Antimicrobial nanofibers were prepared by solubilizing an antimicrobial essential oil (eugenol; 0.75−1.5 wt %) in surfactant micelles (Surfynol 465; 5−10 wt %) to form eugenol-containing microemulsions. Microemulsions were mixed with a nonionic synthetic polymer (poly(vinyl alcohol), PVA; Mw = 130 kDa, degree of hydrolysis ≈ 87%) and solutions subjected to electrospinning to induce nanofiber formation. Solution properties, fiber morphology, and composition of nanofibers were determined. The surface conductivity and viscosity of the polymer solutions increased, while surface tension decreased as both surfactant and eugenol concentration increased. Material deposited on the collector plate consisted primarily of nanofibers with a circular cross section with some surface roughness, although some bead defects were observed. The mean fiber diameters ranged from 57 to 126 nm with fibers having a broad diameter distribution (10−280 nm). The mean diameter of the nanofibers decreased with increasing surfactant concentration and decreasing eugenol concentration. Transmission electron microscopy indicated that microemulsion droplets were homogenously dispersed throughout the nanofibers. Results suggest that electrospun nanofibers may serve as carrier vehicles for microemulsions containing solubilized lipophilic functional compounds such as bioactives, antimicrobials, antioxidants, flavors, and pharmaceuticals.