Toxicity assessment of earthworm exposed to arsenate using oxidative stress and burrowing behavior responses and an integrated biomarker index.

Arsenate (As-V) is a ubiquitous contaminant in soil as a result of excessive use of veterinary drugs and pesticides, causing enormous environmental risks. Multiple biomarkers have been used to assess the ecotoxicity of arsenic, however, the mechanisms of toxicity remain unclear. This paper describes the exposure of the earthworm (Eisenia fetida) to natural soil with different As-V concentrations for 28 days, then biomarkers from oxidative stress and burrowing behavior were quantified to evaluate As-V stress. Dynamic changes in reactive oxygen species (ROS), lipid peroxidation (MDA), adenosine triphosphate (ATP) content and antioxidant enzymes activity (Gpx, SOD, CAT) implied two stages of intensified stress responses and physiological adaptability. The transcriptional expression and regulation of antioxidant enzymes showed different responses. The mRNA expression of sod1 was up-regulated, while that of cat showed no significant change. The related regulators, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1), showed dose-dependent activation, suggesting antioxidant defense induced by Nrf2 signaling. The burrowing behavior after 14-day exposure indicated that As-V inhibited burrowing activity, especially the burrow length and maximum burrow depth. These multiple biomarkers were integrated using a biomarker response index (BRI) model, which showed significant dose-effect relationship especially on day 28, and suggested that ATP was a sensitive and representative biomarker. This study provided evidence that burrowing activity, Nrf2 and HO-1 were useful biomarkers warranting inclusion into the BRI model. Arsenic toxicity was comprehensively understood through redox homeostasis regulation, biochemical and behavioral changes, and these results suggested new strategies for soil pollutants diagnosis.