Barium titanate, perovskite structure is known for its high dielectric constant and piezoelectric properties, which makes it interesting material for fabricating capacitors, transducer, actuator, and sensors. The perovskite crystal structure and lattice vibrations play a crucial role in its piezoelectric and ferroelectric behavior. In the present study, the barium titanate powder was subjected to biofield treatment. Further, the control and treated samples were characterized using X-ray diffraction (XRD) and Fourier transform infrared spectrometer (FT-IR) and Electron spin resonance (ESR). The XRD analysis showed the permanent compressive strain of 0.45% in treated barium titanate powder as compared to control. Furthermore, the biofield treatment has enhanced the density upto 1.38% in barium titanate as compared to control. The FT-IR spectra showed that the stretching and bending vibrations of Ti-O bond in treated BaTiO3 were shifted towards lower frequency as compared to control. The bond length was substantially increased by 0.72 % in treated BaTiO3 as compared to control. The ESR spectra of control and treated BaTiO3 sample showed the g-factor of 2.0; and biofield treatment has substantially changed the width and height of ESR signal in treated BaTiO3 as compared to control. These observations revealed that biofield treatment has significantly altered the crystal structure, lattice strain, and bond vibration of barium titanate.