Combined impact of fishmeal and tetracycline on resistomes in mariculture sediment

Mariculture sediment has been recognized as a major contributor of environmental antibiotic resistance genes (ARGs), which are challenging the treatment of infections worldwide. Both antibiotics and fishmeal are used in aquaculture, and each has the potential to facilitate ARG dissemination, however their combined impact on the sediment resistome and their relative contribution remain unclear. In this study, microcosms were exposed to varying concentrations of tetracycline with or without fishmeal (0.1% wt/wt) for 14 days. Sediment genomic DNA was analyzed using high throughput quantitative PCR and 16S rRNA gene amplicon sequencing to compare the contribution of fishmeal and tetracycline to antibiotic resistomes and bacterial communities in mariculture sediment. Sixty-seven ARGs were detected potentially correlating to resistance for several major antibiotics. Fishmeal, but not the dose of tetracycline, contributed to the significant increase of both ARG abundance and diversity in the sediment. Based on principle coordinate analysis and hierarchical clustering, ARGs were clustered into two groups depending on whether fishmeal was added. Aminoglycoside, macrolide–lincosamide–streptogramin b (MLSb) and tetracycline resistance genes were the most abundant when fishmeal was used, while a significant increase in mobile genetic element (MGE) abundance was also detected (P < 0.05). Meanwhile, bacterial community structures were detected with distinct patterns between the two groups (Adonis, P < 0.05). Using the Mantel test and partial least squares path modeling, we identified that sediment resistomes were significantly correlated with microbial community structures (P < 0.05) which were mainly driven by nutrients in fishmeal. Together our findings suggested that fishmeal plays a more important role than tetracycline in proliferation of ARGs in mariculture sediment. This study may provide new insights into the mitigation of ARG propagation in mariculture operations.