Abstract
Aflatoxins, particularly AFB1, are the most common feed contaminants worldwide, causing significant economic losses to the livestock sector. The current paper describes an outbreak of aflatoxicosis in a herd of 160 male young goat kids (3–4 months), of which 68 young kids succumbed over a period of 25 days after showing neurological signs of abnormal gait, progressive paralysis and head pressing. The haematobiochemical investigation showed reduced haemoglobin, leucocyte count, PCV level, increased levels of AST, ALT, glucose, BUN, creatinine and reduced level of total protein. Grossly, kids had pale mucous membranes, pale and swollen liver; right apical lobe consolidation, and petechiation of the synovial membrane of the hock joints. The microscopic changes were characterized by multifocal hemorrhages, status spongiosus/ vacuolation, vasculitis, focal to diffuse gliosis, satellitosis, and ischemic apoptotic neurons in different parts of the brain and spinal cord. These changes corresponded well with strong immunoreactivity for AFB1 in neurons, glia cells (oligodendrocytes, astrocytes, and ependymal cells) in various anatomical sites of the brain. The higher values of LPO and reduced levels of antioxidant enzymes (Catalase, SOD, GSH) with strong immunoreactivity of 8-OHdG in the brain indicating high level of oxidative stress. Further, the higher immunosignaling of caspase-3 and caspase-9 in the brain points towards the association with intrinsic pathway of apoptosis. The toxicological analysis of feed samples detected high amounts of AFB1 (0.38ppm). These findings suggest that AFB1 in younger goat kids has more of neurotoxic effect mediated through caspase dependent intrinsic pathway.
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The datasets generated in the current study are available from the corresponding author upon reasonable request.
References
Abdelsalam E, el-Tayeb A, Nor Eldin A et al (1989) Aflatoxicosis in fattening sheep. Vet Rec 124:487–488. https://doi.org/10.1136/vr.124.18.487
Ahmed N, El-Rayes SM, Khalil WF et al (2022) Arabic gum could alleviate the aflatoxin B1-provoked hepatic injury in rat: the involvement of oxidative stress, inflammatory, and apoptotic pathways. Toxins 14:605. https://doi.org/10.3390/toxins14090605
Balina A, Kebede A, Tamiru Y (2018) Review on aflatoxin and its impacts on Livestock. J Dairy Veterinary Sci 6:e555685
Bautista-García PB, García-Esquivel N, González-López L et al (2019) Role of apoptosis in human diseases. Austin J Clin Pathol 9:1058
Beers R, Sizer T (1952) Spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195:133–138
Bubert A, Hein I, Rauch M et al (1999) Detection and differentiation of Listeria spp. by a single reaction based on multiplex PCR. Appl Environ Microbiol 65:4688–4692. https://doi.org/10.1128/aem.65.10.4688-4692.1999
Chen J, Chen K, Yuan S et al (2016) Effects of aflatoxin B1 on oxidative stress markers and apoptosis of spleens in broilers. Toxicol Ind Health 32:278–284. https://doi.org/10.1177/0748233713500819
Clarke P, Tyler KL (2009) Apoptosis in animal models of virus-induced disease. Nat Rev Microbiol 7:144–155. https://doi.org/10.1038/nrmicro2071
Couacy-Hymann E, Roger F, Hurard C et al (2002) Rapid and sensitive detection of peste des petits ruminants virus by a polymerase chain reaction assay. J Virol Methods 100:17–25. https://doi.org/10.1016/s0166-0934(01)00386-x
D’Angelo AD, Bellino C, Alborali GL et al (2007) Neurological signs associated with aflatoxicosis in piedmontese calves. Vet Rec 160:698–698. https://doi.org/10.1136/vr.160.20.698
Damiano S, Jarriyawattanachaikul W, Girolami F et al (2022) Curcumin supplementation protects broiler chickens against the renal oxidative stress induced by the dietary exposure to low levels of aflatoxin B1. Front Veterinary Sci 8:1659
Donmez N, Dönmez HH, Keskin E, Kısadere İ (2012) Effects of aflatoxin on some haematological parameters and protective effectiveness of esterified glucomannan in Merino Rams. The Scientific World Journal 2012: 342468. https://doi.org/10.1100/2012/342468
