Jack F. Schijven

Removal of viruses by soil passage: overview of modeling, processes, and parameters.

In this article, the modeling of subsurface virus transport under saturated conditions and the factors that affect adsorption and inactivation are evaluated. Both equilibrium and kinetic adsorption are considered. Equilibrium adsorption is found to be of little significance. Adsorption appears to be mainly kinetically limited. At pH 7 and higher, conditions are generally unfavorable for attachment, but viruses may preferentially attach to a minor surface fraction of soil grains that is positively charged. The relation of pH with surface charge and their effects on sticking efficiencies are evaluated. Dissolved organic matter decreases virus attachment by competition for the same binding sites and thus reduces attachment. Bonded organic matter may provide hydrophobic binding sites for viruses and thus enhance attachment. Dissolved organic matter may disrupt hydrophobic bonds. The enhancing and attenuating effects of organic matter are very difficult to quantify and may be responsible for considerable uncertainty when predicting virus removal. Values of inactivation rate coefficients for attached viruses were calculated using data from some batch studies. Enhanced or reduced inactivation is found to be virus-specific and almost independent of adsorption. Temperature is the most important factor that influences virus inactivation. Probably the inactivation rate coefficients of free and attached viruses change similarly with temperature. Some frequently used bacteriophages are evaluated as model viruses. MS2 and PRD1 meet the requirements for worst-case model viruses, at water temperatures less than about 10°C, at pH 6 to 8, and if the soil does not contain too many hydrophobic sites and not too much multivalent cations. Bacteriophage ϕX 174 may be a relatively conservative model virus, because of its low hy drophobic-ity and stability. Together in a cocktail, these three viruses span a range of properties, like size, surface charge, and hydrophobicity. F-specific RNA bacteriophages (FRNAPHs) may be very useful naturally occurring worst-case viruses. FRNAPHs that are present in surface water or treated wastewater that is used for recharging groundwater, consist of stable and poorly adsorbing viruses. An inventory of parameter values from field studies is made. Attachment appears to be the major process that determines virus removal. Still, only very few data are available on attachment and detachment of viruses under field conditions. Removal of viruses by soil passage, log(C/C 0), appears to decline nonlinearly with distance due to heterogeneities within the soil as well as within the population of transported virus particles. Predictions of virus removal at larger distances are severely overestimated if they are based on removal data from column experiments or from short-distance field studies.

Modeling removal of bacteriophages MS2 and PRD1 by dune recharge at Castricum, Netherlands

Removal of model viruses by dune recharge was studied at a field site in the dune area of Castricum, Netherlands. Recharge water was dosed with bacteriophages MS2 and PRD1 for 11 days at a constant concentration in a 10- by 15-m compartment that was isolated in a recharge basin. Breakthrough was monitored for 120 days at six wells with their screens along a flow line. Concentrations of both phages were reduced about 3 log10 within the first 2.4 m and another 5 log10 in a linear fashion within the following 27 m. A model accounting for one-site kinetic attachment as well as first-order inactivation was employed to simulate the bacteriophage breakthrough curves. The major removal process was found to be attachment of the bacteriophages. Detachment was very slow. After passage of the pulse of dosed bacteriophages, there was a long tail whose slope corresponds to the inactivation rate coefficient of 0.07–0.09 day−1 for attached bacteriophages. The end of the rising and the start of the declining limbs of the breakthrough curves could not be simulated completely, probably because of an as yet unknown process.

Monitoring of Waterborne Pathogens in Surface Waters in Amsterdam, The Netherlands, and the Potential Health Risk Associated with Exposure to Cryptosporidium and Giardia in These Waters

ABSTRACT The water in the canals and some recreational lakes in Amsterdam is microbiologically contaminated through the discharge of raw sewage from houseboats, sewage effluent, and dog and bird feces. Exposure to these waters may have negative health effects. During two successive 1-year study periods, the water quality in two canals (2003 to 2004) and five recreational lakes (2004 to 2005) in Amsterdam was tested with regard to the presence of fecal indicators and waterborne pathogens. According to Bathing Water Directive 2006/7/EC, based on Escherichia coli and intestinal enterococcus counts, water quality in the canals was poor but was classified as excellent in the recreational lakes. Campylobacter, Salmonella, Cryptosporidium, and Giardia were detected in the canals, as was rotavirus, norovirus, and enterovirus RNA. Low numbers of Cryptosporidium oocysts and Giardia cysts were detected in the recreational lakes, despite compliance with European bathing water legislation. The estimated risk of infection with Cryptosporidium and Giardia per exposure event ranged from 0.0002 to 0.007% and 0.04 to 0.2%, respectively, for occupational divers professionally exposed to canal water. The estimated risk of infection at exposure to incidental peak concentrations of Cryptosporidium and Giardia may be up to 0.01% and 1%, respectively, for people who accidentally swallow larger volumes of the canal water than the divers. Low levels of viable waterborne pathogens, such as Cryptosporidium and Giardia, pose a possible health risk from occupational, accidental, and recreational exposure to surface waters in Amsterdam.

