Effects of high grade bentonite on performance, organ weights and serum biochemistry during aflatoxicosis in broilers

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1 Effects of high grade bentonite on performance, organ weights and serum biochemistry during aflatoxicosis in broilers H. C. Indresh, G. Devegowda, S. Wilfred Ruban and M. C. Shivakumar 1. Department of Livestock Production and Management, Veterinary College, Hassan , Karnataka, India; 2. Former Professor & Head, Division of Animal Science, Veterinary College, Hebbal, Bangalore - 24, Karnataka, India; 3. Department of Livestock Products Technology, Veterinary College, Hassan , Karnataka, India; 4. Department of Livestock Production and Management, Veterinary College, Hassan , Karnataka, India Corresponding auor: H. C. Indresh, drindreshpsc@gmail.com Received: , Accepted: , Published online: How to cite is article: Indresh HC, Devegowda G, Ruban SW and Shivakumar MC (2013) Effects of high grade bentonite on performance, organ weights and serum biochemistry during aflatoxicosis in broilers, Vet. World 6(6): , doi: /vetworld Introduction Cereal grains and associated by-products constitute important sources of energy for poultry. However, ere is increasing evidence at global supplies of cereal grains for animal feedstuffs are frequently contaminated wi mycotoxins. It is estimated at as much as 25 percent of e world's cereals are contaminated wi known mycotoxins [1]. Aflatoxins are secondary toxic metabolites, produced by certain strains of fungi, e.g. Aspergillus flavus and Aspergillus parasiticus species, of which aflatoxin B1 (AFB1), is e most toxic among all oer aflatoxins i.e. AFB1, AFB2, AFG1 and AFG2. In poultry, aflatoxin ingestion leads to Aflatoxicosis syndrome at is characterized by retardation in grow, decreased feed consumption and feed conversion efficiency, immunosuppression and e increase of mortality [2]. Co-contamination of cereal grains wi oer mycotoxins produced by different fungal genera, including Fusarium and Aspergillus, Abstract Aim: To evaluate e effect of different levels of High Bentonite on grow performance, organ weight and serum biochemistry in broiler fed on diets containing aflatoxin. Materials and Meods: A total of 360 day-old commercial broiler chicks were divided at random into 8 dietary treatment groups of 42 chicks each having 3 replicates. Dietary levels of aflatoxin (0.5 ppm) and High-grade bentonite (0.5, 0.75 and 1.00%) were tested in a completely randomized design manner, forming a total of 8 dietary treatments each wi ree replicates. Body weight and feed intake were recorded weekly. At 5 wk, six birds from each treatment were sacrificed and liver, kidney, gizzard, pancreas, spleen, bursa of Fabricius and ymus were extracted and weighed. The serum samples were analyzed for total proteins, uric acid, serum albumin, serum globulin and e activities of gamma glutamyl transferase (GGT), alanine amino transferase (ALT) and for antibody titers against Newcastle disease (ND) and infectious bursal disease (IBD) using ELISA technique. Results: A significant (P<0.05) decrease in body weight, feed consumption, relative weights of bursa, ymus, serum protein, anti body titers against NDV and IBDV, and increase in FCR, mortality, relative weight of liver, kidney, and e activity of Gamma Glutamyl Transferase (GGT) was observed. However, e relative weights of gizzard, pancreas, spleen, serum albumin, uric acid and e activity of Alanine Amino Transferase (ALT) were not influenced by inclusion of AF or HGB. Conclusion: The addition of HGB restored e harmful effects of AF on body weight, feed consumption, FCR, mortality, relative weight of liver, kidney, serum protein, IBDV and NDV. Supplementation of high grade bentonite at 1.0 per cent level was found to be beneficial in ameliorating e adverse effects of aflatoxin (AF) in broiler chickens. Keywords: aflatoxin, body weight, feed consumption, high grade bentonite, in-vivo This article is an open access article licensed under e terms of e Creative Commons Attribution License ( org/licenses/by/2.0) which permits unrestricted use, distribution and reproduction in any medium, provided e work is properly cited. has been reported