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AbstractFreshwater biodiversity is increasingly at risk wherever land uses such as agriculture exert multiple stressors that degrade habitat quality. Although stream macroinvertebrates act as bioindicators for monitoring these impacts, their responses are context-specific: examining drivers of community composition is therefore important to understand the results of monitoring efforts. In a primarily agricultural landscape, 15 sites across the Sydenham River watershed, Ontario, Canada, were assessed for in-stream habitat quality and stream macroinvertebrate diversity. We predicted that community assemblage would be driven by differences in surficial geology across branch (east branch versusnorth branch) and catchment position (main stem versustributary). We found that the main stem of the east branch was characterised by significantly higher proportions (P= 0.053) and abundance (P= 0.038) of Ephemeroptera–Plecoptera–Trichoptera (EPT) taxa than north branch sites were, and sites in the east and north branch tributaries were characterised by significantly lower Hilsenhoff Biotic Index (HBI) scores (P= 0.088). Redundancy analysis found that substrate size was the main driver of community composition, generating a model that described these patterns across branch and catchment position. Our findings suggest that EPT abundance and HBI scores were the variables that were most effective at revealing differences in stream communities due to agricultural impacts and that sediment size is an important driver of these patterns.