The continuous and non-prescription use of antibiotics for more than fifty years as a control and treatment of infectious diseases has caused difficulties such as antibiotic resistance and environmental pollution in addition to its side effects on body. Therefore it is necessary to use natural antibiotics as an alternative medicine. Gallnut is one of the natural antibiotics in traditional medicine whith a variety of pharmacological properties such as astringent, anti-diabetic, anesthetic, antiviral, antifungal, antibacterial, larvicidal, anti-in?ammation and wound-healing. The aim of this study was to produce an antibacterial wound dressing based on PVA and natural gallnut. Poly vinyl alcohol was chosen as a substrate due to its good mechanical properties, excellent chemical resistance, biodegradability, easy preparation and film forming ability. Gallnut was incorporated into the poly vinyl alcohol nano fibers by dissolving in PVA solution. In this regard gallnut was dissolved in water: ethyl acetate: acetic acid with 15: 45: 25: 15 w/w ratio and the solution was slowly added to PVA in ethanol (80%) with 10:90 w/w ratios to prepare the electrospinning solution. This emulsion was used to prepare either nanofiber via electrospinning or film by casting. The produced films had both flexibility and traarency properties which are significant parameters in modern wound dressing. To produce nanofiber, electrospinning voltage, working distance and the flow rate were optimized at 14 KV, 13 cm and 0.28 mL/h, respectively. SEM images of produced electrospun fibers showed regular ractangular beads which can be microcapsules. Fibers diameter was calculated about 500 Nano meter containing beads with 1.5 micro meters diameter. Fourier Transform Infrared Spectroscopy (FTIR) confirmed the existence of active compounds inside the nanofiber and also formation of hydrogen bonds between gallnut and PVA. UV-Vis spectrophotometry was used to determine the loading and the releasing rate of the produced nanofiber. The overall loading percent of gallnut, for both nano fibers and film was about 100% which could be related to high hydrophlilicity of the compound and polymer. The antibacterial activity of the gallnut incorporated nanofiber mats was determined by agar diffusion test method of various bacterial species in the presence of gallnut mats. Antibacterial diffusion test result showed 18mm inhibition zone against S.areus for nanofibrous mats containing gallnut which was as good as the antibacterial effect of strong antibiotic, vancomycin standards discs and 20 mm against P. aeroginosa .It was also found that using nanofiber mat had positive effect on antibacterial activity of the produced nano fibers compared to PVA film. It is noticeable that the biodegradability percentage of nano fibers is more than that of film because of nano fibers higher specific surface area. Keywords: gallnut, wound dressing, PVA, antibacterial, nanofiber, emulsion electrospinning