Elevated nitrate (NO3-) concentration in streams and rivers has contributed to environmental problems such as downstream eutrophication and loss of biodiversity. Sycamore Creek in Arizona is nitrogen limited, but previous studies have demonstrated high potential for denitrification, a microbial process in which biologically active NO3- is reduced to relatively inert dinitrogen (N2) gas. Oak Creek is similarly nitrogen limited, but NO3- concentration in reaches surrounded by agriculture can be double that of other reaches. We employed a denitrification enzyme assay (DEA) to compare potential denitrification rate between differing land uses in Oak Creek and measured whole system N2 flux using a membrane inlet mass spectrometer to compare differences in actual denitrification rates at Sycamore and Oak Creek. We anticipated that NO3- would be an important limiting factor for denitrifiers; consequentially, agricultural land use reaches within Oak Creek would have the highest potential denitrification rate. We expected in situ denitrification rate to be higher in Oak Creek than Sycamore Creek due to elevated NO3- concentration, higher discharge, and larger streambed surface area. DEA results are forthcoming, but analysis of potassium chloride (KCl) extraction data showed that there were no significant differences between sites in sediment extractable NO3- on either a dry mass or organic mass basis. Whole-reach denitrification rate was inconclusive in Oak Creek, and though a significant positive flux in N2 from upstream to downstream was measured in Sycamore Creek, the denitrification rate was not significantly different from 0 after accounting for reaeration, suggesting that denitrification does not account for a significant portion of the NO3- uptake in Sycamore Creek. Future work is needed to address the specific factors limiting denitrification in this system.