Silver nanoparticles inhibit microbial growth and have been gaining importance as a solution for numerous biomedical applications. In this study, synthesis of silver nanoparticles is carried out using Bacillus species isolated from silver fabrication area. Nanoparticle Tracking Analysis and Scanning Electron Microscopy revealed the size of particles ranged from 39 to 67.4 nm and spherical shape. XRD spectra revealed nanoparticles are of silver oxide. The anti-bacterial activity of silver oxide nanoparticles of Bacillus species was prominent against the three clinical strains of major Urinary Tract Infection (UTI) causing Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa.