Nutrient enrichment threatens river ecosystem health in urban watersheds, but the influence of urbanization on spatial variation in nutrient concentrations and nutrient limitation of biofilm activity are infrequently measured simultaneously. In summer 2009, we used synoptic sampling to measure spatial patterns of nitrate (NO 3 -), ammonium (NH 4 ? ), and soluble reactive phosphorus (SRP) concentration, flux, and instantaneous yield throughout the Bronx River watershed within New York City and adjacent suburbs. We also quantified biofilm response to addition of NO 3 -, phosphate (PO 4 3-), and NO 3 -? PO 4 3-on organic and inorganic surfaces in the river mainstem and tributaries. Longitudinal variation in NO 3 -was low and related to impervious surface cover across sub-watersheds, but spatial variation in NH 4 ? and SRP was higher and unrelated to sub-watershed land-use. Biofilm respiration on organic surfaces was frequently limited by PO 4 3-or NO 3 -? PO 4 3-, while primary production on organic and inorganic surfaces was nutrientlimited at just one site. Infrequent NO 3 -limitation and low spatial variability of NO 3 -throughout the watershed suggested saturation of biological N demand. For P, both higher biological demand and point-sources contributed to greater spatial variability. Finally, a comparison of our data to synoptic studies of forested, temperate watersheds showed lower spatial variation of N and P in urban watersheds. Reduced spatial variation in nutrients as a result of biological saturation may represent an overlooked effect of urbanization on watershed ecology, and may influence urban stream biota and downstream environments.