This study deals with the chemical composition and antimicrobial activity of essential oils of Vitex agnus castus L. The main constituents of the essential oils were characterized by GC-MS which resulted in the identification of 26 components, representing 100 % of the oil. The dominant compounds in the oil of fruits were trans-caryophyllene (19.17 %), sabinene (18.05 %) and 1,8-cineole (16.13 %), alpha-terpinyl acetate (6.91 %) and dihydroselarene (6.73 %). Antimicrobial activity was tested using the disc diffusion method. According to the inhibition zones, the essential oils were active against all of the tested microorganisms. The essential oils showed the susceptible inhibition zones, but they were less effective against bacterial strains compared to ampicillin and ofloxacin. The organisms most susceptible to these essential oils were Enterococcus faecalis ATCC 29212. However, further studies must be performed to confirm the safety of these oils for use as an antimicrobial agent.