Ozone, hydrogen peroxide, and peroxymonosulfate disinfection of MS2 coliphage in water.

The control of viruses in water is critical to preventing the spread of infectious viral diseases. Many oxidants can inactivate viruses, and this study aims to systematically compare the disinfection effects of ozone (O3), peroxymonosulfate (PMS), and hydrogen peroxide (H2O2) on MS2 coliphage. The effects of oxidant dose and contact time on disinfection were explored, as were the disinfection effects of three oxidizing agents in secondary effluent. The 4-log inactivation of MS2 coliphage required 0.05 mM O3, 0.5 mM PMS, or 25 mM H2O2 with a contact time of 30 min. All three oxidants achieved at least 4-log disinfection within 30 min, and O3 required only 0.5 min. In secondary effluent, all three oxidants also achieved 4-log inactivation of MS2 coliphage. Excitation-emission matrix (EEM) results indicate that all three oxidants removed dissolved organic matter synchronously and O3 oxidized dissolved organic matter more thoroughly while maintaining disinfection efficacy. Considering the criteria of oxidant dose, contact time, and disinfection efficacy in secondary effluent, O3 is the best choice for MS2 coliphage disinfection among the three oxidants.

Efficacy of novel aqueous photo-chlorine dioxide against a human norovirus surrogate, bacteriophage MS2 and Clostridium difficile endospores, in suspension, on stainless steel and under greenhouse conditions.

AIMS: The efficacy of a novel photochemical method for generating chlorine dioxide (photoClO2 ) was evaluated against human noroviruses (HuNoV) surrogate, bacteriophage MS2, and Clostridium difficile endospores. METHODS AND RESULTS: Chlorine dioxide was generated by mixing 1% sodium chlorite with 10 parts-per-million (ppm) Eosin Y and irradiating with a photo-activator-excitable light. PhotoClO2 efficacy was assessed against bacteriophage MS2 and C. difficile endospores in suspension, on hard surfaces and greenhouse conditions under soiled and unsoiled conditions. The estimated effective photoClO2 produced and consumed was 20·39 ± 0·16 ppm at a rate of 8·16 ppm per min in a 1% sodium chlorite solution. In suspension, MS2 phage was reduced by 3·35 and >5·10 log10 PFU per ml in 120 and 90 min, with and without soil, respectively. At the same time, when dried on stainless steel surface, MS2 phage was reduced by >4·53 log10 PFU per carrier in 30 min under both conditions. On the other hand, C. difficile endospores in suspension were reduced by 2·26 and 3·65 log10 CFU per ml in 120 min with and without soiling, respectively. However, on stainless steel surface, maximal reductions of the C. difficile endospores were 0·8 and 1·5 log10 CFU per carrier with and without soiling, respectively, and a maximal reduction of 2·97 log10 CFU per carrier under greenhouse conditions at 24 h. CONCLUSIONS: Overall, photoClO2 showed promise as a technology to control HuNoV contamination on environmental surfaces but requires further optimization and testing against C. difficile endospores. SIGNIFICANCE AND IMPACT OF THE STUDY: Results from this investigation will serve as a model for how to generate and quantify photoClO2 and how to appropriately evaluate this new class of disinfectants against environmentally resilient pathogens: viruses and bacterial endospores.

Modification of Major Contributors Responsible for Latrine Malodor on Exposure to Hypochlorous Acid: The Potential for Simultaneously Impacting Odor and Infection Hazards to Encourage Latrine Use.

