Flow cytogenetic analysis of whole cell clastogenicity of herbicides found in groundwater

Agronomic practices followed in recent years have caused the agrarian sector to depend heavily on agri-chemicals such as herbicides, but herbicides have negative environmental consequences. Their usage has resulted in chemicals being introduced into the groundwater. Although the contaminants are in trace amounts, little is known about their potential clastogenic effects at low concentrations. In the present study, the potential of flow cytometric analysis to detect the whole cell clastogenic properties of low level chemical exposure is examined. Chinese Hamster Ovary (CHO) cells were exposed to three herbicides (atrazine, bentazon and simazine) and two known clastogens (adriamycin and ara-C) at low concentrations for 48 h. Nuclei were isolated from control as well as treatments and analyzed by flow cytometry. The clastogenic property was assessed by measuring the coefficient of variation (CV) of G1 peaks in different treatments. Exposure to known clastogens resulted in increasing CVs with increasing concentrations. Flow cytometry was a very accurate and sensitive technique for the whole cell clastogenic assay. The results indicated that atrazine has the potential to cause whole cell clastogenicity, even at a very low concentration. Bentazon showed indications of clastogenic potential but the increases in CVs observed with this herbicide were not statistically significant. The CVs are unaffected by simazine exposure, thus indicating that simazine does not cause whole cell clastogenesis under short term exposure. Flow cytometry proved to be an excellent tool for assaying the clastogenic potential of agri-chemicals.