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Water contaminated with human and animal enteric pathogens puts public health at serious risk. All countries and regions of the world require highly robust and effective water management and treatment systems to guarantee safe water and protect public health. To this end, we need accurate predictions of the origin of pathogens , how they move through the environment and where they end up.This study is part of a four-year project and aims to develop new bacterial overland transport - BOT models to provide answers to the above questions. The project takes a holistic, quantitative approach to transfer BOT model parameters onto large scales. Small-scale precipitation experiments are conducted in the laboratory and larger-scale experiments are conducted using a rainfall simulation under real environmental conditions. The state-of-the-art combination of quantitative, microbiological, and molecular methods and parameters will provide the scientific basis for more accurate predictions of BOT, which eventually may be extended to viruses and protozoa in the future.

A novel microbial risk analysis of groundwater as part of the Dutch guideline document for Quantitative Microbial Risk Assessment (QMRA) of drinking water consumption encompasses 1) vulnerability assessment of groundwater production sites, 2) calculating the protection zone against microbial contamination to remain below an infection risk of 1/10,000 persons/year, 3) assessment of contamination sources within the protection zone, and 4) QMRA for identified contamination sources. Protection zones are based on a standard virus contamination scenario and may be computed using a required minimal travel time, a hydrological model that includes a first order decay term for virus inactivation and attachment, or the computational tool QMRAwell. QMRAwell is designed for unconfined and (semi)confined sandy aquifers with a forced groundwater gradient due to pumping. QMRAwell can also be used to conduct QMRA.

BackgroundThe COVID 19 pandemic has triggered concerns and assumptions globally about transmission of the SARS-CoV-2 virus via cash transactions.ObjectivesAssess the risk of contracting COVID-19 through exposure to SARS-CoV-2 via cash acting as a fomite in payment transactions.MethodsA quantitative microbial risk assessment was conducted for a worst-case scenario assuming an infectious person at the onset of symptoms, when virion concentrations in coughed droplets are at their highest. This person then contaminates a banknote by coughing on it and immediately hands it over to another person, who might then be infected by transferring the virions with a finger from the contaminated banknote to a facial mucous membrane. The scenario considered transfer efficiency of virions on the banknote to fingertips when droplets were still wet and after having dried up and subsequently being touched by finger printing or rubbing the object.ResultsAccounting for the likelihood of the worst-case scenario to occur by considering 1) a local prevalence of 100 COVID-19 cases/100,000 persons, 2) a maximum of about 1/5th of infected persons transmit high virus loads and 3) the numbers of cash transactions/person/day, the risk of contracting COVID-19 via person-to-person cash transactions was estimated to be much lower than once per 39,000 days (107 years) for a single person. In the general populace, there will be a maximum of 2.6 expected cases/100,000 persons/day. The risk for a cashier at an average point of sale was estimated to be much less than once per 430 working days (21 months).DiscussionThe worst-case scenario is a rare event, therefore, for a single person, the risk of contracting COVID-19 via person-to-person cash transactions is very low. At a point of sale, the risk to the cashier proportionally increases but it is still low.

We developed an innovative approach to estimate the extent of fecal pollution sources for urban river catchments. The methodology consists of 1) catchment surveys complemented by literature data where needed for probabilistic estimates of daily produced fecal indicator (FIBs, E. coli, enterococci) and zoonotic reference pathogen numbers (Campylobacter, Cryptosporidium and Giardia) excreted by human and animal sources in a river catchment, 2) generating a hypothesis about the dominant sources of fecal pollution and selecting a source targeted monitoring design, and 3) verifying the results by comparing measured concentrations of chemical tracers, C. perfringens, and host-associated genetic microbial source tracking (MST) markers in the river, and by multi-parametric correlation analysis. We tested the approach at a study area in Vienna, Austria. The daily produced microbial particle numbers according to the probabilistic estimates indicated that, for the dry weather scenario, the discharge of treated wastewater (WWTP) was the primary contributor to fecal pollution. For the wet weather scenario, 80-99 % of the daily produced FIBs and pathogens resulted from combined sewer overflows (CSOs) according to the probabilistic estimates. When testing our hypothesis in the river, the measured concentrations of the human genetic fecal marker were log10 4 higher than for selected animal genetic fecal markers. Our analyses showed for the first-time statistical relationships between C. perfringens and a human genetic fecal marker (i.e. HF183/BacR287) with the reference pathogen Giardia in river water (Spearman rank correlation: 0.78-0.83, p

BackgroundPrevious reports suggest SARS-CoV-2 transmission risk increases during singing events. From September-October 2020, several clusters of COVID-19 cases among singing events were reported across the Netherlands. Our aim was to investigate whether singing increased SARS-CoV-2 transmission risk during these events.MethodsData from 5 events were retrospectively collected from spokespersons and singing group members via questionnaires. Information was consolidated with the National Notifiable Diseases Surveillance System. Specimens were requested for sequencing for point source and cluster assessment. We described outbreaks in terms of person, place and time and depicted potential SARS-CoV-2 transmission routes. A previously published model (AirCoV2) was used to estimate mean illness risk of 1 person through airborne transmission under various scenarios.ResultsEvents included 9-21 persons (mean: 16), aged 20-89 years (median: 62). Response rates ranged 58-100%. Attack rates were 53-74%. Limited sequencing data was obtained from 2 events. Events lasted 60-150 minutes (singing: 20-120). Rooms ranged 320-3000m3. SARS-CoV-2 transmission likely occurred during all events; with a possible index case identified in 4 events. AirCoV2 showed 86% (54-100%) mean illness risk for 120 minutes of singing, smaller room (300m3), 1 air exchange/hour (ACH), and supershedder presence.ConclusionsDroplet transmission and indirect contact probably caused some cases, but unlikely explain the high attack rates. AirCoV2 indicated that airborne transmission due to singing is possible in case of supershedder presence. Airflow expelling respiratory droplets >1.5m possibly influenced transmission. It is possible that singing itself increased SARS-CoV-2 transmission risk through airborne transmission.SummaryThis outbreak investigation among five singing events with high SARS-CoV-2 attack rates (53-74%) suggested that airflow expelling respiratory droplets >1.5m possibly influenced transmission and it is possible that singing itself increased SARS-CoV-2 transmission risk through airborne transmission.

