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In the present study, copper oxide (CuO) colloidal nanoparticles (NPs) were synthesized using laser ablation of copper pellet immersed in deionized water. Pulsed Nd: YAG laser was used to irradiate the targets at different laser energies and various ablation times. NP suspensions were characterized by UV–Vis spectroscopy, XRD, and TEM. The absorption spectrum exhibited a peak at ~275 nm and another peak with low intensity at ~645 nm. The XRD pattern of the NPs proved the presence of (–111) and (112) planes assigned to the CuO phase. The TEM images showed nearly spherical shape nanoparticles CuO NPs with size of 3–40 nm. The antibacterial activity of CuO NPs was first carried out against four types of bacteria: Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Staphylococcus aureus, by liquid medium method. CuO NPs showed the highest antibacterial activity against E …