Essential oils from the aerial parts of Anacyclus pyrethrum L. were analysed at three developmental stages (vegetative, floral budding and flowering). Oil yield was found to vary depending on the stage of development, and the highest content of oil (0.019% w/w) was obtained at flowering stage. The chemical composition of essential oils studied by GC and GC-MS showed a total of 91 compounds. Whatever the analysed stage is, oxygenated sesquiterpenes were the most abundant group. Their level significantly increased during ripening and varied from 37.1% to 58.6%. The oil showed activity against Candida albicans and Staphylococcus aureus bacteria strains. Thus, they represent an inexpensive source of natural antibacterial substances that may potentially be used in pathogenic systems.