Water as a vehicle of antimicrobial resistance (AMR) is a well-known phenomenon. Therefore, One Health inspired surveillance campaign to detect circulation of antimicrobial resistance through the ecosystem are very often based on investigation of water samples. Bacteria belonging to the genera Staphylococcus and Enterococcus are good models in One Health research because of their ubiquitous occurrence and effortless isolation techniques. In this study, we aimed to compare different types of water sources as potential spreaders of AMR staphylococci and enterococci in the ecosystem. For this reason, we took water samples from the Drava River and small waterholes in a forest area. We hypothesised that rivers could collect diverse bacteria and resistance from the catchment area, while forest waterholes gather bacterial contamination from the local wildlife, thus a less diverse resistant bacterial community characterise them. From 22 and 10 water samples, collected from waterholes and the Drava River active floodplain, we isolated 21 and 13 bacterial strains, respectively. The bacterial community of the river proved more diverse, while waterhole samples contained resistant bacteria to more antimicrobials. In waterhole samples, we detected two methicillin resistant coagulase negative Staphylococcus strains. The most conspicuous difference between the two habitats was the predominance of multidrug resistant S. sciuri in the forest waterholes. We could conclude that water could be a good indicator of AMR contamination of the ecosystem, and large-scale sampling could reveal the exact epidemiological role of different water habitats.