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Our study aimed to determine drivers of longitudinal variation in stream metabolism and benthic macroinvertebrate (BMI) composition and assess concordance of these ecological measures for two Canadian rivers (Rat River and Tobacco Creek). Gross primary production was associated with longitudinal position in both rivers but also with the percentage of the watershed used for agriculture and hydrogeomorphic zone. However, within- and among-zone differences in stream metabolism indicated that longitudinal variation followed a staircase pattern rather than a clinal pattern. BMI composition was associated with network position in both rivers, but hydrogeomorphic zones were only important in Tobacco Creek. Among-zone differences in BMI communities in Tobacco Creek depended on season. Concordance between stream metabolism and BMI composition was not observed within either river despite metabolism and BMI composition being associated with longitudinal position. For these rivers, segment-scale hydrogeomorphic conditions appear to be important modifiers of longitudinal patterns observed at the whole river scale. The lack of concordance between stream metabolism and BMI composition suggests reach-scale processes are driving ecological differences within sampling sites. [ABSTRACT FROM AUTHOR]