Natural chitosan (CS) at four different additions (10, 20, 30, and 40 phr) and particle size ranges (100–140, 140–180, 180–220, and over 260 mesh) are selected to improve the interface adhesion as well as endow a novel antibacterial function to wood flour/polyvinyl chloride (WF/PVC) composites. In the present study, we investigate the interface self-reinforcing ability of CS to composites by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), dynamic mechanical analysis (DMA), mechanical testing instrument, and water absorption behavior test (WB). The antibacterial activity is also estimated by the method of membrane covering test (MCT) using Escherichia coli. The results recorded show that adding 30 phr CS with the particle size of over 260 mesh is considered to be perfect selection to prepare the excellent interfacial self-reinforcing and antibacterial WF/PVC/CS composites. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 39854.