Characterization of Cox-1Nix Fe2O4 nanoparticles (NPs) employing X-ray diffraction and field emission scanning electron microscopy (FESEM) was accomplished simply by chemical coprecipitation, FTIR and finally the Magnetic Resonance Spectrometer (VSM), the singlephase cubic spinel structure in X-ray diffraction. In FESEM, micrographs of Co ferrites nanoparticles that are virtually spherical can be seen with grain sizes of less than 20 nm. Two absorption bands are visible in the FTIR spectrum, with values ranging from 400-600 cm. These beams show that all of the samples have a ferrite spectral composition. Preparation of samples yielded M-H curves because of the tight turn, which means the paper samples are soft magnetic material in the VSM. Antibacterial activity of Cox-1Nix Fe2O4 NPs in limiting the development of isolated pathogenic bacteria Staphylococcus aureus and E. coli was studied to compare their effects with the traditional antibiotics used before. Using Cox-1Nix Fe2O4 nanoparticles, it has been found that the nanoparticles release ions into the environment, which interacts with the group (-SH) of proteins, resulting in the defection of bacteria's cell membranes and the subsequent cell death.