Abstract: The aim of study was to isolate a Bacteriocin producing strain against Gardnerella vaginalis and other bacterial vaginosis causing organisms. For this purpose, about 100 bacteriocin producing strains Lactic acid bacteria were isolated from vaginal swabs of healthy and fecund females and evaluated for their antimicrobial activity. Nine isolates have shown anti-G. vaginalis activity. Out of nine HV 6b isolate have shown significant results against G. vaginalis so it has been selected for further study and it is identified as Lactobacillus fermentum. The antimicrobial activity of all nine strains including HV6b against other bacterial vaginosis associated organisms have also been studied. Mode of action of Bacteriocin produced from Lactobacillus fermentum was studied with scanning electron microscopy which showed the pore formation in the cell wall Gardnerella vaginalis. Antibiotic sensitivity profiles of L. fermentum HV6b was studied results showed that this isolated strain of lactic acid bacteria is resistant to most of antibiotics which have been currently used for treatment of bacterial vaginosis. Antibiotic sensitivity of Gardnerella vaginalis shows resistance of bacteria to Azithromycin, Co-trimoxazole, Metronidazole and Miconazole. Tetracycline and Erythromycin were required in more concentrated dose for inhibition. This study has shown that bacteriocin is a better alternative than antibiotics than the treatment of bacterial vaginosis. Viability loss of Gardnerella vaginalis was observed with different concentration of bacteriocin. Bacteriocin producing L. fermentum HV6b was found to be the responsible strain for formulating topical personal care and efficacy to the therapeutics aimed at prevention and treatment of bacterial vaginosis.