The Effect of Organic and Inorganic Aflatoxin B1 Absorbents on in Vitro Digestibility and Rumen Fermentation Characteristics

Document Type : Ruminant Nutrition


ferdowsi university of mashhad


Introduction Aflatoxins (AF) as secondary metabolites are produced by Aspergillus flavus and Aspergillus parasiticus. The most abundant aflatoxin B1 (AFB1) is toxic and carcinogenic to humans and animals. Utilization of mycotoxin adsorbents are noted as the most practical methods for protection of feed ingredients. The purpose of this study was to investigate the effect of organic and inorganic adsorbents for the adsorption of AFB1 and its effects on dry matter digestibility and rumen fermentation characteristics.
Materials and Methods The experimental diet was a mixture of alfalfa silage and concentrate. Procedure of in vitro batch culture was performed according to the Menke and Steingass procedure. In an anaerobic condition, 30 ml of buffered rumen fluid was dispensed with pipetor pump into a 120-ml serum bottle containing 0.5 g DM of the experimental diet. The content of each bottle was contaminated with 0.5 ppm AFB1. Experimental treatments were: the control diet, commercial bentonite, activated bentonite, activated charcoal, the cell wall of Saccharomyces serevisia with 75% purity, bentonite + activated charcoal + yeast, (0.5 + 0.4 + 0.1 percent), bentonite + activated charcoal + yeast, (0.4 + 0.45 + 0.15 percent) and , bentonite + activated charcoal + yeast(0.3 + 0.5 + 0.2 percent) The amount of absorbents for all treatments was 1% of the experimental diets. All bottles were purged with anaerobic CO2, sealed with rubber stoppers and placed in a shaking water bath for 72 h at 38.6 degree centigrade. The amount of produced gas was recorded at 2, 4, 6, 8, 12, 16, 24, 48 and 72 h of the incubation. At the end of incubation, all the bottles were transferred to refrigerator to stop fermentation, and then opened. After pH measurements 2-ml sample of each filtrate bottle was taken and frozen at 20 degree centigrade after acidification with 2-ml of 0.2 N HCl. The biomass residues were centrifuged at 1000×g for 10 min at 4 degree centigrade. The supernatant in each bottle was decanted and the pellet was dried at 65 degree centigrade to a constant weight for the determination of the residues.
Results and Discussion Absorbents addition increased the amount of produced gas in all treatments in comparison with the control treatment significantly (P


1- Aghashahi, A., A. Nikkhah., S. A. Mirhadi., M. Zahedifar, and H. Mansouri. 2005. Effects of different level of unprocessed bentonite, processed bentonite, and clinoptilolite at different rumen degradable protein level, on ammonia concentration, soluble and digestible protein (in-vitro). Pajouhesh and Sazandegi, 70: 80-90. (In Persian).
2- Aitchison, E. M., J. B. Rowe, and Rix, G. S. 1986. Effect of bentonite clays on rumen fermentation and diet digestibility. Proceedings of the Nutrition Society of Australia, 11: 111-114.
3- Allcroft, R, and R. B. A. Carnaghan. 1963. Toxic product in groundnuts. Biological effects. Chemistry and Industry (Lond.), 9: 50-53.
4- Brown, T. 1996. Fungal diseases and Poultry Diseases. British Poultry Science, 6: 247–260.
5- Buck, W. B, and P. M. Bratich. 1986. Activated charcoal: Preventing unnecessary death by poisoning. Veterinary Medicine, 73: 73-77.
6- Ciegler, A., E. B. Lillehoj., R. E. Peterson, and H. H. Hall. 1966. Microbial detoxification of aflatoxin. Applied Microbiology, 14: 934-939.
7- Conway, H. F., R. A. Anderson, and E. B. Bagley. 1978. Detoxification of aflatoxin contaminated corn by roasting. Cereal Chemistry, 55: 115-117.
8- Cook W. O., J. L. Richard., G. D. Osweiller, and D. W. Trampel. 1986. Clinical and pathologic changes in acute bovine aflatoxicosis: rumen motility and tissue and fluid concentrations of aflatoxins B1 and M1. American Journal of Veterinary Research, 47: 1817-1825.
