Histochemical effects of brodifacoum on rat spleen

In this study, the histochemical effects of Brodifacoum, an anticoagulant used against rodents, on the spleen are examined under a light microscope using CD4 and CD8 histochemical staining methods. A single dose of 0.2 mg Brodifacoum was dissolved in Dimethyl Sulfoxide (DMSO) and was given orally to mature male rats. Spleen samples were collected under ether anesthesia after 24 h, 72 h, 14 days, and 30 days from the rats in the experimental groups and after 14 days from the rats in the control group. In this light microscope study, it was observed that the capsule, white pulp, and red pulp zones in the rat spleen were constructed normally and as their natural structures primary and secondary follicles (germinal center) they were few, and CD4 and CD8 lymphocytes were spherically structured. In the 24 h spleens of the rats, the diameters of germinal centers were expanded and deterioration of the structure of CD4 and CD8 cells was observed. Related to the increase in time (72 h and 14 days) it was determined that primary follicles increased in number and the diameters of germinal centers expanded. In addition to this, after30 days, the rate of CD4:CD8 of the brodifacoum applied rat spleens were approximately the rate of the control group, and the improvement of the structures of the cells was reported as an effect of regeneration. As a result of this study, it was found that Brodifacoum caused immunohistochemical abnormalities in the rat spleen, affected the morphological structure of CD4 and CD8 T lymphocytes and created an immune response in rats. It is thought that the obtained results will be a source for the studies on Brodifacoum.

Keywords:

brodifacoum, Rattus norvegicus, spleen, histochemistry, anticoagulant, CD4 T lymphocyte CD8 T lymphocyte, light microscope,

