Inactivation Mechanism and Efficacy of Grape Seed Extract for Human Norovirus Surrogate.

Proper disinfection of harvested food and water is critical to minimize infectious disease. Grape seed extract (GSE), a commonly used health supplement, is a mixture of plant-derived polyphenols. Polyphenols possess antimicrobial and antifungal properties, but antiviral effects are not well-known. Here we show that GSE outperformed chemical disinfectants (e.g., free chlorine and peracetic acids) in inactivating Tulane virus, a human norovirus surrogate. GSE induced virus aggregation, a process that correlated with a decrease in virus titers. This aggregation and disinfection were not reversible. Molecular docking simulations indicate that polyphenols potentially formed hydrogen bonds and strong hydrophobic interactions with specific residues in viral capsid proteins. Together, these data suggest that polyphenols physically associate with viral capsid proteins to aggregate viruses as a means to inhibit virus entry into the host cell. Plant-based polyphenols like GSE are an attractive alternative to chemical disinfectants to remove infectious viruses from water or food. IMPORTANCE Human noroviruses are major food- and waterborne pathogens, causing approximately 20% of all cases of acute gastroenteritis cases in developing and developed countries. Proper sanitation or disinfection are critical strategies to minimize human norovirus-caused disease until a reliable vaccine is created. Grape seed extract (GSE) is a mixture of plant-derived polyphenols used as a health supplement. Polyphenols are known for antimicrobial, antifungal, and antibiofilm activities, but antiviral effects are not well-known. In studies presented here, plant-derived polyphenols outperformed chemical disinfectants (i.e., free chlorine and peracetic acids) in inactivating Tulane virus, a human norovirus surrogate. Based on data from molecular assays and molecular docking simulations, the current model is that the polyphenols in GSE bind to the Tulane virus capsid, an event that triggers virion aggregation. It is thought that this aggregation prevents Tulane virus from entering host cells.

Influence of point-of-use filters and stagnation on water quality at a preschool and under laboratory conditions.

A local preschool installed NSF/ANSI 42 and 53 certified point-of-use (POU) filters in its classroom sinks and drinking fountains to protect children from the possibility of elevated lead (Pb) levels in drinking water. We examined the effects of these filters during flowing water and immediately following stagnation periods on Pb, chlorine, and bacterial concentrations in the field and the laboratory. Before and after typical school stagnation periods, we collected samples from filtered classroom sinks, a filtered drinking fountain and nearby unfiltered sinks for a year. No unfiltered samples exceeded Illinois State limits of 5 µg/L for Pb in pre-K through 5th grade schools. However, following stagnation periods as short as overnight (14.5 h), over half of post-stagnation filtered samples from classroom sinks exceeded 5 µg/L while post-stagnation unfiltered samples remained below 5 µg/L. Laboratory testing showed no significant increases in Pb with stagnation, suggesting that the preschool classrooms may have had Pb-bearing plumbing downstream of the filters which released Pb into the filtered drinking water. The filters effectively removed free chlorine (99% decrease) in both the preschool and laboratory. Installing the filters had the unintended consequence of significantly increasing the bacterial concentrations (as measured by qPCR) in the preschool's drinking water and in laboratory filter effluent. Legionella pneumophila, Pseudomonas aeruginosa, and Mycobacterium spp. were not detected in pre-stagnation unfiltered and post-stagnation filtered samples. These results suggest that the installation of POU filters be considered as one component of an overall strategy to decrease Pb concentrations in school drinking water that holistically considers the premise plumbing system. A 5-minute flush significantly decreased concentrations of Pb and bacteria in filtered sinks. Replacing Pb-bearing plumbing components downstream of a POU filter may also be needed to combat Pb levels in drinking water.

Inactivation and sensitization of Pseudomonas aeruginosa by microplasma jet array for treating otitis media.

Otitis media (OM), known as a middle ear infection, is the leading cause of antibiotic prescriptions for children. With wide-spread use of antibiotics in OM, resistance to antibiotics continues to decrease the efficacy of the treatment. Furthermore, as the presence of a middle ear biofilm has contributed to this reduced susceptibility to antimicrobials, effective interventions are necessary. A miniaturized 3D-printed microplasma jet array has been developed to inactivate Pseudomonas aeruginosa, a common bacterial strain associated with OM. The experiments demonstrate the disruption of planktonic and biofilm P. aeruginosa by long-lived molecular species generated by microplasma, as well as the synergy of combining microplasma treatment with antibiotic therapy. In addition, a middle ear phantom model was developed with an excised rat eardrum to investigate the antimicrobial effects of microplasma on bacteria located behind the eardrum, as in a patient-relevant setup. These results suggest the potential for microplasma as a new treatment paradigm for OM.

Deposition and aggregation kinetics of rotavirus in divalent cation solutions.

Aggregation kinetics of rotavirus in aqueous solutions and its deposition kinetics on silica surface in the presence of divalent (Ca(2+), Mg(2+)) cations were studied using complementary techniques of time-resolved dynamic light scattering (TR-DLS) and quartz crystal microbalance (QCM). Within a reasonable temporal window of 4 h, aggregation could be observed at levels as low as 10 mM of Ca(2+) and 20 mM of Mg(2+). Attachment efficiencies were always greater in Ca(2+) solutions of the same concentration, and the critical coagulation concentration (CCC) for rotavirus in Ca(2+) solutions was slightly smaller than that in Mg(2+) solutions. No aggregation was detected in Na(+) solution within the temporal window of 4 h. Deposition experiments showed higher attachment coefficients in solutions containing Ca(2+) compared to those obtained in Mg(2+) solution. The classic Derjaguin-Landau-Verwey-Overbeek (DLVO) theory failed to predict both the aggregation behavior of rotavirus and its deposition on silica surface. Besides electrostatic interactions, steric repulsions and specific interactions with divalent cations were important mechanisms in controlling rotavirus deposition and aggregation. Experimental results presented here suggest that rotavirus is not expected to aggregate in groundwater with typical hardness (up to 6 mM Ca(2+)) and rotavirus deposition on silica soil would be more favorable in the presence of Ca(2+) than Mg(2+).