In this study, a new bio-nanocomposite was prepared and characterized with a focus on the formation of hexagonal ZnO and orthorhombic zinc silicate (Zn2SiO3(OH)(2)) phases under ultrasonic irradiation. Chitosan/sepiolite/ZnO and chitosan/silica/ZnO bio-nanocomposites were synthesized using a simple solution method in which extreme physical and chemical conditions created by cavitation within the chitosan solution allowed for the transformation of aqueous Zn(OH)(2) to crystallized ZnO and Zn2SiO3(OF)(2) in room temperature. Both the loading of sepiolite and silica with the zinc precursor significantly influenced the morphology and crystalline structure of the product, however, different zinc compounds were observed. Sepiolite was exfoliated, resulting in a fine, even colloidal solution through ultrasonic dispersion. Exfoliation of sepiolite nanofibers led to the homogeneous dispersion of Zinc in the form of Zn (OH)(2) in chitosan matrix. When the same procedure was conducted using the silica component, a formation of ZnO and Zn2SiO3(OH)(2) was observed, components that were not observed when the procedure was conducted using sepiolite. The average crystalline size of ZnO was calculated as 36 nm for ZnO. In addition, the quantities of crystalline and the ZnO phase volume was determined as 15%. Through zone of inhibition, the silica nanocomposite was discovered to have antibacterial activity. In contrast, the sepiolite compound did not exhibit these properties. We thus hypothesize that HO center dot radicals, formed during ultrasonic irradiation trigger the formation of a silicate ion (SiO32-) and formation of ZnO and Zn2SiO3(OH)(2) species in chitosan/silica/ZnO bio-nanocomposite, which causes to exhibit these antibacterial properties against Gram-negative E. coli. Chemical characterization and dispersion of the structure of the ZnO and Zn2SiO3(OH)(2) phases were done using X-ray diffraction (XRD) and scanning electron microscopy techniques (SEM) with EDAX and X-ray photoelectron spectroscopy (XPS). (C) 2016 Elsevier B.V. All rights reserved.