ABSTRACT:To evaluate the antimicrobial activity of Octenidine (OCT) 0.1%, Chlorhexidine (CHX) 0.2% against bacterial strains of Enterococcus faecalis and Staphylococcus aureus. The strains were inoculated in 7ml of brain heart infusion broth and diluted to reach the concentration equivalent (0.5 McFarland standard). 1ml of organism suspension was contacted with 1ml of each mouthwash and was removed at time interval of 3,5 and 10 minute and plated on Brain Heart Infusion agar. After 72 hours of incubation, colony counts were measured using stereomicroscope. Kruskal Wallis test was conducted on mean number of CFU. Post-hoc tests were conducted by using the Mann Whitney U test and Duncan's-test of multiple comparisons. The results showed that OCT 0.1% was found to be the most effective in substantially reducing total bacterial counts after 3, 5 and 10 min time interval. OCT 0.1% was found to be the most effective in substantially reducing total bacterial counts.