Increasing microbial resistance to chemical antibiotics and their probabilistic side effects cause popularity of medicinal plants, so there is an instantaneous and steady need for novel antimicrobial compounds from plants. As we know, there is no documented proof on antibacterial effects of Pimpinella anisum (PA) hydroalcoholic extract in west of Iran. Gas chromatography mass spectrometry was done to determine chemical composion. As a screen test to discover antibacterial properties of the extract, agar disk and agar well diffusion methods were employed. Macrobroth tube test was performed to specify MIC. The findings show that the most substance found in PA was anethole. The results indicated the MIC and MBC values was 0.031 g/ml for PA except in case of B. subtilis which was 0.062 g/ml. Thus, the present research demonstrates the antibacterial effects of the medical plant on E. coli, S. aureus and B. subtilis, suggesting to use as antibacterial supplement in the developing countries towards the development of new therapeutic agent.