Differences in kinetic metabolomics in Eisenia fetida under single and dual exposure of imidacloprid and dinotefuran at environmentally relevant concentrations

Metabolomic responses of earthworms to neonicotinoids are important for understanding their molecular-level toxicity and assessing their ecological risks, but little is known until now. We investigated impact of imidacloprid (IMI, 52.6 ng/g) and dinotefuran (DIN, 52.5 ng/g) on Eisenia fetida metabolomics under single- and dualcompound exposure scenarios for one to four weeks. Dissimilar metabolites and anti-stress strategies were found for different neonicotinoids and exposure scenarios. Specifically, IMI exposure first increased myo-inositol and UDP-glucuronate associated with transmembrane absorption and transformation to IMI-urea, and then increased glutathione and fourteen amino acids (TCA cycle precursors) to resist stress and replenish energy. In contrast, worms exposed to DIN first prepared TCA cycle intermediates from glucosamine-6-phosphate and amino acids, suppressed urea cycle and DIN transformation, and then alleviated oxidative stress by increasing carnosine, nicotinate-D-ribonucleotide and nicotinamide-beta-riboside. Dual exposure increased four eicosanoids by 1.6-1.9-fold, possibly associated with membrane lipid peroxidation; the amino acids consumed to balance the energy metabolism exhibited a wave-like pattern. This study first systematically revealed the compound/time/ exposure scenario- dependent effects of trace neonicotinoids on earthworm metabolomics and advanced the understanding of their action modes. Neonicotinoid transformation was closely related to worms' metabolic profiles, providing important insights in contaminant fate in soil ecosystems.