Poly vinyl alcohol (PVA), which is a low-volatility, environmentally friendly and water-soluble polymer, is suitable for needleless electro spinning. Chitosan (CS) has drawn much attention due to its excellent antibacterial properties; however, it is hard to needle lessly electro spin CS alone. Thus, a certain amount of CS was added into the electro spinning solution to fabricate PVA nano fibers with antibacterial properties. In this paper, a needleless electro spinning device with spiral disk spinnerets was used to prepare PVA with different concentrations of CS nano fiber membranes. The effects of electro spinning parameters on process stability and anti-water-soluble processing were investigated. The results showed that the electro spinning process was associated with voltage, rotation speed and collection distance. The optimal electro spinning parameters are a voltage of 75 kV, a rotation speed of 11 rpm and a distance of 190 mm. From the comparison and analysis, 5% glutaraldehyde (GA) solution treatment is the superior method to increase the anti-water-solubility of the membrane. The antibacterial properties of PVA/CS nano fiber membranes with CS concentrations ranging from 1.0% to 1.5% were also investigated. The results showed that the inhibition rates of Escherichia coli (E. coli) were all above 99%, and those of Staphylococcus aureus (S. aureus) were all above 90%. Moreover, GA solution treatment has no effect on the antibacterial performance.