Essential oil composition of the aerial parts of Agrimonia pilosa Ledeb growing wild in the central Himalayan region of Uttarakhand, India was analyzed by capillary gas chromatography (GC-FID) and gas chromatography-mass spectrometry (GC-MS). A total of 15 constituents were identified, representing 95.29% of the oil composition. The oil consisted mainly of oxygenated monoterpenes (75.74%) followed by monoterpene hydrocarbons (18.68%). Major constituents identified were methyl myrtenate (71.39%), limonene (11.86 %), α-thujone (5.66%) and myrtenyl acetate (3.14%). The essential oil showed a broad spectrum of antibacterial activity against both the human and plant pathogenic bacteria. Oil showed the highest activity against human pathogenic bacteria K. pneumonia (7.33 mm, MIC 175 µL/mL) and P. aeruginosa (7.00 mm, MIC 175 µL/mL). Plant pathogenic bacteria R. solanacearum (6.70 mm, MIC 175 µL/mL) was found less resistant against the oil. The results showed that oil containing methyl myrtenate a major constituent has potential for treatment of infections caused by these pathogenic bacterial strains.