Sub-lethal effects of soil multiple contamination on the avoidance behaviour of Eisenia fetida

Natural ecosystems are frequently exposed to complex mixtures of different chemicals. However, the environmental risk assessment is mainly based on data from individual substances. In this study, the individual and combined effects on the terrestrial earthworm E. fetida exposed to the anionic surfactant sodium lauryl ether sulphate (SLES) and the pesticides chlorpyrifos (CPF) and imidacloprid (IMI) were investigated, by using the avoidance behaviour as endpoint. Earthworms were exposed to a soil artificially contaminated with five sub-lethal concentrations of each contaminant, both as single substances and in combination of binary and ternary mixtures. Overall results showed that IMI provoked the highest avoidance effect on earthworms, with a concentration value that induced an avoidance rate of 50% of treated organisms (AC50) of 1.30 mg/kg, followed by CPF (AC50 75.26 mg/kg) and SLES (AC50 139.67 mg/kg). The application of the Combination Index (CI) method, indicated that a deviation from the additive response occurred for most of the tested chemical mixtures, leading to synergistic or antagonistic avoidance responses. Synergistic effects were produced by the exposure to the two lowest concentrations of the CPF+IMI mixture, and by the highest concentrations of SLES+CPF and SLES+CPF+IMI mixtures. On the contrary, antagonistic effects were observed at the lowest concentrations of the binary mixtures containing the SLES and at almost all the tested concentrations of the SLES+CPF+IMI mixture (with the exception of the highest tested concentration). These results show that the avoidance test is suitable to assess the detrimental effects exerted on earthworms by chemical mixtures in soil ecosystems and the use of behavioural endpoints can increase the ecological significance of environmental risk assessment procedures.