Open defecation remains a common practice in developing countries and leads to high incidence and prevalence of acute gastroenteritis, which is most often caused by human noroviruses (human NoV). Encouraging the use of toilets and pit latrines is one method of improving sanitation; however, it is often hindered by not only cultural traditions but also from a reluctance to use latrines and toilets due to their odor and impression of uncleanliness. In an effort to establish new means to encourage toilet and latrine use, laboratory experiments tested the ability of hypochlorous acid (HOCl) to modify the malodorous compounds identified in the air in latrines in developing countries (indole, p-cresol, dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS), and butyric acid) and inactivate MS2 bacteriophage, a surrogate for human NoV. After 5 minutes, > 94% of indole, p-cresol, DMDS, and DMTS was modified as determined by high-pressure liquid chromatography in the presence of 100 ppm HOCl. A log10 reduction value (LRV) greater than 6 was seen for MS2 bacteriophage after 5 minutes of exposure to 100 ppm HOCl in solution. Sensory studies indicated that there was a significant difference (P ≤ 0.05) between the untreated and HOCl-treated samples for all five malodorous compounds tested. The findings suggest that introduction of HOCl into the headspace air could encourage latrine and toilet use. Optimization of HOCl dosing in air to accomplish both odor control and reduction of infectious hazards is worthy of further study.

Inactivation of airborne porcine reproductive and respiratory syndrome virus (PRRSv) by a packed bed dielectric barrier discharge non-thermal plasma.

Porcine reproductive and respiratory syndrome virus (PRRSv) is one of the most significant airborne viruses impacting the pork industry in the US. Non-thermal plasmas (NTPs) are electrical discharges comprised of reactive radicals and excited species that inactivate viruses and bacteria. Our previous experiments using a packed bed NTP reactor demonstrated effective inactivation of bacteriophage MS2 as a function of applied voltage and power. The present study examined the effectiveness of the same reactor in inactivating aerosolized PRRSv. A PRRSv solution containing ∼105 TCID50/ml of PRRSv VR2332 strain was aerosolized at 3 ml/min by an air-jet nebulizer and introduced into 5 or 12 cfm air flow followed by NTP exposure in the reactor. Twin impingers upstream and downstream of the reactor collected samples of the virus-laden air flow for subsequent TCID50 assay and qPCR analyses. An optical particle sizer measured upstream and downstream aerosol size distributions, giving estimates of aerosol filtration by the reactor. The results showed that PRRSv was inactivated to a similar degree as MS2 at the same conditions, with the maximum 1.3-log inactivation of PRRSv achieved at 20 kV and 12 cfm air flow rate. The results demonstrate the potential of properly optimized NTPs in controlling PRRSv transmission.

Reduction of Escherichia Coli Using Metal Plates with the Influenced of Applied Low Current and Physical Barrier of Filter Layers.

Although metal contact is known to reduce bacterial growth, the effects of physical barriers and electricity need further investigation. This study examined the bacteria-reducing properties of copper and stainless-steel metal plates with an added electrical current and up to three filter layers on the growth of Escherichia coli (bacteria) and MS2 bacteriophages (virus). When used with a stainless-steel plate, electricity increased bacteria reduction by 39.5 ± 2.30% in comparison with no electricity added, whereas a three-layer physical barrier decreased its efficiency. Copper also reduced the growth of bacteria, by 58.2 ± 8.23%, and the addition of electricity reduced it further (79.5 ± 2.34%). Bacteriophages were also affected by the metal contact. Further experiments showed that MS2 was also reduced by copper, to 82.9 ± 4.5% after 24 h at 37 °C.

The use of bacteriophage MS2 for the development and application of a virucide decontamination test method for porous and heavily soiled surfaces.

AIMS: (i) To develop an analytical method for recovery and quantification of bacteriophage MS2-as a surrogate for foot-and-mouth disease virus-from complex porous surfaces, with and without the presence of laboratory-developed agricultural grime; (ii) to evaluate, with a 4-log dynamic range, the virucidal activity of common biocides for their ability to decontaminate surfaces and hence remediate facilities, following a foreign animal disease contamination incident. METHODS AND RESULTS: An analytical method was developed and optimized for MS2 recovery from simulated agricultural surfaces. The addition of Dey-Engley neutralizing broth to an extraction buffer improved MS2 viability in liquid extracts, with optimal analytical holding times determined as <8 to ≤24 h, depending on matrix. The recovery of MS2 from surface materials decreased in the order: nonporous reference material >grimed porous materials >nongrimed porous materials. In disinfectant testing, two spray applications of pAB were effective against MS2 (≥4-log reduction) on all operational-scale materials. Two per cent citric acid had limited effectiveness, with a ≥4-log reduction observed on a selected subset of grimed concrete samples. CONCLUSIONS: Decontamination efficacy test results can be affected by surface characteristics, extraction buffer composition, analytical holding time and surface-specific organism survivability. Efficacy should be evaluated using a test method that reflects the environmental characteristics of the intended application. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study demonstrate the importance of analytical method verification tests for disinfectant testing prior to application in complex environments.

