A method for direct selection of acid-resistant Bifidobacterium strains was developed by prolonged exposure of human feces to homologous lethal stress conditions. The recovered strains were intrinsically resistant to acidic gastric conditions (pH 2.0) and also showed good tolerance to high concentrations of bile salts and NaCl. Bifidobacterium breve and Bifidobacterium adolescentis were, respectively, the infant- and adult-type bifidobacterial species showing the highest ability to develop an acid-tolerant phenotype. Therefore, this procedure could be applicable to the direct selection of Bifidobacterium strains with improved stability in adverse environments and, probably, contribute to expand the spectra of probiotic species of human origin currently marketed.