Dental diseases are among the most common health problems treated with traditional remedies. The plants used in dental care possess antibacterial activity. In the view of this an effort was made to evaluate the antimicrobial activity of the ethanolic and aqueous extracts of bark of Juglans regia, against oral bacteria, Streptococcus mutans (MTCC 890) and Staphylococcus aureus (MTCC 737). Minimum inhibitory concentration (MIC) 500 μg/ml for ethanolic and aqueous extract were used for this purpose. Different dilutions of these extracts were used to have concentration of 500 μg/ml, 1000 g/ml, 1500 μg/ml, 2000 μg/ml and 2500 μg/ml. In well diffusion method the ethanolic extract at higher concentration 2500 μg/ml showed maximum zone of inhibition 15mm against S.mutans and 12 mm against S.aureus however aqueous extract at same concentration showed maximum zone of inhibition 10 mm against S.mutans and 11 mm against S.aureus. The results of in vitro antimicrobial assay proved that ethanol solvent was more successful in the extraction of phytochemicals from plant samples than aqueous solvent, as ethanol extract showed higher antimicrobial activity than aqueous extract against both the test pathogens. Phytochemical screening of alcoholic extract showed the presence of different types of compounds like alkaloids, phenolics, tannins, flavonoids, steroids, saponins and resins which may contribute for the antimicrobial action of the above medicinal plant. The results of our study may provide the basis for using natural antimicrobial substance for oral hygiene purposes.