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Strains of Bifidobacterium animalis subsp. lactis are extensively incorporated in commercial probiotic preparations due to a number of technological and physiological advantages attributed to them over other members of the same genus. High strain specificity is being reported among microorganisms in their probiotic attributes making it practically difficult to extrapolate the results obtained for one organism to another at the species or even at the strain level. In this context a study was conducted to assess the probiotic properties of two strains of Bifidobacterium animalis subsp. lactis, namely Bifidobacterium animalis subsp. lactis B420 and Bifidobacterium animalis subsp. lactis Bb-12 in terms of their acid, bile salt, lysozyme tolerances, cell surface hydrophobicity and antimicrobial activity. On assessing their tolerance to various stresses, both the strains exhibited remarkably high acid, bile and lysozyme tolerance to the extent that the survivability at pH 2 after 3 hours of exposure was more than 98% and after 12h of exposure at 3% bile more than 80% of the cells were viable. The cell surface hydrophobicity in terms of adhesion to n-hexadecane was found to be more than 85% for both the strains .The antagonistic activity exhibited by the tested bifidobacterial strains were also at par with each other. On statistically analysing the data no significant difference was observed between the two bifidobacterial cultures for any of the tested probiotic attributes substantiating their wide use and popularity as probiotic cultures.