The current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has created a significant threat to global health. While respiratory aerosols or droplets are considered as the main route of human-to-human transmission, secretions expelled by infected individuals can also contaminate surfaces and objects, potentially creating the risk of fomite-based transmission. Consequently, frequently touched objects such as paper currency and coins have been suspected as a potential transmission vehicle. To assess the risk of SARS-CoV-2 transmission by banknotes and coins, we examined the stability of SARS-CoV-2 and bovine coronavirus (BCoV), as surrogate with lower biosafety restrictions, on these different means of payment and developed a touch transfer method to examine transfer efficiency from contaminated surfaces to skin. Although we observed prolonged virus stability, our results, including a novel touch transfer method, indicate that the transmission of SARS-CoV-2 via contaminated coins and banknotes is unlikely and requires high viral loads and a timely order of specific events.

Use of environmental or artificial tracers has been an effective approach to characterize groundwater flow and solute transport, tracking pollutant migration and determine travel time. However, availability of a distinctive number of tracers, variability in interaction with the aquifer matrix, and analytical detection limits are namely few of the significant concerns to be addressed and which led us to focus on employing novel DNA tracers.Besides the quality of being unique, improbably prevalent in nature and environmentally friendly, DNA tracers can be synthesized virtually in infinite numbers of distinct sequences, rendering them a potential candidate for multi-tracer applications for subsurface and groundwater flow characterization. Studies have already demonstrated the potential of DNA tracing in groundwater studies but a blueprint for methodical application and analysis is required.In this study, we investigate the applicability of DNA tracers in determining hydraulic parameters of a natural aquifer, such as, hydraulic conductivity, effective porosity, dispersivity, and travel time, the most significant characters of a matrix, influencing solute or pollutant transport. In addition, we aim to leverage the applicability of the tracers in terms of minimizing the uncertainty in estimating the parameters.In order to capitalize on these advantages of DNA tracers with the aim of addressing the aforementioned objectives, this research focuses on employing multiple dsDNA (ds=double stranded) tracers in a 1.3 m long three-dimensional sand-filled aquifer tank. Under forced-gradient water flow conditions, distinctly sequenced, monodispersed dsDNA tracers are instantaneously injected through injection wells, taking into account different scenarios. The scenarios consider different configurations of injection and sampling strategies. Samples collected periodically were subjected to quantitative polymerase chain reaction (qPCR) for DNA concentration estimation. All the silica-encapsulated DNA particles were comparable in size and surface properties.Individual breakthrough curves from each of the scenarios are carefully analysed for determining water flow and hydraulic properties. In addition, the experiments producing multiple breakthrough curves are cumulatively analysed for obtaining a minimal uncertainty for the parameter estimations.

Background: Rivers are important sources for drinking water supply, however, they are often impacted by wastewater discharges from wastewater treatment plants (WWTP) and combined sewer overflows (CSO). Reduction of the faecal pollution burden is possible through enhanced wastewater treatment or prevention of CSOs. Few methodological efforts have been made so far to investigate how these measures would affect the long-term treatment requirements for microbiologically safe drinking water supply under future changes.Objectives: This study aimed to apply a new integrative approach to decipher the interplay between the effects of future changes and wastewater management measures on the required treatment of river water to produce safe drinking water. We investigated scenarios of climate change and population growth, in combination with different wastewater management scenarios (i.e., no upgrades and upgrades at WWTPs, CSOs, and both). To the best of our knowledge, this is the first study to investigate this interplay. We focussed on the viral index pathogens norovirus and enterovirus and made a cross-comparison with a bacterial and a protozoan reference pathogen (Campylobacter and Cryptosporidium).Methods: We significantly extended QMRAcatch (v1.0 Python), a probabilistic-deterministic model that combines virus fate and transport modelling in the river with quantitative microbial risk assessment (QMRA). To investigate the impact of climatic changes, we used a conceptual semi-distributed hydrological model and regional climate model outputs to simulate river discharges for the period 2035 – 2049. We assumed that population growth leads to a corresponding increase in WWTP discharges. QMRAcatch was successfully calibrated and validated based on a four-year dataset of a human-associated genetic MST marker and enterovirus. The study site was the Danube in Vienna, Austria.Results: In the reference scenario, approx. 98% of the enterovirus and norovirus loads at the study site (median: 1010 and 1013 N/d) originated from WWTP effluent, while the remainder was via CSO events. The required log reduction value (LRV) to produce safe drinking water was 6.3 and 8.4 log10 for enterovirus and norovirus. Future changes in population size, river flows and CSO events did not affect these treatment requirements, and neither did the prevention of CSOs. In contrast, in the scenario of enhanced wastewater treatment, which showed lower LRVs by 2.0 and 1.3 log10, climate-change-driven increases in CSO events had a considerable impact on the treatment requirements, as they affected the main pollution source. Preventing CSOs and installing enhanced treatment at the WWTPs together had the most significant positive effect with a reduction of LRVs by 3.9 and 3.8 log10 compared to the reference scenario.Conclusions: The integrative modelling approach was successfully realised. The simultaneous consideration of source apportionment and concentrations of the reference pathogens were found crucial to understand the interplay among the effects of climate change, population growth and pollution control measures. The approach was demonstrated for a study site representing a large river impacted by WWTP and CSO discharges, but is applicable at other sites to support long term water safety planning.

Alluvial wetlands are important natural habitats and contain valuable drinking water resources. The transport of pathogens via the inflows of river water or the release and runoff from animal faecal deposits into the backwater bodies can pose health risks. The aim of this study was to develop a combined modelling approach for predicting the concentrations and loads of protozoan reference pathogens during floods and rainfall events in an alluvial wetland river. The probabilistic-deterministic model QMRAcatch (v 1.1 python backwater) was newly adapted to account for short-time variations in the flow and microbial transport of alluvial wetlands. The wetland discharge rates, together with the inundated volumes and areas served as input to the model. The latter were determined by means of regression analysis based on results of a 2D hydrodynamic flow model during a flood event. To evaluate the model performance of QMRAcatch, we used concentrations of human, ruminant, pig and bird associated microbial faecal source tracking (MST) markers and E. coli measured in the Danube and in the wetland river from 2010 to 2015. The microbial die-off / degradation was identified to be the most relevant optimization parameter. To obtain this parameter, we conducted a literature survey on the degradation of MST markers in water environments, determined confidence limits of the temperature-dependent rate coefficients, and adjusted them within these limits during the optimization. Scenarios of the different transport pathways of Cryptosporidium and Giardia into the wetland bodies during floods and rainfall events were then simulated. The scenarios showed that the highest loads of Cryptosporidium and Giardia were transported via the main river into the wetland during high flows, followed by the rainfall-induced release from animal faecal deposits, and the resuspension in flooded areas. The combined modelling approach is useful to support the drinking water safety management of alluvial wetlands.Funding source: This work was supported by the Vienna Science and Technology Fund (WWTF) [grant number ESR17-070] and by the European Union and Vienna Water [programme number LE07-13, project name ‘Groundwater Resource Systems Vienna’].

