Hepatoprotective effects of Silymarin in experimental aflatoxicosis in broiler chickens

• Keleş Ömer Faruk Van Yuzuncu Yil University, Faculty of Veterinary, Department of Pathology. Van, Türkiye https://orcid.org/0000-0002-7869-5311
• Zübeyir Huyut Van Yuzuncu Yil University, Faculty of Medicine, Department of Biochemistry. Van, Türkiye https://orcid.org/0000-0002-7623-1492

DOI: https://doi.org/10.52973/rcfcv-e35515

Keywords: Aflatoxicosis, antioxidant, broiler, histopathology, Silymarin

Abstract

Aflatoxicosis, a mycotoxicosis caused by aflatoxins, poses significant economic challenges in animal husbandry and presents serious health risks to humans. The current study aimed to evaluate the protective effects of Silymarin, an antioxidant, against aflatoxicosis in broiler chickens. A total of thirty–two broilers were randomly assigned to four experimental groups: control, Silymarin–treated, Aflatoxin B₁ (AFB₁)–treated, and AFB₁+Silymarin–treated. Broilers in control and Silymarin–treated groups were fed with a diet without Aflatoxin. AFB₁ and AFB₁+Silymarin treated groups received approximately 1 mg of Aflatoxin/broiler/total for 21 days. AFB₁+Silymarin group also received 10 g of Silymarin kg^–1 of food as a supplementation diet for 21 days. Biochemically, liver and serum TAS, SOD, GSH–Px, CAT, and BcL–2 levels in the AFB₁ group were lower than the control, while serum TOS, and serum and liver Caspase–3 levels were high (P<0.001). In addition, in the AFB₁+Silymarin group, liver and serum GSH–Px, SOD and BcL–2, and serum CAT values were significantly higher than the AFB₁ group (P<0.001). Also, in the AFB₁+Silymarin group, there was a slight increase in CAT level in liver samples compared to the AFB₁ group (P>0.05). Furthermore, TOS and Caspase–3 levels of both serum and liver in the AFB₁+Silymarin group were low, while BcL–2 levels were quite high in serum samples compared to the AFB₁ group (P<0.001). Pathologically, the livers were larger and pale in the AFB₁ group. Histopathological analysis of the AFB₁–treated group revealed diffuse hydropic/vacuolar degeneration and fibrosis, characterized by the activation of perisinusoidal myofibroblastic (Ito) cells. These morphological changes were significantly reduced in the AFB₁+Silymarin group. These findings indicate that Silymarin, with its strong antioxidant effect, may be effective against aflatoxicosis in preventing liver damage in broilers.

References

Eskola M, Kos G, Elliott CT, Hajšlová J, Mayar S, Krska R. Worldwide contamination of food–crops with mycotoxins: Validity of the widely cited ‘FAO estimate’of 25%. Crit. Rev. Food. Sci. Nutr. [Internet]. 2020; 60(16):2773–2789. doi: https://doi.org/gncv68 DOI: https://doi.org/10.1080/10408398.2019.1658570

Santos–Pereira C, Cunha SC, Fernandes JO. Prevalent mycotoxins in animal feed: Occurrence and analytical methods. Toxins. [Internet]. 2019; 11(5):290. doi: https://doi.org/ghmsgx DOI: https://doi.org/10.3390/toxins11050290

Emerole GO, Neskovic N, Dixon RL. The detoxication of aflatoxin B1 with glutathione in the rat. Xenobiotica. [Internet]. 1979; 9(12):737–743. doi: https://doi.org/cswzdz DOI: https://doi.org/10.3109/00498257909042342

Essigmann JM, Croy RG, Bennett RA, Wogan GN. Metabolic activation of aflatoxin B1: patterns of DNA adduct formation, removal, and excretion in relation to carcinogenesis. Drug. Metab. Rev. [Internet]. 1982; 13(4):581–602. doi: https://doi.org/b4bjrs DOI: https://doi.org/10.3109/03602538209011088

