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Local site-scale factors explained more variation than watershed-scale factors. Random forest and multiple regression models performed similarly, and the fish MMI-environment relationships were stronger than macroinvertebrate MMI-environment relationships. Among ecoregions, the strongest environmental relationships were observed in the Northern Appalachians and the weakest in the Southern Plains. The fish and macroinvertebrate MMIs were only weakly correlated with each other, and they generally responded more strongly to different groups