In this study, Zinc oxide nanostructures were synthesized via a hydrothermal method by using zinc nitrate hexahydrate and sodium hydroxide as a precursor. Three different annealing temperatures were used to study their effect on ZnO NSs properties. The synthesized nanostructure was characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), Atomic force microscope (AFM), and Fourier Transform Infrared Spectroscopy (FTIR). Their optical properties were studied by using UV -visible spectroscopy. The XRD analysis confirms that all ZnO nanostructures have the hexagonal wurtzite structure with average crystallite size within the range of (30.59 - 34.52) nm. The crystallite size increased due to the incensement of annealing temperature. FESEM analysis indicates that ZnO has hexagonal shape of cylindrical pores, plate-like nanocrystals and Nanorods. AFM analysis shows that the average surface roughness of ZnO Nanostructures increases from 3.96 to 19.1 nm with the increase of annealing temperature. The FTIR peaks indicate successful preparation of ZnO Nanostructures. The FTIR method was used to analyses the chemical bonds which conformed the present of the Zn-O group in the region between (400-500) cm^-1. The UV-visible showed a red shift in the absorption spectra related to the shift in the energy gap related to increase in the particle size.  the band gap energy has been calculated from the optical absorption spectra. The annealing process has been fond more effective on the value of energy gap. As the annealing temperature increases, the value of energy gap, increases as well; from (3.12to 3.22) eV. The prepared Nanostructure is used for antibacterial property. It shows strong antibacterial activity against S. aureus and P.aeuruginosa bacteria by the agar disc diffusion method. The white precipitate of ZnO NSs has superior antibacterial activity on gram-positive (S. aureus) than the gram-negative (P.aeuruginosa) bacteria.