AimsTo identify and characterize the probiotic and functional attributes of bifidobacteria isolated from human sources by invitro methods. Methods and ResultsTwelve isolates of bifidobacteria were obtained from different human sources viz; human milk, infant and adult faeces. The preliminary identification of isolates was done using genus-specific PCR followed by species level identification using 16S rRNA sequencing. All the isolates invariably showed potential probiotic characteristics. Finally, three most promising isolates were subjected to safety evaluation and were found to be safe. These were further evaluated for their potential functional characteristics like, antioxidative, antimutagenic and cholesterol assimilation. The isolates viz; NBIF-5 (7355003%) and NBIF-7 (6406003%) assimilated significantly higher cholesterol than NBIF-2. The maximum antioxidative activity was observed in NBIF-2 (5656 +/- 028% of radical inhibition). High percentage antimutagenicity scores of 5241 +/- 225% and 5368 +/- 198% against sodium azide (NaN3) were shown by NBIF-2 and NBIF-7 respectively. ConclusionsThree strains (NBIF-2, NBIF-5 and NBIF-7) were found to be endowed with appreciable probiotic and functional activities. Significance and Impact of the StudyWith growing demands for probioitcs and bifidobacteria being prominent ones, new strains of diversified origin have the potential to be explored for commercial and functional applications and thus can be added to the existing commercially available strains of this genera.