Combined ecotoxicological effects of different-sized polyethylene microplastics and imidacloprid on the earthworms (Eisenia fetida).

Microplastics (MPs) and pesticides frequently coexist in farmland soil; however, there are relatively few studies on the ecological risk assessment of soil animals attributed to the combined pollution caused by MPs and pesticides. Moreover, the influence of particle size on the combined toxic effects of MPs and pesticides remains poorly understood. In this study, different-sized polyethylene MPs (PE MPs; 10 µm, 500 µm, and 2 mm) were combined with a series of imidacloprid concentrations (IMI; 0.10, 0.50 and 1.00 mg/kg), and earthworms (Eisenia fetida) were exposed to these MP and IMI combinations for 28 d to explore the combined toxic effects and mechanisms. The results showed, compared with IMI or PE MPs exposure alone, the combined exposure of IMI and PE MPs did not substantially increase the acute toxicity of earthworms but significantly inhibited weight increase and induced more serious epidermal damage to earthworms with a size effect; among these 10 µm PE MPs combined with IMI exhibited the strongest toxic effects. In addition, the combined exposure decreased antioxidant enzymes activity and caused oxidative damage in earthworms. Transcriptome results demonstrated most of the treatment combinations affected the ferroptosis pathway, which was further verified by the increase in the total iron content, reactive oxygen species, and malondialdehyde content in earthworms. Combined with the analysis of key signalling pathways, the above results revealed that the combined exposure to IMI and PE MPs showed stronger toxicity to earthworms than exposure to either IMI or MPs alone, which was mediated by the superimposed effect of ferroptosis and oxidative damage. Moreover, the effect was size-dependent, with 10 µm PE MPs combined with IMI exhibiting the strongest toxic effects. This study aimed to provide data to support the ecological risk assessment of soil animals caused by the combined pollution of MPs and coexisting pesticides.