Using sets of behavioral biomarkers to assess short-term effects of pesticide: a study case with endosulfan on frog tadpoles

Mathieu Denoël, Bastien D’Hooghe, G. Francesco Ficetola, Catherine Brasseur, Edwin De Pauw, Jean-Pierre Thomé, Patrick Kestemont

Pesticides and other chemicals often have detrimental effects at environmental concentrations. Many amphibian species are particularly threatened because of their susceptibility but also because wetlands are often polluted. Behavioral assessments of toxicity have the advantage of showing sublethal effects but quantitative measures at varied scales of integrations are rarely considered together. In this study, we aimed at showing that these behavioral endpoints could be differently affected across time and concentrations, and be biomarkers of toxicity. To this end, we tested the effects of an organochlorine pesticide (endosulfan) on amphibians during a standard 96 h test. We evaluated possible lag effects in continuing the analyses after removal of the pesticide. The study was based on 240 tadpoles (4 pesticide treatments: 0.4, 3, 22, and 282 μg/l, 1 control and 1 solvent-control). Abnormal behaviors such as lying and swirling rapidly were exhibited only in the presence of the pesticide. Essential functions such as breathing and feeding were deeply affected by the pesticide: contaminated tadpoles breathed and fed less than control tadpoles. They also moved less and occupied a more central position in the aquariums in the presence of the pesticide. A higher mortality was only found at the highest concentration. These results suggest that endosulfan is toxic to amphibians at environmental concentrations. Behavioral markers showed potential as early warning systems. They should thus be used in complement to other markers to detect sublethal effects only a few days after application of the pesticide and at concentrations where mortality does not occur.