PVA nanofibers incorporated with silver nanoparticles produced from electrospinning have higher surface area to volume ratio, which could be useful for many environmental health and safety purposes such as water disinfection. Although there are several works done regarding the silver nanoparticles and its application as a bactericidal agent, there are no reports available on immobilized silver nanoparticles incorporated in PVA nanofibers and their application against Escherichia coli using growth inhibition in liquid medium, which is the primary cause of many serious diseases associated with contaminated water. We have used electrospinning technique to fabricate PVA/AgNO3 nanofibers and also investigated the effect of several operational parameters such as applied voltage, collection distance, flow rate and some solution parameters like concentration and conductivity on average fiber diameter. Structures were characterized using SEM, TEM and EDX spectroscopy. The antibacterial tests were performed by growing E.coli in liquid LB medium supplemented with pure PVA and PVA/ AgNO3 and results confirmed the antibacterial activity of immobilized PVA nanofibers incorporated with silver nanoparticles while pure PVA showed no inhibition of bacterial growth. Decreasing the ratio of PVA/ AgNO3 by increasing the silver content enhanced the antibacterial efficiency of the fabricated nanofibers.