Dynamic changes in the physicochemical properties of fresh-cut produce wash water as impacted by commodity type and processing conditions.

Organic materials in fresh-cut produce wash water deplete free chlorine that is required to prevent pathogen survival and cross-contamination. This research evaluated water quality parameters frequently used to describe organic load for their fitness to predict chlorine demand (CLD) and chemical oxygen demand (COD), which are major needs identified by the industry-led produce food safety taskforce. Batches of romaine lettuce, iceberg lettuce, or carrot of different cut sizes and shapes were washed in 40 liters of water. Physicochemical properties of wash water including CLD, COD, total organic carbon (TOC), total suspended solids (TSS), total dissolved solids (TDS), turbidity, total sugar content, and pH, were monitored. Results indicate that pH is primarily commodity dependent, while organic load is additionally impacted by cutting and washing conditions. Significant linear increases in COD, TOC, CLD, TDS, and turbidity resulted from increasing product-to-water ratio, and decreasing cut size. Physicochemical parameters, excluding pH, showed significant positive correlation across different cut sizes within a commodity. High correlations were obtained between CLD and COD and between COD and TOC for pooled products. The convenient measurement of TDS, along with its strong correlation with COD and CLD, suggests the potential of TDS for predicting organic load and chlorine reactivity. Finally, the potential application and limitation of the proposed models in practical produce processing procedures are discussed extensively.

Effects of pH Variability on Peracetic Acid Reduction of Human Norovirus GI, GII RNA, and Infectivity Plus RNA Reduction of Selected Surrogates.

With increasing interest in peracetic acid (PAA) as a disinfectant in water treatment processes, this study determined PAA treatment effects on human noroviruses (hNoVs) genotype I (GI) and genotype II (GII) as well as effects on bacteriophage MS2 and murine norovirus (MNV) in relation to pH. Across all pH conditions, PAA achieved between 0.2 and 2.5 log10 reduction of hNoVs over 120 min contact time in buffer solution as measured by reverse transcription-qPCR (RT-qPCR). The PAA treatments produced similar RT-qPCR reductions of MS2 and MNV, in the range of 0.2-2.7 log10. Infectivity assays achieved > 4 log10 reduction of both MS2 and MNV in buffer solution after 120 min contact time. Comparing PAA activity across varying pH, disinfection at pH 8.5, in general, resulted in less reduction of infectivity and molecular signals compared to pH conditions of 6.5 and 7.5. This difference was most pronounced for reductions in infectivity of MNV and MS2, with as much as 2.7 log10 less reduction at pH 8.5 relative to lower pH conditions. This study revealed that PAA was an effective disinfectant for treatment of hNoV GI and GII, MS2 and MNV, with greatest virus reduction observed for MS2 and MNV infectivity. RT-qPCR reductions of MS2 and MNV were lower than concurrent MS2 and MNV infectivity reductions, suggesting that observed hNoV RT-qPCR reductions may underestimate reductions in hNoV infectivity achieved by PAA. Although virus disinfection by PAA occurred at all evaluated pH levels, PAA is most effective at pH 6.5-7.5.

Infectivity reduction efficacy of UV irradiation and peracetic acid-UV combined treatment on MS2 bacteriophage and murine norovirus in secondary wastewater effluent.