Arafa AS, Bloomer RJ, Wilson HR, Simpson CF, Harms RH. Susceptibility of various poultry species to dietary aflatoxin. Br. Poult. Sci. [Internet]. 1981; 22(5):431–436. doi: https://doi.org/cbprn9 DOI: https://doi.org/10.1080/00071688108447906

Rajput SA, Sun L, Zhang N–Y, Khalil MM, Ling Z, Chong L, Wang S, Rajput IR, Bloch DM, Khan FA, Shaukat A, Qi D. Grape seed proanthocyanidin extract alleviates aflatoxinB1–induced immunotoxicity and oxidative stress via modulation of NF–κB and Nrf2 signaling pathways in broilers. Toxins. [Internet]. 2019; 11(1):23. doi: https://doi.org/n337 DOI: https://doi.org/10.3390/toxins11010023

Aly SA, Anwer W. Effect of naturally contaminated feed with aflatoxins on performance of laying hens and the carryover of aflatoxin B1 residues in table eggs. Pak. J. Nutr. [Internet]. 2009; 8(2):181–186. doi: https://doi.org/cmzp63 DOI: https://doi.org/10.3923/pjn.2009.181.186

Kubena LF, Huff WE, Harvey RB, Yersin AG, Elissalde MH, Witzel DA, Giroir LE, Phillips TD, Petersen HD. Effects of a hydrated sodium calcium aluminosilicate on growing turkey poults during aflatoxicosis. Poult. Sci. [Internet]. 1991; 70(8):1823–1830. doi: https://doi.org/n338 DOI: https://doi.org/10.3382/ps.0701823

Doerr JA, Huff WE, Wabeck CJ, Chaloupka GW, May JD, Merkley JW. Effects of low level chronic aflatoxicosis in broiler chickens. Poult. Sci. [Internet]. 1983; 62(10):1971–1977. doi: https://doi.org/n339 DOI: https://doi.org/10.3382/ps.0621971

Loi M, Paciolla C, Logrieco AF, Mulè G. Plant bioactive compounds in pre–and postharvest management for aflatoxins reduction. Front. Microbiol. [Internet]. 2020; 11: 243. doi: https://doi.org/n34b DOI: https://doi.org/10.3389/fmicb.2020.00243

Phillips TD, Kubena LF, Harvey RB, Taylor DR, Heidelbaugh ND. Hydrated sodium calcium aluminosilicate: a high affinity sorbent for aflatoxin. Poult. Sci. [Internet]. 1988; 67(2):243–247. doi: https://doi.org/n34c DOI: https://doi.org/10.3382/ps.0670243

Oguz H, Kurtoglu V. Effect of clinoptilolite on performance of broiler chickens during experimental aflatoxicosis. Br. Poult. Sci. [Internet]. 2000; 41(4):512–517. doi https://doi.org/c4rhpd DOI: https://doi.org/10.1080/713654953

Rosa CA, Miazzo R, Magnoli C, Salvano M, Chiacchiera SM, Ferrero S, Saenz M, Carvalho EC, Dalcero A. Evaluation of the efficacy of bentonite from the south of Argentina to ameliorate the toxic effects of aflatoxin in broilers. Poult. Sci. [Internet]. 2001; 80(2):139–144. doi: https://doi.org/n34d DOI: https://doi.org/10.1093/ps/80.2.139

Ramos AJ, Fink–Gremmels J, Hernández E. Prevention of toxic effects of mycotoxins by means of nonnutritive adsorbent compounds. J. Food. Prot. [Internet]. 1996; 59(6):631–641. doi: https://doi.org/gncv6p DOI: https://doi.org/10.4315/0362-028X-59.6.631

