Lactic acid bacteria (LAB) are integral to human health, particularly in modulating gastrointestinal function and immune responses. Human breast milk, recognized as a vital source of LAB for newborns, contains a diverse array of nutrients and bioactive compounds essential for infant growth and development. This study aimed to isolate and characterize LAB from breast milk samples obtained from lactating donors of varying ages and lactation periods. LAB were isolated using selective media, and primary screening was conducted based on colony morphology, gram characteristics, catalase activity, and motility. Results demonstrated significant variability in LAB distribution among donors, with higher counts observed during early postpartum periods. While mean colony-forming unit (CFU) counts decreased with advancing lactation periods, no direct correlation was found between donor age and LAB counts. The findings underscore the importance of understanding breast milk micro biota dynamics for infant health and development, necessitating further research in this field.