Environmental antibiotics in wastewater disrupt zebrafish embryonic development via Wnt/β-catenin signaling: mechanistic insights and implications for risk assessment

Antibiotic residues in wastewater treatment plant (WWTP) effluents are a matter of global environmental health concern. This study combined global WWTP monitoring data with epidemiological modeling and zebrafish embryo assays to assess developmental risks. A meta-analysis of 121 studies identified amoxicillin, trimethoprim and norfloxacin as high priority contaminants. Ecological modeling using generalized additive models (GAM) revealed a statistical association between effluent antibiotic levels and neonatal disease incidence. Zebrafish exposed to environmentally relevant concentrations exhibited dose-dependent developmental toxicity, including pericardial edema, oxidative stress, and impaired cardiac function. Molecular analysis indicated dysregulation of the Wnt/β-catenin signaling pathway, with Wnt1 upregulation mediating toxic effects. Functional validation using CRISPR interference targeting wnt1 attenuated cardiac malformations and apoptosis, while docking simulations confirmed strong antibiotic-Wnt1 binding. These findings establish a mechanistic link between environmental antibiotic exposure and developmental toxicity and underscore the potential of Wnt1 as a biomarker for ecological risk assessment. This work supports the need for advanced tertiary treatment strategies and molecular-based monitoring frameworks.