Valenzuela–Grijalva NV, Pinelli–Saavedra A, Muhlia–Almazan A, Domínguez–Díaz D, González–Ríos H. Dietary inclusion effects of phytochemicals as growth promoters in animal production. J. Anim. Sci. Biotechnol. [Internet]. 2017; 59:1–17. doi: https://doi.org/n34f DOI: https://doi.org/10.1186/s40781-017-0133-9

Petrovska BB. Historical review of medicinal plants’ usage. Pharmacogn. Rev. [Internet]. 2012; 6(11):1–5. doi: https://doi.org/ghx8xd DOI: https://doi.org/10.4103/0973-7847.95849

Oso AO, Suganthi RU, Manjunatha–Reddy GB, Malik PK, Thirumalaisamy G, Awachat VB, Selvaraju S, Arangasamy A, Bhatta R. Effect of dietary supplementation with phytogenic blend on growth performance, apparent ileal digestibility of nutrients, intestinal morphology, and cecal microflora of broiler chickens. Poult. Sci. [Internet]. 2019; 98(10):4755–4766. doi: https://doi.org/n34g DOI: https://doi.org/10.3382/ps/pez191

Priya K, Preethy J, Usha P, Kariyil BJ, Uma R. Antioxidant effect of Eclipta prostrata (L.) leaf powder in broiler chicken during aflatoxicosis. Pharma. Innov. [Internet]. 2019[cited 12 Jul. 2024]; 8(4):801–803. Available in: https://n9.cl/ru3v3

Pradhan SC, Girish C. Hepatoprotective herbal drug, Silymarin from experimental pharmacology to clinical medicine. Indian J. Med. Res. [Internet]. 2006[cited 10 Jul. 2024]; 124(5):491–504. Available in: https://n9.cl/7mcpk1

Koushki M, Yekta RF, Amiri–Dashatan N. Critical review of therapeutic potential of Silymarin in cancer: A bioactive polyphenolic flavonoid. J. Funct. Foods. [Internet]. 2023; 104(6):105502. doi: https://doi.org/gt29bn DOI: https://doi.org/10.1016/j.jff.2023.105502

Manna SK, Mukhopadhyay A, Van NT, Aggarwal BB. Silymarin suppresses TNF–induced activation of NF–κB, c–Jun N–terminal kinase, and apoptosis. J. Immun. [Internet]. 1999; 163(12):6800–6809. doi: https://doi.org/n34h DOI: https://doi.org/10.4049/jimmunol.163.12.6800

Chand N, Muhammad D, Durrani FR, Qureshi MS, Sahibzada SU. Protective effects of milk thistle (Silybum marianum) against aflatoxin B1 in broiler chicks. Asian–Australasian J. Anim. Sci. [Internet]. 2011; 24(7):1011–1018. doi: https://doi.org/fnrvzk DOI: https://doi.org/10.5713/ajas.2011.10418

Tedesco D, Steidler S, Galletti S, Tameni M, Sonzogni O, Ravarotto L. Efficacy of Silymarin–phospholipid complex in reducing the toxicity of aflatoxin B1 in broiler chicks. Poult. Sci. [Internet]. 2004; 83(11):1839–1843. doi: https://doi.org/gqcjvw DOI: https://doi.org/10.1093/ps/83.11.1839

Hasheminejad SA, Makki OF, Nik HA, Ebrahimzadeh A. The effects of aflatoxin B1 and Silymarin–containing milk thistle seeds on ileal morphology and digestibility in broiler chickens. Vet. Sci. Dev. [Internet]. 2015; 5(2):115–119. doi: https://doi.org/n34j DOI: https://doi.org/10.4081/vsd.2015.6017

Makki OF, Omidi A, Nik HA, Hasheminejad SA, Senjedak SMH. Anti–aflatoxin B1 effects of Shirazi thyme (Zataria multiflora) in broilers: evaluation of performance and liver histopathology. Vet. Sci. Dev. [Internet]. 2016 ;6(1):36–40. doi: https://doi.org/n34k DOI: https://doi.org/10.4081/vsd.2016.6090

