Doramectin induced cytotoxic and genotoxic effects on bovine peripheral lymphocytes and cumulus cells in vitro

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

Acknowledgments

Authors are grateful to the staff of SENASA from Frigorífico Gorina S.A. for providing the bovine ovaries. Juan Mateo Anchordoquy, Juan Patricio Anchordoquy and Noelia Nikoloff contributed equally to this work.

Disclosure statement

The authors declare that there are no conflicts of interest.

Additional information

Funding

This work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica de la República Argentina (PICT BID 1972-2013 and PICT BID 2015-2154), Ministerio de Ciencia, Tecnología e Innovación Productiva de la Nación Argentina; Consejo Nacional de Investigaciones Científicas y Tecnológicas (PIP 112-20130100657); and the Universidad Nacional de La Plata (V246 and V249).