Network analysis reveals significant joint effects of microplastics and tetracycline on the gut than the gill microbiome of marine medaka

Microplastics could accumulate and enrich antibiotics in the aquatic environment. Despite this, the joint effects of microplastics and antibiotics on aquatic organisms are not clear. Here, we investigated the changes of mi-crobial interactions in both gill and gut of marine medaka exposed to polystyrene microbeads (PS) and/or tetracycline for 30 days by using co-occurrence network analysis based on 16S rRNA gene amplicon sequences. We found that the single and combined effects of PS and tetracycline were more profound on the gut than on the gill microbiome. SourceTracker analysis showed that the relative contributions from the gill microbiome to the gut microbiome increased under combined exposure. Moreover, the combined exposure reduced the complexity and stability of the gut microbial network more than those induced by any single exposure, suggesting the synergistic effects of PS and tetracycline on the gut microbiome. The PS and tetracycline combined exposure also caused a shift in the keystone taxa of the gut microbial network. However, no similar pattern was found for gill microbial networks. Furthermore, single and combined exposure to PS and/or tetracycline altered the associa-tions between the gut network taxa and indicator liver metabolites. Altogether, these findings enhanced our understanding of the hazards of the co-occurring environmental microplastics and antibiotics to the fish commensal microbiome.