ORIGINAL_ARTICLE The effect of probiotics on T-2 mycotoxin induced apoptosis in chicken liver tissue In recent years many researchers have described the reduced mycotoxin toxicity caused by probiotic bacteria. Since reduction under gastrointestinal conditions of the bioavailability of mycotoxins by probiotics is not fully investigated in birds, the aim of the study was to determine the effect of probiotic on T-2 mycotoxicosis induced apoptosis in broiler’s liver. For the study, twelve 1-days old broilers were divided equally into T-2 toxin (T2) and probiotic with T-2 (P+T2) groups. From the first experimental day, probiotic Enterococcus faecium DSM 7134 was administered in drinking water to P+T2 group. From the fourth day, T-2 toxin was given for three consecutive days to T2 toxin group. At 8th experimental day chicken were sacrificed, liver was fixed in buffered 10% formalin, embedded into paraffin, slices 5 μm in thickness were cut followed by immunohistochemical staining with polyclonal primary antibodies p21 and p53 (Abcam, UK) according to the manufacturers’ guidelines (IHC kit, Abcam, UK). Blood samples were taken by cardiac puncture to measure liver enzymes. Immunohistochemical staining revealed strong expression of p53 and p21 antibodies in hepatocytes nuclei as well as around blood vessels in T-2 toxin group’s chicken liver tissue. Staining by both antibodies was less intensive in P+T2 group. Enzyme analysis showed significantly increased (p < 0.05) blood aspartate transaminase and alanine transaminase concentrations by 33.87% and 68.03% respectfully in T2 toxin group, while enzyme concentrations were decreased in P+T2 group. The obtained results showed reduced features of liver apoptosis in treatment with probiotic bacteria. https://macvetrev.mk/Files/Article/2020/10.2478/macvetrev-2020-0015/macvetrev-2020-0015.pdf 2020-03-15T09:00:00 55 60 10.2478/macvetrev-2020-0015 T-2 mycotoxin probiotics apoptosis broilers liver Piret Hussar piretut@gmail.com false 1 Department of Anatomy, Faculty of Medicine, University of Tartu, Tartu 50411, Estonia LEAD_AUTHOR Katerina Blagoevska false 2 Deparment of Functional Morphology, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, 1000 Skopje, R. of North Macedonia AUTHOR Tõnu Järveots false 3 Department of Veterinary Bio-and Population Medicine, Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Tartu 51014, Estonia AUTHOR Monika Dovenska false 4 Deparment of Functional Morphology, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, 1000 Skopje, R. of North Macedonia AUTHOR Dimitar Bozinovski false 5 Deparment of Functional Morphology, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, 1000 Skopje, R. of North Macedonia AUTHOR Florina Popovska-Percinic false 6 Deparment of Functional Morphology, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, 1000 Skopje, R. of North Macedonia AUTHOR Richard,  J.L.  (2007).  Some  major  mycotoxins and their mycotoxicoses - an overview. Int J Food Microbiol. 119(1-2): 3-10. PMid:17719115 1 10.1016/j.ijfoodmicro.2007.07.019 Vinderola, G., Ritieni, A. (2014). Role of probiotics against mycotoxins and their deleterious effects. JFR. 4(1): 10-21. 2 10.5539/jfr.v4n1p10 Smith, J.E., Solomons, G., Lewis, C., Anderson, J.G. (1995). Role of mycotoxins in human and animal nutrition and health. Nat Toxins 3, 187-192. PMid:7582615 3 Perkowski, J., Stupera, K., Buskoa, M., Goralb, T., Kaczmarekc, A., Jelend, H. (2012). Differences in metabolomic profiles of the naturally contaminated grain of barley, oats and rye. J. Cereal Science 56(3): 544-551. 4 10.1016/j.jcs.2012.07.012 Zhuang, Z., Yang, D., Huang, Y., Wang, S. (2013). Study on the apoptosis mechanism induced by T-2 toxin. PLoS One 8(12): e83105. PMid:24386148 PMCid:PMC3873290 5 10.1371/journal.pone.0083105 Trufanov, O.V., Kotyk, A.M., Bozhok, L.V. (2008). Effect of probiotic preparation based on Bacillus subtilis (BPS-44) in experimental mycotoxicoses of chickens. Mikrobiol Z. 70(1): 52-58. 