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Stream metabolism is affected by both natural and human-induced processes. While metabolism has multiple implications for ecological processes, relatively little is known about how metabolic rates are influenced by land use in tropical streams. In this study, we assessed the metabolic characteristics and related environmental factors of six streams located in a transition area from Cerrado to Atlantic Forest (São Carlos/Brazil). Three streams were relatively preserved, while three were flowing through more agriculturally and/or urban impacted watersheds. Surface water samples were analyzed for biological and physico-chemical parameters as well as discharge and percentage of canopy cover. Metabolism was determined through the single-station method to estimate gross primary production (GPP), ecosystem respiration (ER) and net ecosystem production (NEP) with BAyesian Single-station Estimation (BASE). Nutrient concentrations tended to be higher in impacted versus preserved streams (e.g., average total phosphorus between 0.028-0.042 mg L-1 and 0.009-0.038 mg L-1, respectively). Average canopy cover varied between 58 and 77%, with no significant spatial or seasonal variation. All streams were net heterotrophic (ER exceeded GPP) in all sampling periods. GPP rates were always lower than 0.7 gO2 m-2 d-1 in all streams and ER varied from 0.6 to 42.1 gO2 m-2 d-1. Linear Mixed-Effect models showed that depth, discharge, velocity and total phosphorus are the most important predictors for GPP. For ER, depth, velocity and canopy cover are significant potential predictors. Canopy cover was the main light limiting factor and influenced stream metabolism. Our findings reinforced the concepts that shifts in the shading effect provided by vegetation (e.g., through deforestation) or changes in discharge (e.g., through land use conversion or water abstractions) can impact freshwater metabolism. Our study suggests that human activities in low latitude areas can alter tropical streams’ water quality, ecosystem function, and the degree of riparian influence. Our data showed that tropical streams can be especially responsive to increases of organic matter inputs leading to high respiration rates and net heterotrophy, and this should be considered to support management and restoration efforts.