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^-1 of NPs for the low (0.3 mg kg^-1) and high (3 mg kg^-1) dose groups, respectively. NPs retention led to the decrease of antioxidant enzyme activity and the accumulation of reactive oxygen species (O₂^- and H₂O₂), 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.

The synergistic impact of microplastics (MPs) and organic pollutants remains poorly understood in the marine environment. This study aimed to assess the toxicity of polypropylene microplastics (PS) and/or di-(2-ethylhexyl) phthalate (DEHP) on marine clams. Both Ruditapes philippinarum and Tegillarca granosa were exposed to PS and DEHP individually and combined at environmentally relevant concentrations for 48 h. The filtration rate, antioxidant enzymes activity, lipid peroxidation, reactive oxygen species accumulation, and histological alterations were evaluated. Our results show that single or co-exposure to MPs and DEHP significantly decreases the filtration rate in both type of clams, but the latter exhibited stronger inhibition effect. Close examination of accumulation of reactive oxygen species and related biomarkers revealed that combined exposure exerts greater oxidative stress in the cells, which causes more serious histopathological damage in the gills of the bivalves. Our study implies that MPs, in synergy with organic pollutants, can be more harmful for marine organisms.