Nanotechnology strategies have been used for delivery and controlled release of antimicrobial drugs. Electrospun nanofibers can be versatile vehicles to incorporate antimicrobials. In this work, poly-epsilon-caprolactone nanofibers functionalized with ketoconazole were produced by electrospinning and tested against filamentous fungi. Ketoconazole-free nanofibers were produced as controls. Functionalized nanofibers showed antifungal activity against Aspergillus flavus, A. carbonarius, A. niger, Aspergillus sp. A29, Fusarium oxysporum and Penicillium citrinum by agar diffusion test. Inhibitory zones ranging from 6 to 44 mm were observed, this larger inhibition was against A. flavus. The nanofibers were incubated in different simulant solutions to evaluate the ketoconazole release, which was only detected in the solution containing 5% (v/v) Tween 20. Electron microscopy images showed the nanofibers with ketoconazole presented mean diameters of 526 nm, and the degradation of the nanofiber structures could be observed by electron microscopy after incubation in simulant solution. Infrared and thermal analyses indicated that ketoconazole was dispersed without chemical interactions with the polycaprolactone matrix. These results suggest that polycaprolactone nanofibers incorporating ketoconazole may be an interesting alternative to control pathogenic fungi. (C) 2016 Elsevier B.V. All rights reserved.