The following study was performed chiefly to analyze the antimicrobial activity of CdO/ZnO-ESM nanocomposites. A unique method was used for the synthesis of CdO/ZnO-ESM nanocomposites using an eggshell membrane as a bio-template which acts as both reducing and stabilizing agents; whereas cadmium nitrate and zinc nitrate were employed as metal precursors. The prepared samples were characterized by the following techniques such as TGA, PXRD, DRS-UV-Visible reflectance, FT-IR, HR-SEM, EDAX, Zeta potential, and Photoluminescence analysis. The prepared samples were subsequently tested for antimicrobial activity against Gram-positive bacteria (Staphylococcus aureus and Bacillus sp.) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Salmonella enterica, and Vibrio sp.) using agar well diffusion method. The minimum inhibitory concentration was determined by the broth microdilution method. The maximum inhibition zone of 24 mm, was observed in Salmonella enterica at a concentration (30 μg/mL) of CdO/ZnO-ESM nanocomposites, while the highest MIC value was observed in Bacillus sp. at a concentration (1.95 μg/mL) of these nanocomposites. The result of this study showed that CdO/ZnO-ESM nanocomposites have an exceptional antimicrobial activity against both Gram-positive and Gram-negative bacteria.