Multiple perspectives reveal the gut toxicity of polystyrene microplastics on Eisenia fetida: Insights into community signatures of gut bacteria and their translocation

The gut is the primary pathway by which soil animals are exposed to microplastics (MPs). However, the gut toxicity of MPs has not been elucidated in earthworms. Herein, we aimed to study the gut toxicity (e.g., gut barrier dysfunction, gut bacterial translocation, and pathogen invasion) of polystyrene microplastics (PS-MPs) on Eisenia fetida and its relationship with gut bacteria. We found that PS-MPs exposure caused gut barrier damage to Eisenia fetida. This damage included apparent injury of gut epithelial cells and significantly lower transcription levels of genes coding for gut tight junction (TJ)-related proteins. We then observed significantly increased levels of bacterial lipopolysaccharide (LPS) and gut bacterial load, indicating the occurrence of gut bacterial translocation and related barrier damage. Subsequently, antibacterial immune responses were activated and accompanied by a failure of the antioxidant defense system, indicating that pathogen invasion might occur. Gut barrier damage could weaken host selective pressures (deterministic process) on gut bacteria, such as particular pathogens. Indeed, members of Proteobacteria, e.g., Aeromonas and Escherichia/Shigella, regarded as potential opportunistic pathogens, were remarkable signatures of groups exposed to PS-MPs. These potential opportunistic gut bacteria were pivotal contributors to gut TJ damage and gut bacterial translocation resulting from PS-MPs exposure. In addition, the gut bacterial networks of PS-MPs exposure groups were more uncomplicated than those of the control group, but more negative interactions were easy to observe. In conclusion, our work sheds light on the molecular mechanism of earthworm gut toxicity caused by PS-MPs exposure and provides a prospective risk assessment of MPs in soil ecosystems.