Aim: The present study was conducted with an aim to assess the antimicrobial activity of Ocimum sanctum (tulsi) extract on Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis.Materials and methods: Tulsi extract with ethanol was prepared using cold extraction method in the present in vitro study. Various concentrations (2, 4, 6, and 8%) were then obtained by dilution with dimethylformamide. A 0.2% chlorhexidine served as the positive control, whereas the negative control was dimeth-ylformamide. Zones of inhibition were measured, each for A. actinomycetemcomitans and P. gingivalis. For comparison within the group and between the study groups, one-way analysis of variance (ANOVA) and Tukey's post hoc tests were used. A statistical significance level of p < 0.05 was established. Results: The 8% concentration of the tulsi extract showed maximum zone of inhibition against A. actinomycetemcomitans and P. gingivalis (40.10 ± 0.90, 33.79 ± 1.82 mm), followed by the 6, 4, and 2% concentrations. The 0.2% chlorhexidine, which was the positive control, had 39.80 ± 1.24 and 32.28 ± 1.28 mm zones of inhibition; dimethylformamide showed 13.55 ± 1.92 and 10.21 ± 2.16 mm zones of inhibition against both the microorganisms. The ANOVA showed highly statistically significant (p < 0.0001) results between and within the groups. The antimicrobial activity of tulsi extract at 6 and 8% concentrations, and 0.2% chlorhexidine against A. actinomycetemcomitans showed statistically significant differences between the groups. The concentration of tulsi extract at 8 and 0.2% chlorhexidine on P. gingivalis showed statistically significant differences between the groups. Conclusion: It was concluded that 8% concentration of O. sanctum (tulsi) extract showed the maximum antimicrobial activity against A. actinomycetemcomitans and P. gingivalis. It is thus recommended that this may be useful as an adjunc-tive to mechanical therapy in the prevention and treatment of periodontal diseases. Clinical significance:O. sanctum (tulsi) is a herb that is abundantly available, easily accessible, economically feasible, and culturally acceptable. Therefore, it is very useful in the management of oral diseases and also for overcoming many barriers that exist for the utilization of dental services, such as affordability, accessibility, availability, and acceptability. Keywords:Aggregatibacter actinomycetemcomitans, Antimicrobial, Ocimum sanctum, Porphyromonas gingivalis.

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