Fedlu M, Seid A, Wollelie A et al (2019) Aflatoxin and its public health significance: a review. J Dairy Vet Sci 12:555837. https://doi.org/10.19080/jdvs.2019.12.555837
Ferretti S, Gatto A, Curatola A et al (2021) Atypical reye syndrome: three cases of a problem that pediatricians should consider and remember. Acta Bio Medica: Atenei Parmensis 92: e2021110.Xu
Frangiamone M, Cimbalo A, Alonso-Garrido M et al (2022) In vitro and in vivo evaluation of AFB1 and ota-toxicity through immunofluorescence and flow cytometry techniques: a systematic review. Food Chem Toxicol 160:112798. https://doi.org/10.1016/j.fct.2021.112798
Fuchs Y, Steller H (2011) Programmed cell death in animal development and disease. Cell 147:742–758. https://doi.org/10.1016/j.cell.2011.10.033
Gagliano N, Donne ID, Torri C et al (2006) Early cytotoxic effects of ochratoxin a in rat liver: a morphological, biochemical and molecular study. Toxicology 225:214–224. https://doi.org/10.1016/j.tox.2006.06.004
Gugliandolo E, Peritore AF, D’Amico R et al (2020) Evaluation of neuroprotective effects of quercetin against aflatoxin B1-intoxicated mice. Animals 10:898. https://doi.org/10.3390/ani10050898
Hashem MA, Mohamed MH (2009) Haemato-biochemical and pathological studies on aflatoxicosis and treatment of broiler chicks in Egypt. Vet Ital 45:323–337
Huang B, Chen Q, Wang L et al (2020) Aflatoxin B1 induces neurotoxicity through reactive oxygen species generation, DNA damage, apoptosis, and S-phase cell cycle arrest. Int J Mol Sci 21:6517. https://doi.org/10.3390/ijms21186517
Ikegwuonu FI (1983) The neurotoxicity of aflatoxin B1 in the rat. Toxicology 28:247–259. https://doi.org/10.1016/0300-483X(83)90121-X
Jahreis S, Kuhn S, Madaj AM et al (2017) Mold metabolites drive rheumatoid arthritis in mice via promotion of IFN-gamma- and Il-17-producing T cells. Food Chem Toxicol 109:405–413. https://doi.org/10.1016/j.fct.2017.09.027
Kebede A (2018) Review on aflatoxin and its impacts on livestock. J Dairy Veterinary Sci 6:555685. https://doi.org/10.19080/jdvs.2018.06.555685
Kraft S, Buchenauer L, Polte T (2021) Mold, mycotoxins and a dysregulated immune system: a combination of concern? Int J Mol Sci 22:12269. https://doi.org/10.3390/ijms222212269
Li S, Blain EJ, Cao J et al (2014) Effects of the mycotoxin nivalenol on bovine articular chondrocyte metabolism in vitro. PLoS ONE 9:e109536. https://doi.org/10.1371/journal.pone.0109536
Lin L, Cao Q, Zhang C et al (2022) Aflatoxin B1 causes oxidative stress and apoptosis in sheep testes associated with disrupting rumen microbiota. Ecotoxicol Environ Saf 232:113225. https://doi.org/10.1016/j.ecoenv.2022.113225
Liu Y, Wang W (2016) Aflatoxin B1 impairs mitochondrial functions, activates Ros generation, induces apoptosis and involves Nrf2 signal pathway in primary broiler hepatocytes. Anim Sci J 87:1490–1500. https://doi.org/10.1111/asj.12550
Loh ZH, Ouwerkerk D, Klieve AV et al (2020) Toxin degradation by rumen microorganisms: a review. Toxins 12(10):664. https://doi.org/10.3390/toxins12100664
Luna LG (1972) Manual of histologic staining methods of the Armed Forces Institute of Pathology, 3rd edn. McGraw-Hill Book Company, New York
Ma J, Liu Y, Guo Y et al (2021) Transcriptional profiling of aflatoxin B1-induced oxidative stress and inflammatory response in macrophages. Toxins 13:401. https://doi.org/10.3390/toxins13060401
Makhlouf MM (2020) Histological and ultrastructural study of AflatoxinB1 induced neurotoxicity in sciatic nerve of adult male albino rats. Ultrastruct Pathol 44:52–60
Malissiova E, Tsakalof A, Arvanitoyannis IS et al (2013) Monitoring aflatoxin M1 levels in ewe’s and goat’s milk in Thessaly, Greece; potential risk factors under organic and conventional production schemes. Food Control 34:241–248. https://doi.org/10.1016/j.foodcont.2013.04.035
Mehrzad J, Fazel F, Pouyamehr N, Hosseinkhani S, Dehghani H (2020) Naturally occurring level of aflatoxin B1 injures human, canine and bovine leukocytes through ATP depletion and caspase activation. Int J Toxicol 39:30–38. https://doi.org/10.1177/1091581819892613
