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Abstract River metabolism and, thus, carbon cycling are governed by gross primary production and ecosystem respiration. Traditionally river metabolism is derived from diel dissolved oxygen concentrations, which cannot resolve diel changes in ecosystem respiration. Here, we compare river metabolism derived from oxygen concentrations with estimates from stable oxygen isotope signatures (δ 18 O 2 ) from 14 sites in rivers across three biomes using Bayesian inverse modeling. We find isotopically derived ecosystem respiration was greater in the day than night for all rivers (maximum change of 113 g O 2  m −2  d −1 , minimum of 1 g O 2  m −2  d −1 ). Temperature (20 °C) normalized rates of ecosystem respiration and gross primary production were 1.1 to 87 and 1.5 to 22-fold higher when derived from oxygen isotope data compared to concentration data. Through accounting for diel variation in ecosystem respiration, our isotopically-derived rates suggest that ecosystem respiration and microbial carbon cycling in rivers is more rapid than predicted by traditional methods.