Denitrifying activity in a sediment from the freshwater part of a polluted estuary in northwest Europe was quantified using two independent approaches. High-resolution N 2 O microprofiles were recorded in sediment cores to which acetylene was added to the overlying water and injected laterally into the sediment. The vertical distribution of the rate of denitrification supported by nitrate uptake from the overlying water was then derived from the time series N 2 O concentration profiles. The rates obtained for the core incubations were compared to the rates predicted by a forward reactive transport model, which included rate expression for denitrification calibrated with potential rate measurements obtained in flowthrough reactors containing undisturbed, 1-cmthick sediment slices. The two approaches yielded comparable rate profiles, with a near-surface, 2-to 3-mm narrow zone of denitrification and maximum in situ rates on the order of 200 to 300 nmol cm ؊3 h ؊1 . The maximum in situ rates were about twofold lower than the maximum potential rate for the 0-to 1-cm depth interval of the sediment, indicating that in situ denitrification was nitrate limited. The experimentally and model-derived rates of denitrification implied that there was nitrate uptake by the sediment at a rate that was on the order of 50 (؎ 10) nmol cm ؊2 h ؊1 , which agreed well with direct nitrate flux measurements for core incubations. Reactive transport model calculations showed that benthic uptake of nitrate at the site is particularly sensitive to the nitrate concentration in the overlying water and the maximum potential rate of denitrification in the sediment.Anthropogenic inputs of nitrogen are a major cause of eutrophication in aquatic environments (39). Denitrification, the bacterial dissimilatory reduction of nitrate to gaseous products, counteracts eutrophication by removing inorganic nitrogen and releasing it to the atmosphere and by decomposing organic matter (34). The emissions resulting from denitrification, however, include nitrous oxide, a powerful greenhouse gas (8). Because of its environmental importance, denitrification has been the subject of numerous studies (for reviews, see references 12, 16, and 34). Oxygen and nitrate concentrations, temperature, pH, and the availability of suitable electron donor substrates (mainly organic carbon compounds) are considered the key factors controlling the occurrence and rate of denitrification (6,22,38).In estuarine and coastal environments, the most intense denitrification often occurs in sediments, where reduced solute transport rates enable the establishment of anaerobic conditions. A number of methods have been developed to quantify denitrification and related nitrogen transformations in sediments; these methods include, among others, benthic flux measurement (17), isotope pairing techniques (32), and microprofiling using nitrous oxide (31) or nitrate microsensors (9,20). The available data show that there are large spatial and temporal variations in denitrification rates in nearsho...