Abstract: Three kinds of volatile oil were extracted from Eucalyptus lemon, Eucalyptus exserta, Eucalyptus urophylla with steam distillation respectively and the components of the oils were analyzed by GC-MS. The antimicrobial effects of the oil against foodborne pathogens were tested using double dilution method and the nitrite was used as positive control substance. The results showed that there were big difference between the chemical composition and content of three kinds of volatile oil. Three kinds of Eucalyptus essential oil had certain inhibition activity on the test bacteria and the antimicrobial activity of lemon eucalyptus essential oil was remarkable. Eucalyptus essential oil had good bacteriostatic effect and had application prospect in food preservation.