The leaf-essential oil from Eucalyptus robusta and E. saligna, the latter in two phenologic stages, were analysed by gas chromatography coupled to mass spectrometry (GC-MS). The major constituent in E. robusta oil was the monoterpene alpha-pinene (73.0% of the total amount). The oil composition of the E. saligna was dependent of the phenologic stage. In the vegetative phase, the major constituents were p-cymene (54.2%) and gamma-terpinene (43.8%), while during the blossoming alpha-pinene became the major constituent followed by p-cymene (22.5%). Additionally, the antimicrobial activity for all three oils was evaluated against Staphylococcus aureus, Escherichia coli and Candida albicans. The E. robusta oil presented the highest growth inhibition for all the microorganisms tested.