Biochemical toxicity, lysosomal membrane stability and DNA damage induced by graphene oxide in earthworms

With the growing production and use of carbon nanomaterials (CNMs), the risk of their releases to the environment has drawn much attention. However, their potential effect on soil invertebrates has not yet been systematically assessed. Herein, the toxic effects of graphene oxide (GO) on earthworms (Eisenia fetida) were thoroughly investigated. Exposure to different doses of GO (0, 5, 10, 20, and 30 g kg^-1) was conducted for 7, 14, 21, and 28 days. The results showed that enzymatic activity was stimulated at the early stages of exposure (7 days and 14 days) and inhibited after 14 days for catalase (CAT) and after 21 days for peroxidase (POD) and superoxide dismutase (SOD), especially at high GO doses. The content of MDA showed an increasing trend over the whole exposure period and was significantly elevated by GO from 21 days except at the dose of 5 g kg^-1on day 21. Lysosomal membrane stability and DNA damage presented dose- and time-dependent relationships. Graphene oxide remarkably decreased lysosomal membrane stability except at the dose of 5 g kg^-1 on day 7. The tail DNA%, tail length and olive tail moment increased with increasing GO dose throughout the exposure duration, reaching maximum values at the end of exposure (28 days). These findings suggest that GO induces oxidative stress and genotoxicity in Eisenia fetida, resulting in lipid peroxidation, decreased lysosomal membrane stability and DNA damage. Therefore, attention should be paid to the potential pollution and risk associated with graphene oxide application. The results can provide valuable information for environmental safety assessment of graphene nanomaterials in soil.