Cytotoxic and genotoxic effects induced by enrofloxacin-based antibiotic formulation Floxagen® in two experimental models of bovine cells in vitro: peripheral lymphocytes and cumulus cells
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The in vitro effect of enrofloxacin (EFZ) was tested on two experimental somatic bovine cells in vitro: peripheral lymphocytes (PLs) and cumulus cells (CCs). The cytotoxicity and genotoxicity of this veterinary antibiotic were assessed using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays, single-cell gel electrophoresis (SCGE) assay, and cytokinesis-block micronucleus cytome (CBMN cyt) assay. Cells were treated during 24 h, and three concentrations were tested (50 μg/mL, 100 μg/mL, 150 μg/mL). When EFZ was tested in PLs, the results demonstrated that the antibiotic was able to induce cell death and DNA damage with all concentrations. In addition, 50 μg/mL and 100 μg/mL EFZ increased frequencies of micronuclei (MNi). On the other hand, the highest EFZ concentration occasioned cellular cytotoxicity in CCs as evidenced by mitochondrial activity alterations. Nevertheless, EFZ was not able to induce DNA damage and MNi in CCs. These results represent the first experimental evidence of genotoxic and cytotoxic effects exerted by EFZ in bovine PLs and CCs.
KeywordsBovine cells MTT SCGE assay CBMN cyt assay Enrofloxacin
We are grateful to the staff of SENASA from Frigorífico Gorina S.A. for providing the bovine ovaries.
This work was supported by grants from the Agencia Nacional de Promoción Científica y Tecnológica de la República Argentina (PICT BID 1972-2013); Ministerio de Ciencia, Tecnología e Innovación Productiva de la Nación Argentina; Consejo Nacional de Investigaciones Científicas y Tecnológicas (PIP 112-20130100657); and Universidad Nacional de La Plata (V246 and V249).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
- Altreuther P (1987) Data on chemistry and toxicology of Baytril. Vet Med Rev 2:87–89Google Scholar
- Babaei H, Roshangar L, Sakhaee E, Abshenas J, Kheirandish R, Dehghani R (2012) Ultrastructural and morphometrical changes of mice ovaries following experimentally induced copper poisoning. Iran Red Crescent Med J 14(9):558–568Google Scholar
- Bautista Garfias CR, Acosta García E, Toledo GI (2000) Evaluación del bioensayo de MTT para determinar la proliferación in vitro de linfocitos de bovino, frescos y congelados Vet. Méx 31(2)Google Scholar
- Botelho RG, Monteiro SH, Tornisielo VL (2015) Veterinary antibiotics in the environment. Chapter 5. IntechGoogle Scholar
- Krisher RL (2004) The effect of oocyte quality on development. J Anim Sci 82:14–23Google Scholar
- Madhuresh KS, Rusha G, Rohit S, Sanjay M, Purbita C, Anil T (2016) Genotoxic impurities evaluation in active pharmaceutical ingredients (API)/drug substance. Pharm Lett 8(12):234–243Google Scholar
- Otero JL, Mestorino N, Errecalde JO (2001) Enrofloxacina una fluoroquinolona de uso exclusive en veterinaria pate II: farmacocinética y toxicidad. Analecta Veterinaria 21(1):42–49Google Scholar
- Prescott JF, Baggot JD, Walker RD (2000) Antimicrobial therapy in veterinary medicine. Iowa State University Press, AmesGoogle Scholar
- Tice R, Strauss GH (1995) The single cell gel electrophoresis/comet assay: a potential tool for detecting radiation-induced DNA damage in humans. Stem Cells 13(1):207–214Google Scholar