The objective of this study was to assess the effect of live yeast (Saccharomyces cerevisiae) supplementation on the populations of specific cellulolytic (Fibrobacter succinogenes and Ruminococcus flavefaciens) and saccharolytic (Streptococcus equinus and Streptococcus bovis) bacteria in the feces of horses fed high-starch and high-fiber diets. Four horses were each fed diets consisting of high fiber with no yeast (HF), high fiber with yeast (HFY), high starch with no yeast (HS), and high starch with yeast (HSY) in a 4 × 4 Latin-square design study. Fresh fecal samples were collected on the last 3 days of each 31-day experimental period and were then assessed, using semiquantitative real-time PCR, for total bacterial load and levels of target bacterial species, relative to the total bacterial load. The most abundant of the target species was F. succinogenes, and the HSY diet resulted in a significant (P = .045) reduction in relative levels of this bacterium. No significant effect (P = .224) of diet was observed in relation to abundance of R. flavefaciens. Results show that diet did not have a significant (P = .068) effect on relative quantities of S. equinus, although there appeared to be a trend for increased levels of this bacterium during feeding of high-starch diets. Numbers of S. bovis were higher (P < .001) when horses were fed HS and HSY diets than when fed the HF and HFY diets. Significant variation in levels of S. equinus (P = .024) and S. bovis (P = .049) was observed between individual horses.