Interaction between functionalized multiwalled carbon nanotubes and MS2 bacteriophages in water.

Fate and transport of carbon nanomaterials can be strongly dependent on the interaction with secondary particulates in the aquatic systems. Bio-particulates in water, e.g., viruses with charged and hydrophobic surface moieties, may profoundly influence the interfacial behavior and hence the environmental fate of nanomaterials (and vice versa). This study systematically evaluates the interfacial interaction of acid-functionalized multiwalled carbon nanotubes (MWNTs) with MS2 bacteriophages, or heteroaggregation behavior of these particulates, under mono- and di-valent cations and with Suwannee River humic acid (SRHA). Results indicate that the highest concentration of MS2 (i.e., MWNT:MS2 of 100:1) renders exceptional stability of MWNTs, even in high salinity conditions. However, at lower MS2 concentrations (i.e., MWNT:MS2 of 1000:1 and 10,000:1), the suppression of MWNT heteroaggregation rate is not as significant. The observed enhanced stability is likely caused by the preferential attachment of the MS2 capsids onto MWNT surfaces, which is mediated by electrostatic attraction (between negatively charged oxygen-containing moieties on MWNTs and positively charged amino acid residues on MS2 surfaces) and/or by MS2 capsids with positive hydropathy index (indicating strong hydrophobicity). Presence of SRHA also shows stability enhancement; however, at lower MS2 concentrations, SRHA dominated the heteroaggregation behavior. These results implicate that preferential interaction between virus capsids (i.e., MS2 and may be other waterborne viruses) and carbonaceous nanomaterials may influence environmental transport of both in aquatic environments.

Influence of algal organic matter on MS2 bacteriophage inactivation by ultraviolet irradiation at 220 nm and 254 nm.

We determined the potential interference of extracellular algal organic matter (EAOM) and intracellular algal organic matter (IAOM) extracted from Microcystis aeruginosa on MS2 bacteriophage inactivation under UV irradiation at two wavelengths (220 and 254 nm). UV irradiation at 220 nm doubled the inactivation rate of MS2 in water containing EAOM than in organic-free phosphate buffered solution. In contrast, EAOM did not change MS2 inactivation by exposure to UV 254 nm. The presence of IAOM did not significantly influence MS2 inactivation by exposure to either UV 254 or UV 220 nm. To achieve 3 log10 inactivation of MS2, UV254 nm required more than double the dose of UV220 nm (45 mJ/cm2 vs. 20 mJ/cm2). Linear correlations between the reduction in infectivity and the reduction in genome copies detected by reverse transcription quantitative polymerase chain reaction suggested that genomic damage is the main mechanism responsible for MS2 inactivation in water containing algal organic matter (AOM) by exposure to UV irradiation. These findings suggest that the presence of AOM did not negatively influence MS2 inactivation by either 220 or 254 nm irradiation, and that a lower UV dose of 220 nm irradiation can be used to achieve the same level of inactivation in water containing AOM.

Influence of water antimicrobials and storage conditions on inactivating MS2 bacteriophage on strawberries.

