Compounds having antimicrobial activity obtained from plant sources are useful because they are new sources against antibiotic resistance. The present study is based on the evaluation of antimicrobial activity of the essential oil of the fresh bark of Betula utilis and to identify the chemical constituents responsible for antimicrobial potential, grown in high altitude of Himalaya, India. The chemical constituents of essential oil were identified by gas chromatography-mass spectrometry (GC-MS). The essential oil was evaluated for the antimicrobial potential against human pathogenic bacteria and fungi by using a micro-dilution assay. GC-MS analysis revealed that the major constituents of the essential oil were geranic acid (11.38 %), β-seleneol (10.98 %), β-linalool (10.91 %), terragon (10.61 %), β-sesquiphellendrene (8.02 %), champacol (6.33 %) and 1,8-cineol (5.49 %). Result showed that B. utilis essential oil has a strong antimicrobial activity against the fungus Candida albicans and Gram (+) and Gram (-) human pathogenic bacteria (MIC: 60.5-240 µg/ml).