Ecotoxicological assessment of the effects of glyphosate and chlorpyrifos in an Argentine soya field

Background, Aim and Scope Continuous application of pesticides may pollute soils and affect non-target organisms. Soil is a complex ecosystem; its components can modulate the effects of pesticides. Therefore, it is recommended to evaluate the potential environmental risk of these compounds in local conditions. We performed an integrated field-laboratory study on an Argentine soya field sprayed with glyphosate and chlorpyrifos under controlled conditions. Our aim was to compare the sensitivity of a series of endpoints for the assessment of adverse effects of the extensive use of these agrochemicals. Materials and Methods A RR soya field in a traditional farming area of Argentina was sprayed with glyphosate (GLY) or chlorpyrifos (CPF) formulations at the commercially recommended rates, according to a randomized complete block design with 3 replicates. In laboratory assays, Eisenia fetida andrei were exposed to soil samples (0-10 cm depth) collected between the rows of soya. Endpoints linked to behavior and biological activity (reproduction, avoidance behavior and bait-lamina tests) and cellular/subcellular assays (Neutral Red Retention Time -- NRRT; DNA damage -- Comet assay) were tested. Field assays included litterbag and bait-lamina tests. Physico/chemical analyses were performed on soil samples. Results GLY reduced cocoon viability, decreasing the number of juveniles. Moreover, earthworms avoided soils treated with GLY. No effects on either reproduction or on avoidance were observed at the very low CPF concentration measured in the soils sampled 10 days after treatment. Both pesticides caused a reduction in the feeding activity under laboratory and field conditions. NRRT was responsive to formulations of CPF and GLY. Comet assay showed significantly increased DNA damage in earthworms exposed to CPF treated soils. No significant differences in DNA migration were observed with GLY treated soils. Litterbag field assay showed no differences between treated and control plots. Discussion The ecotoxicological effects of pesticides can be assessed by monitoring the status of communities in real ecosystems or through the use of laboratory toxicity tests. Litterbag field test showed no influence of the treatments on the organic matter breakdown, suggesting a scarce contribution of soil macrofauna. The bait-lamina test, however, seemed to be useful for detecting the effects of GLY and CPF treatments on the activity of the soil fauna. CPF failed to give significant differences with the controls in the reproduction test and the results were not conclusive in the avoidance test. Although the field population density of earthworms could be affected by multiple factors, the effects observed on the reproduction and avoidance tests caused by GLY could contribute to its decrease, with the subsequent loss of their beneficial functions. Biomarkers measuring effects on suborganism level could be useful to predict adverse effects on soil organisms and populations. Among them, NRRT, a lysosomal destabilization biomarker, resulted in demonstrating more sensitivity than the reproduction and avoidance tests. The Comet assay was responsive only to CPF. Since DNA damage can have severe consequences on populations, it could be regarded as an important indicator to be used in the assessment of soil health. Conclusions Reproduction and avoidance tests were sensitive indicators of GLY exposure, with the former being more labor intensive. Bait-lamina test was sensitive to both CPF and GLY. NRRT and Comet assays revealed alterations at a subcellular level, and could be considered complementary to the biological activity tests. Because of their simplicity, some of these bioassays seemed to be appropriate pre-screening tests, prior to more extensive and invasive testing. Recommendations and Perspectives This study showed deleterious effects of GLY and CPF formulations when applied at the nominal concentrations recommended for soya crops. Further validation is needed before these endpoints could be used as field monitoring tools in Argentine soya soils (ecotoxicological risk assessment -- ERA tools).