New strategies for evaluating imidacloprid-induced biological consequences targeted to Eisenia fetida species and the corresponding mechanisms of its toxicity

Imidacloprid (IMI), a neonicotinoid insecticide, has a wide variety of applications in both agriculture and horticulture. As a result of it massive and repeated use, its traces remained in soil pose severe damage to soil invertebrates, particularly earthworms. Limited information is available regarding the underlying mechanisms of IMI toxicity toward earthworms at the molecular, transcriptional, and cellular levels. Here, Eisenia fetida coelomocytes and key defensive proteins were selected as targeted receptors to explore the toxic mechanisms of oxidative stress-mediated cytotoxicity, genotoxicity, and antioxidant responses induced by IMI stress and the molecular mechanisms underlying the binding of IMI and superoxide dismutase (SOD)/catalase (CAT). Results showed that IMI exposure destroyed the cell membrane integrity of earthworm cells, causing cell damage and cytotoxicity. The intracellular levels of ROS, including ·O2− and H2O2 were induced by IMI exposure, thereby triggering oxidative stress and damage. Moreover, IMI exposure attenuated the antioxidative stress responses (reduced antioxidant capacity and CAT/SOD activities) and caused deleterious effects (enhanced DNA damage, lipid peroxidation (LPO), and protein carbonylation (PCO)) through ROS-mediated oxidative stress pathway. Aberrant gene expression associated with oxidative stress and defense regulation, including CAT, CRT, MT, SOD, GST, and Hsp70 were induced after IMI exposure. Concentration-dependent conformational and structural alterations of CAT/SOD were observed when IMI binding. Also, direct binding of IMI resulted in significant inhibition of CAT/SOD activities in vitro. Molecular simulation showed that IMI preferred to bind to CAT active center through its direct binding with the key residue Tyr 357, while IMI bound more easily to the connecting cavity of two subunits away from SOD active center. In addition, hydrogen bonds and hydrophobic force are the main driving force of IMI binding with CAT/SOD. These findings have implications for comprehensive evaluation of IMI toxicity to soil eco-safety and offer novel strategies to elucidate the toxic mechanisms and pathways of IMI stress. [Display omitted] •IMI destroyed cell membrane integrity and triggered cell injury and cytotoxicity.•IMI caused oxidative stress-mediated DNA damage, LPO, and PCO in E. fetida cells.•IMI binding initiated the structural and conformational changes of CAT/SOD.•IMI preferred to bind to the active center of CAT