to increase e toxicity symptoms in poultry [3]. Among Aflatoxins (AF), AFB1 is e most potent hepatotoxin having carcinogenic and severe performance depressive effects on poultry [4]. At present, one of e more encouraging approaches to make e feed harmless is e addition of non-nutritive and natural adsorbent materials to e contaminated feed in order to selectively bind e mycotoxin during e digestive process. The major advantages of ese adsorbents include low cost, safety, and e ease of addition to animal feed. Layered amino silicates, such as sodium bentonite, have been found effective in counteracting mycotoxins [5-9]. However, e ability of bentonite to bind mycotoxins depends on ph, molecular arrangements, and its geographic region of origin [10]. Considering all ese facts, e present study was undertaken to investigate e ability of high grade bentonite on aflatoxicosis in broilers. Material and Meods Experimental animals: A total of 336-day-old commercial broilers (Vencobb), were procured from a 313

2 Table-1. Percent ingredient composition of basal diet local hatchery, divided at random into 24 replicates of recorded weekly. At 5 wk, six birds from each 14 chicks each having equal number of males and treatment (3 males and 3 females) were humanely females. Each ree such groups were allotted to one of slaughtered as per religious (Halal) meod of e 8 dietary treatments wi due permission from e slaughter by severing e carotid arteries and jugular Animal Eics committee of Veterinary College, veins and liver, kidney, gizzard, pancreas, spleen, bursa Bangalore. of Fabricius and ymus were collected and weighed. The weights were adjusted to one kg live weight and Experimental design and test diets: Dietary levels of e treatment means were calculated. AF (0.5 ppm) and High-grade bentonite (0.5, 0.75 and Blood was collected in non-heparinized tubes 1.00%) were tested in a completely randomized design from six birds in each treatment (3 males and 3 manner, forming a total of 8 dietary treatments each females) by puncturing e brachial vein during 5 wi ree replicates. Basal diet was formulated and week of age. Serum was collected after 8 to 10 hours as compounded to meet e nutrient requirements of per e standard procedures [13] and was stored at commercial broilers during e starter (0-3 wks) and 200C for subsequent analysis. The individual serum finisher (4-5 wks) phases (Table-1). samples were analyzed for total proteins, uric acid, AF were produced using e pure culture of serum albumin, serum globulin and e activities of Aspergillus parasiticus MTCC 1894 (Microbial Type gamma glutamyl transferase (GGT), alanine amino Culture Collection and Gene Bank, IMT, Chandigarh, India) grown on potato dextrose agar. Then AF transferase (ALT) using automatic analyzer produced on relevant Medias were extracted [11] and (Boehringer Mannheim Hitachi 704 automatic quantified by in layer chromatography [12]. analyzer, Japan). The meodology and e set of Individual ingredients were screened for mycotoxin reagents used in respect of each parameter were as per content prior to compounding e feed. Compounded e recommendations of e manufacturer of e feed was analyzed for e presence of AF before analyzer system. Data are presented as means of each including e rice and wheat culture materials, and en treatment. The serum samples were assayed for e diets were prepared by incorporating required antibody titers against Newcastle disease (ND) and quantities (0.5 ppm) of rice/wheat culture containing infectious bursal disease (IBD) using ELISA technique AF into e diet. To e basal diet High-grade bentonite at five weeks of age. Treatment-wise means of titers at 0.5, 0.75 and 1.00 percent were added to prepare e were calculated. different experimental diets. Compounded experimen- Statistical analysis: The results were subjected to onetal diets were analyzed for mycotoxin content to way analysis of variance. Means were compared using counter check e required levels. Chicks were fed ad Tukey meod [14]. The level of significance was libitum. Feeding of test diets commenced at zero day of tested (P<0.05). age and continued till e termination of experiment at five weeks of age. Results and Discussion Data collection: Body weight and feed intake were Grow performance: Birds fed on aflatoxin 314

