Testing the Potential Clastogenic/Cytotoxic Effects of Pesticide CALYPSO 480 SC

The detection of chromosomal damage serves as a tool for the verification of the genotoxic effects of chemical substances in vitro. We used conventional cytogenetic analysis in order to test for the potential genotoxic action of the insecticide thiacloprid (the active ingredient in commercial preparation CALYPSO 480 SC). The test cultures of bovine lymphocytes obtained from the peripheral blood were incubated with the insecticide in concentrations of: 30, 120, 240 and 480 μg.ml−1 for 24 and 48 hours. After 24 hours of incubation, we observed that the increasing concentrations resulted in a significant (P < 0.05; P < 0.01) increase in the frequency of DNA damage. Our experiments showed the presence of aberrations of a non-stable type (chromatid and chromosome breakage). The conventional chromosome analysis was supplemented with fluorescence in situ hybridization for the detection of numeric and stable structural aberrations. Whole chromosome probes for bovine chromosomes 1, 5 and 7 (BTA 1, BTA 5 and BTA 7) were used in the experiments

Analysis of Sister Chromatid Exchanges and Proliferation of Human Peripheral Blood Lymphocytes Exposed to Epoxiconazole

The potential genotoxic/cytotoxic effect of epoxiconazole was evaluated by means of sister chromatid exchanges (SCE) following the 24 and 48 h in vitro exposure of human peripheral blood lymphocytes to epoxiconazole at concentrations of: 5, 10, 25, 50 and 100 μg. ml–1. Dimethyl sulphoxide (DMSO), used as an epoxiconazole solvent, was used as a negative control and mitomycine (MMC) as a positive control. After the 24-hour exposure, we failed to observe a significant increase in SCE frequencies in comparison with the negative control, however, the concentrations of 10—100 μg.ml–1 caused a significant decrease in the proliferation index (PI; P < 0.001). Also, the 48-hour exposure produced no significant alterations in the SCE frequencies in comparison with the control. At epoxiconazole concentrations ranging from 10 to 50 μg.ml–1 we recorded a moderate to strong, dose-dependent inhibition of PI (P < 0.05; P < 0.01; P < 0.001), while at the highest dose (100 μg.ml–1) the reduction in PI compared to the control was less pronounced (P < 0.05). The reduction in PI at the concentration range of 10—100 μg.ml–1 depended on the number of cells in the M1, M2 and M3 phases of the cell cycle per total number of 100 evaluated metaphases. Our results indicated a significant cytotoxic or cytostatic effect on human peripheral blood lymphocytes

Detection of Chromosomal Breaks Induced by Thiacloprid in Human Lypmhocytes and Detection of Double-Strand Breaks Based on γH2AX Histone Phosphorylation

Thiacloprid, a neonicotinoid insecticide, is widely used to control various species of pests in the current agriculture of today. The potential genotoxic effects of thiacloprid on human peripheral blood lymphocytes were investigated in vitro by chromosome aberrations (CA), and double-strand breaks (DSB), which were detected by the phosphorylation of γH2AX histone. Human peripheral blood lymphocytes were exposed to 30, 60, 120, 240, 480 µg.ml−1 doses for the last 24 and 48 hours of culture. Thiacloprid increased CA at the concentrations of 240, 480 μg.ml−1 (P < 0.05), but these results did not confirm genotoxicity. The mitotic index (MI) was important to us; it served as a basis for the confirmation of the cytotoxicity of this insecticide. During 48 hours of culture, at the concentration of 480 µg.ml−1, its value rapidly decreased (0.42) (P < 0.001), which did not allow us to analyse the results because of the high cytotoxic response

Assessment of Potential Clastogenic Effect of the Insecticide Mospilan® 20SP in Human Peripheral Blood Lymphocytes After in Vitro Exposure

Acetamiprid, that is known as the commercial formulation Mospilan® 20SP is the part of the neonicotinoid insecticide group and is widely used against various pests. In our study we assessed the potential clastogenic effects of Mospilan® in human peripheral blood lymphocytes in vitro using a chromosome aberration test. The lymphocytes were treated with acetamiprid in the concentration range of 5, 10, 25 and 50 µg.ml−1 for 24 and 48 h. After 24 h exposure, the insecticide induced statistically significant higher levels of chromosome aberrations from the concentration of 10 µg.ml−1 (P < 0.05 and P < 0.001) and a significant decrease in mitotic index (MI) at the concentrations of 25 and 50 µg.ml−1 (P < 0.05 and P < 0.01), respectively. After a 48 h exposure, we found a dose dependent increase in the percentage of chromosome aberrations at all concentrations (P < 0.05; P < 0.01 and P < 0.001) and a decrease in MI at concentrations of 25 and 50 µg.ml−1 (P < 0.05 and P < 0.01). Our results indicated that neonicotinoid insecticide formulations containing acetamiprid may have potential cytotoxic and genotoxic effects

Analysis of Chromosomal Damage Caused by Acetamiprid

Different chemicals can have genotoxic effects on the body, as confirmed by chromosome damage detection. Using conventional cytogenetic analysis and fluorescence in situ hybridization, we tested the extent of chromosome damage caused by the acetamiprid-based insecticide Mospilan 20SP on bovine peripheral blood lymphocytes at concentrations of, 2.5, 5, 25 and 50 µg.ml−1 after a 24 h incubation period. During the experiment, the presence of unstable aberrations—chromosomal and chromatid breaks and gaps—were detected by conventional cyto-genetic analysis. With increasing insecticide concentrations, we observed a statistically significant increase in chromosome damage frequency after 24 hours of exposure. Fluorescence in situ hybridization was used to detect stable structural aberrations; whole-chromosome painting probes for bovine chromosomes 1 and 7 (BTA 1 and BTA 7) were used for this purpose. As a result of exposure to the insecticide, neither BTA 1/BTA 7 translocations nor other types of translocations were observed

Micronucleus Assay in Environmental Biomonitoring

Nowadays many chemicals are widely used in agriculture to ensure high crop yields or in veterinary/human medicine to cure diseases. After their improper usage they may contaminate the environment, persist in it and adversely affect both the target and/or the non-target organisms. One of the ways to detect the occurrence of chemicals in the environment is to assess their impact on aquatic and farm animals; both are directly or indirectly exposed via their feed and water. The micronucleus assay is a standardly used cytogenetic test for the simultaneous detection of clastogenic and aneugenic agents. Additionally, cytotoxic effects are also assessed by analysing the proliferation changes using the cytokinesis-blocked proliferation index. The occurrence of micronuclei is analysed in many types of cells like the peripheral blood cells, bone marrow or cell lines according to standards for micronuclei detection. The analysis of published results has shown that the micronucleus assay is, together with the chromosomal aberration test, one of the most often used test in genotoxicity assessment. Its results have contributed to reassessing the use of multiple chemicals available on the market. Moreover, it is a compulsory test before approving the chemical/ pesticide for the market