Background: The vaginal microbiome harbors diverse communities of microorganisms, known as vaginal flora which has an important impact on women s health as well as that of their newborns Bacteria that colonize the vaginal microbiota of women play an important role in health and homostasis that exist in a dynamic balance and establishing with each other and with the host. In healthy reproductive- aged women, the vaginal microbiome,generally shows a predominance of Lactobacillus genus,and most women display the prevalence of one species among L., crispatus, L., iners, L., jensenii and L., gasseri. Lactobacilli promote the maintenance of the vaginal homoestasis and prevent the colonization and growth of adverse microorganisms,especialy pathogens. Methodology: 150 high vaginal swabs were collected from 100 specimen from women suffering from vaginitis and 50 specimen from healthy women. Each specimen were cultured on UTI chromogenic agar under aerobic conditions and on MRS agar under anaerobic conidion Antibiotics susceptibility performed according to NCCLS (National Committee for Clinical Laboratory Standereds).Biofiilms formation were assayed by TCP. Results: Results revealed that a total 150 specimens, 90 were gave positive cultured with single bacterial growth on UTI chromogenic agar as following: Escherichia. coli (42/90), Staphylococcus aureus (27/90), Enterobacter (13/90), Klebsiella pneumonia (8/90), 30 specimens were gave mixed culture and 30 others specimens were no growth. Out of 150 specimens also cultured on MRS agar,30 specimens were gave positive culture with pure bacterial isolates of Lactobacillus spp. 20 isolates of Lactobacillus (10 isolates from healthy women and 10 isolates from infected women with vaginitis) were selected to study the effect of antibiotics. It has been found that all Lactobacillus isolated from healthy women specimens resistant for chloromphincol and amikiacin. 50% resistant for ceftriaxone, 40% resistant for ciproflaxcin, gentamycin, and trimethoprim-sulfamethoxazole, 30% resistant for nitrofurantoin, 20% resistant for levofloxacin, and all Lactobacillus isolates sensitive to imipenem while all Lactobacillus isolates of women with vaginitis were sensitive to all antibiotics used in this study. Also 20 Lactobacillus isolates were selected to study the ability of them to produce biofilm formation. The Lactobacillus isolates from healthy women produce strong and moderate biofilm former 50%, while Lactobacillus isolates from vaginitis women produce (0%, 40%, 40%, 20%) strong, moderate, weak and non-biofilm former respectively. Conclusion: Most vaginal infections are treated by antibiotics but frequent recurrences and chronic infections are common due to the adverse effect on the indigenous lactobacilli . Probiotics have the potential to optimize, maintain and restore the microflora of the vaginal ecosystem hence the alternative approach to reducing vaginal infections and promoting consumer health by using biotherapoeutic with pre- and probiotic.The types of antibiotics used to treat vaginitis must not to kill the beneficial bacteria (lactobacilli) that help in preservation of vaginal health and ecosystem as being one of probiotic bacteria . [ABSTRACT FROM AUTHOR]