3 Table-3. Effect of aflatoxin wi or wiout High grade bentonite (HGB) on organ weights (g/ kg live wt.) and lymphoid organ weights of broiler chickens at 5 wk of age (Mean+ SE). contaminated feed (0.5 ppm) showed a significant bentonite against AF induced organ weight increase in (P<0.05) decrease in body weight, feed intake and broilers were in accordance wi [20,21,30]. Compared increased FCR. The result of present investigation was wi control, relative weights of gizzard and pancreas in accordance to e observations of [15-19]. were not affected eier in AF fed group, control and The addition of 0.75 and 1.0 per cent HGB HGB supplemented groups. The relative ymus significantly restored e body weight, feed intake and (38.99%) and bursal weights (31.36%) were FCR (Table-2). The present results relating to e use of significantly (P<0.05) lower at 0.5 ppm inclusion of Bentonite in AF containing diets is in agreement wi AF group (Table-3) compared to control diet. A Similar e findings of [20-22] who opined at improvement reduction in relative weights of ymus and bursa wi in body weight upon bentonite inclusion may be due to AF were reported by [4, 17, 27, 28]. eir montmorillonites content at bind irreversibly Relative weights of ymus and bursal were not wi toxin in e gut and ereby preventing absorption altered by supplementation of varying levels of HGB of e toxin across e intestinal wall, providing a compared to control diets. Compared wi control, prophylactic action. relative weight of spleen was not affected eier in AF High mortality rate of per cent was fed group or HGB supplemented groups. The observed in group fed wi diet containing 0.5 ppm AF. protective effects of HGB may be due to sequestration Similar mortality rate in broilers due to AF toxin were of AF in e gut which reduces bioavailability of e AF reported by several workers [17, 23-26] Mortality rate toxin. was reduced considerably in control as well as in e Serum Biochemistry: The Serum antibody titres against groups supplemented wi 0.5, 0.75 and 1.0 per cent ND and IBD vaccination were significantly (P<0.05) HGB. who reported. The results were in accordance depressed by aflatoxin which were restored wi e wi e findings of [20], who reported a significant inclusion of 1.0 per cent HGB (Table-4). These reduction in mortality of broilers upon addition of 0.5 findings agree wi e previous reports of decreased per cent bentonite to 0.5 ppm aflatoxin containing antibody titres against ND and IBD in AF fed broilers diets. [31, 32, 33, 34]. The decreased antibody titers could be Organ weights: The relative weights of liver (19.56%), due to inhibition of DNA and protein synesis by and kidney (18.38%) increased significantly (P<0.05) aflatoxin rough impairment of amino acid transport wi addition of 0.5 ppm AF (Table-3), whereas and m-rna transcription resulting in lowered level of improvement was observed wi dietary inclusion of antibody production [35] and 1.0 per cent HGB. The increase in liver and The results clearly demonstrated e protective kidney weights were in accordance wi e findings of effects of bentonite at 1.0 per cent inclusion to aflatoxin [17-19, 27, 28]. The increase in liver weight could be diet in chickens. The findings of present study were attributed to increased lipid deposition in liver due to comparable to e reports of [36] who reported an impaired fat metabolism [29]. The protective effects of improved antibody titres in chickens fed on aflatoxin

4 Table-5. Effect of aflatoxin wi or wiout High grade bentonite (Ashafeed) on levels of serum biochemical parameters and serum enzyme activities of broilers at 5 wk of age (Mean+ SE). The serum concentration of total protein In vitro Binding Ability of Mycotoxin Binder in Commercial (38.37%) significantly (P<0.05) decreased by addition Broiler Feed. African J. Agri. Res. 4(2): Mohamadi, H. and Alizadeh, M. (2010) A Study of e of aflatoxin (Table-5), but was elevated to normal level Occurrence of Aflatoxin M1 in Dairy Products Marketed in wi e inclusion of 1.0 per cent HGB. Serum Urimia, Iran. J. Agri. Sci. Tech. 12: concentration of uric acid and