β-Glucans obtained from barley, seaweed, bacteria, and mushroom sclerotia were incubated with pure cultures of Bifidobacterium infantis, Bifidobacterium longum, and Bifidobacterium adolescentis for a 24 h batch fermentation to evaluate their bifidogenic effect with inulin as the positive control. The pH value in all culture media was decreased by 0.5-1.5 units. All β-glucans supported the growth of the three bifidobacteria with B. infantis, having a relatively larger increase in populations (3-4 log(10) colony forming units). B. infantis produced almost double the amount of total short-chain fatty acids (SCFAs) than the other two bifidobacteria. The SCFA profile of B. infantis had a relatively higher proportion of propionic and butyric acid but less acetic acid than the other bifidobacteria. The utilization of all the β-glucans isolated from different sources regardless of their differences in glycosidic linkages and molecular weight by all three bifidobacteria was comparable to that of inulin.