Peracetic acid (PAA) is a strong oxidant/bactericide that has been applied in various industries (e.g., food processing, pharmaceuticals, medical device sterilization, etc.) as a disinfectant. There is increasing interest in using PAA for wastewater disinfection because it does not form halogenated byproducts, and no post-treatment quenching is required. Previous studies have demonstrated good efficiency in controlling bacteria in wastewater, but limited information is available for viruses, especially those hosted by mammals (e.g., norovirus). Therefore, a study on the infectivity reduction of murine norovirus (MNV) was undertaken to evaluate the disinfection efficacy of PAA or UV alone and in combination with UV irradiation in undisinfected secondary effluent from a municipal wastewater reclamation facility (MWW) and phosphate buffer solution (PBS) at pH 7. Experiments employing MS2 bacteriophage were also performed in parallel for comparison purposes. MS2 infectivity reduction was found to be lower than MNV infectivity reduction for each condition studied - PAA, PAA + UV, and UV disinfection. These data suggest that MS2 may not be an appropriate surrogate to accurately predict the reduction of MNV infectivity. UV irradiation, in a dose range of 5-250 mJ/cm2, provided linear log inactivation (-log (N/N0)) with a regression slope (cm2mJ-1) of 0.031-0.034 and 0.165-0.202 for MS2 and MNV, respectively. UV irradiation provided similar inactivation for MS2 and MNV in both suspensions (PBS or MWW). Low infectivity reduction of MS2 was observed when PAA was used alone at a practical dose of 1.5 mg/L and below. A greater reduction of both MNV and MS2 was observed in PAA disinfection experiments using PBS as the microbial suspension medium, than in secondary effluent. Similar results were observed in PAA + UV experiments, in which greater synergistic effects were found in PBS than in MWW. Results of OH radical formation experiments suggest the presence of radical scavengers in MWW, which resulted in less opportunity for MNV and MS2 to encounter OHradicals. This study also demonstrated that the type of water can have a substantial impact on wastewater disinfection when employing PAA or PAA + UV treatment due to the matrix effect and the presence of radical scavengers, respectively. The results from this study could be employed to aid in the conceptual design of PAA and UV disinfection facilities, especially when norovirus is the organism of concern.

Impacts of virus processing on human norovirus GI and GII persistence during disinfection of municipal secondary wastewater effluent.

Noroviruses cause significant global health burdens and waterborne transmission is a known exposure pathway. Chlorination is the most common method of disinfection for water and wastewater worldwide. The purpose of this study was to investigate the underlying causes for discrepancies in human norovirus (hNoV) resistance to free chlorine that have been previously published, and to assess hNoV GI and GII persistence during disinfection of municipal secondary wastewater (WW) effluent. Our results reveal that choice of hNoV purification methodology prior to seeding the viruses in an experimental water matrix influences disinfection outcomes in treatment studies. Common hNoV purification processes such as solvent extraction and 0.45-µm filtration were ineffective in removing high levels of organics introduced into water or wastewater samples when seeding norovirus positive stool. These methods resulted in experimental water matrices receiving an additional 190 mg/L as Cl2 of 15-s chlorine demand and approximately 440 mg/L as Cl2 of 30-min chlorine demand due to seeding norovirus positive stool at 1% w/v. These high organic loads impact experimental water chemistry and bias estimations of hNoV persistence. Advanced purification of norovirus positive stool using sucrose cushion ultracentrifugation and ultrafiltration reduced 15-s chlorine demands by 99% and TOC by 93% for loose (i.e. unformed diarrhea) stools. Using these methods, hNoV GI and GII persistence was investigated during free chlorination of municipal WW. A suite five of kinetic inactivation models was fit to viral reverse transcription-qPCR reduction data, and model predicted CT values for 1, 2, and 3 log10 reduction of hNoV GI in municipal WW by free chlorine were 0.3, 2.1, and 7.8 mg-min/L, respectively. Model predicted CT values for reduction of hNoV GII in WW were 0.4, 2.0, and 7.0 mg-min/L, respectively. These results indicate that current WW treatment plant disinfection practices employing free chlorine are likely protective for public health with regards to noroviruses, and will achieve at least 3-log reduction of hNoV GI and GII RNA despite previous reports of high hNoV resistance.

Reduction of Human Norovirus GI, GII, and Surrogates by Peracetic Acid and Monochloramine in Municipal Secondary Wastewater Effluent.

