Changes in nutrient bioavailability due to increased loading of dissolved organic matter (DOM) may impact boreal freshwaters. Yet, the relative bioavailability of carbon (C), nitrogen (N), and phosphorus (P) associated with terrestrial DOM remains poorly understood. We applied short-term bioassays with natural bacterial inocula to determine seasonal variation in bioavailable organic nutrient pools from four boreal headwater streams in northern Sweden. Experiments were designed to exhaust bioavailable nutrients associated with DOM by inducing limiting conditions when all required resources except for the targeted nutrient (C, N, or P) are provided in excess. We hypothesized that the supply of different bioavailable nutrients to streams would reflect seasonal variations in terrestrial demand, hydrology, and temperature. The delivery of bioavailable DOM-associated resources from the four streams were, on average, 2%, 11%, and 38% of the total dissolved organic C, N, and P, respectively, emphasizing the relatively low C bioavailability in these DOM-rich waters. Bioavailable N : P ratios peaked in autumn for all sites, with lower values in winter and spring. Both in terms of relative (% of total) and absolute bioavailable organic nutrient concentrations, the seasonal pattern was characterized by systematically high values for the autumn period. Furthermore, links between bioavailable resources and temperature and hydrology varied across sites, time periods, and the different elements. Thus, elevated concentrations of bioavailable organic resources in autumn suggest the potential for leaf fall, as well as late season storms that rewet dry soils, to serve as considerable sources of C, N, and P to boreal aquatic ecosystems.