Integration of transcriptomics and metabolomics reveals the responses of earthworms to the long-term exposure of TiO nanoparticles in soil.

Titanium dioxide nanoparticles (nTiO) are widely used and their environmental occurrence has raised concerns about the potential toxicity to biota. However, few studies have investigated the effect of long-term exposure to nTiO on soil invertebrates. This study therefore for the first time investigated the long-term (120 days) effect of nTiO (0, 5, 50, and 500 mg/kg) on the phenotypes, transcriptomic, and metabolomic profiles of earthworm (Eisenia fetida) in soil. The results showed that the long-term exposure to nTiO did not significantly affect the growth, reproduction, and Ti content of earthworms. However, the antioxidant system and the transcriptomic and metabolomic profiles of earthworms were significantly affected. The superoxide dismutase (SOD) activity and the reduced glutathione/oxidized glutathione (GSH/GSSG) ratio significantly decreased under the 500 mg/kg nTiO treatment. The metabolomics analysis showed that glycine and pyroglutamic acid contents involved in the GSH metabolism were significantly altered under the 500 mg/kg treatment. Moreover, transcriptomics and metabolomics data revealed that the long-term exposure to nTiO affected the synthesis of carbohydrates, proteins, and lipids. However, the transcriptomics results indicated that the genes involved in ribosome biogenesis in eukaryotes pathway and TGF-beta signaling pathway were upregulated, which could explain why the growth and reproduction of earthworms were apparently not affected by the nTiO exposure. The combination of transcriptomics and metabolomics reveals the global responses that cannot be observed by conventional toxicity endpoints, facilitating the assessment of long-term ecological effect of engineered nanoparticles in the environment.