Jahanian E, Mahdavi AH, Asgary S, Jahanian R. Effects of dietary inclusion of Silymarin on performance, intestinal morphology and ileal bacterial count in aflatoxin–challenged broiler chicks. J. Anim. Physiol. Anim. Nutr. [Internet]. 2017; 101(5):e43–e54. doi: https://doi.org/n34m DOI: https://doi.org/10.1111/jpn.12556

Suchý P, Straková E, Kummer V, Herzig I, Písaříková V, Blechová R, Mašková J. Hepatoprotective effects of milk thistle (Silybum marianum) seed cakes during the chicken broiler fattening. Acta Vet. Brno. [Internet]. 2008; 77(1):31–38. doi: https://doi.org/cfhhsg DOI: https://doi.org/10.2754/avb200877010031

Amiridumari H, Sarir H, Afzali N, Fanimakki O. Effects of milk thistle seed against aflatoxin B1 in broiler model. J. Res. Med. Sci. [Internet]. 2013 [cited 22 Jul. 2024]; 18(9):786. Available in: https://n9.cl/xyauzq

Banu GS, Kumar G, Murugesan AG. Ethanolic leaves extract of Trianthema portulacastrum L. ameliorates aflatoxin B1 induced hepatic damage in rats. Indian J. Clin. Biochem. [Internet]. 2009; 24(3):250–256. doi: https://doi.org/dz3x53 DOI: https://doi.org/10.1007/s12291-009-0047-5

Youssef SF, Sayed–Elahl RMH, Mohamed MHA, El–Gabry HE, El–Halim A, Hassan AH, Eshera AA. Supplementing growing quail diets with Silymarin and curcumin to improve productive performance and antioxidant status and alleviate aflatoxin b1 adverse effects during the summer season. Egypt. J. Agric. Res. [Internet]. 2022; 100(4):529–539. doi: https://doi.org/n34p DOI: https://doi.org/10.21608/ejar.2022.155457.1263

Ortatatli M, Oğuz H, Hati̇poğlu F, Karaman M. Evaluation of pathological changes in broilers during chronic aflatoxin (50 and 100 ppb) and clinoptilolite exposure. Res. J. Vet. Sci. [Internet]. 2005; 78(1):61–68. doi: https://doi.org/ckvn7t DOI: https://doi.org/10.1016/j.rvsc.2004.06.006

Li S, Muhammad I, Yu H, Sun X, Zhang X. Detection of Aflatoxin adducts as potential markers and the role of curcumin in alleviating AFB1–induced liver damage in chickens. Ecotoxicol. Environ. Saf. [Internet]. 2019; 176:137–145. doi: https://doi.org/gwhnrg DOI: https://doi.org/10.1016/j.ecoenv.2019.03.089

Wang XH, Li W, Wang XH, Han MY, Muhammad I, Zhang XY, Sun XQ, Cui XX. Water–soluble substances of wheat: a potential preventer of aflatoxin B1–induced liver damage in broilers. Poult. Sci. [Internet]. 2019;98 (1): 136–149. doi: https://doi.org/gtgzmw DOI: https://doi.org/10.3382/ps/pey358

Uyar A, Yener Z, Dogan A. Protective effects of Urtica dioica seed extract in aflatoxicosis: histopathological and biochemical findings. Br. Poult. Sci. [Internet]. 2016; 57(2):235–245. doi: https://doi.org/ghzkfs DOI: https://doi.org/10.1080/00071668.2015.1129664

Karaman M, Ozen H, Tuzcu M, Çİğremİş Y, Onder F, Ozcan K. Pathological, biochemical and haematological investigations on the protective effect of α–lipoic acid in experimental aflatoxin toxicosis in chicks. Br. Poult. Sci. [Internet]. 2010; 51(1):132–141. doi: https://doi.org/bg66gd DOI: https://doi.org/10.1080/00071660903401839

Enzan H, Hara H, Himeno H, Iwamura S, Saibara T, Onishi S, Yamamoto Y. Immunohistochemical identification of Ito cells and their myofibroblastic transformation in adult human liver. Virchows Arch. [Internet]. 1994; 424(3): 249–256. doi: https://doi.org/cpqgs8 DOI: https://doi.org/10.1007/BF00194608

Jubb KVF. Pathology of Domestic Animals. 3th ed. Orlando (FLA, USA): Academic Press; 1985. 557 p.

