BACKGROUND: Periodontal pathogens play an important role in etiology and pathogenesis of periodontitis. Microbiological examination of sub gingival plaque is used at the present time in etiological research as well as in clinical treatment of periodontitis to select the appropriate antibiotic agent if indicated. Pine tar has been used for the treatment of various skin diseases. So the study was done to evaluate the effect of Pine Tar oil on bacteria isolated from periodontitis patients.METHODS: Plaque samples from volunteer patients were collected using sterile paper points. Robertson's Cooked Meat (RCM) medium was used for the transportation and cultivation of aerobic, microaerophilic, and anaerobic microorganisms.RESULTS: The result suggests the use of Pine tar oil for topical application in periodontal diseases. Disc diffusion analysis was sufficient enough to illustrate that 75 μl tar oil solution produced growth inhibition of microbial strains.CONCLUSION: Pine tar oil has become one of the important areas of research both in pharmaceutical and periodontal research, hence in vivo studies has to be carried out with various form of pine tar.