Very little is known about the growth physiology and metabolic niche of the human oral isolate Selenomonas sputigena. The objective of this study was to devise a minimal medium for comparing growth rates and fermentation of rumen Selenomonas ruminantium strains with S. sputigena. When anaerobically grown on a minimal glucose medium containing yeast extract as the only chemically undefined component, S. sputigena produced acetate, propionate, and succinate while S. ruminantium strains produced primarily lactate. When strains were compared (P < 0.05) for each carbon source that yielded growth, rumen strain HD4 grew faster than all other strains on glucose, cellobiose and glycerol while strain GA192 grew faster on trehalose. Rumen strains GA192, PC18, and HD4 grew faster on mannitol than rumen strains D and GA31. S. sputigena grew faster on lactate (0.38 ± 0.04) than any of the S. ruminantium strains. The minimal medium developed in this study should be useful for jurmer physiological studies on fermentation and metabolism in S. sputigena.