Efficacy of Neutral Electrolyzed Water for Inactivation of Human Norovirus

Abstract

Human norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Persistence on surfaces and resistance to many conventional disinfectants contribute to widespread transmission of norovirus. We examined the efficacy of neutral electrolyzed water (NEW; pH 7) for inactivation of human NoV GII.4 Sydney in suspension (ASTM method 1052-11) and on stainless steel surfaces (ASTM method 1053-11) with and without an additional soil load. The impact of the disinfectant on viral capsid was assessed using reverse transcriptase quantitative PCR (RT-qPCR; with an RNase pretreatment), SDS-PAGE, transmission electron microscopy, and a histo-blood group antigen (HBGA) receptor-binding assay. These studies were done in parallel with those using Tulane virus (TuV), a cultivable human NoV surrogate. Neutral electrolyzed water at 250 ppm free available chlorine produced a 4.8- and 0.4-log₁₀ reduction in NoV genome copy number after 1 min in suspension and on stainless steel, respectively. Increasing the contact time on surfaces to 5, 10, 15, and 30 min reduced human NoV genomic copies by 0.5, 1.6, 2.4, and 5.0 log₁₀ and TuV infectious titers by 2.4, 3.0, 3.8, and 4.1 log₁₀ PFU, respectively. Increased soil load effectively eliminated antiviral efficacy regardless of testing method and virus. Exposure to NEW induced a near complete loss of receptor binding (5 ppm, 30 s), degradation of VP1 major capsid protein (250 ppm, 5 min), and increased virus particle aggregation (150 ppm, 30 min). Neutral electrolyzed water at 250 ppm shows promise as an antinoroviral disinfectant when used on precleaned stainless steel surfaces.IMPORTANCE Norovirus is the leading cause of acute viral gastroenteritis worldwide. Transmission occurs by fecal-oral or vomitus-oral routes. The persistence of norovirus on contaminated environmental surfaces exacerbates its spread, as does its resistance to many conventional disinfectants. The purpose of this research was to evaluate the antinoroviral efficacy of neutral electrolyzed water (NEW), a novel chlorine-based disinfectant that can be used at reduced concentrations, making it more environmentally friendly and less corrosive than bleach. An industrial-scale electrochemical activation device capable of producing relatively stable electrolyzed water at a wide pH range was used in this study. Experiments showed that 250 ppm NEW effectively eliminated (defined as a 5-log₁₀ reduction) human norovirus GII.4 Sydney (epidemic strain) on clean stainless steel surfaces after a 30-min exposure. Supporting studies showed that, like bleach, NEW causes inactivation by disrupting the virus capsid. This product shows promise as a bleach alternative with antinoroviral efficacy.

Keywords: disinfection; environmental contamination; norovirus; public health; surrogate; virus inactivation.