The microbiome of the vagina is characterized by a community bacteria playing important roles in the overall health status of the female genital tract. This study was conducted to isolate and characterize bacteria from the female genital tract and as well evaluate the antibiotics susceptibility pattern of the vaginal bacterial isolates. For this purpose, a total of fifty (50) vaginal swab samples were collected (using sterile swab sticks) from females attending a tertiary institution in Port Harcourt, Rivers State, Nigeria, and subjected to standard bacteriological analysis. Antibiotics sensitivity analysis was carried out using the modified Kirby Bauer disc diffusion method. A total of 160 bacterial isolates were obtained from the subjects of different age brackets in the study population. Molecular identification based on the nucleic acid sequence of the bacterial isolates revealed the isolates to be Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Bacillus flexus and Lysinibacillus macrolides. The result further showed that Escherichia coli was the most occurring bacterial isolate. Also, female subjects within the age bracket 21-23 years recorded the highest number of bacterial isolates (67) and 24-26 years had the least number of bacterial isolates (36). The antibiotic sensitivity analysis revealed that Escherichia coli and Staphylococcus aureus were resistant to 50% of the antibiotics tested, whereas Klebsiella pneumoniae was resistant to all (100% of) the antibiotics tested. The study has revealed that the vaginal microbiome of healthy female subjects is characterised by diverse species of bacteria, including opportunistic bacterial pathogens. The study therefore, recommended that regular screening for bacterial vaginosis as well as personal hygiene, sensitization programs to improve knowledge of women, should be encouraged.