Developmental toxicity and transcriptome analysis of zebrafish (Danio rerio) embryos following exposure to chiral herbicide safener benoxacor

Benoxacor, a chiral herbicide safener for S-metolachlor, has been detected in streams. However, the potential risk this poses to aquatic ecosystems is not clear. This study used zebrafish (Danio rerio) embryos as a model to assess the enantioselective toxicity of benoxacor and its effects on biological activity and development from 2 h to 96 h post-fertilization (hpf). Results showed that benoxacor had negative effects on hatchability, malformations, and mortality. Compared to either individual enantiomer, embryos exposed to Rac-benoxacor had higher acute and developmental toxicities, glutathione S-transferase (GST) and glutathione peroxidase (GPx) enzyme activities, and nrf 2 expression levels. They also had lower superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GR) enzyme activity and krt 17, tbx 16, osx, cat, bcl 2, bax, and ifn expression levels. High-throughput RNA sequencing revealed that Rac-benoxacor had a greater effect on gene regulation than either enantiomer. Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses demonstrated that changes in oxidoreductase activity, cellular lipid metabolic process, and catalytic activity related genes may be due to the enantioselective effects of benoxacor isomers. These results suggest that the ecotoxicology data and safety knowledge about the effects of chiral benoxacor on zebrafish should be considered in future environmental risk evaluation.