Foodborne illnesses caused by norovirus contaminated fresh produce remain a food safety concern worldwide. In the present study, the impacts of commercial and home processing conditions of strawberries were evaluated for inactivation of the MS2 bacteriophage. MS2 was used as a surrogate of norovirus and was spot inoculated onto strawberries to achieve 6.6 log PFU/g. The inoculated strawberries were washed with tap water, electrolyzed water, or 50 ppm chlorine for 90 s prior to and after storage. After initial washing, the strawberries were separately stored at -20 °C and -80 °C for 30 days. Change in MS2 populations on strawberries was evaluated by plaque assay method on day 1, 15, and 30 for -20 °C and -80 °C groups. The results showed that washing strawberries prior to storage resulted in a significant decrease (approximately 1 log PFU/g) of MS2 population regardless of the treatment (p < 0.05). Frozen storage had minor effects on inactivating MS2, which resulted in approximately a 0.5 log PFU/g reduction at the end of storage. Washing frozen berries in electrolyzed water or 50 ppm chlorine on day 30 resulted in an additional 1 log PFU/g decrease in MS2 compared to water alone. These results suggest that washing strawberries with a chemical antimicrobial prior to and post frozen storage may enhance microbial safety.

Free Chlorine Disinfection of Full-Scale MBR Effluent to Achieve 5-Log Virus Inactivation.

A series of full-scale testing was performed at the City of Lathrop Consolidated Treatment Facility to determine the "concentration times time" (Ct) value for free chlorine disinfection of nitrified membrane bioreactor (MBR) effluent to achieve more than 5-log virus inactivation. The full-scale testing consisted of tracer study, chlorine decay study, and virus seeding study. The virus seeding study was performed at a flow rate of 1 million gal/d (3800 m3/min), which corresponded to a theoretical contact time of 117 minutes in the chlorine contact basin. It was found that Ct values as small as 3 mg·min/L were sufficient to achieve 5-log inactivation of MS2 bacteriophage in this study, which is comparable to the results of previous bench- and pilot-scale free chlorine disinfection studies on nitrified MBR effluent (3 to 40 mg·min/L), as well as those of pilot- and full-scale studies on granular media-filtered nitrified effluent (2 to 22 mg·min/L).

A new approach to testing the efficacy of drinking water disinfectants.

New disinfection procedures are being developed and proposed for use in drinking-water production. Authorising their use requires an effective test strategy that can simulate conditions in practice. For this purpose, we developed a test rig working in a flow-through mode similar to the disinfection procedures in waterworks, but under tightly defined conditions, including very short contact times. To quantify the influence of DOC, temperature and pH on the efficacy of two standard disinfectants, chlorine and chlorine dioxide, simulated use tests were systematically performed. This test rig enabled quantitative comparison of the reduction of four test organisms, two viruses and two bacteria, in response to disinfection. Chlorine was substantially more effective against Enterococcus faecium than chlorine dioxide whereas the latter was more effective against the bacteriophage MS2, especially at pH values of >7.5 at which chlorine efficacies already decline. Contrary to expectation, bacteria were not generally reduced more quickly than viruses. Overall, the results confirm a high efficacy of chlorine and chlorine dioxide, validating them as standard disinfectants for assessing the efficacy of new disinfectants. Furthermore, these data demonstrate that the test rig is an appropriate tool for testing new disinfectants as well as disinfection procedures.

Mechanism of Virus Inactivation by Cold Atmospheric-Pressure Plasma and Plasma-Activated Water.

Viruses cause serious pathogenic contamination that severely affects the environment and human health. Cold atmospheric-pressure plasma efficiently inactivates pathogenic bacteria; however, the mechanism of virus inactivation by plasma is not fully understood. In this study, surface plasma in argon mixed with 1% air and plasma-activated water was used to treat water containing bacteriophages. Both agents efficiently inactivated bacteriophages T4, Φ174, and MS2 in a time-dependent manner. Prolonged storage had marginal effects on the antiviral activity of plasma-activated water. DNA and protein analysis revealed that the reactive species generated by plasma damaged both nucleic acids and proteins, consistent with the morphological examination showing that plasma treatment caused the aggregation of bacteriophages. The inactivation of bacteriophages was alleviated by the singlet oxygen scavengers, demonstrating that singlet oxygen played a primary role in this process. Our findings provide a potentially effective disinfecting strategy to combat the environmental viruses using cold atmospheric-pressure plasma and plasma-activated water.IMPORTANCE Contamination with pathogenic and infectious viruses severely threatens human health and animal husbandry. Current methods for disinfection have different disadvantages, such as inconvenience and contamination of disinfection by-products (e.g., chlorine disinfection). In this study, atmospheric surface plasma in argon mixed with air and plasma-activated water was found to efficiently inactivate bacteriophages, and plasma-activated water still had strong antiviral activity after prolonged storage. Furthermore, it was shown that bacteriophage inactivation was associated with damage to nucleic acids and proteins by singlet oxygen. An understanding of the biological effects of plasma-based treatment is useful to inform the development of plasma into a novel disinfecting strategy with convenience and no by-product.