The impact of temperature on the inactivation of enteric viruses in food and water: a review

Temperature is considered as the major factor determining virus inactivation in the environment. Food industries, therefore, widely apply temperature as virus inactivating parameter. This review encompasses an overview of viral inactivation and virus genome degradation data from published literature as well as a statistical analysis and the development of empirical formulae to predict virus inactivation. A total of 658 data (time to obtain a first log10 reduction) were collected from 76 published studies with 563 data on virus infectivity and 95 data on genome degradation. Linear model fitting was applied to analyse the effects of temperature, virus species, detection method (cell culture or molecular methods), matrix (simple or complex) and temperature category (<50 and ≥50°C). As expected, virus inactivation was found to be faster at temperatures ≥50°C than at temperatures <50°C, but there was also a significant temperature–matrix effect. Virus inactivation appeared to occur faster in complex than in simple matrices. In general, bacteriophages PRD1 and PhiX174 appeared to be highly persistent whatever the matrix or the temperature, which makes them useful indicators for virus inactivation studies. The virus genome was shown to be more resistant than infectious virus. Simple empirical formulas were developed that can be used to predict virus inactivation and genome degradation for untested temperatures, time points or even virus strains.

Exposure assessment for swimmers in bathing waters and swimming pools.

Bathing water compliant with bathing water legislation may nevertheless contain pathogens to such a level that they pose unacceptable health risks for swimmers. Quantitative Microbiological Risk Assessment (QMRA) can provide a proper basis for protective measures, but the required data on swimmer exposure are currently limited or lacking. The objective of this study was to collect exposure data for swimmers in fresh water, seawater and swimming pools, i.e. volume of water swallowed and frequency and duration of swimming events. The study related to swimming in 2007, but since the summer of 2007 was wet and this might have biased the results regarding surface water exposure, the study was repeated relating to swimming in 2009, which had a dry and sunny summer. Exposure data were collected through questionnaires administered to approximately 19 000 persons representing the general Dutch population. Questionnaires were completed by 8000 adults of whom 1924 additionally answered the questions for their eldest child (< 15 years). The collected data did not differ significantly between 2007 and 2009. The frequency of swimming and the duration of swimming were different for men, women and children and between water types. Differences between men and women were small, but children behaved differently: they swam more often, stayed in the water longer, submerged their heads more often and swallowed more water. Swimming pools were visited most frequently (on average 13-24 times/year) with longest duration of swimming (on average 67-81 min). On average, fresh and seawater sites were visited 6-8 times/year and visits lasted 41-79 min. Dependent on the water type, men swallowed on average 27-34 ml per swimming event, women 18-23 ml, and children 31-51 ml. Data on exposure of swimmers to recreational waters could be obtained by using a questionnaire approach in combination with a test to measure mouthfuls of water for transformation of categorical data to numerical data of swallowed volumes of water. Previous assumptions on swimmer exposure were replaced with estimates of exposure parameters, thus reducing uncertainty in assessing the risk of infection with waterborne pathogens and enabling identification of risk groups. QMRA for Cryptosporidium and Giardia was demonstrated based on data from previous studies on the occurrence of these pathogens in recreational lakes and a swimming pool.

Removal of microorganisms by deep well injection

The removal of bacteriophages MS2 and PRD1, spores of Clostridium bifermentans R5 and . Escherichia coli WR1 by deep well injection into a sandy aquifer, was studied at a pilot field site in the southeast of the Netherlands. Injection water was seeded with the microorganisms for 5 days. Breakthrough was monitored for 93 days at 4 monitoring wells with their screens at a depth of about 310 m below surface. Within the first 8 m of soil passage, concentrations of MS2 and PRD1 were reduced by 6 log , that of R5 spores by 5 log and that of WR1 by 7.5 log . 10 10 10

Long-Term Inactivation Study of Three Enteroviruses in Artificial Surface and Groundwaters, Using PCR and Cell Culture

ABSTRACT Since the transmission of pathogenic viruses via water is indistinguishable from the transmission via other routes and since the levels in drinking water, although significant for health, may be too low for detection, quantitative viral risk assessment is a useful tool for assessing disease risk due to consumption of drinking water. Quantitative viral risk assessment requires information concerning the ability of viruses detected in drinking water to infect their host. To obtain insight into the infectivity of viruses in relation to the presence of virus genomes, inactivation of three different enteroviruses in artificial ground and surface waters under different controlled pH, temperature, and salt conditions was studied by using both PCR and cell culture over time. In salt-peptone medium, the estimated ratio of RNA genomes to infectious poliovirus 1 in freshly prepared suspensions was about 100. At 4°C this ratio was 103 after 600 days, and at 22°C it was 104 after 200 days. For poliovirus 1 and 2 the RNA/infectious virus ratio was higher in artificial groundwater than in artificial surface water, but this was not the case for coxsackievirus B4. When molecular detection is used for virus enumeration, it is important that the fraction of infectious virus (based on all virus genomes detected) decays with time, especially at temperatures near 22°C.

