Feed additives shift gut microbiota and enrich antibiotic resistance in swine gut

Antibiotic resistance genes (ARGs) are emerging environmental contaminants posing a threat to public health. Antibiotics and metals are widely used as feed additives and could consequently affect ARGs in swine gut. In this study, high-throughput quantitative polymerase chain reaction (HT-qPCR) based ARG chip and next-generation 16S rRNA gene amplicon sequencing data were analyzed using multiple statistical approaches to profile the antibiotic resistome and investigate its linkages to antibiotics and metals used as feed additives and to the microbial community composition in freshly collected swine manure samples from three large-scale Chinese pig farms. A total of 146 ARGs and up to 1.3×10¹⁰ total ARG copies per gram of swine feces were detected. ARGs conferring resistance to aminoglycoside, macrolide-lincosamide-streptogramin B (MLSB) and tetracycline were dominant in pig gut. Total abundance of ARGs was positively correlated with in-feed antibiotics, microbial biomass and abundance of mobile genetic elements (MGEs) (P<0.05). A significant correlation between microbial communities and ARG profiles was observed by Procrustes analysis. Network analysis revealed that Bacteroidetes and Firmicutes were the most dominant phyla co-occurring with specific ARGs. Partial redundancy analysis indicated that the variance in ARG profiles could be primarily attributed to antibiotics and metals in feed (31.8%), gut microbial community composition (23.3%) and interaction between feed additives and community composition (16.5%). These results suggest that increased levels of in-feed additives could aggravate the enrichment of ARGs and MGEs in swine gut.