Zinc oxide nanoparticles were prepared and subsequently deposited onto the surface of the cotton fiber by ultrasonic irradiation. The optical, structure and morphology of the coated and un-coated cotton were examined by UV, fourier transform infrared spectroscopy, X-ray diffraction analysis (XRD) and scanning electron microscope (SEM)/Energy Dispersive X-ray analysis. XRD analysis revealed the presence of the crystalline metal oxide of hexagonal phase with an average crystallite size of 12 nm. These nanoparticles are probably physically adsorbed onto the cotton fiber surface. SEM analysis showed a distribution of ZnO nanorod assemblies of various diameters and lengths physically adsorbed onto the cotton fiber surface may take place. The ZnO-cotton fiber nano-composite were tested against Escherichia coli (gram negative) and Staphylococcus aureus (gram positive) cultures, and showed a significant antimicrobial activity.