Aged microplastics change the toxicological mechanism of roxithromycin on Carassius auratus: Size-dependent interaction and potential long-term effects

Size effects of microplastics have received extensive attention for their influence on other pollutants and harm to organisms. In this study, we investigated the uptake, elimination, tissue distribution and potential toxicity mechanism of roxithromycin (ROX) in the presence of 0.5, 5 and 50 μm of aged microplastics (AMPs) in Carassius auratus. The results showed that AMPs promoted the ROX bioaccumulation of various tissues in a size-dependent manner. AMPs and ROX significantly induced superoxide dismutase and catalase activities of liver and gut, and inhibited acetylcholinesterase activities of brain. The coexistence of smaller AMPs exacerbated pathological abnormalities in liver, gill and brain induced by ROX, while larger AMPs caused more intestinal damage. Moreover, high-throughput 16S rRNA gene sequencing indicated that the abundance of Proteobacteria in 0.5 μm AMPs and ROX joint treatments and Firmicutes and Bacteroidota in 50 μm AMPs and ROX joint treatments were significantly raised (p < 0.05). Metabolomics revealed that AMPs and ROX had a size-dependent long-term effect on gut microbial metabolites, which was mainly related to galactose metabolism, amino acid metabolism and primary bile acid biosynthesis pathways after a 7-day elimination, respectively. These results provide important insights into the relationship between the size effect of AMPs and interaction mechanism of AMPs and coexisting pollutants on aquatic organisms.