Health risks of chlorothalonil, carbendazim, prochloraz, their binary and ternary mixtures on embryonic and larval zebrafish based on metabolomics analysis

Abstract

Chlorothalonil (CTL), carbendazim (CBZ), and prochloraz (PCZ) are fungicides widely used in many countries. The use of these fungicides raises concerns because they are often applied together or used in the same agricultural area. However, the toxicity of these fungicides or mixtures, especially to aquatic organisms, has received limited attention. Here, embryonic and larval zebrafish were exposed to indicated concentrations of CTL, CBZ, and PCZ and their binary (CTL+CBZ, CTL+PCZ and CBZ+PCZ) and ternary (CTL+CBZ+PCZ) mixtures for 24 h. Based on metabolomics analysis, we observed that hundreds of metabolites were altered, and glycolysis metabolism and amino acid metabolism were the two most affected pathways. Interestingly, a total of 9 and 26 metabolites changed significantly in embryos and larvae treated with all fungicides, respectively. Among these altered metabolites, 2-aminoadipic acid (2-AAA) levels increased significantly in all groups, indicating that 2-AAA potentially represents a useful biomarker for evaluating the toxicity of fungicides. Furthermore, the joint effects of CTL+PCZ on embryos and larvae, especially on amino acid metabolism, were weaker than those in other groups, but combined treatment did not influence individual fungicidal activity. Data acquired from metabolomics provided important insight for understanding the mechanism by which fungicides or their mixtures affect zebrafish.