Abstract: In this study, antimicrobial activity and ethnomedicinal uses of 40 medicinal plants along with medicinal properties has been reported from Similipal Biosphere Reserve, Orissa, India. Aqueous extracts of different parts of the plant (leaf, stem, bark, bulb, fruit and root) reported to have medicinal properties were tested for their antimicrobial activity against gram positive and gram negative human pathogenic bacteria such as Staphylococcus aureus MTCC 1144, Bacillus licheniformis MTCC 7425, Bacillus brevis MTCC 7404, Bacillus subtilis MTCC 7164, Staphylococcus epidermidis MTCC 3615, Streptococcus aureus (Lab. isolate), Pseudomonas aeruginosa MTCC 1034, Escherichia coli MTCC 1089, Vibrio cholerae (Lab. isolate), Shigella flexneri (Lab. isolate) and one fungal species Candida krusei (Lab. isolate). Results showed that 23 out of 40 medicinal plants have antimicrobial activity of which, 14 medicinal plants have outstanding antimicrobial activity. Prominent species with antimicrobial activity are Urginea indica (bulb), Croton roxburghii (bark), Melastoma malabathricum (leaf), Diospyros melanoxylon (bark), Pterospermum acerifolium (leaf), Nyctanthes arbortristis (bark), Oroxylum indicum (bark), Agava sisalana (leaf), Clausena excavate (root) Vitex negundo (leaf), Glycyrrhiza glabra (fruit), Enhydra fluctuans (leaf), Hemidesmus indicus (leaf) and Flemingia nana (root) with inhibition zones more than 20 mm where as 9 other plants were found to have moderate antimicrobial activity with inhibition zones of less than 20 mm. Rest 17 plants did not show any antimicrobial activity.