BackgroundEvidence for indoor airborne transmission of SARS-CoV-2 is accumulating. If SARS-CoV-2 also spreads via aerosols, this has implications for measures taken to limit transmission.ObjectivesThe aim of this study is to assess exposure to airborne SARS-CoV-2 particles from breathing, speaking, coughing and sneezing in an indoor environment.MethodsAn exposure assessment model was developed to estimate numbers of SARS-CoV-2 particles in aerosol droplets, expelled during breathing, speaking, coughing and sneezing by an infected person in an unventilated indoor environment, and subsequent inhalation by one or more persons. Scenarios encompass a range of virus concentrations, room sizes and exposure times.ResultsThe calculated total volume of expelled aerosol droplets was highest for a sneeze, followed by a cough and speaking for 20 minutes, and lastly breathing for 20 minutes. A few to as much as tens of millions of virus particles were expelled. Exposure probability strongly depends on the viral concentration in mucus, as well as on the scenario. Exposure probabilities were generally below 1% at a virus concentration in mucus below 105 per mL for all scenarios, increasing steeply at different higher concentrations. According to nose / throat swab data collected from patients, 75%, 50% and 5% of infected individuals carry an estimated number of SARS-CoV-2 per mL mucus of at least 105, 106 and 108, respectively.DiscussionExposure to SARS-CoV-2 via aerosols generated during breathing, speaking, coughing and sneezing in an unventilated indoor environment is possible. This study forms a basis to estimate probabilities of exposure to SARS-Cov-2 by airborne transmission in indoor spaces. As long as it is uncertain what fraction of the airborne virus particles is infectious and as long as a dose response relation is lacking, it is recommended to be precautious.

Aquifer microbial water quality evaluations are often performed by collecting groundwater samples from monitoring wells. While samples collected from continuously pumped sources are seldom disputed as representative of the aquifer, natural biofilm present in the vicinity of well screens may introduce unwanted microbial artefacts in monitoring wells that are only periodically sampled. The need for well water purging to obtain samples void of these artefacts has been widely recognized. However, purging methods are not standardized; many approaches presume that physico-chemical water quality stability achieved through the removal of 3 to 5 well volumes is indicative of the stability of target analytes. Using a data set collected from a shallow unconfined aquifer in Southern Ontario, Canada, the need for using dedicated approaches that account for the time-dependent nature of microbial water quality changes was demonstrated. Specifically, the utility of adenosine triphosphate (ATP) as a rapid, field-ready biochemical indicator of microbial water quality stability was investigated. This work shows that ATP concentrations reflect time-limited (bio)colloid transport processes that are consistent with other microbial water quality parameters monitored, but different from commonly measured physical and chemical water quality indicators of well purging adequacy. ATP concentrations occasionally fluctuated even after 3 or 4 h of purging, indicating that microbial artefacts attributable to biofilms in the vicinity of the well screen can still persist. The recurrence of characteristic ATP patterns in each well was systematically examined through the novel application of dynamic time warping (DTW), a nonparametric time series analysis approach. These patterns are believed to be linked with seasonal hydrogeological conditions, which warrant consideration in the design and interpretation of subsurface microbial water quality investigations.

Microbial water quality evaluations are essential for determining the vulnerability of subsurface drinking water sources to fecal pathogen intrusion. Rather than directly monitor waterborne pathogens using culture- or enumeration-based techniques, the potential of assessing bacterial community using 16S rRNA gene amplicon sequencing to support these evaluations was investigated. A framework for analyzing 16S rRNA gene amplicon sequencing results featuring negative-binomial generalized linear models is demonstrated, and applied to bacterial taxa sequences in purge water samples collected from a shallow, highly aerobic, unconfined aquifer. Bacterial taxa relevant as indicators of fecal source and surface connectivity were examined using this approach. Observed sequences of Escherichia, a genus suggestive of fecal source, were consistently detected but not confirmed by culture-based methods. On the other hand, episodic appearance of anaerobic taxa sequences in this highly aerobic environment, namely Clostridia and Bacteroides, warrants further investigation as potential indicators of fecal contamination. Betaproteobacteria sequences varied significantly on a seasonal basis, and therefore may be linked to understanding surface-water groundwater interactions at this site. However, sequences that are often encountered in surface water bodies (Cyanobacteria and Flavobacteriia) were notably absent or present at very low levels, suggesting that microbial transport from surface-derived sources may be rather limited. This work demonstrates the utility of 16S rRNA gene amplicon sequencing for contextualizing and complementing conventional microbial techniques, allowing for hypotheses about source and transport processes to be tested and refined.

Some viruses cause tumor regression and can be used to treat cancer patients; these viruses are called oncolytic viruses. To assess whether oncolytic viruses from animal origin excreted by patients pose a health risk for livestock, a quantitative risk assessment (QRA) was performed to estimate the risk for the Dutch pig industry after environmental release of Seneca Valley virus (SVV). The QRA assumed SVV excretion in stool by one cancer patient on Day 1 in the Netherlands, discharge of SVV with treated wastewater into the river Meuse, downstream intake of river water for drinking water production, and consumption of this drinking water by pigs. Dose–response curves for SVV infection and clinical disease in pigs were constructed from experimental data. In the worst scenario (four log10 virus reduction by drinking water treatment and a farm with 10,000 pigs), the infection risk is less than 1% with 95% certainty. The risk of clinical disease is almost seven orders of magnitude lower. Risks may increase proportionally with the numbers of treated patients and days of virus excretion. These data indicate that application of wild‐type oncolytic animal viruses may infect susceptible livestock. A QRA regarding the use of oncolytic animal virus is, therefore, highly recommended. For this, data on excretion by patients, and dose–response parameters for infection and clinical disease in livestock, should be studied.

Microorganisms constitute an almost exclusive form of life in the earth's subsurface environment (not including caves), particularly at depths exceeding the soil horizon. While of broad interest to ecology and geology, scientific interest in the fate and transport of microorganisms, particularly those introduced through the anthropogenic environment, has focused on understanding the subsurface environment as a pathway for human pathogens and on optimizing the use of microbial organisms for remediation of potable groundwater. This special section, inspired by the 2014 Ninth International Symposium for Subsurface Microbiology, brings together recent efforts to better understand the spatiotemporal occurrence of anthropogenic microbial groundwater contamination and the fate and transport of microbes in the subsurface environment: in soils, deep unsaturated zones, and within aquifer systems. Work includes field reconnaissance, controlled laboratory studies to improve our understanding of specific fate and transport processes, and the development and application of improved mechanistic understanding of microbial fate and transport processes in the subsurface environment. The findings confirm and also challenge the limitations of our current understanding of highly complex microbial fate and transport processes across spatiotemporal scales in the subsurface environment; they also add to the increasing knowledge base to improve our ability to protect drinking water resources and perform in situ environmental remediation.

