Empirical assessments of the type and strength of stream fish habitat associations can advance understanding of functional diversity and promote effective conservation.

The ability to accurately quantify biodiversity is fundamental to understanding ecological trends, identifying drivers of declines, and selecting effective conservation options. Scientists and resource managers have grappled with what metrics best show relevant biodiversity patterns and are still practical enough to aid on-the-ground resource conservation. Our purpose is to construct empirically derived, functional habitat guilds for prairie stream fish, then recommend future directions for constructing and using diversity metrics that aid field-based conservation. Working in the Upper Neosho River, KS, USA, we used univariate methods, cluster analysis, non-metric multi-dimensional scaling, and an analysis of similarity to functionally group stream fish taxa. The 11 most abundant fish species grouped into seven ecological guilds: riffle specialist, pool specialist, riffle generalist, pool generalist, riffle–run generalist, pool–run generalist, and generalist. Combining the habitat type and strength of association added ecological accuracy to our species groups. Employing multiple statistical methods increased confidence and generality in our grouping results. Moving forward will require a coordinated, coalition-driven, conservation-related strategy on which researchers and practitioners collaborate to synthesize diverse empirical results, organize general principles of structure and function, and balance accuracy with practicality.