Physical, chemical, hydrologic, and biologic factors affecting nitrate (NO3 −) removal were evaluated in three agricultural streams draining orchard/dairy and row crop settings. Using 3-d “snapshots” during biotically active periods, we estimated reach-level NO3 − sources, NO3 − mass balance, in-stream processing (nitrification, denitrification, and NO3 − uptake), and NO3 − retention potential associated with surface water transport and ground water discharge. Ground water contributed 5 to 11% to stream discharge along the study reaches and 8 to 42% of gross NO3 − input. Streambed processes potentially reduced 45 to 75% of ground water NO3 − before discharge to surface water. In all streams, transient storage was of little importance for surface water NO3 − retention. Estimated nitrification (1.6–4.4 mg N m−2 h−1) and unamended denitrification rates (2.0–16.3 mg N m−2 h−1) in sediment slurries were high relative to pristine streams. Denitrification of NO3 − was largely independent of nitrification because both stream and ground water were sources of NO3 − Unamended denitrification rates extrapolated to the reach-scale accounted for <5% of NO3 − exported from the reaches minimally reducing downstream loads. Nitrate retention as a percentage of gross NO3 − inputs was >30% in an organic-poor, autotrophic stream with the lowest denitrification potentials and highest benthic chlorophyll a, photosynthesis/respiration ratio, pH, dissolved oxygen, and diurnal NO3 − variation. Biotic processing potentially removed 75% of ground water NO3 − at this site, suggesting an important role for photosynthetic assimilation of ground water NO3 − relative to subsurface denitrification as water passed directly through benthic diatom beds.