Currently, the number of reported cases of recreational- water-related Vibrio illness in the Netherlands is low. However, a notable higher incidence of Vibrio infections has been observed in warm summers. In the future, such warm summers are expected to occur more often, resulting in enhanced water temperatures favoring Vibrio growth. Quantitative information on the increase in concentration of Vibrio spp. in recreational water under climate change scenarios is lacking. In this study, data on occurrence of Vibrio spp. at six different bathing sites in the Netherlands (2009-2012) were used to derive an empirical formula to predict the Vibrio concentration as a function of temperature, salinity, and pH. This formula was used to predict the effects of increased temperatures in climate change scenarios on Vibrio concentrations. For Vibrio parahaemolyticus, changes in illness risks associated with the changed concentrations were calculated as well. For an average temperature increase of 3.7 °C, these illness risks were calculated to be two to three times higher than in the current situation. Current illness risks were, varying per location, on average between 10(-4) and 10(-2) per person for an entire summer. In situations where water temperatures reached maximum values, illness risks are estimated to be up to 10(-2) and 10(-1) . If such extreme situations occur more often during future summers, increased numbers of ill bathers or bathing-water-related illness outbreaks may be expected.

To estimate the exposure of local residents to substances emitted by livestock farms, we applied a dispersion model to calculate the air concentrations in the surroundings following from these emissions. At several livestock farms, indoor air measurements were performed to determine emission strengths, while ambient measurements were carried out to compare with model results. Measured substances were particulate matter (PM), endotoxins and micro-organisms. The dispersion model only simulated PM concentrations, which were used as a proxy to determine the dispersion concentrations of endotoxins and micro-organisms. For the living micro-organisms, the process of inactivation has to be taken into account. Here we describe the followed methodology and preliminary results.

The goal of this research was to investigate the difference in behavior of hydrophilic and hydrophobic colloids during transport in two-phase flow, in general, and their attachment and remobilization characters, in particular. Experiments were performed in a hydrophobic polydimethylsiloxane (PDMS) micromodel. Water and fluorinert-FC43 were used as the two immiscible liquids. Given the fact that PDMS is a hydrophobic material, fluorinert was the wetting phase and water was the nonwetting phase in this micromodel. As model colloids, we used hydrophilic polystyrene carboxylate-modified microspheres (dispersible in water) and hydrophobic fluorous-modified silica microspheres (dispersible in fluorinert) in separate experiments. Using a confocal laser scanning microscope, we directly observed fluid distribution and colloid movement within pores of the micromodel. We also obtained concentration breakthrough curves by measuring the fluorescent intensities in the outlet of the micromodel. The breakthrough curves during steady-state flow showed that the colloid attachment rate is inversely related to the background saturation of the fluid in which the colloids were dispersed. Our visualization results showed that the enhanced attachment of hydrophilic colloids at lower water saturations was due to the retention at the fluorinert-water interface and fluorinert-water-solid contact lines. This effect was observed to be much less in the case of hydrophobic colloids (dispersed in fluorinert). In order to explain the colloids behavior, we calculated interaction potential energies of colloids with PDMS surfaces, fluid-fluid interfaces, and fluid-fluid-solid contact lines. Also, balance of forces that control colloid, including DLVO, hydrodynamic, and surface tension forces, were determined. Our calculations showed that there is a stronger repulsive energy barrier between hydrophobic colloids and fluorinert-water interface and solid-fluid interface, compared with the hydrophilic colloids. Moreover, hydrophobic colloids were seen to aggregate due to strong attractive forces among them. These aggregates had even less tendency to attach to various interfaces, due to an increase in the corresponding energy barrier. For the colloid retention at fluid-fluid-solid contact lines, we found that the role of DLVO interactions was less important than capillary forces. During transient events, we found that attached colloids become remobilized. The colloids deposited on the solid-fluid interface were detached by the moving fluid-fluid-solid contact lines. But, this happened only when the liquid containing colloids was being displaced by the other liquid. We simulated breakthrough curves using a model based on a coupled system of equations for two-phase flow, advection-dispersion of colloids, adsorption to and desorption from fluid-fluid interfaces and fluid-solid interfaces. Very good agreements were obtained among measured breakthrough curves, visualization results, and numerical modeling.

Riverbank fi ltration is considered an effi cient method for removing contaminants from infi ltrated surface water in the subsurface. Despite indications that changing water temperatures affect the biochemical and biological mediated removal processes of contaminants, the impact of temperature induced fl uid viscosity and density effects on contaminant removal during riverbank fi ltration is not well understood. This paper investigates the viscosity and density effects associated with seasonal changes in groundwater temperature on virus and dissolved organic carbon (DOC) removal during riverbank fi ltration. Hypothetical aquifer and fl ood wave scenarios were assumed. Data on groundwater temperature were taken from an Austrian fi eld site of the River Danube recorded during 2010/2011. Based on removal rates taken from previously published fi eld experiments, virus and DOC transport was simulated for highly permeable gravel, fi ne gravel and fi ne sandy gravel material. Our simulations indicate that for DOC and a wide range of virus types the viscosity and density effects induced by water temperature changes can counteract with temperature dependent decay and inactivation rates. For particular situations, however, such as for receding fl oods during colder periods, our simulations indicate that fl uid viscosity and density effects can result in a net de- crease in the virus removal effi ciency during colder periods. Persistent types of viruses (e.g. polio 1 or HAV ) can be reduced less effectively and may travel by up to 25 % faster during warmer than during colder periods. Our simula- tions indicate that viscosity and density effects induced by temperature changes should be considered for studying and simulating virus or DOC removal and transport during riverbank fi ltration. The effects may be important spe- cifi cally at fi eld sites with a high river-aquifer exchange and large variations in groundwater temperature.

