Nitrate uptake may be improved in regulated floodplain rivers by increasing hydrological connectivity to backwaters. We examined summer nitrate uptake in a series of morphologically similar backwaters on the Upper Mississippi River receiving flow-regulated nitrate loads via gated culverts. Flows into individual backwaters were held constant over a summer period but varied in the summers of 2003 and 2004 to provide a range of hydraulic loads and residence times (??). The objectives were to determine optimum loading and ?? for maximum summer uptake. Higher flow adjustment led to increased loading but lower ?? and contact time for uptake. For highest flows, ?? was less than 1 day resulting in lower uptake rates (Unet, < 300 mg m-2 day-1), low uptake efficiency (U% < 20%) and a long uptake length (Snet > 4000 m). For low flows, ?? was greater than 5 days and U% approached 100%, but Unet was 200 mg m-2 day-1. Snet was < half the length of the backwaters under these conditions indicating that most of the load was assimilated in the upper reaches, leading to limited delivery to lower portions. Unet was maximal (384-629 mg m-2 day-1) for intermediate flows and ?? ranging between 1 and 1.5 days. Longer Snet (2000-4000 m) and lower U% (20-40%) reflected limitation of uptake in upper reaches by contact time, leading to transport to lower reaches for additional uptake. Uptake by ???10 000 ha of reconnected backwaters along the Upper Mississippi River (13% of the total backwater surface area) at a Unet of ???630 mg m-2 day-1 would be the equivalent of ???40% of the summer nitrate load (155 mg day-1) discharged from Lock and Dam 4. These results indicate that backwater nitrate uptake can play an important role in reducing nitrate loading to the Gulf of Mexico. Copyright ?? 2008 John Wiley & Sons, Ltd.