Nanoplastics induces oxidative stress and triggers lysosome-associated immune-defensive cell death in the earthworm Eisenia fetida

Nanoplastics (NPs) are increasingly perceived as an emerging threat to terrestrial environments, but the adverse impacts of NPs on soil fauna and the mechanisms behind these negative outcomes remain elusive. Here, a risk assessment of NPs was conducted on model organism (earthworm) from tissue to cell. Using palladium-doped polystyrene NPs, we quantitatively measured nanoplastic accumulation in earthworm and investigated its toxic effects by combining physiological assessment with RNA-Seq transcriptomic analyses. After a 42-day exposure, earthworm accumulated up to 15.9 and 143.3 mg kg of NPs for the low (0.3 mg kg) and high (3 mg kg) dose groups, respectively. NPs retention led to the decrease of antioxidant enzyme activity and the accumulation of reactive oxygen species (O and HO), which reduced growth rate by 21.3 %-50.8 % and caused pathological abnormalities. These adverse effects were enhanced by the positively charged NPs. Furthermore, we observed that irrespective of surface charge, after 2 h of exposure, NPs were gradually internalized by earthworm coelomocytes (∼0.12 μg per cell) and mainly amassed at lysosomes. Those agglomerations stimulated lysosomal membranes to lose stability and even rupture, resulting in impeded autophagy process and cellular clearance, and eventually coelomocyte death. In comparison with negatively charged nanoplastics, the positively charged NPs exerted 83 % higher cytotoxicity. Our findings provide a better understanding of how NPs posed harmful effects on soil fauna and have important implications for evaluating the ecological risk of NPs.