In this work, a unique nanocomposite film based on poly (vinyl alcohol)/silver (PVA/Ag) has been synthesized using solution casting technique and the samples were exposed to gamma radiation for different doses 0, 25, 50 and 75 kGy. Ultraviolet-Visible spectroscopy, the characteristic Surface Plasmon Resonance (SPR) band placed at around 427 nm. FTIR analysis shows the formation of chemical bonding/conjugation between the Ag nanoparticles and PVA chains. X-ray diffraction analysis shows that Ag metal is present in face-centered cubic (fcc) crystal structure. The morphology of particles was studied using Field Emission Scanning Electron Microscopy (FESEM) it shows the nanoparticles are in spherical shape. The presence of Ag was confirmed using Energy Dispersive X-ray Spectroscopy (EDX) elemental analysis. Transmission Electron Microscopy (TEM) shows the particles were nano size and monodispersed. The antibacterial activities of the PVA/Ag nanocomposites were obtained using the zone inhibition method. The samples were tested against Gram-positive bacteria: Bacillus Subtilis, Staphylococcus aureus, and Gram-negative bacteria: Escherichia coli, Klebsiella pneumoniae. This study reveals that the irradiated PVA/Ag nanocomposites film shows good antibacterial activity against the Gram-positive and Gramnegative bacteria.