The objective of this work was to investigate and obtain quantitative relations for the effects of Ca 2 + concentration on virus removal in saturated soil and to compare the experimental findings with predictions of the DLVO theory. In order to do so, a systematic study was performed with a range of calcium concentrations corresponding to natural field conditions. Experiments were conducted in a 50-cm column with clean quartz sand under saturated conditions. Inflow solutions were prepared by adding CaCl 2, NaCl and NaHCO 3 to de-ionized water. Values of pH and ionic strength were fixed at 7 and 10 mM, respectively. Bacteriophage PRD1 was used as a conservative model virus for virus removal. The samples were assayed using the plaque forming technique. Attachment, detachment and inactivation rate coefficients were determined from fitting breakthrough curves. Attachment rate coefficients were found to increase with increasing calcium concentration. Results were used to calculate sticking efficiency, for which an empirical formula as a function of Ca 2 + was developed. Numerical solutions of the Poisson–Boltzmann equation were obtained to evaluate the effect of Ca 2 + on the double-layer interactions between quartz and PRD1. Based on these results, the DLVO interaction energies were calculated. It turned out that the experimental findings cannot be explained with the distance profiles of the DLVO interaction. The discrepancy between theory and experiment can be attributed to underestimation of the van der Waals interactions, chemisorption of Ca 2 + onto the surfaces, or by factors affecting the double-layer interactions, which are not included in the Poisson–Boltzmann equation. When abruptly changing from inflow solution containing Ca 2 + to a Ca 2 + -free solution, pronounced mobilization of viruses was observed. This indicates virus removal is not irreversible and that chemical perturbations of the groundwater can cause a burst of released viruses.

Quantitative microbiological risk assessment was used to quantify the risk associated with the exposure to Legionella pneumophila in a whirlpool. Conceptually, air bubbles ascend to the surface, intercepting Legionella from the traversed water. At the surface the bubble bursts into dominantly noninhalable jet drops and inhalable film drops. Assuming that film drops carry half of the intercepted Legionella, a total of four (95% interval: 1-9) and 4.5×10(4) (4.4×10(4) - 4.7×10(4) ) cfu/min were estimated to be aerosolized for concentrations of 1 and 1,000 legionellas per liter, respectively. Using a dose-response model for guinea pigs to represent humans, infection risks for active whirlpool use with 100 cfu/L water for 15 minutes were 0.29 (∼0.11-0.48) for susceptible males and 0.22 (∼0.06-0.42) for susceptible females. A L. pneumophila concentration of ≥1,000 cfu/L water was estimated to nearly always cause an infection (mean: 0.95; 95% interval: 0.9-∼1). Estimated infection risks were time-dependent, ranging from 0.02 (0-0.11) for 1-minute exposures to 0.93 (0.86-0.97) for 2-hour exposures when the L. pneumophila concentration was 100 cfu/L water. Pool water in Dutch bathing establishments should contain

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.

Objectives of this work are to investigate effects of pH and ionic strength (IS) on virus transport in saturated soil and to develop a quantitative relationship for these effects. A series of 50-cm column experiments with clean quartz sand under saturated conditions and with pH values of 5, 6, 7, 8, and IS values of 1, 10, and 20 mM were conducted. Bacteriophage PRD1 was used as a model virus. Applying a one-site kinetic model, attachment, detachment, and inactivation rate coefficients were determined from fitting breakthrough curves using the software package Hydrus-1D. Attachment rate coefficients increased with decreasing pH and increasing IS, in agreement with DLVO theory. Sticking efficiencies were calculated from the attachment rate coefficients and used to develop an empirical formula for sticking efficiency as a function of pH and IS. This relationship is applicable under unfavorable conditions for virus attachment. We compared sticking efficiencies predicted by the empirical formula with those from field and column experiments. Within the calibrated range of pH and IS, the predicted and observed sticking efficiencies are in reasonable agreement for bacteriophages PRD1 and MS2. However, the formula significantly overestimates sticking efficiencies for IS higher than 100 mM. In addition, it performs less well for viruses with different surface reactivity than PRD1 and MS2. Effects of pH and IS on detachment and inactivation rate coefficients were also investigated but the experimental results do not allow constraining these parameters with sufficient certainty.

Our study aimed to assess the accumulation of bacteriophages in sandy and clayey fresh water sediments. All of the 24 natural fresh water sediments were positive for somatic and F-specific phages, though their concentrations in the overlying water were undetectable in 1 and 11 samples, respectively, out of 24, corresponding to 4 and 46% for somatic and F-specific phages, respectively. Based on the sediment-to-water ratios, F-specific phages accumulate over 100 times more than the somatic coliphages in clayey sediments. Inactivation of bacteriophages in clayey and sandy sediments over a 1-month period at 15°C was negligible. Our data suggest that persistence of deposited viruses in fresh water sediments leads to accumulation and the findings call for additional investigations on the fate of entrapped pathogenic viruses.

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.

Though coliform bacteria are used worldwide to indicate faecal pollution of groundwater, the parameters determining the transport of Escherichia coli in aquifers are relatively unknown. To investigate the effect of dissolved organic carbon (DOC) on the attachment of E. coli to saturated goethite-coated sand, we carried out column experiments with E. coli with and without humic acid (HA) in monovalent and divalent salt solutions. To characterize sorption of DOC and attachment of E. coli , we measured the pH of the influent and effluent, the cation concentrations and the zeta potential of particles. Depending on the chemistry of the E. coli suspension, the normalized breakthrough concentrations were over 80 times higher in columns treated with HA compared with columns not treated with HA. However, this difference was not constant: there were time-dependent variations in attachment of E. coli to the collector surface, and in the chemical composition of the bacterial suspension. Reduction in removal occurred because HA altered the surface charge of the collector and also sterically hindered E. coli . In addition, reduction of removal in a CaCl 2 bacterial suspension was probably caused by site-blocking mechanisms between HA and Ca 2+ ions. Our results indicate that in the presence of DOC, the concept of geochemical heterogeneity in explaining attachment of biocolloids has limited relevance.

The two most significant processes controlling virus mobility in the subsurface environment are virus attachment and inactivation. In particular, models that predict subsurface virus transport are highly sensitive to inactivation. Virus inactivation is known to depend on temperature as well as hydrochemical conditions. The aim of the current work was to study the effects of temperature and hydrochemical conditions on the inactivation of bacteriophage PRD1 as a model virus, and to develop a quantitative relation for these effects. Series of batch experiments under controlled temperature were conducted, for a range of conditions: 9.5 °C and 12 °C, pH4 - pH8, sodium concentrations of 1, 10 and 20 mM, and calcium concentrations of 0.5, 1.5, and 3 mM. By multivariate regression analysis, a joint log-square model was developed that describes the inactivation rate of PRD1 as a function of these hydrochemical conditions. This model approximates two rate and Weibull models and accounts for the observed non-linear inactivation at increased pH and salt concentrations. Model predictions are within ±0.4 log10 (0.4-2.5 times) virus concentration reduction. The nature of the log-square model does not allow extrapolation of virus inactivation beyond the experimental conditions. Inactivation rate of PRD1 was found to increase with increasing temperature and increasing sodium and calcium concentrations, and to be lowest between pH 6.5 and pH 7.5. Within the studied conditions, the developed log-square model may be applied at field scale for predicting inactivation during subsurface transport of viruses.

