Results/Conclusions Our results suggest that human-related impacts at the watershed scale drove patterns of diversity and species richness in more developed landscapes, as represented by our OH streams. In ID, where anthropogenic stressors were markedly less pronounced, regional and reach scale factors, were more important for determining fish assemblage characteristics. Specifically, in both regions (ID and OH), we found that patterns in assemblage richness and diversity were largely driven by variables describing stream size (e.g., drainage area and stream order). Regional factors including mean annual precipitation and latitude, in addition to local factors (e.g., geomorphic characteristics) explained a substantial portion of the variation observed in assemblage richness and diversity. In contrast, watershed-scale variables, especially pertaining to urban and exurban land use, were the predominant predictors of OH fish assemblage characteristics. In conclusion, our results offer evidence that human-induced changes to the physical environment may significantly alter the influence of scale-related factors considered critical in shaping stream fish communities.