Submicrometric and nanometric poly(lactic acid)/poly(vinyl pyrrolidone) (PLA/PVP) fibrous membranes containing 0, 5, 10, 15, and 20 wt % PVP, with or without 20 wt % Copaiba oil (Copaifera sp.), were produced by solution blow spinning (SBS), using polymer injection rate of 120 lL min 21 , gas pressure of 2.4 kPa, working distance of 20 cm, and collector rotation of 200 rpm. The morphological, thermal, and spectroscopic properties of these membranes were characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetry (TG), and Fourier transform infrared spectroscopy (FTIR). A method for the evaluation of membrane microbial barrier properties based on resazurin colorimetric method was proposed. Results showed that the addition of both PVP and Copaiba oil produced thicker fibers; otherwise, there was no effect on morphology. Thermal analyses (TG and DSC) indicated the immiscible nature of polymer blends produced, also confirmed by the spectroscopic studies. Antimicrobial barrier properties were related to the antimicrobial effect of Copaiba oil, combined with it hydrophobic nature. The hydrophilic nature of PVP favored degradation of fiber mats, impairing barrier property when higher concentrations of PVP were added. Results indicate that produced spun mats can potentially be used in applications such as wound dressing.