Generally, two different types of fructose polymer are found in nature. One is inulin, whose fructosyl residues are linked mainly by a ${eta}-(2,1)-linkage$, while the other is high-molecular-weight levan, whose fructosyl residues are linked mainly by a ${eta}-(2,6)-linkage$. In contrast to the extensive studies on the prebiotic properties of inulin, there has been no report on the effect of levan on the large bowel microflora in viva. Therefore, to examine whether dietary levan can be used as a prebiotic, Sprague-Dawley male rats were fed one of two diets for 3 weeks: 1) basal diet plus sucrose; 2) basal diet plus 10% (wt/wt) levan. The cecal bowel mass, cecal and colon short-chain fatty acids (SCFAs), pH, and microflora were then compared. The intake of the levan-containing diet significantly increased the total cecal weight and wall weight. The analyses of the SCFAs in the cecal and colonic contents revealed that levan was converted into acetate, butyrate, and lactate, which resulted in acidic conditions. The intake of levan also significantly increased the total number of microorganisms by 5-fold and lactic acid-producing bacteria (LAB) 30-fold in the feces. Accordingly, the current work shows that levan can be used as a prebiotic for stimulating the growth of LAB in an animal model.