Here, silver oxide nanoparticles were made using the chemical reduction approach. We expect that the resulting nano-AgOparticles have a bactericidal effect on their mentioned Gram-positive and Gram-negative bacteria. We looked at the structural and morphological characteristics of nano-AgO particles, which are well known to be very toxic to both gram-negative and gram-positive microorganisms, including drug-resistant bacteria. The created nano-AgO particles were examined using a variety of methods, including Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), Fourier Transforms Infrared spectroscopy (FTIR), and X-ray Diffraction spectroscopy (XRD). The characterization results for AgO NPs demonstrate aggregates of grains with a constrained size range, with individual particle radii ranging from 100 to 200 nm. The bactericidal activity of nano-AgO particles against Pseudomonas aeruginosa (P. aeruginosa), Streptococcus pyogenes (S. pyogenes), and Klebsiella pneumoniaewas proven by well diffusion results within a 17 mm zone of inhibition.