At the request of the U.S. Bureau of Land Management, we performed a study during April–July 2010 to characterize mercury (Hg), monomethyl mercury (MMeHg), and other geochemical constituents in sediment, water, and biota at the Clyde Gold Mine and the Elgin Mercury Mine, located in neighboring subwatersheds of Sulphur Creek, Colusa County, California. This study was in support of a Comprehensive Environmental Response, Compensation, and Liability Act - Removal Site Investigation. The investigation was in response to an abatement notification from the California Central Valley Regional Water Quality Control Board to evaluate the release of Hg from the Clyde and Elgin mines. Samples of water, sediment, and biota (aquatic macroinvertebrates) were collected from sites upstream and downstream from the two mine sites to evaluate the level of Hg contamination contributed by each mine to the aquatic ecosystem. Physical parameters, as well as dissolved organic carbon, total Hg (HgT), and MMeHg were analyzed in water and sediment. Other relevant geochemical constituents were analyzed in sediment, filtered water, and unfiltered water. Samples of aquatic macroinvertebrates from each mine were analyzed for HgT and MMeHg. The presence of low to moderate concentrations of HgT and MMeHg in water, sediment, and biota from the Freshwater Branch of Sulphur Creek, and the lack of significant increases in these concentrations downstream from the Clyde Mine indicated that this mine is not a significant source of Hg to the watershed during low flow conditions. Although concentrations of HgT and MMeHg were generally higher in samples of sediment and water from the Elgin Mine compared to the Clyde Mine, concentrations in comparable biota from the two mine areas were similar. It is likely that highly saline effluent from nearby hot springs contribute more Hg to the West Fork of Sulphur Creek than the mine waste material at the Elgin Mine.