In this study, we examine the biological processes involved in ammonia and nitrate removal in a bioretention system characterized by low infiltration rates and long drainage times. The system removed 33% of influent nitrate and 56% of influent total nitrogen. While influent ammonia concentrations were low (<0.3 mg/L), the bioretention cell also removed ammonia produced within the treatment system. Soil cores collected from the bioretention cell were analyzed for total 16S rDNA and both nitrification and denitrification genes (amoA, nirS, nirK, norB, and nosZ) using quantitative PCR. Total bacterial 16S rDNA levels in the surface layer were similar to those in very sandy soils. Gene counts for both nitrification and denitrification genes decreased as a function of depth in the media, and corresponded to similar changes in total 16S rDNA. The abundance of denitrification genes was also positively correlated with the average inundation time at each sampling location, as determined by modeling of stormwater data from a three-year period. These results suggest that both nitrification and denitrification can occur in bioretention media. Time of saturation, filter medium, and organic carbon content can all affect the extent of denitrification in bioretention systems.