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The chemical composition of the essential oils from aerial parts of two taxa of Centaurea belonging to subgenus Acrolophus, Centaurea diffusa Lam. and C. micrantha Hoff. ssp. melanosticta (Lange) Dostàl, respectively collected in Croatia and Spain, were evaluated by GC and GC-MS. The main components of C. diffusa were hexadecanoic acid (31.1%), (Z,Z)-9,12-octadecadienoic acid (10.7%) and damascenone isomers (6.4%), whereas hexahydrofarnesyl acetone (27.8%), hexadecanoic acid (8.3%) and caryophyllene oxide (6.4%) were the most abundant components of C. micrantha ssp. melanosticta. The oils showed good antibacterial and antifungal activities against some microorganisms that infest historical art works.