Microbiota-Driven Metabolic Alterations Induced by BPA, TDCPP and PFOA in an Ex Vivo Human Fecal Fermentation Model

The gut microbiome is increasingly recognized as a key contributor to chemical toxicity. Endocrine-disrupting chemicals (EDCs) such as bisphenol A (BPA), tris(1,3-dichloro-2-propyl) phosphate (TDCPP), and perfluorooctanoic acid (PFOA) are widespread environmental contaminants with the potential to affect host health. To characterize microbiota-specific response to these compounds, we employed an ex vivo fecal fermentation model using samples from healthy adult donors. Fecal slurries were exposed to BPA, TDCPP and PFOA (75 μM) for up to 24 h under anaerobic conditions. Targeted LC-MS/MS quantified parent compounds over time, while untargeted metabolomics profiled microbial metabolic alterations at 4 and 24 h. TDCPP levels decreased similarly in fecal and abiotic controls, suggesting a nonmicrobial loss (e.g., instability or adsorption), whereas PFOA levels remained stable across donors. Untargeted metabolomics revealed compound- and time-dependent perturbations, with PFOA eliciting the strongest metabolic shifts. A curated set of 124 annotated metabolites indicated disruptions in bile acid transformation short-chain fatty acid production, nucleotide turnover, redox balance, and phytochemical catabolism. Several altered metabolites have been previously linked to immunomodulatory processes, suggesting potential implications for host-microbiota interactions. Overall, this study demonstrates the utility of ex vivo fermentation systems for assessing microbiota-mediated metabolic responses to xenobiotics and highlights the relevance of incorporating microbiome-related end points into chemical risk assessment.