Multi-omics analysis reveals the synergistic and intergenerational toxicity of Cr(VI) and chlorfenapyr on zebrafish: disruption of gut-liver axis and mitochondrial impairment

Concurrent hexavalent chromium [Cr(VI)] and chlorfenapyr contamination in aquatic environments poses risks to environmental organisms. A parental zebrafish (Danio rerio) exposure model with multi-omics analyses was used to elucidate molecular and transgenerational effects. Combined exposure decreased intestinal tight junction transcription (zo-1, and occludin), caused morphological damage, and activated pro-inflammatory cytokines (il-1β, il-6, il-8, and TNF-α). 16S rRNA sequencing revealed reduced butyrate-producing bacteria (e.g., Coprococcus), while untargeted metabolomics showed declines in phospholipid precursors (choline, and glycerophosphocholine), implicating disrupted gut barrier metabolism. Hepatic transcriptomics identified downregulation of oxidative phosphorylation components (atp5l, atp5mc3b, atp5po, and ndufs7) and antioxidant enzymes (gpx, and mn-sod), correlating with decreased ATP, mitochondrial membrane potential, and increased apoptosis. F₁ offspring exhibited aberrations in gene transcriptions associated with oxidative phosphorylation and inflammation, suggesting the presence of intergenerational toxicity. These results highlighted the gut-liver axis and mitochondrial function as targets for molecular biomarkers and inform eco-safety regulation addressing chemical mixture hazards.