The bifidogenic effect of an infant formula supplemented with inulin and fructooligosaccharides (4.0 g/l) was examined clinically and in vitro, and compared that of mature breast milk. In a 28-day clinical study, fecal samples of 21 infants, divided into two groups: one receiving the infant formula and the other breast milk, were microbiologically and biochemically examined. In the in vitro investigation, microbiological and biochemical changes in the infant formula and breast milk induced by the action of bifidobacteria isolated from infant feces were examined. There were no significant differences in the fecal numbers of lactobacilli, total aerobes, anaerobes or yeasts and fungi. In contrast, the bifidobacteria numbers in the stools increased significantly during the study in the infants receiving the supplemented formula. The comparative in vitro test showed that the bifidogenic effect was similar for infant formula and breast milk in terms of the number of bifidobacteria. Consumption of infant formula with added inulin and fructooligosaccharides stimulated the bifidogenic effect, both clinically and in vitro. The in vitro test can quickly and objectively determine the bifidogenic effect of infant formula and indicate their quality. However, a clinical test is necessary to determine the acceptance and biological value of infant formula.