The present study examined the mode of action of a patented Saccharomyces cerevisiae yeast hydrolysate product (YHP) on the fermentation of bovine rumen in vitro. Three experiments were conducted. Fresh fluid from rumen-cannulated dairy cows was used as an inoculum to ferment a mixture of grass silage and concentrate feed with or without YHP. The first two experiments were batch fermentations of 12–24 h duration while the third experiment was a semi-continuous fermentation of six days. Production of gas, concentration of short chain fatty acids (SCFAs), microbial cell density and pH were measured from the fermentation medium as a function of time. In experiment 1, YHP dose-dependently stimulated the production of gas, and increased the density of microbial cells and concentration of SCFAs. Experiment 2 studied the effect of YHP on the ruminal fermentation using three ratios of concentrate feed to grass silage (25:75, 50:50, and 75:25). Both YHP and the elevated proportion of concentrate in the feed mixture significantly increased the production of gas, microbial populations and SCFAs, including propionic acid, by the ruminal microbiota. In experiment 3, YHP increased the concentration and relative proportion of propionic acid in the fermentation medium. YHP stimulated the rate of microbial fermentation of bovine ruminal microbiota, indicated by the effects on gas and SCFA production and microbial mass in these experiments. In particular, YHP increased the production of propionic acid. These results, which were likely due to modulation of microbial community by YHP, suggest that YHP enhances bovine ruminal fermentation and may thus improve the performance of these animals.