Probiotic potential of two bifidobacterial strains isolated from feces of healthy adults and identified as Bifidobacterium animalis subsp. lactis was evaluated using in vitro testing. The analyzed strains were able to ferment a broad spectrum of carbohydrates, produced bioactive exopolysaccharides, demonstrated high survival rate in model GIT conditions, under heat and oxidative stresses, inhibited growth of a wide range of pathogenic and opportunistic bacteria, and proved to be safe for biotechnological application. Based on the complex phenotypic characteristics tested, Bifidobacterium animalis subsp. lactis may be regarded as prospective probiotic cultures.