To assess the effects of urbanization on assemblages (fish, invertebrate, and algal), physical habitat, and water chemistry, we investigated the relations among varying intensities of basin urbanization and stream ecology in three metropolitan areas: the humid northeastern United States around Boston, Massachusetts; the humid southeastern United States around Birmingham, Alabama; and the semiarid western United States around Salt Lake City, Utah. A consistent process was used to develop a multimetric urban intensity index (UII) based on locally important variables (land-use/land-cover, infrastructure, and socioeconomic variables) in each study area and a common urban intensity index (CUII) based on a subset of five variables common to all study areas. The UII was used to characterize 30 basins along an urban gradient in each metropolitan area. Study basins were located within a single ecoregion in each of the metropolitan areas. The UII, ecoregions, and site characteristics provided a method for limiting the variability of natural landscape characteristics while assessing the magnitude of urban effects. Conditions in Salt Lake City (semiarid climate and water diversions) and Birmingham (topography) required nesting sites within the same basin. The UII and CUII facilitated comparisons of aquatic assemblages response to urbanization across different environmental settings. ?? 2005 by the American Fisheries Society.