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Updated: Jun 9, 2026
An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
Published on: May 13, 2019
Micronucleus frequency in sheep lymphocytes after in vitro exposure to fungicide tolylfluanid
Irena Sutiaková1, Natália Kovalkovicová, Jaroslav Legáth
1Department of Biology, University of Presov, Presov, Slovakia. sutiakova@gmail.com
Abstract:
The fungicide tolylfluanid (N -dichlorofluoromethylthio-N', N -dimethyl -N -p -tolylsulfamide), was investigated by cytokinesis-block micronucleus assay. Tolylfluanid at the lowest concentration (1 x 10(- 6)mol L(- 1))did not influence significantly the frequency of micronuclei in sheep lymphocyte cultures in comparison with control (32.33 +/- 3.51/1000 binucleated cells versus 30.33 +/- 2.82/1000 binucleated cells in dimethylsulfoxide (DMSO) control, P = 0.44). Higher tolylfluanid concentrations (1 x 10(- 4) and, 1 x 10(- 5) mol L(- 1)) resulted in a significant dose-dependent increase in the number of micronuclei in comparison with control (74.00 +/- 13.00/1000 binucleated cells and 52.67 +/- 10.12/1000 binucleated cells versus 30.33 +/- 2. 82/1000 binucleated cells in DMSO control, P = 0.005 and 0.02, respectively, ANOVA followed by Tukey test P < 0.05). Many of the treated cells also possessed multiple micronuclei. Tolylfluanid did not affect the nuclear division index at all treatment concentrations. Our in vitro results thus demonstrate that tolylfluanid had a significant genotoxic effect at only the highest concentration tested.
Insights
The fungicide tolylfluanid showed genotoxic effects in sheep lymphocytes, significantly increasing micronuclei at higher concentrations. Lower concentrations did not impact cell cultures, indicating a dose-dependent genotoxic response.
Area of Science:
- Environmental toxicology
- In vitro toxicology
- Genotoxicity testing
Background:
- Fungicides are widely used in agriculture, necessitating toxicological assessments.
- Tolylfluanid is a common fungicide with potential environmental and health impacts.
- Understanding the genotoxic potential of pesticides is crucial for risk assessment.
Purpose of the Study:
- To evaluate the in vitro genotoxicity of the fungicide tolylfluanid.
- To determine the effects of tolylfluanid on micronucleus formation in sheep lymphocytes.
- To assess the dose-dependent genotoxic potential of tolylfluanid.
Main Methods:
- Cytokinesis-block micronucleus assay was employed.
- Sheep lymphocyte cultures were treated with varying concentrations of tolylfluanid.
- Micronuclei frequency and nuclear division index were analyzed.
Main Results:
- Tolylfluanid did not significantly increase micronuclei at the lowest concentration (1 x 10(-6) mol L(-1)).
- Higher concentrations (1 x 10(-4) and 1 x 10(-5) mol L(-1)) caused a significant, dose-dependent increase in micronuclei.
- Multiple micronuclei were observed in treated cells, but the nuclear division index remained unaffected.
Conclusions:
- Tolylfluanid exhibits significant genotoxic effects in vitro at higher concentrations.
- The fungicide's genotoxicity is dose-dependent, with minimal impact at low levels.
- Further investigation into the in vivo effects and mechanisms of tolylfluanid genotoxicity is warranted.
