Abstract
The aim of this study was to evaluate the genotoxic and cytotoxic effects of amitraz (AMZ) on the primary culture of bovine cumulus cells (CC) and oocyte nuclear maturation. Cytotoxicity was evaluated by assessing mitochondrial activity with the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Genotoxicity was estimated using the alkaline single cell gel electrophoresis (SCGE) assay. Apoptosis was detected with the Annexin V-affinity assay. The in vitro maturation test was performed in bovine oocytes. To understand AMZ action, glutathione content, superoxide dismutase enzyme activity, and lipid peroxidation were evaluated in CC. Results showed that AMZ lethal concentration (LC 5024h) for bovine CC was 32.55 μg/mL (MTT assay). A 25 μg/mL induced late apoptosis and necrotic cells (p < 0.05); however, DNA damage was decreased at the same concentration (SCGE assay; p < 0.05). A decrease in metaphase II was observed at 25 μg/mL, and degenerate oocytes were observed at 15 and 25 μg/mL (p < 0.05). None of the oxidative stress parameters evaluated showed significant differences. This study contributes to a better understanding of AMZ in this model, suggesting its potential cytotoxicity and impact on bovine reproduction.
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The datasets used during the current study are available from the corresponding author on reasonable request.
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Acknowledgments
We are grateful to the staff of Frigorífico Gorina S.A. for providing the bovine ovaries and Dr. Sebastian J. Picco for making Amitraz (Biogénesis Bagó, Argentina) available. Thanks are also due to A. Di Maggio for manuscript correction and edition.
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This work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica de la República Argentina (PICT BID 2016-3727) Ministerio de Ciencia, Tecnología e Innovación Productiva de la Nación Argentina.
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NN created the methodology, conducted the investigation, made the formal analysis, and wrote the original draft. ACCM created the methodology, conducted the investigation, made the formal analysis, and wrote the original draft. MF participated in the investigation and made the data curation. CCF provided the research resources, reviewed, and edited the manuscript.
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•The viability curve of CC treated with AMZ for 24 h has an abrupt drop indicating that there is no linear response to the dose.
•AMZ damage in CC seems to be irreparable due to increase late apoptosis and necrosis
• AMZ-induced CC damage appears to be unrelated to reactive oxygen species
• AMZ increases oocyte damage and delays its maturation, which could cause reproductive alterations
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Nikoloff, N., Carranza Martin, A.C., Fabra, M.C. et al. Amitraz induced cytotoxic effect on bovine cumulus cells and impaired oocyte maturation. Environ Sci Pollut Res 28, 29188–29199 (2021). https://doi.org/10.1007/s11356-021-12670-x
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DOI: https://doi.org/10.1007/s11356-021-12670-x