In this study, we examined the chemical compositions of essential oil and tested the efficacy of oil and organic extracts of Cestrum nocturnum L. against food-borne pathogens. The chemical compositions of the oil was analysed by gas chromatography-mass spectrometry (GC-MS). Forty-seven compounds representing 93.28% of the total oil were identified. The oil [5 μL of 1:5 (v/v) dilution of oil with methanol] and organic extracts of hexane, chloroform, ethyl acetate and methanol (300 μg per disc) of C. nocturnum displayed a great potential of antibacterial activity against Staphylococcus aureus (ATCC 6538 and KCTC 1916), Listeria monocytogenes (ATCC 19166 and ATCC 15313), Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa KCTC 2004, Salmonella typhimurium KCTC 2515 and Escherichia coli ATCC 8739. Also the oil had strong detrimental effect on the viable count of the tested bacteria. The results obtained from this study may contribute to the development of new antimicrobial agents with potential applications in food industries as natural preservatives to control food-borne pathogens.