Some organic solvents are extremely toxic to living organisms by virtue of their ability to partition into and disrupt the normal functioning of biological membranes. In recent years, several bacteria have been discovered that are more tolerant to these toxic solvents than most microorganisms. Using enrichment procedures, we have isolated new organic solvent-tolerant bacteria from both hyrdocarbon-contaminated and pristine soil samples. These organisms were characterized by several different experimental procedures including description of their cellular physiology, 16S rDNA homology, organic solvent tolerance range, and survival after solvent exposure. The results indicate that gram-positive bacteria can be isolated from the environment that are as tolerant to toxic organic solvents, if not more so, than the most organic solvent-tolerant gram-negative bacteria.