Health risks of sulfentrazone exposure during zebrafish embryo-larvae development at environmental concentration

Knowledge about the negative effects and mechanism of sulfentrazone (SUL) on aquatic early life stages is still limited. Here we investigated the lethal and sub-lethal effects of SUL during zebrafish embryo-larvae development. Results demonstrated that the 96 h and 120 h-LC₅₀ of SUL to embryonic zebrafish was 2.02 mg/L, and the 30 d-LC₅₀ was 0.899 mg/L after embryos exposed to SUL for 30 d. High concentrations of SUL delayed yolk sac absorption, disordered the hatching and heart rate during zebrafish embryonic stage, while 0.0100-0.100 mg/L SUL had no phenotypic changes on embryonic development, but decreased the body weight of larvae after 30 d exposure. RNA-seq identified 321, 394 and 727 differentially expressed genes in larvae after embryos exposed to 0.0100 mg/L, 0.0400 mg/L and 0.400 mg/L SUL for 30 d, found that the transcriptional profiles involved in heart development and endocrine disruption were simultaneously influenced by different concentrations of SUL, such as adrenergic signaling in cardiomyocytes, cardiac muscle contraction, cell adhesion molecules and steroid biosynthesis. Biochemical analysis showed that SUL increased the levels of E₂, T₃ and TSH, induced the activities of mitochondrial complex IV, cytochrome c oxidase, Ca²⁺-ATPase, total Na⁺K⁺-ATPase and Ca²⁺Mg²⁺-ATPase, and decreased ATP formation after embryos exposed to SUL for 5 d and 30 d. Further comprehensive analysis demonstrated that SUL caused more significantly alteration on the transcript, level or activity of the key elements involved in heart development and endocrine disruption after 30 d exposure, indicated long-term SUL exposure might cause more negative effects on zebrafish at doses below the presumed no-observed-adverse-effect level during early life development. The results inferred the environmental concentration of SUL might cause potential cardiac and endocrine health risk in zebrafish later life stages, also facilitated a better understanding of the sub-lethal effects and molecular mechanism of SUL on aquatic organism.