Loss of ecological integrity due to excessive suspended sediment in rivers and streams is a major cause of water quality impairment in the USA. Current assessment protocols for development of sediment total maximum daily loads (TMDLs) lack a means to link temporally variable sediment transport rates with specific losses of ecological functions as loads increase. In order to accomplish this linkage assessment, a functional traits-based approach was used to correlate site occurrences of 17 fish species traits in three main groups (preferred rearing habitat, trophic feeding guild, and spawning behavior) with suspended sediment transport metrics. The sediment transport metrics included concentrations, durations, and dosages for a range of exceedance frequencies; and mean annual suspended sediment yields (SSY). In addition, this study in the Northwestern Great Plains Ecoregion examined trait relationships with three environmental gradients: channel stability, drainage area, and elevation. Potential stressor responses due to elevated suspended sediment concentration (SSC) levels were correlated with occurrences of five traits: preferred pool habitat; feeding generalists, omnivores, piscivores, and nest-building spawners; and development of ecologically based TMDL targets were demonstrated for specific SSC exceedance frequencies. In addition, reduced site occurrences for preferred pool habitat and nest-building spawners traits were associated with unstable channels and higher SSY. At an ecoregion scale, a functional traits assessment approach provided a means to quantify relations between biological impairment and episodically elevated levels of suspended sediment, supporting efforts to develop ecologically based sediment TMDLs.