The objective of this study was to characterize human norovirus (hNoV) GI and GII reductions during disinfection by peracetic acid (PAA) and monochloramine in secondary wastewater (WW) and phosphate buffer (PB) as assessed by reverse transcription-qPCR (RT-qPCR). Infectivity and RT-qPCR reductions are also presented for surrogate viruses murine norovirus (MNV) and bacteriophage MS2 under identical experimental conditions to aid in interpretation of hNoV molecular data. In WW, RT-qPCR reductions were less than 0.5 log10 for all viruses at concentration-time (CT) values up to 450 mg-min/L except for hNoV GI, where 1 log10 reduction was observed at CT values of less than 50 mg-min/L for monochloramine and 200 mg-min/L for PAA. In PB, hNoV GI and MNV exhibited comparable resistance to PAA and monochloramine with CT values for 2 log10 RT-qPCR reduction between 300 and 360 mg-min/L. Less than 1 log10 reduction was observed for MS2 and hNoV GII in PB at CT values for both disinfectants up to 450 mg-min/L. Our results indicate that hNoVs exhibit genogroup dependent resistance and that disinfection practices targeting hNoV GII will result in equivalent or greater reductions for hNoV GI. These data provide valuable comparisons between hNoV and surrogate molecular signals that can begin the process of informing regulators and engineers on WW treatment plant design and operational practices necessary to inactivate hNoVs.

Inactivation of Human Norovirus Genogroups I and II and Surrogates by Free Chlorine in Postharvest Leafy Green Wash Water.

Human noroviruses (hNoVs) are a known public health concern associated with the consumption of leafy green vegetables. While a number of studies have investigated pathogen reduction on the surfaces of leafy greens during the postharvest washing process, there remains a paucity of data on the level of treatment needed to inactivate viruses in the wash water, which is critical for preventing cross-contamination. The objective of this study was to quantify the susceptibility of hNoV genotype I (GI), hNoV GII, murine norovirus (MNV), and bacteriophage MS2 to free chlorine in whole leaf, chopped romaine, and shredded iceberg lettuce industrial leafy green wash waters, each sampled three times over a 4-month period. A suite of kinetic inactivation models was fit to the viral reduction data to aid in quantification of concentration-time (CT) values. Results indicate that 3-log10 infectivity reduction was achieved at CT values of less than 0.2 mg · min/liter for MNV and 2.5 mg · min/liter for MS2 in all wash water types. CT values for 2-log10 molecular reduction of hNoV GI in whole leaf and chopped romaine wash waters were 1.5 and 0.9 mg · min/liter, respectively. For hNoV GII, CT values were 13.0 and 7.5 mg · min/liter, respectively. In shredded iceberg wash water, 3-log10 molecular reduction was not observed for any virus over the time course of experiments. These findings demonstrate that noroviruses may exhibit genogroup-dependent resistance to free chlorine and emphasize the importance of distinguishing between genogroups in hNoV persistence studies.IMPORTANCE Postharvest washing of millions of pounds of leafy greens is performed daily in industrial processing facilities with the intention of removing dirt, debris, and pathogenic microorganisms prior to packaging. Modest inactivation of pathogenic microorganisms (less than 2 log10) is known to occur on the surfaces of leafy greens during washing. Therefore, the primary purpose of the sanitizing agent is to maintain microbial quality of postharvest processing water in order to limit cross-contamination. This study modeled viral inactivation data and quantified the free-chlorine CT values that processing facilities must meet in order to achieve the desired level of hNoV GI and GII reduction. Disinfection experiments were conducted in industrial leafy green wash water collected from a full-scale fresh produce processing facility in the United States, and hNoV GI and GII results were compared with surrogate molecular and infectivity data.

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.

UV-induced effects on toxicity of model disinfection byproducts.

UV (Ultraviolet)-based treatment has been demonstrated to be effective for removal of some disinfection byproducts (DBPs) and to be beneficial for reduction of genotoxicity and cytotoxicity in chlorinated water. However, to a large extent, UV-induced effects on chemistry and toxicology have been treated as a black box, in the sense that little or no UV dose-dependent behavior has been reported. To address this issue, the effects of UV254 irradiation on 1,4-dibenzoquinone (BQ), 2,6-dichloro-1,4-benzoquinone (DCBQ), and chlorocreatinine (Cl-Cre) as model DBPs were examined, both in terms of photodegradation and cytotoxicity. These compounds have been identified as DBPs that are relevant in swimming pool settings; however, these compounds will be relevant in other water treatment settings, including drinking water production and wastewater reuse. UV254 irradiation was shown to promote photodecay of all three compounds. However, for BQ and DCBQ, the corresponding cytotoxicity of the UV-irradiated samples remained essentially unchanged, even when the compound was completely photodegraded. These results indicate that the photodegradation products of BQ and DCBQ carry similar cytotoxicity as their respective parent compounds. On the other hand, UV254-irradiation of Cl-Cre yielded a decrease in cytotoxicity that correlated with photodechlorination of Cl-Cre. These experiments also demonstrated a reduction in cytotoxicity in connection with photodechlorination of an N-chlorinated organic compound. Overall, the results of these experiments indicate the importance of defining products of UV photodecay processes, both in terms of chemistry and toxicity; these attributes are expected to be important in many UV-based applications, including potable water production, water reuse, and recreational water settings.

Comparative Inactivation of Murine Norovirus and MS2 Bacteriophage by Peracetic Acid and Monochloramine in Municipal Secondary Wastewater Effluent.

Chlorination has long been used for disinfection of municipal wastewater (MWW) effluent while the use peracetic acid (PAA) has been proposed more recently in the United States. Previous work has demonstrated the bactericidal effectiveness of PAA and monochloramine in wastewater, but limited information is available for viruses, especially ones of mammalian origin (e.g., norovirus). Therefore, a comparative assessment was performed of the virucidal efficacy of PAA and monochloramine against murine norovirus (MNV) and MS2 bacteriophage in secondary effluent MWW and phosphate buffer (PB). A suite of inactivation kinetic models was fit to the viral inactivation data. Predicted concentration-time (CT) values for 1-log10 MS2 reduction by PAA and monochloramine in MWW were 1254 and 1228 mg-min/L, respectively. The 1-, 2-, and 3-log10 model predicted CT values for MNV viral reduction in MWW were 32, 47, and 69 mg-min/L for PAA and 6, 13, and 28 mg-min/L for monochloramine, respectively. Wastewater treatment plant disinfection practices informed by MS2 inactivation data will likely be protective for public health but may overestimate CT values for reduction of MNV. Additionally, equivalent CT values in PB resulted in greater viral reduction which indicate that viral inactivation data in laboratory grade water may not be generalizable to MWW applications.

Ultraviolet-induced effects on chloramine and cyanogen chloride formation from chlorination of amino acids.

Ultraviolet (UV)-based treatment is commonly used to augment chlorination in swimming pools. However, the effects of combined application of UV254/chlorine on disinfection byproduct (DBP) formation are incompletely defined. To examine this issue, experiments were conducted with amino acids (l-arginine, l-histidine, and glycine) that are representative of those introduced to swimming pools via human body fluids. For each precursor, stepwise experiments were conducted with chlorination and UV254 exposure, with/without post-chlorination. Net formation and decomposition of chloramines and cyanogen chloride (CNCl) were measured for a range of chlorine/precursor (Cl/P) molar ratios and UV254 doses. Substantial production of NH2Cl from l-arginine and l-histidine was observed at Cl/P = 1.0 and 2.0 when post-chlorination was applied to UV254-irradiated samples. These results suggested a mechanism of rapid N-chlorination, followed by cleavage of NH3 by UV254 irradiation. CNCl formation was observed from UV254-irradiated samples of l-arginine and l-histidine when Cl/P = 2.0 and 3.0, as well as from glycine for Cl/P ≤ 1. Structurally related precursor compounds were examined for CNCl formation potential in chlorination/UV experiments. CNCl formation was promoted by UV254 exposure of chlorinated imidazole and guanidine compounds, which suggested that these groups contributed to CNCl formation. The results have implications with respect to the application of chlorine and UV for water treatment in swimming pools and other settings, such as water reuse and advanced oxidation processes.

Effects of UV 254 irradiation on residual chlorine and DBPs in chlorination of model organic-N precursors in swimming pools.