The impact of chlorine and heat on the infectivity and physicochemical properties of bacteriophage MS2.

Enteric viruses and bacteriophages are exposed to various inactivating factors outside their host, and among them chlorine and heat are the most commonly used sanitizer in water industry and treatment in the food industry, respectively. Using MS2 phages as models for enteric viruses, we investigated the impact of free chlorine and heat on their physicochemical properties. Free chlorine was first evaluated alone. No increase in either capsid permeability or hydrophobicity was observed. The negative surface charge slightly increased suggesting molecular changes in the capsid. However, a weakening of the capsid by chlorine was suggested by differential scanning fluorimetry. This phenomenon was confirmed when chlorination was followed by a heat treatment. Indeed, an increase in the inactivation of MS2 phages and the permeability of their capsids to RNases was observed. More interestingly, an increase in the expression of hydrophobic domains at the phage surface was observed, but only for phages remaining infectious. The chlorine-caused weakening of the capsid suggested that, for an optimal use, the oxidant should be followed by heat. The increased permeability to RNases and the expression of hydrophobic domains may contribute to the development or improvement of molecular methods specific for infectious enteric viruses.

Improved virus inactivation using a hot bubble column evaporator (HBCE).

An improved hot bubble column evaporator (HBCE) was used to study virus inactivation rates using hot bubble-virus interactions in two different conditions: (1) using the bubble coalescence inhibition phenomenon of monovalent electrolytes and (2) with reducing the electrostatic repulsive forces between virus and bubble, by the addition of divalent electrolytes. It is shown that the continuous flow of (dry) air, even at 150-250 °C, only heats the aqueous solution in the bubble column to about 45°-55 °C and it was also established that viruses are not significantly affected by even long term exposure to this solution temperature, as confirmed separately from water bath experiments. Hence, the effects observed appeared to be caused entirely by collisions between the hot air bubbles and the virus organisms. It was also established that the use of high air inlet temperatures, for short periods of time, can reduce the thermal energy requirement to only about 25% (about 114 kJ/L) of that required for boiling (about 450 kJ/L).

Evaluation of hospital-grade disinfectants on viral deposition on surfaces after toilet flushing.

BACKGROUND: Past studies have shown that infectious aerosols created during toilet flushing result in surface contamination of the restroom. The goals of this study were to quantify viral contamination of surfaces in restrooms after flushing and the impact of disinfectants added to the toilet bowl prior to flushing on reducing surface contamination. METHODS: The degree of contamination of surfaces in the restroom was assessed with and without the addition of coliphage MS2 to the toilet bowl before flushing. The bowl water and various surfaces in the restroom were subsequently tested for the presence of the virus. RESULTS: The toilet bowl rim, toilet seat top, and toilet seat underside were contaminated in all trials without a disinfectant added to the bowl water before flushing. All disinfectants significantly reduced concentrations on surfaces when the contact time was ≥15 minutes. Hydrogen peroxide resulted in very little reduction of virus in the toilet bowl (<1 log10). Peracetic acid and quaternary ammonium had the greatest log reductions on virus in the organic matter in the toilet. CONCLUSIONS: Toilet flushing resulted in extensive contamination of surfaces within the restroom. Addition of disinfectant to the toilet bowl prior to flushing reduced the level of contamination in the bowl and fomites after flushing.

Effects of solution chemistry on the sunlight inactivation of particles-associated viruses MS2.