Though coliform bacteria are used worldwide to indicate fecal pollution of groundwater, the parameters determining the transport of Escherichia coli in aquifers are relatively unknown. We evaluated the occurrence of both straining and attachment of E. coli ATCC25922 in columns of ultra-pure, angular, saturated quartz sand. The column experiments were conducted over a wide range of porous medium sizes, column heights, input concentrations, and pore water flow velocities. Straining and attachment were examined by modelling the breakthrough curves (with HYDRUS 1D). In addition, model output was compared with measured strained and attached bacteria via column extrusion experiments (in which sand was extruded from the column and placed in excess water) and flow reversal experiments (in which the pore water flow direction was reversed, thereby dislodging strained bacteria). Our model consisted of an attachment rate coefficient and a straining rate coefficient; both of these decreased with transport distance. The straining rate coefficient also decreased in a Langmuirian way, in response to the filling of available pore space, which in turn depended on influent bacteria concentration, quartz grain diameter, and transport distance. The maximum strained fraction was 25-30% of total bacteria mass applied to the column; the maximum attached fraction was 30-35%. The fit between modelled and measured (strained and attached) bacteria masses was acceptable, as was the sensitivity of the model output to fitted parameter values. Our results lead to a new description for the time-dependent mass balance of strained bacteria, which entails using three fitting parameters. The results also imply that column experiments in combination with retention profiles (or various column lengths) are not enough to explain the retention processes in a column. Column extrusion and flow reversal experiments provide vital additional information on the occurrence and magnitude of straining. Our straining model could be of assistance in evaluating the importance of straining and in incorporating the straining process in bacteria transport modelling.

The decimal elimination capacity (DEC) of slow sand filtration (SSF) for Cryptosporidium parvum was assessed to enable quantitative microbial risk analysis of a drinking water production plant. A mature pilot plant filter of 2.56 m2 was loaded with C. parvum oocysts and two other persistent organisms as potential surrogates; spores of Clostridium perfringens (SCP) and the small-sized (4–7 μm) centric diatom (SSCD) Stephanodiscus hantzschii. Highly persistent micro-organisms that are retained in slow sand filters are expected to accumulate and eventually break through the filter bed. To investigate this phenomenon, a dosing period of 100 days was applied with an extended filtrate monitoring period of 150 days using large-volume sampling. Based on the breakthrough curves the DEC of the filter bed for oocysts was high and calculated to be 4.7 log. During the extended filtrate monitoring period the spatial distribution of the retained organisms in the filter bed was determined. These data showed little risk of accumulation of oocysts in mature filters most likely due to predation by zooplankton. The DEC for the two surrogates, SCP and SSCD, was 3.6 and 1.8 log, respectively. On basis of differences in transport behaviour, but mainly because of the high persistence compared to the persistence of oocysts, it was concluded that both spores of sulphite-reducing clostridia (incl. SCP) and SSCD are unsuited for use as surrogates for oocyst removal by slow sand filters. Further research is necessary to elucidate the role of predation in Cryptosporidium removal and the fate of consumed oocysts.

Drinking water distribution networks are vulnerable to accidental or intentional contamination events. The objective of this study was to investigate the effects of seeding duration and concentration, exposure pathway (ingestion via drinking of water and tooth brushing and inhalation by taking a shower) and pathogen infectivity on exposure and infection risk in the case of an intentional pathogenic contamination in a drinking water distribution network. Seeding of a pathogen for 10 min and 120 min, and subsequent spreading through a drinking water distribution network were simulated. For exposure via drinking, actual data on drinking events and volumes were used. Ingestion of a small volume of water by tooth brushing twice a day by every person in the network was assumed. Inhalation of contaminated aerosol droplets took place when taking a shower. Infection risks were estimated for pathogens with low (r = 0.0001) and high (r = 0.1) infectivity. In the served population (48 000 persons) and within 24 h, about 1400 persons were exposed to the pathogen by ingestion of water in the 10-min seeding scenario and about 3400 persons in the 120-min scenario. The numbers of exposed persons via tooth brushing were about the same as via drinking of water. Showering caused (inhalation) exposure in about 450 persons in the 10-min scenario and about 1500 in the 120-min scenario. Regardless of pathogen infectivity, if the seeding concentration is 10(6) pathogens per litre or more, infection risks are close to one. Exposure by taking a shower is of relevance if the pathogen is highly infectious via inhalation. A longer duration of the seeding of a pathogen increases the probability of exposure.

Given the complex hydrologic dynamics of water catchments and conflicts between nature protection and public water supply, models may help to understand catchment dynamics and evaluate contamination scenarios and may support best environmental practices and water safety management. A catchment model can be an educative tool for investigating water quality and for communication between parties with different interests in the catchment. This article introduces an interactive computational tool, QMRAcatch, that was developed to simulate concentrations in water resources of Escherichia coli, a human-associated Bacteroidetes microbial source tracking (MST) marker, enterovirus, norovirus, Campylobacter, and Cryptosporidium as target microorganisms and viruses (TMVs). The model domain encompasses a main river with wastewater discharges and a floodplain with a floodplain river. Diffuse agricultural sources of TMVs that discharge into the main river are not included in this stage of development. The floodplain river is fed by the main river and may flood the plain. Discharged TMVs in the river are subject to dilution and temperature-dependent degradation. River travel times are calculated using the Manning–Gauckler–Strickler formula. Fecal deposits from wildlife, birds, and visitors in the floodplain are resuspended in flood water, runoff to the floodplain river, or infiltrate groundwater. Fecal indicator and MST marker data facilitate calibration. Infection risks from exposure to the pathogenic TMVs by swimming or drinking water consumption are calculated, and the required pathogen removal by treatment to meet a health-based quality target can be determined. Applicability of QMRAcatch is demonstrated by calibrating the tool for a study site at the River Danube near Vienna, Austria, using field TMV data, including a sensitivity analysis and evaluation of the model outcomes.

The transport of viruses in unsaturated porous media has been a subject of great interest in recent years because of the enhanced removal of these microorganisms compared with saturated conditions. We studied the transport of bacteriophages MS2 and fX174, used as surrogate pathogenic viruses, at various water contents and solution chemistries in terms of pH and ionic strength (IS). The objectivewas to explore the interaction of viruses with the solid–water interfaces (SWI) and air–water interfaces (AWI) for a range of conditions. The experimental data were fitted with a transport model to determine the adsorption (attachment and detachment rate) parameters. Our results show that the retention of viruses in the soil column increases as water saturation decreases when the chemical conditions are favorable for adsorption(pH7andrelativelyhighIS). Ouranalysis indicatesthatthe enhanced retention of fX174 viruses at lower water contents is caused by increased attachment to the SWI and that retention by the AWI is not significant. Results obtained from a first series of experiments (pH 9 and low IS) showed insignificant attachment of MS2 viruses to both the SWI and the AWI. The MS2 breakthrough data for a second series of experiments (pH 7 and high IS) did not allow us to separate out the role of the AWI. Although attachment of MS2 viruses to the AWI cannot be ruled out in our experiments, we suspect that the increased retention of this phage under unsaturated condition was also due to enhanced attachment to the SWI. Increased attachment to the SWI underunsaturated conditionsisattributedtoincreasedmass transferof viruses to the SWI due to a reduced diffusion length at lower water contents. Our results demonstrate that if there is any attachment to the AWI, it is reversible. When unfavorable conditions occur for attachment to the SWI, the attached viruses may be detached by moving solid–water–air contact lines (SWA).

Divers may run a higher risk of infection with waterborne pathogens than bathers because of more frequent and intense contact with water that may not comply with microbiologic water quality standards for bathing water. In this study we aimed to estimate the volume of water swallowed during diving as a key factor for infection risk assessment associated with diving. Using questionnaires, occupational and sport divers in the Netherlands were asked about number of dives, volume of swallowed water, and health complaints (nausea, vomiting, diarrhea, and ear, skin, eye, and respiratory complaints). Occupational divers, on average, swallowed 9.8 mL marine water and 5.7 mL fresh surface water per dive. Sport divers swallowed, on average, 9.0 mL marine water; 13 mL fresh recreational water; 3.2 mL river, canal, or city canal water; and 20 mL water in circulation pools. Divers swallowed less water when wearing a full face mask instead of an ordinary diving mask and even less when wearing a diving helmet. A full face mask or a diving helmet is recommended when diving in fecally contaminated water. From the volumes of swallowed water and concentrations of pathogens in fecally contaminated water, we estimated the infection risks per dive and per year to be as high as a few to up to tens of percents. This may explain why only 20% of the divers reported having none of the inquired health complaints within a period of 1 year. It is highly recommended that divers be informed about fecal contamination of the diving water.

Protection zones of shallow unconfined aquifers in The Netherlands were calculated that allow protection against virus contamination to the level that the infection risk of 10−4 per person per year is not exceeded with a 95% certainty. An uncertainty and a sensitivity analysis of the calculated protection zones were included. It was concluded that protection zones of 1 to 2 years travel time (206–418 m) are needed (6 to 12 times the currently applied travel time of 60 days). This will lead to enlargement of protection zones, encompassing 110 unconfined groundwater well systems that produce 3 × 108 m3 y−1 of drinking water (38% of total Dutch production from groundwater). A smaller protection zone is possible if it can be shown that an aquifer has properties that lead to greater reduction of virus contamination, like more attachment. Deeper aquifers beneath aquitards of at least 2 years of vertical travel time are adequately protected because vertical flow in the aquitards is only 0.7 m per year.The most sensitive parameters are virus attachment and inactivation. The next most sensitive parameters are grain size of the sand, abstraction rate of groundwater, virus concentrations in raw sewage and consumption of unboiled drinking water. Research is recommended on additional protection by attachment and under unsaturated conditions.

To elucidate the parameters determining the transport of Escherichia coli in aquifers, the attachment of E. coli in low concentrations to column sediments was investigated. The sediments comprised 0.18–0.50 mm quartz sand, grains coated with goethite, calcite grains or grains of activated carbon (AC), in varying fractions ( λ = 0 , 0.05, 0.1, 0.2, 0.4, 0.7, 1.0) and all of similar diameter to the quartz sand. The weighted sum of favourable and unfavourable sticking efficiencies ( α total ) showed that upon increasing the fraction of favourable mineral grains ( λ ) there was an initial rapid increase, which then slowed down. This was most pronounced in the AC experiments, followed by the calcite experiments and then the goethite experiments. We ascribe this non-linear relation to surface charge and hydrophobic heterogeneity of the E. coli population.

Effects of climate change in The Netherlands in the 21st century on the microbiological quality of water for drinking water production and recreation were evaluated. The following was concluded: increased temperature leads to significant additional inactivation of enteric pathogens in surface waters with residence times of more than a month, but not in waters with residence times of up to ten days. Increased precipitation, runoff and storm water overflow lead to more peak concentrations of waterborne pathogens in surface water. Peak concentrations strongly determine the infection risk through drinking water consumption. Drought lowers and increased precipitation elevates groundwater tables, but an effect on the risk of groundwater contamination with waterborne pathogens is not clear. Climate effects are not noticeable near a groundwater well, where the groundwater table and flow rate are mainly determined by the pumping rate. Exposure of recreants to waterborne pathogens that can grow in the water is expected to increase due to increased recreation and increased growth opportunities of these pathogens. Due to warmer summers, pathogens, like amoeba, that have not caused problems up to date in The Netherlands, may now emerge in recreational waters.

In a recent field study on dune recharge, bacteriophages MS2 and PRD1 were found to be removed 3 log10 over the first 2.4 m and only 5 log10 over the next 27 m. To understand the causes of this nonlinear removal, column experiments were carried out under conditions similar to the field: same recharge water, temperature (5 +/- 3 degrees C) and pore water velocity (1.5 m day(-1)). Soil samples were taken along a streamline between the recharge canal and the first monitoring well. Bacteriophage phiX174 was included for comparison. The high initial removal in the field was found not to be due to heterogeneity of phage suspensions but to soil heterogeneity. Phage removal rates correlated strongly positively with soil organic carbon content, and relatively strongly positively with silt content and the presence of ferric oxyhydroxides. Soil organic carbon content, silt content and the presence of ferric oxyhydroxides were found to decrease exponentially with travel distance. Removal rates of phiX174 were found to be 3-10 times higher than those of MS2 and PRD1 due to the lower electrostatic repulsion that the less negatively charged phiX174 experiences. It is suggested that the high initial removal in the field is due to the presence of favorable sites for attachment formed by ferric oxyhydroxides that decrease exponentially with travel distance. Similar removal rates may be found at both laboratory and field scale. However, due to local variations at field scale detailed knowledge on soil heterogeneity may be needed to enable a reliable prediction of removal.

