The chemical composition of the hydrodistilled leaf essential oil from Pulicaria stephanocarpa Balf. Fil was determined by GC-MS analysis, and its antimicrobial, antioxidant and anticholinesterase (AChE) activities were evaluated. Eighty-three compounds were identified representing 97.2% of the total oil. (E)-Caryophyllene 13.4%, (E)-nerolidol 8.5%, caryophyllene oxide 8.5%, alpha-cadinol 8.2% spathulenol 6.8% and tau-cadinol 4.7%, were the main components. Antimicrobial activity of the oil, evaluated using the disc diffusion and broth dilution methods, demonstrated the highest susceptibility on Gram-positive bacteria and Candida albicans. The free radical scavenging ability of the oil was assessed by the DPPH assay to show antiradical activity with IC50 of 330 microg/mL. Moreover, the oil revealed an AChE inhibitory activity of 47% at a concentration of 200 microg/mL using Ellman's method.