Measures of species diversity are valuable tools for assessing ecosystem health. However, most assessments have addressed individual sites or regional taxon pools, with few comparisons of differences in assemblage composition within or among regions. We examined the effects of anthropogenic disturbance on local richness (α diversity) and species turnover (β diversity) of benthic macroinvertebrates in small streams within and between 2 ecoregions (Northern Piedmont vs Southeastern Plains ecoregions) of the Patuxent River basin (Maryland, USA). Regional species pools did not differ between ecoregions (Piedmont  =  166 taxa, Plains  =  162 taxa); however, local richness was lower in the Plains (mean  =  17.4 taxa/stream) compared to the Piedmont (mean  =  22.2 taxa/stream). When streams were categorized into disturbance classes (low, medium, high), local richness did not differ among categories for either region. However, at the entire Patuxent scale, local richness tended to decrease with % impervious cover in a watershed. Variation in species composition, analyzed with nonmetric multidimensional scaling (nMDS), differed significantly between Piedmont and Plains streams, and Plains streams had higher β diversity than Piedmont streams. When partitioned by disturbance category and region, β diversity differed only between the low-disturbance sites (Plains > Piedmont). Relationships between β diversity and environmental variables varied by region. β diversity was weakly negatively related to % row-crop cover in a watershed at the entire Patuxent scale. For the Piedmont region, β diversity tended to decrease with % forest, % pasture, and % row-crop cover in a watershed. Such negative relationships between β diversity and landuse variables indicate a possible homogenization of the assemblage. The incongruence between diversity measures and composition measures, together with differing effects of anthropogenic land use on β diversity in the 2 regions, emphasizes the need to incorporate both α and β diversity and regional environmental factors in conservation/land management studies.