Peroxide Disinfectionof Vesicle-Cloaked Murine NorovirusClusters: Vesicle Membranes Protect Viruses from Inactivation

Vesicle-cloaked virus clusters, also known as viral vesicles, exhibit higher infectivity than free viruses and demonstrate persistence in the environment as well as resilience against disinfection. These emerging pathogens represent significant, yet often underestimated, health risks. Our study investigated peroxide disinfection of murine norovirus vesicles, a surrogate for human norovirus vesicles, and elucidated disinfection mechanisms. Peracetic acid, a neutral peroxide, rapidly inactivated murine norovirus vesicles. In contrast, negatively charged peroxides, i.e., peracetate and peroxymonosulfate, exhibited restricted effectiveness in inactivating murine noroviruses within vesicles. The largely intact viruses cloaked within vesicles remained infectious and retained their ability to replicate upon vesicle lysis triggered by mechanical forces, enzymatic activity, or chemical reactions following disinfection. Peroxides primarily targeted vesicle/viral proteins, particularly amino acid residues such as cysteine and methionine, without affecting the viral ORF2 gene fragment or vesicle structures. Disruption of viral internalization, rather than binding, plays a key role in infectivity loss. This work highlights the protective role of vesicle membranes and emphasizes the need for innovative disinfection approaches to effectively target viruses cloaked within vesicles.