Ultraviolet (UV) irradiation is commonly applied as a secondary disinfection process in chlorinated pools. UV-based systems have been reported to yield improvements in swimming pool water and air chemistry, but to date these observations have been largely anecdotal. The objectives of this investigation were to evaluate the effects of UV irradiation on chlorination of important organic-N precursors in swimming pools. Creatinine, L-arginine, L-histidine, glycine, and urea, which comprise the majority of the organic-N in human sweat and urine, were selected as precursors for use in conducting batch experiments to examine the time-course behavior of several DBPs and residual chlorine, with and without UV(254) irradiation. In addition, water samples from two natatoria were subjected to monochromatic UV irradiation at wavelengths of 222 nm and 254 nm to evaluate changes of liquid-phase chemistry. UV(254) irradiation promoted formation and/or decay of several chlorinated N-DBPs and also increased the rate of free chlorine consumption. UV exposure resulted in loss of inorganic chloramines (e.g., NCl(3)) from solution. Dichloromethylamine (CH(3)NCl(2)) formation from creatinine was promoted by UV exposure, when free chlorine was present in solution; however, when free chlorine was depleted, CH(3)NCl(2) photodecay was observed. Dichloroacetonitrile (CNCHCl(2)) formation (from L-histidine and L-arginine) was promoted by UV(254) irradiation, as long as free chlorine was present in solution. Likewise, UV exposure was observed to amplify cyanogen chloride (CNCl) formation from chlorination of L-histidine, L-arginine, and glycine, up to the point of free chlorine depletion. The results from experiments involving UV irradiation of chlorinated swimming pool water were qualitatively consistent with the results of model experiments involving UV/chlorination of precursors in terms of the behavior of residual chlorine and DBPs measured in this study. The results indicate that UV(254) irradiation promotes several reactions that are involved in the formation and/or destruction of chlorinated N-DBPs in pool settings. Enhancement of DBP formation was consistent with a mechanism whereby a rate-limiting step in DBP formation was promoted by UV exposure. Promotion of these reactions also resulted in increases of free chlorine consumption rates.

Ozone and UV254 radiation for municipal wastewater disinfection.

Bench-scale experiments were conducted with municipal wastewater effluent samples to examine the feasibility of combined application of ozone and ultraviolet (UV) radiation for disinfection. Effluent samples displayed rapid initial ozone demand, which promotes ozone transfer but diminishes disinfection efficacy. Ozone doses up to 10 mg/L yielded only trace quantities of residual ozone; despite the fact that initial ozone demand was never exceeded, quantifiable (though variable) inactivation of E. coli was observed, along with modest improvements of UV transmittance. Results from collimated beam experiments demonstrated that compliance with effluent discharge permit limitations could be achieved consistently with a UV254 dose of 12.4 mJ/cm2 at a pre-ozonation dose of 2 to 3 mg/L. In the absence of pre-ozonation, consistent compliance was observed at a UV dose of 16.5 mJ/cm2. No evidence of synergism between ozone and UV254 radiation was found in the measured inactivation responses of E. coli.

Disinfection by-product dynamics in a chlorinated, indoor swimming pool under conditions of heavy use: national swimming competition.

Anecdotal evidence suggests that water quality in chlorinated, indoor pools deteriorates under conditions of heavy use. However, data to define these dynamics have not been reported. To address this issue, a study was performed in which water chemistry was monitored in a chlorinated, indoor pool before and during a national swimming competition, a period of heavy, intense use. NCl3 concentration was observed to double after the first day, and increased by a factor of 3-4 over the 4 days of competition. CNCHCl2 and CH3NCl2 concentrations both increased by a factor of 2-3 during the course of the meet, while CHCl3 concentration showed only a modest increase during this same period. Diurnal patterns of NCl3, CH3NCl2 and CHCl3 concentrations were observed, and these patterns appeared to depend on the Henry's law constant of the compound. Urea concentration showed a diurnal pattern, superimposed on a trend of steady increase during each day of the competition; however, the diurnal pattern of urea behavior could not be explained by reactions with chlorine, as the urea-free chlorine reaction is relatively slow. It is more likely that the overnight decrease in urea concentration was attributable to mixing of surface water with water in the deeper parts of the pool. The findings of this study provide an indication of the changes in pool water chemistry that take place in a chlorinated, indoor pool under heavy use conditions.