Flow variability plays an important role in structuring lotic communities, yet comparatively little is known about processes governing assemblage dynamics in stream ecosystems with stable environmental conditions, such as spring-fed rivers. Volcanic spring-fed rivers (hereafter spring-fed rivers) occur in geologically active landscapes of the western USA and around the globe. We sampled invertebrate assemblages and quantified primary productivity and habitat characteristics of spring-fed and runoff rivers in northern California over 4 seasons. We predicted that abiotic factors would be more stable and nutrient availability greater and that invertebrate density would be greater and diversity lower in spring-fed than in runoff rivers. Runoff rivers exhibited high variability in discharge and temperature, whereas spring-fed rivers were relatively stable with high naturally occurring nutrient levels. On average, NO3− and PO43− concentrations were 40× greater in spring-fed than in runoff rivers. Spring-fed rivers supported nearly 7 to 16× greater densities of invertebrates than runoff systems, depending on season. However, invertebrate species richness was greater in runoff rivers in all seasons. Spring-fed river invertebrate assemblages were strongly correlated with elevated nutrient concentrations and basal C sources, whereas runoff assemblages were associated with discharge variability and median substrate size. We suggest that strong differences in abiotic variability between spring-fed and runoff rivers play an important role in determining invertebrate assemblage structure. Because spring-fed rivers exhibit more stable temperatures throughout the year and lower temperatures during the summer than runoff rivers, they may provide essential refugia for coldwater taxa in a warming climate.