The environmental DNA (eDNA) method is being increasingly applied in various environments. Although eDNA undergoes rapid degradation in aqueous environments, it has been detected in streams up to 10 km downstream from its source. As environmental bacteria can uptake free DNA, transfer their genetic traits, and amplify, there is a potential risk that they, rather than a target aquatic species, could become a source of measured eDNA. This study examined whether bacteria with incorporated fish DNA could be such a source by investigating the detectability of fish DNA generated by bacteria inhabiting river water and riverbed sediment. We attempted to detect common carp (Cyprinus carpio) eDNA in stream water and sediment samples and the DNA of common carp produced by bacterial colonies (Escherichia coli, total coliform, and heterotrophic bacteria) cultured from the samples. The eDNA was detected in the environmental samples but the carp DNA from the targeted bacteria was rarely detected in both water and riverbed sediment samples. Our results suggest that the risk of bacterium-induced false positive detection for fish eDNA is negligible.