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Many factors determine the resistance properties of a Bacillus spore to heat, chemical and physical processing, including thick proteinaceous coats, peptidoglycan cortex and low water content, high levels of dipicolinic acid (DPA), and divalent cations in the spore core. Recently, attention has been focused on non-thermal inactivation methods based on high pressure, ultrasonic, high voltage electric fields and cold plasmas for inactivating Bacillus spores associated with deterioration in quality and safety. The important chemical sporicides are glutaraldehyde, chorine-releasing agents, peroxygens, and ethylene oxide. Some food-grade antimicrobial agents exhibit sporostatic and sporicidal activities, such as protamine, polylysine, sodium lactate, essential oils. Surfactants with hydrophilic and hydrophobic properties have been reported to have inactivation activity against spores. The combined treatment of physical and chemical treatment such as heating, UHP (ultra high pressure), PEF (pulsed electric field), UV (ultraviolet), IPL (intense pulsed light) and natural antimicrobial agents can act synergistically and effectively to kill Bacillus spores in the food industry.