Evaluation of the cytotoxic and genotoxic effects of cyprodinil in vitro

Introduction. Cyprodinil is a broad-spectrum systemic fungicide of the anilinopyrimidine class. Available literature data on the genotoxicity of this pesticide are contradictory.

The aim of the work is to study the cytotoxic and genotoxic effects of ciprodinil.

Material and methods. The mutation activity of cyprodinil was assessed in the Ames test. The level of DNA fragmentation in peripheral blood lymphocytes was determined by conducting alkaline gel electrophoresis of individual cells (DNA comet assay). Cell viability was assessed using a TC20 cell counter (BioRad). The ADAMII LS automated fluorescent cell analyzer (Nano Entek) was used to determine the pathways of cell death.

Results. Cyprodinil has a cytotoxic effect on S. typhimurium bacterial cells starting from a concentration of 1.6 mg/ cup under metabolic activation conditions and at concentrations of 0.5 mg/cup and higher in the absence of the S9 mixture. No mutagenic effect was detected in the Ames test. The level of DNA breaks in peripheral blood lymphocytes (%, Tail Intensity) increased compared to the control after their cultivation in a medium with cyprodinil at a concentration of 100 µg/ml in the absence of metabolic activation. In this case, 70 to 95% of the cells died by apoptosis and necrosis.

Limitations. The study is limited to in vitro cytotoxicity and genotoxicity assessment of cyprodinil only.

Conclusion. Cyprodinil does not have a genotoxic effect. The observed increase in the level of DNA breaks in peripheral blood lymphocytes in vitro in the presence of cyprodinil is a consequence of the general toxic effect, which can lead to the destruction of cellular structures, the release of enzymes, including endonucleases, causing DNA breaks.

Compliance with ethical standards. The study was approved by the Ethics Committee of the Federal Scientific Centre of Hygiene named after F.F. Erisman, Rospotrebnadzor (Protocol No. 1, September 29, 2020). All participants gave informed voluntary written consent to participate in the study.

Authors contribution:
Gorenskaya O.V. – collecting and processing of data; statistical analysis; analysis of the results; writing the text;
Egorova O.V. – concept and design of the study; collecting and processing of data; analysis of the results; writing the text;
Averyanova N.S., Kotnova A.P. – the collection of data;
Ilyushina N.A. – concept and design of the study; collecting and processing of data; analysis of the results; writing the text; editing.
All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version.

Acknowledgements. The authors express their gratitude to Bioline LLC and personally to Eduard Mingazov for the opportunity to conduct the study on the assessment of cytotoxic effects using the automatic fluorescent cell analyzer ADAMII LS, Nano Entek, obtained under a free lease agreement.

Conflict of interest. The authors declare no conflict of interest.

Funding. The study had no sponsorship.

Accepted: October 9, 2024 / Revised: November 8, 2024 / Received: November 12, 2024 / Published: December 28, 2024