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Doramectin induced cytotoxic and genotoxic effects on bovine peripheral lymphocytes and cumulus cells in vitro
Authors:Juan Mateo Anchordoquy  Juan Patricio Anchordoquy  Noelia Nikoloff  Rocío Gambaro  Gisel Padula  Analía Seoane
Affiliation:1. Instituto de Genética Veterinaria “Ing. Fernando N Dulout” (UNLP-CONICET-CONICET LA PLATA), Facultad de Ciencias Veterinarias – UNLP, IGEVET, La Plata, Buenos Aires, Argentina;2. Facultad de Ciencias Naturales y Museo – UNLP, Plata, Buenos Aires, La Argentina
Abstract:The effect of doramectin (DOR) was tested on two experimental somatic bovine cells in vitro: peripheral lymphocytes (PL) and cumulus cells (CC). The cytotoxicity and genotoxicity of DOR were assessed using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, single cell gel electrophoresis assay (SCGE) and cytokinesis-block micronucleus cytome (CBMN Cyt) assay. Both cells were treated with three concentrations of DOR (20, 40, 60?ng?mL–1) for 24?h. The results obtained from PL demonstrated that DOR was able to induce cytotoxic effect and DNA damage with all concentrations tested. Additionally, DOR increased micronuclei (MNi) frequency and nuclear buds (NBuds) with 20, 40, 60?ng?mL–1, and nucleoplasmic bridges (NPBs) only with 40?ng?mL–1. On the other hand, the three concentrations of DOR were not able to induce cytotoxic effect and DNA damage using SCGE in the bovine CC. Nevertheless, the two higher concentrations of DOR (20, 40?µg mL–1) significantly increased the frequency of micronucleus formation in bovine CC. These results represent the first experimental evidence of genotoxic and cytotoxic effects exerted by DOR on bovine PL and CC.
Keywords:Bovine peripheral lymphocytes  bovine cumulus cells  doramectin  MTT  comet assay  CBMN Cyt assays
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