The synthesis of metal and semiconductor nanoparticles is an expanding research area due to the potential applications in the development of novel technologies. Especially, biologically synthesized nanomaterial has become an important branch of nanotechnology. The present work, described the synthesis of zinc oxide nanoparticles (ZnO NPs) using leaf aqueous extract of Azadirachta indica (L.) and its antimicrobial activities. The nanoparticles was obtain characterized by UV-Vis spectroscopy, Photoluminescence (PL), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscope (SEM) analysis, Energy dispersive X-ray analysis (EDAX), Field emission scanning electron microscopy (FESEM) and Atomic force microscope (AFM) analysis. In this study we also investigated antimicrobial activity of green synthesized ZnO NPs. The results depicted concentration of ZnO NPs was increased (50, 100, 200 mu g/mL) and also increase in antimicrobial activities was due to the increase of H2O2 concentration from the surface of ZnO. However, green synthesized ZnO NPs was more potent than Bare ZnO and leaf of A. indica. Finally concluded the zinc oxide nanoparticles exhibited an interesting antimicrobial activity with both Gram positive and Gram negative bacterial and yeast at micromolar concentration. (C) 2015 Elsevier B.V. All rights reserved.