Aquifer sediments from Norman, Oklahoma, were used to study the potential for microbial reduction of Cr(VI) to Cr(III). Black, clay-like sediments rapidly reduced Cr(VI) in both autoclaved and viable microcosms, indicating an abiotic mechanism. Lightcolored sandy sediments slowly reduced Cr(VI) only in viable microcosms, indicating a biological process. Cr(VI) reduction in these sediments had a pH optimum of 6.8 and temperature optima of 22°C and 50°C. Nearly complete inhibition of Cr(VI) reduction was observed when sandy sediments were shaken in the presence of oxygen. The addition of nitrate but not sulfate, selenate, or ferrous iron to sandy sediments inhibited Cr(VI) reduction. When electron acceptors were supplied in combinations with Cr(VI), reduction of Cr(VI) was greatest in the absence of nitrate. No loss of sulfate and no production of Fe(II) occurred in the presence of Cr(VI). The addition of molybdate to the microcosms did not affect Cr(VI) reduction in sandy sediments until very high concentrations (40 times the Cr[VI] concentration) were used. Interestingly, the addition of bromoethanesulfonic acid in amounts less than, or slightly greater than, the Cr(VI) concentration partially inhibited Cr(VI) reduction in sandy sediments. In the absence of this bacterial inhibitor, the sandy sediments produced methane. A methanogenic enrichment capable of reducing Cr(VI) during growth was obtained from sandy sediments. However, the enrichment produced methane only when Cr(VI) was absent, indicating that a shift in electron flow from methane production to Cr(VI) reduction may have occurred. These studies showed that Cr(VI) reduction in sandy aquifer sediments is a biologically mediated, anaerobic process that is inhibited by oxygen and partially inhibited by nitrate. The lack of sulfate reduction and sulfide production, as well as a lack of inhibition of Cr(VI) reduction by molybdate, argues against an indirect mechanism for Cr(VI) reduction, in which the sulfide produced during sulfate reduction would chemically reduce Cr(VI). Rather, Cr(VI) reduction may be mediated by a community of microorganisms that ordinarily use methanogenesis as the terminal electron-accepting process.