Biological N2 fixation hypothetically could balance the N∶P supply ratio in lakes and reservoirs, causing P to be the ultimate limiting element. However, the efficiency and time scale of this mechanism has been poorly studied and remains largely unknown. We compared the relative rates of N supply to the epilimnion of 3 reservoirs of similar age and morphology but with differing external nutrient inputs. Our objective was to determine if N2-fixation rates in these reservoirs exceeded other internal N supply pathways during the growing season, a situation that would suggest that these systems are still evolving in terms of nutrient-limitation status. Phytoplankton N2-fixation rates ranged from 3.4 to 16 g N m−2 y−1 in the reservoirs. These rates are among the highest N2-fixation rates reported in the scientific literature. Measureable N2 fixation also occurred in sediments in contact with reservoir epilimnia (0.85–1.5 g N m−2 y−1). Sediment N2-fixation rates exceeded rates of N regeneration from epilimnetic sediments (0.10–0.24 g N m−2 y−1), a result suggesting that much of this fixed N either accumulates seasonally in sediments or is lost via coupled nitrification–denitrification. NH4 flux from hypolimnia (0.13–1.8 g N m−2 y−1) and lower metalimnia (∼0.00–1.8 g N m−2 y−1) was a substantial internal N supply to epilimnia during summer stratification. However, phytoplankton N2-fixation rates always exceeded rates of N input from these internal sources. These findings suggest that, even though the annual cycle of ecosystem N inputs via N2 fixation has occurred over many years or decades in these reservoirs, reactive N accumulation has not been sufficient to alleviate seasonal N deficiency and lead to perpetual P limitation of phytoplankton.