Kinetic modeling of virus transport at the field scale.

Bacteriophage removal by soil passage in two field studies was re-analyzed with the goal to investigate differences between one- and two-dimensional modeling approaches, differences between one- and two-site kinetic sorption models, and the role of heterogeneities in the soil properties. The first study involved removal of bacteriophages MS2 and PRDI by dune recharge, while the second study represented removal of MS2 by deep well injection. In both studies, removal was higher during the first meters of soil passage than thereafter. The software packages HYDRUS-ID and HYDRUS-2D, which simulate water flow and solute transport in one- and two-dimensional variably saturated porous media, respectively, were used. The two codes were modified by incorporating reversible adsorption to two types of kinetic sites. Tracer concentrations were used first to calibrate flow and transport parameters of both models before analyzing transport of bacteriophages. The one-dimensional one-site model did not fully describe the tails of the measured breakthrough curves of MS2 and PRD1 from the dune recharge study. While the one-dimensional one-site model predicted a sudden decrease in virus concentrations immediately after the peaks, measured data displayed much smoother decline and tailing. The one-dimensional two-site model simulated the overall behavior of the breakthrough curves very well. The two-dimensional one-site model predicted a more gradual decrease in virus concentrations after the peaks than the one-dimensional one-site model, but not as good as the one-dimensional two-site model. The dimensionality of the problem hence can partly explain the smooth decrease in concentration after peak breakthrough. The two-dimensional two-site model provided the best results. Values for k(att2) and k(det2) could not be determined at the last two of four monitoring wells, thus suggesting that either a second type of kinetic sites is present in the first few meters of dune passage and not beyond the second monitoring well, or that effects of soil heterogeneity and dimensionality of the problem overshadowed this process. Variations between single collector efficiencies were relatively small, whereas collision efficiencies varied greatly. This implies that the nonlinear removal of MS2 and PRD1 is mainly caused by variations in interactions between grain and virus surfaces rather than by physical heterogeneity of the porous medium. Similarly, a two-site model performed better than the one-site model in describing MS2 concentrations for the deep well injection study. However, the concentration data were too sparse in this study to have much confidence in the fitted parameters.

Bacteriophages and clostridium spores as indicator organisms for removal of pathogens by passage through saturated dune sand

In a field study on the efficiency of dune recharge for drinking water production, bacteriophage MS2 was shown to be removed 8 log(10) by passage through the dune sand. The question of whether pathogenic viruses would be removed as much as MS2 was studied by comparing complete breakthrough curves of MS2 with those of the human viruses Coxsackievirus B4 (CB4) and Poliovirus 1 (PV1) in laboratory columns. The columns were designed to closely simulate the field conditions: same sand, water, porewater velocity and temperature. Employing a two-site kinetic model to simulate breakthrough curves, attachment/detachment to two types of kinetic sites as well as inactivation of free and attached viruses were evaluated. It was found that attachment to only one of the sites is of significance for determining overall removal. At field scale, removal of the less negatively charged PV1 was extrapolated to be about 30 times greater than that of MS2, but removal of CB4 would be only as much as that of MS2. Also, removal of spores of Clostridium perfringens D10, a potential surrogate for Cryptosporidium oocysts, was studied. The attachment rate coefficient of the spores was 7.5 times greater than that of MS2. However, this does not imply that the removal of the spores is 7.5 times greater than that of MS2. Due to negligible inactivation in combination with detachment of previously attached spores, the actual removal rate of the spores depends on the duration of contamination and eventually all spores will break through. Provided no irreversible attachment or physical straining occurs, this may also be the case for other persistent microorganisms, like oocysts of Cryptosporidium.

QMRAspot: a tool for Quantitative Microbial Risk Assessment from surface water to potable water.

In the Netherlands, a health based target for microbially safe drinking water is set at less than one infection per 10,000 persons per year. For the assessment of the microbial safety of drinking water, Dutch drinking water suppliers must conduct a Quantitative Microbial Risk Assessment (QMRA) at least every three years for the so-called index pathogens enterovirus, Campylobacter, Cryptosporidium and Giardia. In order to collect raw data in the proper format and to automate the process of QMRA, an interactive user-friendly computational tool, QMRAspot, was developed to analyze and conduct QMRA for drinking water produced from surface water. This paper gives a description of the raw data requirements for QMRA as well as a functional description of the tool. No extensive prior knowledge about QMRA modeling is required by the user, because QMRAspot provides guidance to the user on the quantity, type and format of raw data and performs a complete analysis of the raw data to yield a risk outcome for drinking water consumption that can be compared with other production locations, a legislative standard or an acceptable health based target. The uniform approach promotes proper collection and usage of raw data and, warrants quality of the risk assessment as well as enhances efficiency, i.e., less time is required. QMRAspot may facilitate QMRA for drinking water suppliers worldwide. The tool aids policy makers and other involved parties in formulating mitigation strategies, and prioritization and evaluation of effective preventive measures as integral part of water safety plans.