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Hydrological dynamics drive the transition of antibiotic resistance genes between particle-attached and free-living lifestyles in a deep freshwater reservoir

Despite the growing awareness of antibiotic resistance genes (ARGs) spreading in the environment, there is a knowledge gap on the fate and transport of ARGs in particle-attached (PA) and free-living (FL) lifestyles in deep freshwater ecosystems experiencing seasonal hydrological changes. Here, we examined the ARG profiles using high-throughput quantitative PCR in PA and FL lifestyles at four seasons representing two hydrological seasons (i.e., vertical mixing and thermal stratification) in the Shuikou Reservoir (SR), Southern China. The results indicated that seasonal hydrological dynamics were critical for influencing ARGs in PA and FL fractions, and the transition of ARGs between the two lifestyles. Although both PA and FL ARG profiles were likely to be shaped by horizontal gene transfer, PA and FL ARGs had different responses to the changes in physico-chemicals (e.g., nutrients and dissolved oxygen) caused by seasonal hydrological dynamics. The particle-associated niche (PAN) index revealed that there were 94 non-conservative ARGs (i.e., no preferences for PA and FL), 23 conservative ARGs that preferred PA lifestyle, and 16 conservative ARGs for FL lifestyle. A sharp decline in the number of conservative ARGs in stratified seasons suggests a hydrological dynamics-dependent transition of ARGs between two lifestyles. Remarkably, the conservative ARGs (in PA or FL lifestyle) were more closely related to bacterial OTUs in their preferred lifestyle compared to their counterpart lifestyle, suggesting a lifestyle-dependent ARG enrichment. Altogether, these findings enhance our understanding of the role of seasonal hydrological changes in the dissemination of ARGs in different size fractions in deep aquatic ecosystems.