Assessment of genome damage in a population of Croatian workers employed in pesticide production by chromosomal aberration analysis, micronucleus assay and Comet assay

The widespread use of pesticides suggests that the evaluation of their genotoxicity should be extended using the different assays available. In the present study we used two standard cytogenetic methods (chromosomal aberration analysis and micronucleus assay) and the Comet assay as a relatively new and powerful technique. The study included 10 workers occupationally exposed to a complex mixture of pesticides (atrazine, alachlor, cyanazine, 2,4-dichlorophenoxyacetic acid, malathion) during their production and 20 control subjects with no history of exposure to any physical or chemical agents. For the chromosomal aberration analysis, whole blood was cultivated for 48 h, whereas for the micronucleus assay, whole blood was cultivated for 72 h. For the comet assay whole blood was embedded in agarose on a microscope slide, lysed with detergent, denaturated and subjected to alkaline electrophoresis. Damage to DNA was evaluated by measuring tail length and calculating the tail moment. A significantly increased number of chromatid and chromosome breaks, as well as the presence of dicentric chromosomes and chromatid exchanges in exposed subjects compared with control subjects (P < 0.05), was found. There was also a statistically significant difference in frequency and distribution of micronuclei between the two groups examined. In the exposed subjects the Comet assay showed a statistically significant (P < 0.001) increase in DNA migration. Results suggest that long-term occupational exposure to pesticides could cause genome damage in somatic cells and therefore may represent a potential hazard to human health. Copyright (C) 2002 John Wiley Sons, Ltd.