Fish scale gelatin, as a reducing agent and stabilizer, has been used to prepare silver nanoparticles (AgNPs) and fish scale gelatin/agar composite films with AgNPs loading (0.04-0.2%, dry weight). AgNPs were characterized by UV-Vis spectroscopy (UV-Vis), transmission microscopy (TEM), X-ray diffraction (XRD) to check the physicochemical properties. AgNPs were spherical and homogeneous with an average particle size of 9.3 nm. With the addition of AgNPs, the color of fish scale gelatin/agar composite films turned yellow, the transparency reduced, and the absorption of ultraviolet and visible light enhanced. Besides, the elongation at break, water-resistance and water vapor barrier properties of the films were significantly enhanced with the addition of AgNPs; however, the thickness and tensile strength had no change. FTIR and TG analyses showed that Ag NPs chemically interacted with fish scale gelatin and agar, and the thermal stability of the composite films was improved to some extent. Finally, the films incorporated with Ag NPs exhibited strong antibacterial activity against tested pathogens bacteria. Importantly, AgNPs and fish scale gelatin-agar composite films are compatible with the utility of AgNPs at a very low silver concentration in antibacterial packaging.