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The rate and mechanism of nitrate removal along and between groundwater flow paths were investigated using a series of well nests screened in an unconfined sand and gravel aquifer. Intensive agricultural activity in this area has resulted in nitrate concentrations in groundwater often exceeding drinking water standards. Both the extent and rate of denitrification varied depending on the groundwater flow path. While little or no denitrification occurred in much of the upland portions of the aquifer, a gradual redox gradient is observed as aerobic upland groundwater moves deeper in the aquifer. In contrast, a sharp shallow redox gradient is observed adjacent to a third-order stream as aerobic groundwater enters reduced sediments. An essentially complete loss of nitrate concurrent with increases in excess N2 provide evidence that denitrification occurs as groundwater enters this zone. Electron and mass balance calculations suggest that iron sulfide (e.g., pyrite) oxidation is the primary source of electrons for denitrification. Denitrification rate estimates were based on mass balance calculations using nitrate and excess N2 coupled with groundwater travel times. Travel times were determined using a groundwater flow model and were constrained by chlorofluorocarbon-based age dates. Denitrification rates were found to vary considerably between the two areas where denitrification occurs. Denitrification rates in the deep, upland portions of the aquifer were found to range from <0.01 to 0.14 mM of N per year; rates at the redoxcline along the shallow flow path range from 1.0 to 2.7 mM of N per year. Potential denitrification rates in groundwater adjacent to the stream may be much faster, with rates up to 140 mM per year based on an in situ experiment conducted in this zone.