9- Dalvi, R. R, and C. McGowan. 1984. Experimental induction of chronic aflatoxicosis in chickens by purified aflatoxin B1 and its reversal by activated charcoal, Phenobarbital and reduced glutathione. Poultry Science, 63: 485-491.
10- Dawson, K. A., J. Evans, and M. Kudupoje. 2001. Understanding the Adsorption Characteristics of Yeast Cell Wall Preparations Associated with Mycotoxin Binding. Nottingham University Press, Nottingham, UK.
11- Devegowda, G., M. V. L. N. Raju., A. Nazar, and H. V. L. N. Swamy. 2000. Effect of rumen fluid on in vitro aflatoxin binding capacity of different sequestering agents and in vivo release of the sequestered toxin. Animal Feed Science and Technology, 98: 241–255.
12- Diaz, D., W. Hagler., B. Hopkins, and L. Whitlow. 2003. Aflatoxin binders I: In vitro binding assay for aflatoxin B1 by several potential sequestering agents. Mycopathologia, 156: 223-226
13- Fehr, P. M, and J. Delage. 1970. Effect de l’aflatoxine sur les fermentations dans le rumen. C.R. Academia des Science (Series D), 270: 550-553.
14- Fenn, P. D, and R. A. Leng. 1989. Wool growth and sulfur amino acid entry rate in sheep fed roughage based diets supplemented with bentonite and sulfur amino acids. Australian Journal of Agricultural Research, 40: 889-896.
15- Frenkel, M. 1974. Surface acidity of montmorillonites. Clays Clay Miner, 22: 435-441.
16- Galvano, F., A. Pietri., T. Bertuzzi., M. Bognanno., L. Chies., A. Angelis, and M. Galvano. 1997. Activated carbons: in vitro affinity for aflatoxin B1 and relation of adsorption ability to physicochemical parameters. Journal of Food Protection, 60: 985-991.
17- Greeballah, H., E. L. Obeid., A. Faisal, H. Nabil, and H. Bashir. 2005. In vitro effect of some insecticides on rumen fluid fermentation. Journal of Agricultural Science, 3: 80-89.
18- Hamilton, P. B. 1985. Factors influencing activity of fungi and anti- fungal agents in poultry feed in Tricothecenes and Other Mycotoxins. John Wiley and Sons Ltd, New York.
19- Helal, F, and K. Abdel-Rahman. 2010. Productive performance of lactating ewes fed diets supplementing with dry yeast and bentonite as feed additives. World Journal of Agricultural Sciences, 6: 489-498
20- Helferich, W. G., W. N. Garrett., D. P. H. Hsieh, and R. L. Baldwin. 1986. Feedlot performance and tissue residues of cattle consuming diets containing aflatoxins. Journal of Animal Science, 62: 691-696.
21- Ivan, M. 1992. Effects of Bentonite and monensine on selected elements in the stomach and liver of Fauna-Free and faunated sheep. Journal of Dairy Science, 75: 201-208.
22- Jacques, K. A., D. E. Axe., T. R. Harris, and D. L. Harmon. 1986. Effect of sodium bicarbonate and sodium bentonite on digestion, solid and liquid flow, and ruminal fermentation characteristics of forage sorghum silage-based diets fed to steers. Journal of Animal Science, 63: 923-932.
23- Jiang, Y. H., H. J. Yang, and P. Lund. 2012. Effect of aflatoxin B1 on in vitro ruminal fermentation of rations high in alfalfa hay or ryegrass hay. Animal Feed Science and Technology, 175: 85-89.
24- Jiang, Y. H., P. Wang., H. J. Yang, and Y. Chen. 2014. The efficacy of bamboo charcoal in comparison with smectite to reduce the detrimental effect of aflatoxin B1 on in vitro rumen fermentation of a hay-rich feed mixture. . Animal Feed Science and Technology, 175: 85-89.
25- Kutz, R. E., J. D. Sampson., L. B. Pompeu., D. R. Leduox., J. N. Spain, and M. Vazquez-Anon. 2009. Efficacy of Solis, NovasilPlus, and MTB-100 to reduce aflatoxin M1 levels in milk of early to mid-lactation dairy cows fed aflatoxin B1. Journal of Dairy Science, 92: 3959-3963
26- Lindemann, M. D., D. J. Blodgett., E. T. Kornegay, and G. G. Schurig. 1993. Potential amelioratiors of aflatoxicosis in weanling/growing swine. Journal of Animal Science, 71: 171–178.
