Dissolution of the Silurian-Devonian aquifer in the Lake Huron Basin has produced several karst formations in the bedrock (sinkholes), through which groundwater emerges onto the lake floor. During September 2003, we explored a recently discovered submerged sinkhole ecosystem (55 m × 40 m × ˜1 m) located at a depth of 93 m with a remotely operated vehicle (ROV) equipped with a conductivity-temperature-depth (CTD) system, an acoustic navigational system, a video camera, and a water sampling system. In addition to two morphotypes of benthic mats, a 1-2 m thick visibly cloudy near-bottom nepheloid-like layer (sinkhole plume) with a strong hydrogen sulfide odor prevailed just above the seepage area of clear water. Relative to lake water, water samples collected within the sinkhole plume were characterized by slightly higher (by 4°C) temperatures, very high levels of chloride (up to 175 mg l−1) and conductivity (1,700 μS cm−1), as well as extremely high concentrations of sulfate (1,400 mg l−1), phosphorus (3 mg l−1) and particulate organic matter (400 mg C l−1). Compared to background lake water, sinkhole plume water was characterized by approximately twofold lower C:N ratios and tenfold higher levels of dissolved organic carbon, bacterial biomass as well as heterotrophic bacterial production. Significant uptake of 14C-bicarbonate in dark incubations provided preliminary evidence for occurrence of chemosynthesis, possibly mediated by specialized Bacteria and Archea present in this submerged sinkhole ecosystem in the Laurentian Great Lakes.