Occurrence and Prevalence of Antimicrobial Resistance in Urban Karst Groundwater Systems Based on Targeted Resistome Analysis

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Antimicrobial resistance (AMR) is a global crisis threatening human, animal, and environmental health. The natural environment, specifically water resources, has been recognized as a reservoir and dissemination pathway for AMR; however, urban karst aquifer systems have been overlooked. This is a concern as these aquifer systems provide about 10% of the global population with drinking water; yet, the urban influence on the resistome in these vulnerable aquifers is often overlooked. This study used high-throughput qPCR to determine the occurrence and relative abundance of urban-related antimicrobial resistant genes (ARG) in a developing urban karst groundwater system in Bowling Green, KY. Ten sites throughout the city were sampled weekly and analyzed for 85 ARGs, as well as seven microbial source tracking (MST) genes for human and animal sources, providing a spatiotemporal understanding of the resistome in urban karst groundwater. To further understand AMR in this environment, potential drivers (landuse, karst feature type, season, source of fecal pollution) were considered in relation to the resistome relative abundance. The MST markers highlighted a prominent human influence to the resistome in this setting. The concentration of targeted genes varied between the sample weeks, but all targeted ARGs were prevalent throughout the aquifer regardless of karst feature type or season, with high concentrations captured for sulfonamide (sul1), quaternary ammonium compound (qacE), and aminoglycoside (strB) antimicrobial classes. Higher prevalence and relative abundance were detected during the summer and fall seasons, as well as at the spring features. Linear discriminant analysis suggested that karst feature type had higher influence on AMR compared to season and the source of fecal pollution had the least influence. This study is one of the first to define an urban karst groundwater resistome. These findings can contribute to the development of effective management and mitigation strategies for AMR.



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