Heated scallop-shell (HSS) nano-particles, prepared using a wet grinding mill, and microparticles were examined for their antibacterial activity against vegetative bacterial cells and spores. The median diameters of the nano-particles and micro-particles were approximately 20 nm and 30 µm, respectively. The antibacterial activity of HSS against Escherichia coli increased with an increase in concentration, regardless of particle size; however, the antibacterial activity of the nano-particles was much higher than that of micro-particles. The sporicidal activity of the nano-particles was also much higher than that of micro-particles, with HSS nano-particles able to kill Bacillus subtilis spores. A reduction of more than three orders of magnitude for B. subtilis spores was confirmed following a 30 min treatment at 5 mg/ml and 60℃, showing that the combination of HSS nano-particle treatment with mild heating was particularly effective for controlling bacterial spores.