Disassociating the independent effects of flow and water quality on the ecology of flowing waters is an overarching goal in water resource science needed to improve the efficacy of watershed management. However, the interrelatedness of these gradients and their subsequent alteration due to land use change has constrained progress made on this front. The objective of this study was to use benthic macroinvertebrate and fish assemblage data to characterize flow-ecology relationships that were unchanged by water quality impacts across two southeastern landscapes in the USA to help detect ecological change driven by flow alteration. General linear latent models were used to identify taxa that were responsive to high or low flow metrics and water quality gradients. Bayesian hierarchical generalized additive models were then developed using these indicator taxa and three biological metrics to identify flow-specific relationships that were unaffected by water quality impacts. Three low flow-specific relationships were identified, illustrating how potential agricultural or urban impacts to hydrology reduced stream biological health. Importantly, flow-ecology relationships developed using indicator taxa in this study effectively captured hydrology-specific impacts while biological metrics typical of state monitoring and assessment programs did not. Therefore, developing flow-specific biological metrics is a critical step when developing management strategies targeting flow alteration. Implementing standardized frameworks such as the one characterized here can limit contradictory findings and improve streamflow enhancement and restoration project efficacy. These low flow-specific relationships will enhance managers' capacity to develop environmental flow standards, monitor their success, and better understand urban and agricultural impacts on stream assemblages.