In the present work, zinc oxide (ZnO) and silver (Ag) doped ZnO nanostructures are synthesized using a hydrothermal method. Structural quality of the products is attested using X-ray diffraction, which confirms the hexagonal wurtzite structure of pure ZnO and Ag-doped ZnO nanostructures. XRD further confirms the crystallite orientation along the c-axis, (101) plane. The field emission scanning electron microscope study reveals the change in shape of the synthesized ZnO particles from hexagonal nanoparticles to needle-shaped nanostructures for 3 wt% Ag-doped ZnO. The optical band gaps and lattice strain of nanostructures is increased significantly with the increase of doping concentration of Ag in ZnO nanostructure. The antimicrobial activity of synthesized nanostructures has been evaluated against the gram-positive human pathogenic bacteria, Staphylococcus aureus via an agarose gel diffusion test. The maximum value of zone of inhibition (22 mm) is achieved for 3 wt% Ag-doped ZnO nanostructure and it clearly demonstrates the remarkable antibacterial activity.