[Purpose] The aim of this study was to evaluate the effect of pulsed high intensity neodymium-doped yttrium aluminum garnet (Nd: YAG) laser on staphylococcus aureus (S. aureus) and pseudomonas aeruginosa (P. aeruginosa) bacterial growth, which cause many health problems and establish which doses are effective in bacterial inhibition. [Materials and Methods] Five samples of S. aureus and five samples of P. aeruginosa were prepared in the microbiology lab, one used as control sample and the other 4 samples acted as experimental samples. The experimental samples received pulsed high intensity Nd: YAG laser with a total dose of 500, 600, 700 and 800 joules. The primary measures are colony count and the percentage decrease in colony count, the colony count was assessed at baseline and after 24 h of laser application. [Result] There was significant decrease in colony count and the percentage decrease in colony count after pulsed high intensity Nd: YAG laser application in all experimental samples of S. aureus and P. aeruginosa after 24 h of application for all doses (500, 600, 700 and 800 j) as compared with the control sample, with the most effect in higher doses of pulsed high intensity Nd: YAG laser than lower doses in both types of bacteria. [Conclusion] pulsed high intensity Nd: YAG laser was found to be an effective modality for inhibition of S. aureus and P. aeruginosa growth after a single application.