albumin were not 3. Hagler, W. M., Tyczkowska, K. and Hamilton, P. B. (1984) affected eier in AF fed group or HGB supplemented Simultaneous Occurrence of Deoxynivalenol, Zearalenone and Aflatoxin in 1982 Scabby Wheat from Midwestern groups (Table 5). Reduced levels of serum total protein, United States. Appl. Environ. Microbiol.47: serum uric acid and serum albumin in AF fed diets were 4. Devegowda, G. (2002) Mycotoxins: Economic risks and reported earlier [23, 27, 29]. eir control. In: Handbook of Poultry Nutrition. Published The activity of serum GGT significantly by American Soybean Association, pp Smi, J. E. and Ross, K. (1991) The Toxigenic Aspergilli. (P<0.05) increased in AF fed group (Table-5). The In: "Mycotoxins and Animal Foods", (Eds.): Smi, J. E. and addition of graded levels of HGB to AF containing diet Henderson, CRC Press, Boca Raton, PP did not show significant reduction in e activity of 6. Santurio, J. M., Mallmannl, C. A., Rosa, A. P., Appel, G., serum GGT. Activity of serum ALT was not affected Heer, A., Dageforde, S. and Bottcher, M. (1999) Effect of Sodium Bentonite on e Performance and Blood Variables eier in AF fed group or control and HGB of Broiler Chickens Intoxicated wi Aflatoxin. Brit. Poultry supplemented groups (Table-5). The increased serum Sci. 40: enzyme activity as observed during aflatoxicosis is 7. Hagler, W. M., Jr. Grimes, J. and Fairchild, L. (1992) Effects interpreted as e sequalae of hepatocyte degeneration of Astra-Ben 20 on Broiler Chicks Exposed to AFB1 or T-2 Toxin. Nor Carolina State University. Poultry Abstract. and subsequent leakage of enzymes into e circulation 16: 123. [29, 37]. 8. Rosa, C. A. R., Miazoo, R., Magnoli, C., Salvano, M., Chicchiera, S. M., Ferrero, S., Saenz, M., Carvalho, E. C. Q. Conclusions and Dalcero, A. (2001) Evaluation of e Efficacy of Grow performance, organ weights and serum Bentonite from e Sou Argentina to Ameliorate e Toxic Effects of Aflatoxin in Broilers. Brit. Poultry Sci. 32: 100- profile of broilers were significantly (P<0.05) altered 108. by e aflatoxin, indicating eir adverse effects on e 9. Eralsan, G., Essz, D., Akdogan, M., Sahindokuyucu, F. and general heal of broilers. Supplementation of high Altrintas, L. (2005) The Effects of Aflatoxin and Sodium grade bentonite at 1.0 per cent level was found Bentonite and Alone on Some Blood Electrolyte Levels in beneficial in ameliorating e adverse effects of AF in Broiler Chickens. Turk Vet. Hayvanclk Dergisi. 29: Vieira, S.L. (2003) Nutritional implication of mould broiler chickens. development in feed stuffs and alternatives to reduce e Auors contribution mycotoxin problem in poultry feeds. World's Poultry Science Journal, 59: HCI carried out e experiment. GD designed e 11. Rukmini, C. and R.V. Bhat. (1978) Occurrence of T- 2 toxin experiment, guided during e experiment. SWR and in Fusarium infested sorghum from India. J. of Agri. Food Chemist. 26: MCS helped in analysis of data, drafted and revised e 12. AOAC. (1995) Official Meods of Analysis.16 Edn. manuscript. All auors read and approved e final Association of Official Analytical Chemists, Arlington, VA. manuscript. 13. Calnek, B.W., Barnes, H.J., Beard, C.W., Reid, W.M. and yoder, Jr. H.W. (1992) Diseases of Poultry, 9 Edn., Wolfe Acknowledgements Publishing, Ltd., USA. 14. Snedecor, G.W. and W.G.Cochran. (1989) Statistical The auors are ankful to The Dean, Veterinary meods. Six ed. Iowa State University Press. Ames. Owa. College, KVAFS University, Bangalore for providing 15. Swamy, H.V.L.N. and Devegowda, G. (1998) Ability of e basic facilities for e conduct of e research. mycosorb to counteract aflatoxicosis in commercial broilers. Indian J. Poult. Sci. 33: Competing interests 16. Sapcota, D., Islam, R. and Bamah, K.K. (2005) Effect of dietary meomin as a counteracting agent in experimental Auors declare at ey have no competing interests. aflatoxicosis of broilers. Ind. J. Poult. Sci. 40(2): References 17. Raju, M.V.L.N. and Devegowda, G. (2000) Influence of esterified glucomannan on performance and organic 1. Manafi, M., Narayana Swamy, H. D. and Pirany, N. (2009) morphology, serum biochemistry and haematology in 316

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