The Homestake gold mine in Lead, South Dakota, USA is rich in sulfur and sulfur compounds, as well as iron and a variety of metals. We wondered if the bacterial genus Shewanella could be isolated from mine materials, since this genus uses a variety of metals and elemental sulfur as terminal electron acceptor under anaerobic conditions. Using anaerobic reduction of elemental sulfur as the screening tool, we obtained three facultatively anaerobic isolates under mesophilic conditions that appeared to reduce elemental sulfur anaerobically upon repeated subculture. However, none appeared to be Shewanella based on partial small subunit rDNA sequencing. One was a Pseudomonas fluorescens biotype G ; while the other two were Bacillus strains. All three strains could grow at 27 oC and 37 oC, but not at 50 oC or 70 oC. The P. fluorescens strain produced an iron-binding siderophore, while the Bacillus strains did not. The two Bacillus strains produced carboxymethylcellulase, but the pseudomonad did not. The extent of the ability of the strains to use other external electron acceptors and to decompose lignocellulosic materials is being examined further.