Enicostema axillare (Lam.). Raynal, syn. E.littorale Blume (Family) Gentinaceae is a perennial herb found throughout the greater part of India. Locally it is known as Chota chirayita and used in indigenous medicines in the treatment of fevers and as bitter tonic and forms one ingredients of many hypoglycemic marketed formulations. In the present study in vitro antimicrobial activity (Minimum Inhibitory Concentration by Micro-titre plate method of Chloroform, Ethyl acetate, Methanol, Petroleum ether extract of whole plant has been evaluated. Four Bacterial Species and two fungal strains used for study are: Staph aureus, Pseudomonas aeroginosa, Salmonella typhi, Shigella sonnei, and two fungal strains are: Aeromonas hydrophila, Candida albicans. It was observed that Ethyl acetate, Methanolic extract showed prominent antimicrobial activity against all micro organisms.