The compositional heterogeneity of biotic assemblages among sites, or β-diversity, regulates the relationship between local and regional species diversity across scales. Recent work has suggested that increased harshness of environmental conditions tends to reduce β-diversity by decreasing the importance of stochastic processes in structuring assemblages. We investigated the effect of nutrient enrichment on the compositional heterogeneity of lake benthic invertebrate assemblages in Ireland at both local (within-lake) and regional (among-lake) scales. At local scales, we found that the compositional heterogeneity of benthic assemblages was related inversely to the extent of nutrient enrichment (as indicated by measurements of water column total phosphorus, total nitrogen, and chlorophyll a), after effects of lake morphology (i.e., surface area, connectivity, and depth of sampling) and alkalinity were accounted for. At regional scales, we found that nutrient-rich lakes had significantly more homogenous benthic assemblages than nutrient-poor lakes, over and above the effect of alkalinity and across a similar range of lake morphologies. These findings have profound implications for global aquatic biodiversity, as the homogenization of benthic assemblages at both local and regional scales may have important and unpredictable effects on whole aquatic ecosystems, with potentially considerable ecological and evolutionary consequences.