Menami M, Yoshikawa H (1979) Simplifies assay method of superoxide dismutase activity of clinical use. Clin Chim Acta 92:337–342
Mohsenzadeh MS, Hedayati N, Riahi-Zanjani B, Karimi G (2016) Immunosuppression following dietary aflatoxin B1 exposure: a review of the existing evidence. Toxin Reviews 35:121–127. https://doi.org/10.1080/15569543.2016.1209523
Motawee MM, Bauer J, McMahon DJ (2009) Survey of aflatoxin M1 in cow, Goat, Buffalo and Camel Milks in Ismailia-Egypt. Bull Environ Contam Toxicol 83:766–769. https://doi.org/10.1007/s00128-009-9840-3
Okoye JOA, Asuzu IU, Gugnani HC (1988) Paralysis and lameness associated with aflatoxicosis in broilers. Avian Pathol 17:731–734. https://doi.org/10.1080/03079458808436490
Panneum S, Rukkwamsuk T (2017) Diagnosis of caprine arthritis encephalitis virus infection in dairy goats by Elisa, PCR and viral culture. Pol J Vet Sci 20:347–353. https://doi.org/10.1515/pjvs-2017-0042
Parmar H, Sharma P, Anerao I et al (2016) Aflatoxin B1 induced developmental neurotoxicity in RIR egg. Adv Appl Sci Res, 7, pp.23 – 7
Pauletto M, Giantin M, Tolosi R Curcumin mitigates AFB1-induced hepatic toxicity by triggering cattle antioxidant and anti-inflammatory pathways: A whole transcriptomic in vitro study. Antioxidants 9:1059.Plaser ZA, Cushman LL, Johnson BC et al (2020) (1966). Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Analytical Biochemistry 16: 359 – 364
Peles F, Sipos P, Győri Z et al (2019) Adverse effects, transformation and channeling of aflatoxins into food raw materials in livestock. Front Microbiol 10:2861
Petrova T, Orellana C, Jelic V et al (2020) Cholinergic dysfunction, neurodegeneration, and amyloid-beta pathology in neurodegenerative diseases. Psychiatry Research: Neuroimaging 302:111099. https://doi.org/10.1016/j.pscychresns.2020.111099
Qureshi H, Hamid SS, Ali SS et al (2015) Cytotoxic effects of aflatoxin B1 on human brain microvascular endothelial cells of the blood-brain barrier. Med Mycol 53:409–416. https://doi.org/10.1093/mmy/myv010
Raj HG, Kohli E, Rohil V et al (2001) Acetoxy-4-methylcoumarins confer differential protection from aflatoxin B1-induced micronuclei and apoptosis in lung and bone marrow cells. Mutat Research/Genetic Toxicol Environ Mutagen 494:31–40. https://doi.org/10.1016/s1383-5718(01)00176-0
Rathod P, Gangadhar K, Gangane G et al (2017) Effect of aflatoxin on haematological and biochemical alteration in broilers. Int J Environ Sci Technol 6:824–831
Reddy L, Odhav B, Bhoola K (2006) Aflatoxin B1-induced toxicity in hepg2 cells inhibited by carotenoids: morphology, apoptosis and DNA damage. Biol Chem 387:87–93. https://doi.org/10.1515/bc.2006.012
Rezaei SA, Dalir-Naghadeh B (2006) Evaluation of antioxidant status and oxidative stress in cattle naturally infected with Theileria annulata. Vet Parasitol 142:179–186
Richard SA, Manaphraim NYB, Kortei NK (2020) The novel neurotoxic and neuroimmunotoxic capabilities of aflatoxin B1 on the nervous system: a review. Adv Bioscience Clin Med 8:1–8. https://doi.org/10.7575/aiac.abcmed.v.8n.3p.1
Rushing BR, Selim MI (2019) Aflatoxin B1: a review on metabolism, toxicity, occurrence in food, occupational exposure, and detoxification methods. Food Chem Toxicol 124:81–100. https://doi.org/10.1016/j.fct.2018.11.047
Samarajeewa U, Arseculeratne SN, Tennekoon GE (1975) Spontaneous and experimental aflatoxicosis in goats. Res Vet Sci 19:269–277. https://doi.org/10.1016/s0034-5288(18)33501-x
Sawale GK, Ghosh RC (2008) Effect of dietary aflatoxin B1 on haemato-biochemical profile in laying hen. Indian J Anim Sci 78(3):283–284
Singh K, Saso K (2019) Oxidative stress: role and response of short guanine tracts at genomic locations. Int J Mol Sci 20:4258. https://doi.org/10.3390/ijms20174258
Tabatabaei M, Abdollahi F (2018) Isolation and identification of Mannheimia haemolytica by culture and polymerase chain reaction from Sheep’s pulmonary samples in Shiraz. Iran Veterinary World 11:636–641. https://doi.org/10.14202/vetworld.2018.636-641
