Disintegration of Extremely Recalcitrant Bacillus cereus Spores via In Situ Layer‐By‐Layer Oxidative Disruption

ABSTRACT Spores with a unique multi‐layered stress‐resistant defense system exhibit extreme tolerance to conventional disinfection processes, leading to widespread survival and persistent public health risks. Herein, we develop a layer‐by‐layer oxidative disruption technology to achieve efficient control of highly recalcitrant Bacillus cereus spores in medical wastewater. The reactive chlorine species (dominated by Cl ₂ · ⁻ , >96%) and hydroxyl radicals are in situ generated from the wastewater matrix under coupling electrochemical oxidation and UV irradiation, which can mediate progressively layered damage to spores from the outer coat to the inner core, ultimately driving irreversible spore inactivation. The proteomic perturbation of membrane and cytoplasmic proteins is further validated via quantitative proteomic analysis. Accordingly, 6 log spores can be completely disrupted within 15 min, with a specific energy consumption of only 5.62 kWh/m ³ . The system maintains excellent operational stability during 720 h of continuous operation in practical applications, with disinfection byproduct concentrations well below the internationally accepted limits.