The aim of this study was to evaluate the antimicrobial property of essential oil extracted from the leaves ofOcimum americanumagainst oral bacteria related to periodontal disease. Three species of periodontal pathogens includingPorphyromonas gingivalisW50,Prevotella intermediaATCC 25611 andFusobacterium nucleatumATCC 25586 were included in the study. Agar diffusion was performed initially to screen the antimicrobial activity ofO.americanumessential oil. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were then determined using the Millipore Membrane method. The result showed that in the agar diffusion, essential oil extracted fromOcimum americanumexhibited antimicrobial activity against all test bacteria with the zone of inhibition ranging from 24 to 30 mm. The MIC values againstP. gingivalisandP. intermediawere 0.35 mg/mL whereas that ofF. nucleatumwas 0.70 mg/mL. The MBC values againstP. gingivalisandP. intermediawere 0.70 mg/mL whereas that ofF. nucleatumwas 1.4 mg/mL. In conclusion,O.americanumessential oil has an antimicrobial activity which may be a beneficial component of oral health care products to control or prevent periodontal disease by reducing these bacteria in the oral cavity.