The aim of this study was to produce ultrafine fibers from zein incorporated with a complex of eucalyptus essential oil (EEO) and beta-cyclodextrin (beta-CD) with antimicrobial properties by electrospinning technique. The EEO was characterized by chemical composition and antimicrobial tests against three Gram positive and four Gram negative bacteria. The inclusion complex (IC) was prepared with beta-CD and EEO by co-precipitation technique and added at different concentrations in zein polymer solution using aqueous ethanol as solvent. The morphology, thermal properties, functional groups, and antimicrobial activity against L. monocytogenes and S. aureus of the ultrafine fibers were evaluated. The composite membranes containing 24% IC exhibited a greater reduction of growth as compared to the fibers without addition of IC. For L. monocytogenes the growth reduction was 28.5% and for S. aureus it was 24.3%. The electrospun IC-beta-CD/EEO composite membranes are promising for use in antimicrobial applications, such as food packaging. (C) 2017 Elsevier B.V. All rights reserved.