Mixture toxicity of four commonly used pesticides at different effect levels to the epigeic earthworm, Eisenia fetida

- Clothianidin was the most toxic of all the tested pesticides to the earthworm.
- Synergism of the quarternary mixture was evident throughout the effect levels.
- Combination index method could more accurately predict the combined toxicity.
- Chemical mixtures with synergistic effects should be given attention.

As commonly used pesticides, chlorpyrifos (CPF), fenobucarb (FEN), clothianidin (CLO) and acetochlor (ACE) are widely applied on crops worldwide. In this study, the combined toxicities of their binary, ternary and quaternary mixtures were evaluated using the earthworm Eisenia fetida as test organism. Mixture toxicities were studied using the combination index (CI) method and visualized by isobolograms, and then data were compared with traditional concentration addition (CA) and independent action (IA) models. Two binary mixtures of CPF+FEN and FEN+ACE, two ternary mixtures of CPF+CLO+FEN and CPF+FEN+ACE, and quaternary mixture of CPF+FEN+ACE+CLO exhibited a clear synergistic effect. The CI method was compared with the classical models of CA and IA, indicating that the CI method could accurately predict the combined toxicities of the chemicals. The results indicated that it was difficult to predict combined effects of these pesticides from mode of action alone because of existence of complicated synergistic and antagonistic responses. More attention should be paid to the potential synergistic effects of chemicals interactions, which might cause serious ecological problems.