Surface waters are susceptible to contamination as they are exposed to environmental events, such as storms and rainfall, which bring in surface runoffs from surrounding animal farms, wildlife, infrastructure, and sewage discharges into waterways. This study was initiated in order to investigate the prevalence and diversity of pathogenic and commensal bacteria, namely Salmonella, E. coli, and Enterococcus, present in surface water of the Upper Oconee watershed, GA and provide a basic understanding of the occurrence and persistence of antimicrobial resistance (AR) in these bacteria. Bacteria of interest were isolated and identified to species (Enterococcus), serotype (Salmonella), and phylogroup (E. coli) levels and characterized using serological and molecular methods. Upon identifying AR phenotypes, AR bacteria were tested for the presence of AR genes and mobile genetic elements (MGEs) responsible for the AR phenotypes using molecular tools, such as PCR and sequencing. Isolates that exhibited multidrug resistance and novel AR phenotypes were whole genome sequenced for an in-depth study for the characterization of the vehicles for the dissemination of AR genes. The results of this study showed that surface waters are highly contaminated with Salmonella, E. coli, and Enterococcus, a substantial portion of which is human-associated. Moreover, AR bacteria are widely distributed within the watershed, and both persisting and emerging AR strains, including resistance to newer drugs, have been detected. The continuing spread of AR throughout the watershed is of growing environmental and public health concern. The surveillance of the prevalence and diversity of bacteria in environmental water is needed not only to monitor the bacterial contamination of the water that is used for recreational activities, irrigation, and drinking, but also to enable the tracking of the sources and fates of the bacteria. Furthermore, the role of surface water in the emergence and spread of AR needs to be established to completely understand the AR development and its impact on human health.