Epoxiconazole disturbed metabolic balance and gut microbiota homeostasis in juvenile zebrafish

Epoxiconazole (EPX) is a broad-spectrum fungicide extensively used in agricultural pest control. Emerging evidence suggests that EPX can adversely affect different endpoints in non-target organisms. Here, the toxicity of EPX was assessed using earlier developmental stage of zebrafish as a model to investigate its effects on metabolism and intestinal microbiota after 21 days of exposure. Our findings indicated that EPX exposure resulted in physiological alterations in juvenile zebrafish, including increase in triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL), and glycose (Glu). Nile red staining demonstrated enhanced lipid accumulation in juvenile, accompanied by a marked upregulation in the expression of genes associated with TG synthesis. Moreover, EPX led to alterations in amino acids and carnitines levels in 21 dpf (days post fertilization) zebrafish. We also observed that EPX disrupted intestinal barrier function in juvenile zebrafish, manifested by decreasing mucus secretion and changing in genes related to tight junctions. Moreover, for a more comprehensive analysis of the intestinal microbiota in 21 dpf zebrafish, the intestine tissues were dissected under a microscope for 16S rRNA sequencing analysis. The results revealed that EPX altered the structure and abundance of intestinal microflora in zebrafish, including decreased alpha diversity indices and shifted in bacteria at phylum and genus levels. Notably, the correlation analysis demonstrated strong associations between alterations in various pathogenic bacterial genera and levels of amino acids and carnitines. Overall, these findings confirm that the fungicide EPX promotes metabolic disorders and alterations in the intestinal micro-environment in 21 dpf zebrafish, shedding light on the toxicologic effects of chemicals to aquatic organisms during the development stage.