Fate of antibiotic resistance genes and their associations with bacterial community in livestock breeding wastewater and its receiving river water

Large amounts of antibiotics are currently used in livestock breeding, which is the main driving factor contributing to the occurrence, spread and proliferation of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in the environment. In this study, high-throughput sequencing based metagenomic approaches were employed to characterize the tempo-spacial changes of antibiotic resistome, bacterial community and their correlations in pig farming wastewater and its receiving river. A total of 194 ARG subtypes within 14 ARG types were detectable in all the samples, and their total relative abundance increased in the river water after receiving wastewater discharge, while decreased in the downstream river water. Network analysis showed that 25.26% ARGs within the same type or among the different types showed higher incidences of non-random co-occurrence. The wastewater discharge evidently increased bacterial diversity and induced bacterial community shift in the receiving river water. The genera of Treponema, Prevotella, Pseudomonas, Bacteroides, Oscillibacter and Acholeplasma dominated in the wastewater samples and almost disappeared in the receiving river water, but bacterial pathogens Clostridium difficile and Arcobacter butzleri still occurred in the receiving water. Correlation analysis and host analysis consistently showed that the changes in the abundances of several key genera like Prevotella and Treponema were significantly and positively correlated with the antibiotic resistome alteration. Variation partitioning analysis indicated that bacterial community played a more important role in the resistome alteration than mobile genetic elements. This study may help to understand the correlations among antibiotic resistome, microbiota and environmental conditions in the wastewater-receiving river water.