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The study aimed to indicate the presence of milk microbiome in both lactating and nonlactating women, in addition to investigate some of relations between related parameters. A total of 90 women (20-35 years) were selected for providing samples of breast milk after birth for different periods. Culturing, isolation, staining, and genetic identification was applied. After PCR and Sequencing for 60 isolates, the possibility of identifying 54 of them illustrated that; the most frequent isolated bacteria were Enterococcus spp with 26.7%, followed by Staphylococcus spp 11.1%, Leuconostoc spp 10.0%, Bacillus spp 3.3%, and 2.2% for both, Proteous spp and Providencia spp. While the lowest isolated bacteria were Exiguobacterium aquaticum with 1.1%. Other non-identified organisms were Candida in 10%, mixed growth for bacteria and Candida 15.6%, and non-identified bacteria as 15.6%. A significant differences was found between type of lactation at P. value < 0.05, but there was non-significant differences between age groups, mode of delivery (birth way), use of medication and lactation stages.