Studies were initiated to determine the release behavior of Cryptosporidium oocysts and Giardia cysts from dairy calf manure to waters of various salinities. Experiments were conducted by sprinkling a particular aqueous solution over a manure disk and collecting the runoff water. Effluent concentrations of manure and (oo)cysts were initially several orders of magnitude below their starting concentration in the manure, after continued application of water the concentrations gradually decreased, and then exhibited persistent concentration tailing. Solution salinity significantly affected the shape and magnitude of the manure and (oo)-cyst concentration curves. Increases in solution salinity tended to decrease the manure and (oo)cyst concentrations at a particular time. This was attributed to a stabilization of manure by compression of the double layer thickness between negatively charged components of the manure phase. Calculated release efficiencies of the (oo)cysts (relative to manure release) also decreased with increasing solution salinity. Experimental observations indicate that only the surface layer of manure was depleted of finer manure materials and (oo)cysts and that the manure will act as a long-term source of contamination. A conceptual model to describe and predict manure and (oo)cyst release rates and cumulative loading for the various solution salinities was proposed and applied to the experimental data. The calibrated model yielded a reasonable description of the experimental results.

Virus removal from groundwater by soil passage often appears to be much higher during the first few metres due to the presence of more favorable sites for attachment than thereafter. A model is presented which interprets virus removal as a function of collision efficiencies αbeta; and αλl, inactivation rate coefficient μll and rate parameter γ. Initial high removal is determined by αβ, which decreases exponentially at a rate γ to a constant base removal rate that is determined by αλ and μl. A hypothetical worst case was simulated to calculate the travel distance and time required for 9 log10 protection against virus contamination of groundwater wells in anoxic sandy aquifers. Unfavorable conditions for attachment were assumed. Virus was constantly leaking from a sewage pipe lying at the groundwater table. Mixing reduced virus concentration by 3.1 to 4.0 log10. For an additional 5.0 to 5.9 log10 protection against virus contamination by attachment and inactivation, residence times of about three to seven times longer than the current guideline of 60 days are needed, depending on abstraction rates, aquifer thickness and grain size of the sand.

Modelling the discharge of parasitic protozoa into surface water and the dispersion in rivers and streams gives insight into the contribution of the different sources of environmental contamination and in the transmission of these organisms from the point of discharge to drinking water abstraction points and bathing sites. We tested the applicability of emission (PROMISE) and dispersion (WATNAT) models developed for chemical pollutants to describe the environmental behaviour of Cryptosporidium and Giardia in the Netherlands. The annual load of Cryptosporidium and Giardia in domestic wastewater was 3.2×10 13 and 3.8×10 14 , respectively. The majority (85%) of the Cryptosporidium oocysts was discharged with effluent of wastewater treatment plants, while the majority (82%) of the Giardia cysts was discharged with untreated wastewater discharges and sewer overflows. The estimated annual import through the river Rhine and Meuse was 3.2×10 14 Cryptosporidium oocysts and 2.1×10 15 Giardia cysts, of which the river Rhine contributed 87 and 66%, respectively. This outweighed the total load of the discharges of treated and untreated wastewater in the Netherlands. The combination of PROMISE and WATNAT predicted concentrations of Cryptosporidium and Giardia in surface water that were in the same order of magnitude as the concentrations that were observed at 5 of the 6 sites compared. At a site with primarily agricultural contamination, the models predicted concentrations that were 1 10 log-unit lower than the observed concentrations. This is a first step in the direction of a quantitative description of the transmission cycle of Cryptosporidium and Giardia through water. The use of these models combines observational occurrence data and experimental data from laboratory survival studies into a single integrated description. The description needs further improvement by incorporation of agricultural run-off and increasing the number and time frame of input monitoring data.

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

Experiments with batch suspensions, recirculating columns and flow-through columns have been carried out involving a sandy soil and five bacteriophages: MS2, PRD1, φX174, Qβ and PM2. In batch and recirculating column experiments, attachment and detachment rate coefficients were determined by fitting a two-parameter (attachment and detachment) model. In general, attachment and detachment rate coefficients were not found to be significantly different between the two kinds of experiments. There was one exception, however: MS2 appeared to detach faster in the presence of strong advective flow. In the case of flow-through column experiments, it is shown that a two-site model, with adsorption to equilibrium and kinetic sites, fits the breakthrough curves of all the phages, except PM2, satisfactorily. A one-site kinetic model was found to be appropriate for phage PM2. A small proportion of bacteriophages MS2, PRD1, and Qβ adsorbed to equilibrium sites, whereas a large proportion of φX174 adsorbed to equilibrium sites. Such a distinction between adsorption to equilibrium and kinetic sites cannot be made in the case of batch or recirculating column experiments. Kinetic attachment rate coefficients were found to be significantly higher for the bacteriophages with presumably stronger negative charge. This may be ascribed to the presence of multivalent cations. Under these conditions, bacteriophage φX174 appears to behave more conservatively than more negatively charged viruses, and may then be a better choice as a relatively conservative tracer for virus transport through the subsurface.

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.

Climate change may impact waterborne and foodborne infectious disease, but to what extent is uncertain. Estimating climate-change-associated relative infection risks from exposure to viruses, bacteria, or parasites in water or food is critical for guiding adaptation measures. We present a computational tool for strategic decision making that describes the behavior of pathogens using location-specific input data under current and projected climate conditions. Pathogen-pathway combinations are available for exposure to norovirus, Campylobacter, Cryptosporidium, and noncholera Vibrio species via drinking water, bathing water, oysters, or chicken fillets. Infection risk outcomes generated by the tool under current climate conditions correspond with those published in the literature. The tool demonstrates that increasing temperatures lead to increasing risks for infection with Campylobacter from consuming raw/undercooked chicken fillet and for Vibrio from water exposure. Increasing frequencies of drought generally lead to an elevated infection risk of exposure to persistent pathogens such as norovirus and Cryptosporidium, but decreasing risk of exposure to rapidly inactivating pathogens, like Campylobacter. The opposite is the case with increasing annual precipitation; an upsurge of heavy rainfall events leads to more peaks in infection risks in all cases. The interdisciplinary tool presented here can be used to guide climate change adaptation strategies focused on infectious diseases.