Per- and polyfluoroalkyl substance (PFAS) mixtures induce gut microbiota dysbiosis and metabolic disruption in silkworm (Bombyx mori L.).

Mixed legacy and emerging per- and polyfluoroalkyl substances (PFASs) are commonly found in soil and dust; however, the potential toxicity of PFAS mixtures (mPFASs) in insects is unknown. Using 16S rRNA gene sequencing and transcriptome sequencing (RNA-Seq), we evaluated the adverse effects of mPFASs on silkworms, a typical lepidopteran insect. After exposure to mPFASs, the silkworm midgut was enriched with high levels of PFASs, which induced histopathological changes. The composition of the midgut microbiota was significantly affected by mPFAS exposure, and functional predictions revealed significant disruption of some metabolic pathways. RNA-seq analysis revealed that mPFASs significantly changed the transcription profiles. Functional enrichment analysis of the differentially expressed genes also revealed that biological processes related to metabolic pathways and the digestive system were significantly affected, similar to the results of the gut microbiota analysis, suggesting that mPFAS exposure had an adverse effect on the metabolic function of silkworms and may further affect their normal growth. Finally, the significant correlation between abundance changes in the gut microbiota and metabolism/digestion-related genes further highlighted the role of the gut microbiota in mPFAS-related processes affecting the metabolic functions of silkworms. To our knowledge, this study is the first to evaluate the toxic effects of mPFASs in insects and provide basic data for further PFAS toxicity investigations in insects and comprehensive ecological risk assessments of mPFASs.