In this work, the fabrication conditions of anodic aluminum oxide (AAO) templates with different pore size diameters were achieved using oxalic, tartaric, and phosphoric acidic electrolytes. Silver (Ag) nanostructures (NSs) were embedded in AAO template by simple hydrothermal and photoreduction methods. Moreover, titanium dioxide (TiO2) NSs (nanowires) was deposited into these porous templates by sol-gel method. FESEM results suggested that Ag nanofishstars, nanonecklaces (NNs), and TiO2 nanowires (NWs) like structures were grown in AAO pores with high-order and -aspect ratios. An anti-adhesive method was used to estimate the nano-size effect of the structures for enhancing antibacterial mechanism against Pseudomonas aeruginosa Gram-negative bacterium, and Staphylococcus aureus Gram-positive bacterium. In this study, the inhibition percentages of the Ag NNs/AAO membrane were 86.4, and 77.4%, respectively whereas that of the Ag film on glass substrate were 65, and 53.9%. Moreover, the inhibition percentages of the TiO2 NWs/AAO membrane were 85.9, and 70.1%, on the other hand, the TiO2 film on glass substrate were 60.3, and 45.2%. Results proved that the high porosity of the AAO template improved the contact-killing and release-killing actions of nanoparticles against biofilms.