The thermal treatment method was employed to achieve higher homogeneity of calcium ferrite (CaFe2O4) and Poly (vinyl alcohol) (PVA) nanocomposites. The influences of phase transformation on physical and biological properties of calcined specimens were investigated by various experimental techniques including X-ray diffraction (XRD), Transmission Electron Microscopy (TEM), high resolution Field emission scanning electron microscope (FESEM) and Fourier transform infrared spectroscopy (FT-IR). Heat treatment was conducted at temperatures between 723 and 923 K, so that a phase transformation occurred from cubic to orthorhombic spinel structure at 923 K. The chemical analysis of the PVA/CaFe2O4 nanocomposite was performed by energy dispersion X-ray analysis (EDXA), demonstrated the PVA/CaFe2O4 nanocomposites contained the elements of C, Ca, Fe, and O. The formed nanocomposites exhibited ferromagnetic behaviors which were confirmed by using a vibrating sample magnetometer (VSM). The calcined specimens were carried out to an antimicrobial or antifungal test.