Humane acute testing with tadpoles for risk assessment of chemicals: Avoidance instead of lethality.

In spite of the sensitivity of amphibians to contamination, data from fish have been commonly used to predict the effects of chemicals on aquatic life stages. However, recent studies have highlighted that toxicity data derived from fish species may not protect all the aquatic life stages of amphibians. For pesticide toxicity assessment (PTA), EFSA has highlighted that more information on lethal toxicity for the aquatic life stages of amphibians is still needed to reduce uncertainties. The current review aims to propose a test with amphibians based on spatial avoidance, as a more humane alternative method to the lethality tests for chemicals. A review of lethal toxicity tests carried out with amphibians in the period between 2018 and 2021 is presented, then we discuss the suitability of using fish toxicity data as a surrogate to predict the effects on more sensitive amphibian groups. The possible differences in sensitivity to chemicals may justify the need to develop further tests with amphibian embryos and larvae in order to reduce uncertainties. A new test is proposed focused on the avoidance behaviour of organisms fleeing from contamination to replace lethal tests. As avoidance indicates the threshold at which organisms will flee from contamination, a reduction in the population density, or its disappearance, at the local scale due to emigration is expected, with ecological consequences analogous to mortality. Avoidance tests provide an ethical advantage over lethal tests as they respect the concepts of the 3 Rs (mainly Refinement), reducing the suffering of the organisms.