The effects of dietary β-glucan, obtained from bacterial fermentation, on the intestinal mass, short chain fatty acids, lactate production and pH in Sprague-Dawley (SD) rats were evaluated. SD rats fed with 0% (control group), 1% or 5% β-glucan supplemented diets (w/w) for 3 weeks. The presence of β-glucan in the diets resulted in a significant increase in colonic contents in a dose dependent manner. The amount of short chain fatty acids increased in rats fed β-glucan diets. Rats fed the 5% β-glucan diets had higher levels of acetate, propionate and butyrate by 1.8, 1.7 and 3.0 fold of the control group in the cecum, and 2.2, 2.9 and 3.1 fold of the control group in the colon, respectively. The β-glucan diets also significantly increased the levels of cecal and colonic lactate by 1.4~3.4 fold, when compared to the control diet, indicating that dietary β-glucan stimulated the growth of lactic acid bacteria within the intestine. These results suggest that dietary β-glucan, by providing short chain fatty acids and reducing the cecal and colonic pH, may be beneficial in improving gut health, and provide evidence for the use of β-glucan as a dietary supplement for human consumption.