The toxic effect of triclosan and methyl-triclosan on biological pathways revealed by metabolomics and gene expression in zebrafish embryos

The omnipresence of antimicrobial triclosan (TCS) and by-products in aquatic environments is a threat to aquatic organisms. Traditionally, the adverse effects of TCS and its by-products have been evaluated by examining the phenotypic output relevant to predicting acute toxicity rather than studying the perturbation of biological pathways. Identifying alterations in the key pathways and molecular mechanisms caused by toxic chemicals helps researchers assess the ecological risks of TCS and its by-products to aquatic environments. In this study, we used metabolomics and reverse transcription qPCR to investigate the adverse effects of a wide range of concentrations of triclosan and its derivative methyl-triclosan (MTCS), ranging from relatively low environmentally relevant levels (ng/L) to high-dose concentrations (sublethal concentration), on zebrafish (Danio rerio) embryos. The metabolism and transcriptome analysis revealed changes in the metabolite and transcripts expression of zebrafish embryos after 96 h exposure at 30 μg/L and 300 μg/L of TCS, 400 μg/L of MTCS and the TCS/MTCS mixture (30 μg/L TCS + 3 μg/L MTCS and 300 μg/L TCS + 30 μg/L MTCS). Significant dysregulations in the expression of the urea transporter (UT), glucose-6-phosphate dehydrogenase (G6PD), alanine transaminase (ALT), glutamate dehydrogenase (GDH), phosphoglucomutase (PGM), and fatty acid synthase (FASN), together with changes in alanine, urea, glucose, 6-phosphogluconalactone, and palmitic acid were observed in the TCS, MTCS, and TCS/MTCS treatments. Particularly, the MTCS treatment group showed fold changes in the mRNA expression of nitrogen metabolism, energy metabolism, and fatty acid synthesis, indicating a disruption of the zebrafish embryos' biological pathways. The changes in the metabolites and gene expressions induced by the TCS, MTCS and the TCS/MTCS mixture treatment demonstrate the pathway changes in starch and sucrose metabolism, nitrogen metabolism, fatty acid synthesis, and phenylalanine, tyrosine and tryptophan biosynthesis. Therefore, our study provides better insights into the risks of the parental compound (TCS) and its by-product (MTCS), as well as the perturbation in biological pathways induced by these two compounds in aquatic environments.