Dissolved organic matter (DOM) is the largest pool of organic matter in aquatic ecosystems and is a primary substrate for microbial respiration in streams. However, understanding the controls on DOM processing by microbes remains limited, and DOM decay rates remain largely unconstrained. Many DOM decay rates are quantified with bioassays in dark bottles, which may underestimate DOM decay in streams because these bioassays do not include a benthic zone and do not account for abiotic factors of DOM loss, such as photodegradation and volatilization. We measured decay of labile and semi-labile DOM over 3 d in experimental streams and bottle bioassays. Incubations included 3 types of labile DOM (algal, light-degraded soil, and light-degraded plant leachates) and 2 types of semi-labile DOM (plant and soil leachates). We also quantified decay rates when labile and semi-labile DOM were mixed to test for non-additive effects, or priming, of semi-labile DOM by labile DOM. We converted dissolved organic carbon (DOC) decay rates to half-lives and uptake velocities and compared these metrics to previous studies that quantified DOC loss in bioassays or real streams. Percent DOC lost over time, or biodegradable DOC, was greater in experimental streams than in bioassays. DOC decay rates and uptake velocities did not differ between bioassays and experimental streams but were lower than in real streams. Mixing of labile and semi-labile DOM resulted in both positive and negative non-additive effects. Consistent non-additive effects were difficult to quantify because decay rates were not constant over the course of each incubation, as shown by faster decay rates calculated over the first 6 h of incubation compared to \u3e70 h. Decay rates of leachates from natural substrates (e.g., algae and soil) incubated over short periods of time (hours–days) are needed for models that aim to quantify organic matter transformation in aquatic ecosystems with short residence times, such as rivers and streams