Dohnal V, Wu Q, Kuča K. Metabolism of aflatoxins: key enzymes and interindividual as well as interspecies differences. Arch. Toxicol. [Internet]. 2014; 88(9):1635–1644. doi: https://doi.org/f6d3w4 DOI: https://doi.org/10.1007/s00204-014-1312-9

Gowda NKS, Rajendran US, Malathi V, Anjanappa R. Effectiveness of hydrated sodium calcium alumino silicate for preventing aflatoxin toxicity in sheep. Anim. Nutr. Feed. Technol. 2006; 6(2):243–250.

Benkerroum N. Chronic and acute toxicities of aflatoxins: Mechanisms of action. Int. J. Environ. Res. Public. Health. [Internet]. 2020; 17(2):423. doi: https://doi.org/grg6bc DOI: https://doi.org/10.3390/ijerph17020423

Prakoso YA, Puspitasari, Rini CS, Aliviameita A, Salasia SIO, Kurniasih, Ikram AFD, Walalangi B, Utama KP, Al Huda MF, Su’udiyah NA. The role of Sauropus androgynus (L.) Merr. leaf powder in the broiler chickens fed a diet naturally contaminated with aflatoxin. J. Toxicol. [Internet]. 2018; 1:2069073. doi: https://doi.org/n34v DOI: https://doi.org/10.1155/2018/2069073

Manafi M, Hedayati M, Yari M. Effectiveness of rosemary (Rosmarinus officinalis L.) essence on performance and immune parameters of broilers during aflatoxicosis. Adv. Life Sci. [Internet]. 2014 [cited 22 Jul. 2024]; 4(3):166–173. Available in: https://goo.su/VOmv

Yarru LP, Settivari RS, Antoniou E, Ledoux DR, Rottinghaus GE. Toxicological and gene expression analysis of the impact of aflatoxin B1 on hepatic function of male broiler chicks. Poult. Sci. [Internet]. 2009; 88(2):360–371. doi: https://doi.org/c9dwrc DOI: https://doi.org/10.3382/ps.2008-00258

Valko M, Leibfritz D, Moncol J, Cronin MTD, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease. Int. J. Biochem. Cell. Biol. [Internet]. 2007; 39(1):44–84. doi: https://doi.org/bdqwsb DOI: https://doi.org/10.1016/j.biocel.2006.07.001

Towner RA, Qian SY, Kadiiska MB, Mason RP. In vivo identification of aflatoxin–induced free radicals in rat bile. Free. Radic. Res. [Internet]. 2003; 35(10):1330–1340. doi: https://doi.org/chzvvk DOI: https://doi.org/10.1016/j.freeradbiomed.2003.08.002

Ajiboye TO, Yakubu MT, Oladiji AT. Lophirones B and C prevent aflatoxin B1–induced oxidative stress and DNA fragmentation in rat hepatocytes. Pharm. Biol. [Internet]. 2016; 54(10):1962–1970. doi: https://doi.org/n35f DOI: https://doi.org/10.3109/13880209.2015.1137603

Umaya SR, Vijayalakshmi YC, Sejian V. Exploration of plant products and phytochemicals against aflatoxin toxicity in broiler chicken production: Present status. Toxicon. [Internet]. 2021; 200:55–68. doi: https://doi.org/gt33qw DOI: https://doi.org/10.1016/j.toxicon.2021.06.017