Electrospinning was used to produce poly(vinyl alcohol) (PVA)-based nanofibers containing nerolidol (NER) as the antimicrobial agent. The scanning electron microscopy results indicated the formation of homogeneous fibers in the absence of NER. However, its incorporation into the fiber prevented the complete solvent evaporation, ultimately leading to the coalescence of the fibers. It became more evident as the amount of NER increased from 0 to 50% m/v. The incorporation of NER also decreased the crystallinity of the fibers from 10 to ca. 6.5%. However, this decrease was not proportional to the concentration of NER. The contact angle results confirmed the presence of NER molecules on the surface of the fibers especially due to the increase of the contact angle from 0 (PVA/NER0) to 27.8º (PVA/NER50). The fibrous mats exhibited good antimicrobial activity, being even more efficient than the control in some cases, suggesting their potential for wound-dressing purposes.