The inactivation efficacy of bacteriophage MS2 by simulated sunlight irradiation was investigated to understand the effects of MS2 aggregation and adsorption to particles in solutions with different components. Kaolinite and Microcystis aeruginosa were used as model inorganic and organic particles, respectively. Lower pH and di-valent ions (Ca2+) were main factors on the aggregation and inactivation of MS2. In the presence of both particles, there was no significant impact on the MS2 inactivation efficacy by kaolinite (10-200mM) or Microcystis aeruginosa (102-105Cells/mL) in 1mM NaCl at pH 7. However at lower pH 3, MS2 aggregates formed in the particle-free and kaolinite-containing solutions, caused lower inactivation since the outer viruses of aggregation protect the inner viruses. In addition, more MS2 adsorbed on Microcystis aeruginosa at lower pH (3 and 4). Microcystis aeruginosa would act as a potential photosensitizer for ROS production to inactivate the adsorbed MS2, since extracellular organic matter (EOM) of Microcystis aeruginosa was detected in this study, which has been reported to produce ROS under solar irradiation. At pH 7, Na+ had no effect on the inactivation of MS2, because MS2 was stable and dispersed even at 200mM Na+. MS2 aggregated and adsorbed on particles even at 10mM Ca2+ and led to lower inactivation. Kaolinite cannot offer enough protection to adsorbed MS2 as aggregation and Microcystis aeruginosa acts as potential photosensitizer to produce ROS and inactivate the adsorbed MS2 at high concentration of Ca2+. In particle-free solution, SRNOM inhibited MS2 inactivation by shielding the sunlight and coating MS2 to increase its survival.

Analogies and differences among bacterial and viral disinfection by the photo-Fenton process at neutral pH: a mini review.

Over the last years, the photo-Fenton process has been established as an effective, green alternative to chemical disinfection of waters and wastewaters. Microorganisms' inactivation is the latest success story in the application of this process at near-neutral pH, albeit without clearly elucidated inactivation mechanisms. In this review, the main pathways of the combined photo-Fenton process against the most frequent pathogen models (Escherichia coli for bacteria and MS2 bacteriophage for viruses) are analyzed. Firstly, the action of solar light is described and the specific inactivation mechanisms in bacteria (internal photo-Fenton) and viruses (genome damage) are presented. The contribution of the external pathways due to the potential presence of organic matter in generating reactive oxygen species (ROS) and their effects on microorganism inactivation are discussed. Afterwards, the effects of the gradual addition of Fe and H2O2 are assessed and the differences among bacterial and viral inactivation are highlighted. As a final step, the simultaneous addition of both reagents induces the photo-Fenton in the bulk, focusing on the differences induced by the homogeneous or heterogeneous fraction of the process and the variation among the two respective targets. This work exploits the accumulated evidence on the mechanisms of bacterial inactivation and the scarce ones towards viral targets, aiming to bridge this knowledge gap and make possible the further application of the photo-Fenton process in the field of water/wastewater treatment.

Evaluation of Chlorine Treatment Levels for Inactivation of Human Norovirus and MS2 Bacteriophage during Sewage Treatment.