Tao W, Li Z, Nabi F et al (2021) Penthorum chinense Pursh compound ameliorates AFB1-induced oxidative stress and apoptosis via modulation of mitochondrial pathways in broiler chicken kidneys. Front Veterinary Sci 8:750937. https://doi.org/10.3389/fvets.2021.750937
Tata AM, Velluto L, D’Angelo C et al (2014) Cholinergic system dysfunction and neurodegenerative diseases: cause or effect? CNS & neurological Disorders-DrugTargets. 13:1294–1303. https://doi.org/10.2174/1871527313666140917121132
Townsend KM, Frost AJ, Lee CW et al (1998) Development of PCR assays for species- and type-specific identification of Pasteurella multocida isolates. J Clin Microbiol 36:1096–1100. https://doi.org/10.1128/jcm.36.4.1096-1100.1998
Tulayakul P, Sakuda S, Dong KS, Kumagai S (2005) Comparative activities of glutathione-S-transferase and dialdehyde reductase toward aflatoxin B1 in livers of experimental and farm animals. Toxicon 46:204–209. https://doi.org/10.1016/j.toxicon.2005.03.023
Van Kuppeveld FJ, van der Logt JT, Angulo AF et al (1992) Genus and species-specific identification of mycoplasmas by 16S rRna amplification. Appl Environ Microbiol 58:2606–2615. https://doi.org/10.1128/aem.58.8.2606-2615.1992
Virdis S, Scarano C, Spanu V et al (2014) A survey on aflatoxin M1 content in sheep and goat milk produced in Sardinia Region, Italy (2005–2013). Italian J Food Saf 3:4517. https://doi.org/10.4081/ijfs.2014.4517
Wang WJ, Xu ZL, Yu C et al (2017) Effects of aflatoxin B1 on mitochondrial respiration, Ros generation and apoptosis in broiler cardiomyocytes. Anim Sci J 88:1561–1568. https://doi.org/10.1111/asj.12796
Wu LL, Chiou CC, Chang PY et al (2004) Urinary 8-ohdg: a marker of oxidative stress to DNA and a risk factor for cancer, atherosclerosis and diabetics. Clin Chim Acta 339:1–9. https://doi.org/10.1016/j.cccn.2003.09.010
Yang XJ, Lu HY, Li ZY et al (2012) Cytochrome P450 2013 mediates aflatoxin B1-induced cytotoxicity and apoptosis in human bronchial epithelial cells. Toxicology 300:138–148. https://doi.org/10.1016/j.tox.2012.06.010
Yang Q, Jiang Y, Fu S et al (2021) Protective effects of Ulva Lactuca polysaccharide extract on oxidative stress and kidney injury induced by D-galactose in mice. Mar Drugs 19:539. https://doi.org/10.3390/md19100539
Yassein SN, Zghair ZR (2012) Study of toxicity and pathogenicity of aflatoxin B1 and G1 in mice. Al-Anbar J Veterinary Sci 5:23
Yilmaz S, Kaya E, Kisacam MA (2017) The effect on oxidative stress of aflatoxin and protective effect of lycopene on aflatoxin damage. Aflatoxin-Control Anal Detect Health Risks 30:67–90. https://doi.org/10.5772/intechopen.69321
Zain ME (2011) Impact of mycotoxins on humans and animals. J Saudi Chem Soc 15:129–144. https://doi.org/10.1016/j.jscs.2010.06.006
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The authors are thankful to the Director, ICAR-Indian Veterinary Research Institute, Izatnagar, India for providing necessary facilities to carry out this work.
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Monalisa Sahoo and Rajendra Singh: contributed to the study conception and design; methodology: Jigarji Chaturji Thakor, Pradeep Kumar, Santawana Palai, Pratima Behera; Formal analysis and prepared the Figures: Sushmita Patra, Jagannath Prasad Tripathy, Ramakanta Acharya, Nihar Ranjan sahoo; Investigation carried out by Monalisa Sahoo, Pawan Kumar, Pradeep Kumar, Bhavani Puvala; resources were provided by Rajendra Singh, Karampal singh, Ajay Kumar; the first draft of the manuscript was written by Monalisa sahoo, and Rajendra Singh. All authors reviewed the submitted manuscript.
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Sahoo, M., Thakor, J.C., Kumar, P. et al. AFB1 induced free radicals cause encephalopathy in goat kids via intrinsic pathway of apoptosis: pathological and immunohistochemical confirmation of non-hepatic neuroaflatoxicosis. Vet Res Commun (2023). https://doi.org/10.1007/s11259-023-10216-9
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DOI: https://doi.org/10.1007/s11259-023-10216-9