Essential oil from the aerial parts of Lavandula multifida L. (Lamiaceae), used in the Moroccan traditional medicine, was extracted by hydrodistillation and analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). 34 constituents, representing 95.25 % of the total oil were identified. The major components at the species level were Carvacrol (47.62%), ?- bisabolene (9.01%), and Dodecyl Acrylate (8.37%), Linalol (7.42%), Menthone (4.98%), ?- Caryophyllene (3.34%), ?- Pinene (3.21%). Antibacterial activity of this oil was tested against human pathogenic bacteria: gram-negative and gram-positive bacteria by the agar diffusion method. The minimum inhibitory concentrations (MIC) of the oil were determined by the microdilution technique. The oil showed significant inhibitory activity against the bacteria, S. Aureus (14.330 ± 0.577 mm), B. subtilis (12.670± 0.577 mm), P. mirabilis (12± 1 mm), P. vulgaris (11.67± 0.577 mm), L. innocua (10.660 ± 0.577 mm), L. monocytogenes (8.667± 0.577 mm), E. coli CECT (9.667± 0.577 mm) and E. coli K12 (9.333 ± 0.577 mm). No inhibitory activity was observed against the bacterium Ps. aeruginosa.