This study examined the inactivation of human norovirus (HuNoV) GI.1 and GII.4 by chlorine under conditions mimicking sewage treatment. Using a porcine gastric mucin-magnetic bead (PGM-MB) assay, no statistically significant loss in HuNoV binding (inactivation) was observed for secondary effluent treatments of ≤25 ppm total chlorine; for both strains, 50 and 100 ppm treatments resulted in ≤0.8-log10 unit and ≥3.9-log10 unit reductions, respectively. Treatments of 10, 25, 50, and 100 ppm chlorine inactivated 0.31, 1.35, >5, and >5 log10 units, respectively, of the norovirus indicator MS2 bacteriophage. Evaluation of treatment time indicated that the vast majority of MS2 and HuNoV inactivation occurred in the first 5 min for 0.2-µm-filtered, prechlorinated secondary effluent. Free chlorine measurements of secondary effluent seeded with MS2 and HuNoV demonstrated substantial oxidative burdens. With 25, 50, and 100 ppm treatments, free chlorine levels after 5 min of exposure ranged from 0.21 to 0.58 ppm, from 0.28 to 16.7 ppm, and from 11.6 to 53 ppm, respectively. At chlorine treatment levels of >50 ppm, statistically significant differences were observed between reductions for PGM-MB-bound HuNoV (potentially infectious) particles and those for unbound (noninfectious) HuNoV particles or total norovirus particles. While results suggested that MS2 and HuNoV (measured as PGM-MB binding) behave similarly, although not identically, both have limited susceptibility to chlorine treatments of ≤25 ppm total chlorine. Since sewage treatment is performed at ≤25 ppm total chlorine, targeting free chlorine levels of 0.5 to 1.0 ppm, these results suggest that traditional chlorine-based sewage treatment does not inactivate HuNoV efficiently.IMPORTANCE HuNoV is ubiquitous in sewage. A receptor binding assay was used to assess inactivation of HuNoV by chlorine-based sewage treatment, given that the virus cannot be routinely propagated in vitro Results reported here indicate that chlorine treatment of sewage is not effective for inactivating HuNoV unless chlorine levels are above those routinely used for sewage treatment.

Disinfection of bacteriophage MS2 by copper in water.

Households that lack piped water supply are often forced to meet water needs by storing in the home, leaving water vulnerable to contamination by viruses. Storage in copper containers can potentially prevent this type of contamination, but the inactivation kinetics of viruses by copper need to be described to make appropriate storage recommendations. This work characterized inactivation kinetics of bacteriophage MS2 as a surrogate for enteric viruses by dissolved ionic copper in water. Reduction of MS2 increased with increasing doses of copper. At 0.3 mg/L, there was a 1.8-log10 reduction of MS2 within 6 h. At 1 and 3 mg/L, 2-2.5 log10 inactivation could be achieved between 6 and 24 h. Parameters for the Chick-Watson, Hom, and One Hit-Two Population models of inactivation were calculated and evaluated, all of which demonstrated strong goodness-of-fit and predictability at various contact times. Copper inactivates MS2 under controlled conditions at doses between 0.3 and 3 mg/L. Although requiring longer contact times than conventional disinfectants, it is a candidate for improving the safety of stored drinking water.

Minimizing Bias in Virally Seeded Water Treatment Studies: Evaluation of Optimal Bacteriophage and Mammalian Virus Preparation Methodologies.

One key assumption impacting data quality in viral inactivation studies is that reduction estimates are not altered by the virus seeding process. However, seeding viruses often involves the inadvertent addition of co-constituents such as cell culture components or additives used during preparation steps which can impact viral reduction estimates by inducing non-representative oxidant demand in disinfection studies and fouling in membrane assessments. The objective of this study was therefore to characterize a mammalian norovirus surrogate, murine norovirus (MNV), and bacteriophage MS2 at sequential stages of viral purification and to quantify their potential contribution to artificial oxidant demand and non-representative membrane fouling. Our results demonstrate that seeding solvent extracted and 0.1 micron filtered MNV to ~105 PFU/mL in an experimental water matrix will result in additional total organic carbon (TOC) and 30 min chlorine demand of 39.2 mg/L and 53.5 mg/L as Cl2, respectively. Performing sucrose cushion purification on the MNV stock prior to seeding reduces the impacts of TOC and chlorine demand to 1.6 and 0.15 mg/L as Cl2, respectively. The findings for MNV are likely relevant for other mammalian viruses propagated in serum-based media. Thus, advanced purification of mammalian virus stocks by sucrose cushion purification (or equivalent density-based separation approach) is warranted prior to seeding in water treatment assessments. Studies employing bacteriophage MS2 as a surrogate virus may not need virus purification, since seeding MS2 at a concentration of ~106 PFU/mL will introduce only ~1 mg/L of TOC and ~1 mg/L as Cl2 of chlorine demand to experimental water matrices.