Juniperus excelas is an important flowering medicinal plant belonging to the Cupressaceae family. Essential oil was obtained from fresh fruit of J. excelas by hydro-distillation in a Clevenger apparatus and analysed by gas chromatography–mass spectrometry (GC–MS). The chemical constituents of the essential oil were identified by their mass spectra, retention time and retention indices. The yield was 0.27%. We identified 48 chemical compounds accounting for 89.74% of the composition. The major chemical components were α-terpinene (23.85%), limonene (23.42%), fenchene (6.57%), camphene (6%), δ-3-carene (4.17%), 4-terpineol (2.93%), germacrene B (2.21%), myrcene (1.96%), α-pinene (1.77%), β-pinene (1.53%) and abietatriene (1.13%). The antimicrobial activity of the essential oil of J. excelas was determined against one Gram-positive and two Gram-negative foodborne pathogenic bacteria, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. No activity was detected.