The extraction yield of the essential oil (EO) extracted by hydrodistillation from the cones of Tunisian Cupressus sempervirens L. was of 0.518%. The chemical composition was analyzed by GC-MS. Results showed that this essential oil was mainly composed of monoterpene hydrocarbons (65%) with í µí»¼-pinene as the major constituent (47.51%). Its antioxidant activity was ascertained by evaluating the total antioxidant capacity and also by evaluating its inhibitory effect against DPPH and ABTS radicals. In addition, it showed a strong antioxidant power against the DPPH (IC 50 = 151 í µí¼‡g/mL) and ABTS (IC 50 = 176.454 í µí¼‡g/mL) radicals scavenging. Moreover, its antibacterial activity was tested against different species of pathogenic bacteria (three Gram-positive and eight Gram-negative bacteria). The bacterial strains susceptible to the evaluated oil were Bacillus subtilis, Escherichia coli, Klebsiella oxytoca, Morganella morganii, Shigella, and Vibrio cholerae.