6 Blagoevska, K., Ristoski, T., Dodovski, A., Blagoevski, A., Stojkovski, V., Ilieski, V., Pendovski, L., Petkov, V., Dovenska, M., Popovska- Percinic, F. (2014). Probiotic effect on broilers' liver morphology in T-2 mycotoxicosis. Proceedings of the XXXth Congress of the European Association  of Veterinary Anatomists, 23-26 July 2014, Cluj- Napoca, Romania, AHE 43 (1): pp. 24. 7 10.1111/ahe.12127 Ferrer, M., Maynes, L., Maries, J., Meca, G. (2015). Influence of prebiotics, probiotics and protein ingredients on mycotoxin bioaccessibility. Food Funct. 6(3): 987-994. PMid:25673154 8 10.1039/C4FO01140F Kabak, B., Dobson, A. D. W. (2009). Biological strategies to counteract the effects of mycotoxins. J Food Prot. 72(9): 2006-2016. 9 10.4315/0362-028X-72.9.2006 El-Nezami, H., Mykkänen, H., Kankaanpää, P., Salminen, S., Ahokas, J. (2000). Ability of Lactobacillus and Propionibacterium strains to remove aflatoxin  B  from  the  chicken  duodenum. J Food Prot. 63(4): 549-552. PMid:10772225 10 10.4315/0362-028X-63.4.549 Michael, B., Yano, B., Sellers, R.S., Perry, R., Morton, D., Roome, N., Johnson, J.K., Schafer, K. (2007). Evaluation of organ weights for rodent and non-rodent toxicity studies: A review of regulatory guidelines and a survey of current practices. Toxicol Pathol. 35(5): 742-750. PMid:17849357 11 10.1080/01926230701595292 Nyblom, H., Björnsson, E., Smiren, M., Aldenborg, F., Almer, S., Olsson, R. (2006). The AST/ALT ratio as an indicator of cirrhosis in patients with PBC. Liver Int. 26(7): 840-845. PMid:16911467 12 10.1111/j.1478-3231.2006.01304.x Peters, J.M., Boyd, E.M. (1966). Organ weights and water levels of the rat following reduced food intake. J Nutr. 90(4): 354-360. PMid:5954817 13 10.1093/jn/90.4.354 Hussar, P., Järveots, T., Pendovski, L., Blagoevska, K., Popovska-Percinic, F. (2018).  T-2 mycotoxin induced apoptosis in broiler's liver tissue. Papers on Anthropology XXVII (1): 9-16. 14 10.12697/poa.2018.27.1.01 Turker, L., Gumus, S. (2009). A theoretical study on vomitoxin and its tautomers. J Hazard Mater. 163, 285-294. PMid:18657904 15 10.1016/j.jhazmat.2008.06.087 Li, Y., Wang, Z., Beier, R.C., Shen, J., De Smet, D., De Saeger, S., Zhang, S. (2011). T-2 toxin, a trichothecene mycotoxin: review of toxicity, metabolism, and analytical methods. J Agric Food Chem. 59(8): 3441-3453. PMid:21417259 16 10.1021/jf200767q El- Deiry, W.S., Tokino, T., Velculescu, V.E., Levy, D.B., Parsons, R., Trent, J.M., et al. (1993). WAF1, a potential mediator of p53 tumor suppression. Cell 75(4): 817-825. 17 10.1016/0092-8674(93)90500-P Waldman, T., Kinzler, K.W., Vogelstein, B. (1995). p21 is necessary for the p53-mediated G1 arrest in human cancer cells. Cancer Research 55(22): 5187-5190. 18 Hill, C., Guarner, F., Reid, G., Gibson, G.R., Merenstein, D.J., Pot, B., et al. (2014). Expert consensus document. The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol. 11(8): 506-514. PMid:24912386 19 10.1038/nrgastro.2014.66 Rijkers, G.T., De Wos, W.M., Brummer, R.J., Morelli, L., Corhier, G., Marteau, P. (2011). Health benefits and health claims of probiotics: Bridging science and marketing. Br J Nutr. 106(9): 1291-1296. PMid:21861940 20 10.1017/S000711451100287X Edrington, T.S., Kubena, L.F., Harvey, R.B., Rottinghaus, G.E. (1997). Influence of a superactivated charcoal on the toxic effects of aflatoxin or T-2 toxin in growing broilers. Poult Sci. 76, 1205-1211. PMid:9276881 21 10.1093/ps/76.9.1205 Hashem, M.A., Mohamed, M.H. (2009). Hemato-biochemical and pathological studies on aflatoxicoses and treatment of broiler chicks in Egypt. Veterinaria Italiana 45(2): 323-337. 22