Infections caused by Staphylococcus aureus and Klebsiella pneumoniae are becoming a common health issue throughout the world. Medicines that have been used to treat infections caused by these organisms are becoming obsolete due to the emergence of drug resistance at an alarming rate. Recently, the use of green synthesized nanoparticles has been gaining momentum as an alternative approach to combating microbial infections, since traditional methods for the synthesis of metal nanoparticles involve the use of toxic chemicals and the formation of harmful by-products. In this study, an aqueous extract of the medicinal plant Prunella vulgaris is used for the synthesis of silver oxide nanoparticles (AgONPs). The formation of AgONPs is confirmed by UV-visible spectroscopy in the wavelength range of 200-400 nm. X-ray analysis of AgONPs shows crystallite size of the particles to be 36.9 nm. Scanning electron microscopic image shows the synthesized AgONPs are seemed to be spherical and oval in morphology with an average size of 55-182 nm, whereas FTIR spectroscopy of AgONPs reveal the presence of functional group. Finally, AgONPs are tested to be effective against S. aureus and k. Pneumoniae.