia of food sciences and nutrition(second edition 2003)Ardic M, Karakaya Y, Atasever M, Adiguzel G, 2009. Aflatoxin M1 levels of Turkish white brined cheese. Food Control 20:196-9.
9. Bhat, R.V. and Vashanti,S. (2003). Mycotoxins and food safety risk in developing countries. Food safety in food security and food trade. Retrieved May 2009 from http/www.fpri.org/2020/focus10/focus 03.pdf
10.Boudra H, Barnouin J, Dragacci S, Morgavi P, 2007. Aflatoxin M1 and ochratoxin A in raw bulk milk from French dairy herds. J Dairy Sci 90:3197-201.
11.Boutrif E. (1998). Prevention of Aflatoxin in Pistachios. Food, Nutrition and Agriculture – World Health Organization Food Safety, FAO.
12. Davis, N. D., U. L. Diener, and V. P. Agnihotri. 1967. Production of aflatoxin B1 and G1 in a chemically defined medium. Mcyopathol. Mycol. Appl. 31:251-256.
13. Diener, U. L., and N. D. Davis. 1969. Aflatoxin formation by Aspergillus .flavus. pp. 13-54. In L. A. Goldblatt (ed.) Aflatoxin. Academic Press, New York.
14. EFSA Panel on Contaminants in the Food Chain 2007 EFSA J. 446 1–127.
15. Fallah A, Jafari T, Fallah A, Rahnama M, 2009. Determination of Aflatoxin M1 levels in Iranian white and cream cheese. Food Chem Toxicol 47:1872-5.
16. FAO (2004).Aflatoxin and Fumonisins Contamination of Sun-dried Sweet Potato Chips in Kahama District, Tanzania.FAO Statistics.Food and Agriculture Organization Rome, Italy.
17. Gong Y.Y, Cardwell K, Hounsa A; (2004). Determinants of Aflatoxin Exposure in Young Children from Benin and Togo, West Africa: The Critical Role of Weaning. International Journal of Epidemiology. 32(4):556-562.
18. Gong Y.Y, Cardwell K, Hounsa A;(2004). Determinants of Aflatoxin Exposure in young children from benin and togo west Africa: the critical of Afaltoxin role of weaning.International Journal of epidemiology. 32(4):556-562)
19. Gürbay A, Aydin G, Girgin B, Engin S, 2006. Assessment of aflatoxin M1 levels in milk in Ankara, Turkey. Food Control 17:1-4
20. Hassan, L. G., and Umar, K. J. (2007). Nutritional value of balsaru apple (Momordica balsa Mine L)
21. Hell K, Cardwell K, Setamou M. and Poehling H.M (2000). The influence of storage practices on aflatoxin contamination in maize in four agroecological zones of Benin, West Africa. Journal Stored Prod Res; 36:365–82.
22. Hussain I, Anwar J, 2008. A study on contamination of aflatoxin M1 in raw milk in the Punjab province of Pakistan. Food Control 19:393-5. [Google Scholar]
23. Hussain I, Anwar J, Asi M, Munawar M, Kashif M, 2010. Aflatoxin M1 contamination in milk from five dairy species in Pakistan. Food Control 21:122-4
24. Jarvis, B. 1971. Factors affecting the production of mycotoxins. J. Appl. Bacterial. 34:199-213.
25.Kensler,T.W., Roebuck, B.D.,Wogan, G.N. and Groopman, J.D (2011). Aflatoxin: A 50-Year Odyseey of mechanistic and translational toxicological sci.120 (1):28-48.
26. Kiermeier. F. 1977. Significance of aflatoxins in the dairy industry. Annual Bulletin. International Dairy Federation. Document 98.
27. Kiessling, K.H. (1986).Biochemical mechanism of action of mycotoxins.Pure& Applied Chemistry, 58, 327-338.
28. Kissell, L., S. Davidson, B. A. Hopkins, G. W. Smith, and L. W. Whitlow. 2013. Effect of experimental feed additives on aflatoxin in milk of dairy cows fed aflatoxin-contaminated diets. J. Anim. Physiol. Anim. Nutr. 97:694-700.