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Walther, B., Geduhn, A., Schenke, D. & Jacob, J. Exposure of passerine birds to brodifacoum during management of Norway rats on farms. Science of the Total Environment, 762, (2021), 144160. https://doi.org/10.1016/j.scitotenv.2020.144160
Albert, C.A., Wilson, L.K., Mineau, P., Trudeau, S. & Elliott, J.E. Anticoagulant rodenticides in three owl species from western Canada, 1988-2003. Archives of Environ. Cont. and Toxicology, 58, (2010), 451–459. https://doi.org/10.1007/s00244-009-9402-z
Chu, Y.-J., Lin, J.-H. & Hung, D.-Z. Oral administration of injectable vitamin K1 in Brodifacoum intoxication. BioMedicine, 12, (2022), 47–49. https://doi.org/10.37796/2211-8039.1305
Mosterd, J.J. & Thijssen, H.H.W. The long-term effects of the rodenticide, brodifacoum, on blood coagulation and vitamin K metabolism in rats. British Journal of Pharmacol, 535, (1991), 531–535. https://doi.org/10.37796/221-039.1305
Elliott, J.E., Hindmarch, S., Albert, C.A., Emery, J., Mineau, P. & Maisonneuve, F. Exposure pathways of anticoagulant rodenticides to nontarget wildlife. Environ. Monit. Assess., 186, (2014). https://doi.org/10.1007/s10661-013-3422-x
Nosal, D.G., van Breemen, R.B., Haffner, J.W., Rubinstein, I. & Feinstein, D.L. Brodifacoum pharmacokinetics in acute human poisoning: implications for estimating duration of vitamin K therapy. Toxicol. Commun., 5, (2021), 69–72. https://doi.org/10.1080/24734306.2021.1887637
Breckenridge, A.M., Cholerton, S., Hart, J.A., Park, B.K. & Scott, A.K. A study of the relationship between the pharmacokinetics and the pharmacodynamics of the 4-hydroxycoumarin anticoagulants warfarin, difenacoum and brodifacoum in the rabbit. British Journal of Pharmacology, 84, (1985), 81–91.
Eason, C.T., Murphy, E.C., Wright, G.R.G. & Spurr, E.B. Assessment of risks of brodifacoum to non-target birds and mammals in New Zealand. Ecotoxicology, 11, (2002), 35–48. https://doi.org/10.1023/a:1013793029831
Tosh D.G., McDonald R.A., Bearhop S., Lllewellyn N.R., Fee S., Sharp E.A., Barnett E.A., Shore R.F. Does small mammal prey guild affect the exposure of predators to anticoagulant rodenticides?. Environmental Pollution, 159, (2011), 3106–3112. https://doi.org/10.1016/j.envpol.2011.03.028
Stansley, W., Cummings, M., Vudathala, D. & Murphy, L.A. Anticoagulant rodenticides in red-tailed hawks, Buteo jamaicensis, and great horned owls, Bubo virginianus, from New Jersey, USA, 2008–2010. Bulletin of Environmental Contamination and Toxicology Toxicol., 92, (2014), 6–9. https://doi.org/10.1007/s00128-013-1135-z
Mebius, R.E. & Kraal, G. Structure and function of the spleen. Nature Reviews Immunology, 5, (2005), 606–616. https://doi.org/10.1038/nri1669
Cesta, M.F. Normal structure, function, and histology of the spleen. Toxicologic Pathology, 34, (2006), 455–465. https://doi.org/10.1080/01926230600867743
Fabrizio, V.A., Rodriguez-Sanchez, M.I., Mauguen, A., Dahi, P.B., Doubrovina, E., O’Reilly, R.J. & Prockop, S.E. Adoptive therapy with CMV-specific cytotoxic T lymphocytes depends on baseline CD4+ immunity to mediate durable responses. Blood Advances 5.2 (2021), 496-503. https://doi.org/10.1182/bloodadvances.2020002735
Ademokun, A.A. & Dunn-Walters, D. Immune Responses: Primary and Secondary. eLS, (2010). https://doi.org/10.1002/9780470015902.a0000947.pub2
Langeveld, M., Gamadia, L.E. & Ten Berge, I.J.M. T-lymphocyte subset distribution in human spleen. European Journal of Clinical Investigation, 36, (2006), 250–256. https://doi.org/10.1111/j.1365-362.2006.01626.x
Falchetti, R., Lanzilli, G., Casalinuovo, I.A., Gaziano, R., Palamara, A.T., Di Francesco, P., Ravagnan, G. & Garaci, E. Splenic CD4+and CD8+T cells from influenza immune mice concurrently produce in vitro IL2, IL4, and IFN-γ. Cellular Immunology, 170, (1996), 222–229. https://doi.org/10.1006/cimm.1996.0155
Hadler, M.R. & Buckle, A.P. Forty five years of anticoagulant rodenticides—past, present and future trends. Proceedings of the Fifteenth Vertebrate Pest Conference (1992), 149–155.
Whiteland, J.L., Nicholls, S.M., Shimeld, C., Easty, D.L., Williams, N.A. & Hill, T.J. Immunohistochemical detection of T-cell subsets and other leukocytes in paraffin-embedded rat and mouse tissues with monoclonal antibodies. Journal of Histochemistry and Cytochemistry, 43, (1995), 313–320. https://doi.org/10.1177/43.3.7868861
Murphy, K M., Reiner, S.L. The lineage decisions of helper T-cells. Nature Reviews Immunology, 2, (2002), 933–944. https://doi.org/10.1038/nri954
Chen, W., Jin, W., Hardegen, N., Lei, K.J., Li, L., Marinos, N., McGrady, G. & Wahl, S.M. Conversion of peripheral CD4+CD25− naive T-cells to CD4+CD25+ regulatory T-cells by TGF-β induction of transcription factor foxp3. Journal of Experimental Medicine, 198, (2004), 1875–1886. https://doi.org/10.1084/jem.20030152
Jin, E., Li, S., Ren, M., Hu, Q., Gu, Y., & Li, K. Boron affects immune function through modulation of splenic T-lymphocyte subsets, cytokine secretion, and lymphocyte proliferation and apoptosis in rats. Biological Trace Element Research, 178, (2017), 261–275. https://doi.org/10.1007/s12011-017-0932-3
Omar, H.E.-D.M., Saad Eldien, H.M., Badary, M.S., Al-Khatib, B.Y. & AbdElgaffar, S.K. The immunomodulating and antioxidant activity of fucoidan on the splenic tissue of rats treated with cyclosporine A. The Journal of Basic and Applied Zoology, 66, (2013), 243–254. https://doi.org/10.1016/j.jobaz.2013.05.003
Saad, A.J. & Jerrells, T.R. Flow cytometric and immunohistochemical evaluation of ethanol‐induced changes in splenic and thymic lymphoid cell populations. Alcoholism: Clinical and Exper. Research, 15, (1991), 796–803. https://doi.org/10.1111/j.1530-0277.1991.tb00603.x
Neishabouri, E., Hassan, Z.M. & Ostad, S. Humoral and cellular immunomodulation induced by propoxure in C57-bl/6 mice. Iranian Journal of Pharmaceutical Research, 3, (2004), 41–45. https://doi.org/10.22037/ijpr.2010.295
Falchetti, R., Lanzilli, G., Casalinuovo, I.A., Gaziano, R., Palamara, A.T., Di Francesco, P., Ravagnan, G. & Garaci, E. Splenic CD+ and CD8+T cells from influenza immune mice concurrently produce in vitro IL2, IL4, and IFN-. Cellular Immunology, 170, (1996), 222–229. https://doi.org/10.1006/cimm.1996.0155
Fujitani, T., Tada, Y. & Yoneyama, M. Chlorpropham-induced splenotoxicity and its recovery in rats. Food and Chem. Toxicology, 42, (2004), 1469–1477. https://doi.org/10.1016/j.fct.2004.04.008
Valchev, I., Binev, R., Yordanova, V. & Nikolov, Y. Anticoagulant rodenticide intoxication in animals - A review. Turkish Journal of Veterinary & Animal Sciences, 32, (2008), 237–243.
Maamar, H., Mallem, L. & Boulakoud, M.S. The effect of the anticoagulant rodenticide “Brodifacoum” on the bioindicators parameters in male rabbit. Annals of Biological Research, (2013), 53-61.