Proper packaging is an essential issue in seafood safety leads to the preservation of food quality and extending the shelf life. In this study, fabrication and characterization of LDPE/Ag/TiO2 and LDPE/Ag+Cu/TiO2 nanocomposite flms for application in Nile Tilapia packaging were evaluated. After investigation of scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), disk difusion test, minimum inhibitory concentration, and minimum bactericidal concentration, the antimicrobial efect of produced nanocomposite flms was evaluated on the Tilapia fsh samples stored at 4 °C and −20 °C for 5 and 10 days. Chemical features (pH, protein and fat concentrations, and free fatty acid profle) of the Tilapia fsh covered with the produced nanocomposites and nanoparticle migration were also assessed. EDS and SEM techniques confrmed the presence of nanoparticles on the polymer surface and their relative homogeneity. The antibacterial tests of LDPE/Ag+Cu/ TiO2 nanocomposite flm exhibited strong antibacterial activities against Escherichia coli and Listeria monocytogenes bacteria (P=0.000). The microbiological tests revealed that Ag+Cu-contained flm had signifcantly higher antimicrobial efcacy on the Nile Tilapia samples (P<0.05). The overall result of the series of chemical experiments showed that Tilapia samples packed in Ag+Cu flm had the least changes in chemical properties compared to fresh samples (P<0.05 for pH, fat concentration, and free fatty acid profle). Besides, the migration of Ag and Cu nanoparticles from the flm to Tilapia samples was slightly (the amount of Ag and Cu release was <2.0 µg/Kg and <10 µg/Kg, respectively). From the obtained results, it could be concluded that a flm containing 2.5% silver, 2.5% copper, and 5% titanium dioxide nanoparticles had the most signifcant antimicrobial efect on the Nile Tilapia fsh. Therefore, using LDPE/Ag+Cu/TiO2 nanocomposite flm can be a promising approach to create active packaging in the seafood industry.