Abundance of antibiotic resistance genes and their association with bacterial communities in activated sludge of wastewater treatment plants: Geographical distribution and network analysis

Wastewater treatment plants (WWTPs) are deemed reservoirs of antibiotic resistance genes (ARGs). Bacterial phylogeny can shape the resistome in activated sludge. However, the co-occurrence and interaction of ARGs abundance and bacterial communities in different WWTPs located at continental scales are still not comprehensively understood. Here, we applied quantitative PCR and Miseq sequence approaches to unveil the changing profiles of ARGs (sul1, sul2, tetW, tetQ, tetX), intI1 gene, and bacterial communities in 18 geographically distributed WWTPs. The results showed that the average relative abundance of sul1and sul2 genes were 2.08 × 10^-1 and 1.32 × 10^-1 copies/16S rRNA copies, respectively. The abundance of tetW gene was positively correlated with the Shannon diversity index (H'), while both studied sul genes had significant positive relationship with the intI1gene. The highest average relative abundances of sul1, sul2, tetX, and intI1 genes were found in south region and oxidation ditch system. Network analysis found that 16 bacterial genera co-occurred with tetW gene. Co-occurrence patterns were revealed distinct community interactions between aerobic/anoxic/aerobic and oxidation ditch systems. The redundancy analysis model plot of the bacterial community composition clearly demonstrated that the sludge samples were significant differences among those from the different geographical areas, and the shifts in bacterial community composition were correlated with ARGs. Together, these findings from the present study will highlight the potential risks of ARGs and bacterial populations carrying these ARGs, and enable the development of suitable technique to control the dissemination of ARGs from WWTPs into aquatic environments.