RT Journal Article SR Electronic T1 A Scenic River in Rural Ohio Shows Elevated Antibiotic Resistance Genes, Including Acinetobacter Tetracycline and Macrolide Resistance, Downstream of Wastewater Treatment Plant Effluent JF bioRxiv FD Cold Spring Harbor Laboratory SP 2021.04.26.441562 DO 10.1101/2021.04.26.441562 A1 April Murphy A1 Daniel Barich A1 Siobhan Fennessy A1 Joan L. Slonczewski YR 2021 UL http://biorxiv.org/content/early/2021/04/27/2021.04.26.441562.abstract AB Rivers in rural areas receive continual influx of wastewater carrying antibiotics originally administered to humans and livestock. The entry of antibiotic resistance genes (ARGs) into aquatic systems has been documented for large municipal wastewater treatment plants, but there is less study of the impact of smaller plants that are situated on small rural rivers. We sampled water metagenomes for ARG and taxa composition from the Kokosing State Scenic River, a small rural river in Knox County, Ohio. Samples were obtained upstream, a few meters downstream, and 6 km downstream from the effluent release of the Mount Vernon wastewater treatment plant (WWTP). Through all seasons, the metagenome just downstream of the WWTP effluent showed a substantial elevation of at least 15 different ARGs, including 6 ARGs from Acinetobacter baumannii such as msrE, mphE (macrolide resistance) and tet(39) (tetracycline resistance). The ARGs most prevalent near the effluent pipe persisted 6 km downstream. The taxa distribution near the effluent showed elevation of Acinetobacter species as well as gut-associated taxa, Bacteroides and Firmicutes. The ARG levels and taxa prevalence showed little dependence on seasonal chlorination of effluent. Nitrogen and phosphorus were elevated near the effluent pipe but had no consistent effect on ARG levels. We show that in a rural river microbiome, year-round wastewater effluent substantially elevates ARGs including those of multidrug-resistant A. baumanii.IMPORTANCE Antibiotic resistance is a growing problem worldwide, with frequent transmission between pathogens and environmental organisms. Rural rivers support recreational use by people unaware of inputs from treated wastewater. In our study, the river water proximal to wastewater effluent shows increased prevalence of multidrug-resistant Acinetobacter baumanii, an opportunistic pathogen of concern for hospitals but also widespread in natural environments. Our work highlights the importance of wastewater effluent in management of environmental antibiotic resistance.