In this study, chemical composition, antioxidant, antimicrobial and antiospasmodic activities of the essential oil of the berries Juniperus excelsa Bieb. subsp. excelsa were evaluated in vitro. GC and GS-MS analysis of the oil resulted in the identification of 25 compounds, representing 97.2 % of the oil; alpha-pinene (46.1 %) was the main component. Essential oil showed remarkable antimicrobial activity against the test microorganisms. The sample was also subjected to a screening for its possible antioxidant activity by using 2,2-diphenyl-1- picrylhydrazyl (DPPH) and beta-carotene-linoleic acid assays. It was exhibited moderate antioxidant activity in beta-carotene-linoleic acid test system. In rat ileum, J. excelsa essential oil effect was directly investigated on spontaneous contractions. It has been found that amplitude and frequency did not change significantly at all dose concentrations. Amplitude has been inhibited % 25 and frequency has been inhibited % 20 at 1 mg/mL.