27- Menke, K. H, and H. Steingass. 1988. Estimation of the energetic feed value obtained from chemical analysis and in vitro gas production using rumen fluid. Animal Research and Development, 7: 28-55.
28- Mojtahedi, M., M. Danesh Mesgaran., S. A. Vakili, and E. Abdi Ghezeljeh. 2013. Effect of esterified glucomannan on carryover of aflatoxin from feed to milk in lactating holstein dairy cows. Annual Review & Research in Biology, 3:76-82.
29- Mokhtarpur, A., A. A. Naserian., R. Valizadeh, and A. M. Tahmasbi. 2012. Effect of pistachio silage products processed with polyethylene glycol and urea on phenolic compounds and in vitro gas production and performance of Holstein dairy cows. Iranian Journal of Animal Science Research, 4: 55-61. (In Persian).
30- Morgavi, D. P., H. Boudra, and J. P. Jouany. 2008. Consequences of mycotoxins in ruminant production. Animal Feed Science and Technology, 137: 201-212
31- Moschini, M., A. Gallo., G. Piva, and F. Masoero. 2008. The effects of rumen fluid on the in vitro aflatoxin binding capacity of different sequestering agentsand in vivo release of the sequestered toxin. Animal Feed Science and Technology, 147: 292-309.
32- Motoi, Y. 1988. Present status on animal disease due to low fiber intake. Japanese Grassland Science, 10: 24-31.
33- Osuji, P. O., I. V. Nsahlai, and H. Khalili. 1993. Feed Evaluation. Ilca manual (International Livestock Center for Africa), Addis Ababa, Ethiopia.
34- Raju, M. V. L. N, and G. Devegowda. 2000. Influenceof esterified glucomannan on performance and organ morphology, serum biochemistry and hematology in broilers exposed to individual and combined mycotoxicosis. British Poultry Science, 41: 640-650.
35- Ramos, A. J., J. Fink-Gremmels, and E. Hernandez. 1996. Prevention of toxic effects of mycotoxins by means of nonnutritive adsorbent compounds. Journal of Food Protection, 59: 631-641.
36- Saleh, M. S, and A. B. Bonf. 2000. Bentonite supplementation to concentrate for lactating buffaloes. Egypt Journal of Nutrient Feeds, 6:67-78
37- Sarr, A. B., B. A. Clement, and T. D. Phillips. 1991. Molecular mechanism of aflatoxin B1 chemisorption by hydrated sodium calcium aluminosilicate. Toxicologist, 197: 91-97
38- SAS. SAS/STAT User's guide Statistics, Version 9.1. SAS Inst., Inc., Cary, NC.
39- Theodorou, M. K., B. A. Williams., M. S. Dhanoa., A. B.McAllan, and J. France. 1994. A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Animal Feed Science and Technology, 48: 185-197.
40- Valizadeh, R., M. Ghadami, and F. Mellati. 2011. Determining the chemical composition and nutritional value of Eurotia ceratoides using plastic bags and gas production. Iranian Journal of Animal Science Research, 3: 159-169. (In Persian).
41- Wallace, R. J, and C. J. Newbold. 1991. Effect of bentonite on fermentation in the rumen simulation technique and rumen ciliate protozoa. Journal of Agricultural Science, 116: 163-168.
42- Westlake, K., R. I. Mackie, and M. F. Dutton. 1989. In vitro metabolism of mycotoxins by bacterial protozoal and ovine ruminal fluid preparations. Animal Feed Science and Technology, 25: 169-178.
43- Williams, P. P., J. D. Robbins., J. Gutierrez, and R. E. Davis. 1963. Rumen bacterial and protozoa responses to insecticide substrates. Applied Microbiology, 9: 405-409.
44- Williams, A. G, and S. E. Withers. 1993. Changes in the rumen microbial population and its activities during the fermentation period after the reintroduction of ciliate protozoa in to rumen of defaunated sheep. Canadian Journal of Microbiology, 31: 61-69.
45- Yildiz, A. O., S. S. Parlatand, and I. Yildirim. 2004. Effect of dietary addition of live yeast (Saccharomyces cerevisiae) on some performance parameters of adult Japanese quail (Coturnix japonica) induced by aflatoxicosis. Revue Medicine Veterania, 155: 38-41.