AbstractAimBiological communities are the result of a stepwise spatiotemporal filtering process, driven by large‐scale historical and local contemporary determinants. The biogeographical pattern and species richness of North American fishes are predominantly determined by historical processes of past glaciations and postglacial dispersal and by contemporary environmental and ecological processes. We evaluated the effects of postglacial dispersal through glacial Lake Agassiz and habitat heterogeneity, represented by lake surface area, on contemporary freshwater fish species richness patterns of northwestern Ontario lakes.LocationNorthwestern Ontario, Canada.TaxonFreshwater fishes.MethodsApplying the theory of island biogeography and species–area curves, we examined the effects of isolation from the past dispersal corridor of glacial Lake Agassiz and habitat heterogeneity on species richness across 264 contemporary lakes. While controlling for correlations among the predictor variables, generalized linear models were constructed between species richness, as the response variable and the explanatory variables of lake elevation and surface area, and connection to the dispersal corridor.ResultsDifferential cover by glacial Lake Agassiz led to variation in fish species richness across contemporary lakes and it is higher in lakes that were covered by Lake Agassiz relative to basins remaining outside of the boundaries of the glacial lake. Lake surface area is the strongest predictor of species richness, while lake elevation is the strongest factor predicting isolation.Main ConclusionsHabitat heterogeneity and postglacial colonization have led to differences in fish richness within the same geographical region, which increases with lake surface area and decreases with elevation. These are likely driven by greater niche diversity facilitating the assembly of more diverse communities and isostatic rebound and fluctuating levels of Lake Agassiz isolating lakes at high elevations from the dispersal route earlier during the colonization process, respectively. These patterns underscore the importance of incorporating historical and environmental community determinants in biodiversity studies.