Integrated ecotoxicological assessment of the complex interactions between chlorpyrifos and glyphosate on a non-target species Cnesterodon decemmaculatus (Jenyns, 1842)

Abstract Pesticide mixtures are frequent in freshwaters systems around the world, threatening the biota exposed to these conditions. The aim of this study was to determine the single and joint effect of two widely used pesticides in southern South America on a widely distributed fish species. In a 96-hour assay, individuals of Cnesterodon decemmaculatus were exposed to 0.84 nL/L and 8.4 nL/L of Clorfox and 0.2 mg/L and 2 mg/L of Roundup Max, commercial formulations of chlorpyrifos and glyphosate, respectively. Also, there were four mixture treatments with all the possible combinations of both pesticides. A multi-level approach was carried out to assess their effects covering the following relevant biomarkers: behavior (immobile time, line crossings and average speed), somatic conditions (Fulton condition factor and hepatosomatic index), serum parameters (cortisol levels, lactate dehydrogenase (LDH), and creatine phosphokinase activity (CPK)), brain and muscle acetylcholinesterase and cytological characteristics (micronuclei frequency and nuclear abnormalities in erythrocytes). Our results showed that Clorfox exposures affect behavioral parameters, serum cortisol, and nuclear characteristics of erythrocytes. Roundup Max affects only the cortisol levels whereas mixture treatments have an effect on behavioral parameters, cortisol levels, LDH and CPK activities, and nuclear characteristics of erythrocytes. Potentiation was the main interaction at the lowest concentrations of both pesticides whereas antagonism occurred at the highest concentrations of both pesticides. These results are highly significant since they arise from an integrated ecotoxicological assessment at several levels of biological organization but even more important is that the potentiated effects of the mixtures we registered are environmentally relevant concentrations.