29. Lamanaka, B.T., H C., Vicente. R.S.F. and Taniwaki, M.H. (2007).Aflatoxigenic Fungi and aflatoxin occurrence in sultan and dried figs commercialized in Brazil.Food control,18:454-457 leave. J. Nutr., 5: 522-529.
30. Lewis L.; Mary Onsongo; Henry Njapau; AbdikherMohamudDahiye. (2005), Aflatoxin contamination of commercial maize products during an outbreak of acute aflatoxicoisis in Eastern and central Kenya.
31. Lie, J. L., and E. H. Marth (1968). Aflatoxin formation by Aspergillusflavus and Aspergillus parasiticus in casein substrate at different pH values. J. Dairy Sci. 51:1743-1747.
32. Ma, Z. X., F. X. Amaro, J. J. Romero, O. G. Pereira, K. C. Jeong, and A. T. Adesogan. 2017. The capacity of silage inoculant bacteria to bind aflatoxin B1 in vitro and in artificially contaminated corn silage. J. Dairy Sci. 100:7198-7210.
33. Maki, C. R., A. D. Thomas, S. E. Elmore, A. A. Romoser, R. B. Harvey, H. A. Ramirez-Ramirez, and T. D. Phillips. 2016. Effects of calcium montmorillonite clay and aflatoxin exposure on dry matter intake, milk production, and milk composition. J. Dairy Sci. 99:1039-1046.
34. Omar (2013).Mycotoxin and Food Safety in Deveoping Countries,Edition; first, Chapter;3
35. Prandini A, Tansini G, Sigolo S, Filippi L, Laporta M and Piva G 2009 Food Chem. Toxicol. 47 984–91
36. Prestka, J.J and Bondy, G.S (1994).Immunotoxic effect of mycotoxins in J.D, Miller and H.I, Trenholm Eds. Mycotoxins in Grains, Compounds other than aflatoxins. St. Paul, M.N. Lagan Press. 339-359.
37. Reddy, J .V. and Waliyar, F. (2009).Properties of aflatoxin and its producing fungi. International crop research Institute for the semi-arid tropics. Retrived, May 23 2009, from http.springerlink.com/t732j7178h8305.

38. Sadeghi N, Oveisi M, Jannat B, Hajimahmoodi B, Bonyani H, Jannat F, 2010. Incidence of aflatoxin M1 in human breast milk in Tehran, Iran. Food Control 20:75-8
39. Splensley, P.C (1963). Aflatoxin, the active principle in Turkey X diseases. Endeavour 22:75-79.
40. Strosnider H, Azziz-Baumgartner E, Banziger M, Bhat RV, Breiman R, Brune MN, DeCock K, Dilley A, Groopman J, Hell K, et al. Workgroup report: public health strategies for reducing aflatoxin exposure in developing countries. Environ Health Perspect. 2006; 114:1898–1903
41. Sulzberger, S. A., S. Melnichenko, and F. C. Cardoso. 2017. Effects of clay after an aflatoxin challenge on aflatoxin clearance, milk production, and metabolism of Holstein cows. J. Dairy Sci. 100:1856-1869
42. Van Egmond, H.P. (1989).Current situation on regulations for mycotoxins. Overview of tolerances and status of standard methods of sampling and analysis. Food Addit Contam. 6(2): 139-88.
43. Wagacha, J.M. and Muthomi, J.W. (2008).Mycotoxins problem in Africa: Current Status, Implication to food safety, health and possible management strategies. International Journal of Food Microbiology 124:1-2.
44. Widiastuti R, Maryam R, Bahri S and Firmansyah R 2006 Pros. Semin. Nas. Teknol. Peternak. dan Vet. 2006 239–43