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Chlorpyrifos [O,O-diethyl-O-(3,5,6-trichloro-2-pyridyl)phosphorothioate] is a widely used non-systemic organophosphorus insecticide frequently detected in surface waters around the world. The goal of this study is to evaluate the acute and chronic effects of this insecticide on Rana dalmatina tadpoles. To assess the sensitivity of this species, the LC50 value (i.e. the concentration at which 50% of tadpoles die) was determined after 96 h. Our results showed that 5.174 mg L(-1) chlorpyrifos caused 50% mortality in tadpoles at Gosner stage 25. Chronic toxicity tests were also conducted to evaluate the sublethal effects of chlorpyrifos; tadpoles were exposed to three ecologically relevant concentrations (0.025, 0.05 and 0.1 mg L(-1)) in static renewal tests from Gosner stage 25 (tadpoles shortly after hatching) until completed metamorphosis (Gosner stage 46). No significant reduction was observed in survival, larval growth (mass), snout-vent length, stage development or number metamorphosed. In contrast, chlorpyrifos exhibited significant chronic toxic effects on larval development, manifested as the appearance of abnormalities, including tail flexure, skeletal and muscle defects in later stages of development in tadpoles exposed to all tested concentrations. We also evaluated the chronic effects of chlorpyrifos on gill morphology and ultrastructure. Tadpoles were sacrificed after 8 days and 30 days of exposure. Observations by both scanning (SEM) and transmission electron microscopy (TEM) showed considerable morphological and ultrastructural changes. The main gill effects recorded were mucous secretion, epithelium detachment and a degeneration phenomenon. Comparing these results with our previous findings, we demonstrate that the first effect of chlorpyrifos on R. dalmatina is gill alteration, thus supporting the role of a morphological approach in toxicological studies.

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