Dispersion of silica-encapsulated DNA magnetic particles in a homogeneous sand tank.

In this study, we focused on the 3D dispersion of colloids. To our knowledge, we were the first to do so. Thereto, we injected silica encapsulated DNA tagged superparamagnetic particles (SiDNAmag) in a homogeneous coarse grain sand tank. At four downstream locations, SiDNAmag concentrations were determined as a function of time. Longitudinal and transverse dispersivity values and associated uncertainties of SiDNAmag were determined using Monte Carlo modelling approach. The parameter associated uncertainties of hydraulic conductivity as well as of the effective porosity estimated from SiDNAmag breakthrough curves were statistically similar to those estimated from salt tracer breakthrough curves. Further, the SiDNAmag dispersivity uncertainty ranges were then statistically compared with the salt tracer (NaCl, and fluorescein) dispersivities. Our results indicated that time to rise, time of peak concentration and shape of the breakthrough curves of SiDNAmag were similar to those of the salt tracer breakthrough curves. Despite the size difference between the salt tracer molecules and SiDNAmag, size exclusion did not occur, probably due to the large pore throat diameter to SiDNAmag diameter ratio. The median longitudinal dispersivity (αL) of salt tracer and SiDNAmag were 4.9 and 5.8 × 10-4 m, respectively. The median ratio of horizontal and vertical transverse dispersivities to αL, (αTH /αL and αTV /αL, respectively), for salt tracer and SiDNAmag ranged between 0.52 and 0.56. Through the statistical tests, we concluded that the longitudinal and traverse dispersivities of SiDNAmag were not statistically significantly different from salt tracer in 3 dimensions and could be used to characterize the dispersive properties of the medium we used. Our work contributes to a better understanding of 3D dispersion of SiDNAmag in saturated porous media.

QMRA of Ralstonia solanacearum in potato cultivation: Risks associated with irrigation water recycled through managed aquifer recharge.

Agricultural aquifer storage recovery and transfer (ASTR) stores excess fresh water for later reuse in irrigation. Moreover, water quality improves because chemical pollutants and pathogens will be removed by degradation and attachment to the aquifer material. The source water may contain the bacterial plant pathogen Ralstonia solanacearum which causes plant infections and high yield losses. We used quantitative microbial risk assessment (QMRA) to investigate the removal of R. solanacearum during ASTR to predict infection risks of potato plants after irrigation with the recovered water. Laboratory experiments analyzed the ASTR treatment by investigating the bacterial die-off in the water phase and the removal by attachment to the aquifer sediment. Die-off in the water phase depends on the residence time and ranged between 1.3 and 2.7 log10 after 10 or 60 days water storage, respectively. A subpopulation of the bacteria persisted for a prolonged time at low concentrations which may pose a risk if the water is recovered too early. However, the natural aquifer sand filtration proofed to be highly effective in removing R. solanacearum by attachment which depends on the distance between injection and abstraction well. The high removal by attachment alone (18 log10 after 1 m) would reduce bacterial concentrations to negligible numbers. Upscaling to longer soil passages is discussed in the paper. Infection risks of potato plants were calculated using a dose-response model and ASTR treatment resulted in negligible infection risks of a single plant, but also when simulating the irrigation of a 5 ha potato field. This is the first QMRA that analyzed an agricultural ASTR and the fate of a plant pathogen focusing on plant health. QMRA is a useful (water) management tool to evaluate the treatment steps of water reclamation technologies with the aim to provide safe irrigation water and reduce risks disseminating plant diseases.

Quantitative Microbial Risk Assessment of Contracting COVID-19 Derived from Measured and Simulated Aerosol Particle Transmission in Aircraft Cabins.

BACKGROUND: SARS-CoV-2 can be effectively transmitted between individuals located in close proximity to each other for extended durations. Aircraft provide such conditions. Although high attack rates during flights were reported, little was known about the risk levels of aerosol transmission of SARS-CoV-2 in aircraft cabins. OBJECTIVES: The major objective was to estimate the risk of contracting COVID-19 from transmission of aerosol particles in aircraft cabins. METHODS: In two single-aisle and one twin-aisle aircraft, dispersion of generated aerosol particles over a seven-row economy class cabin section was measured under cruise and taxi conditions and simulated with a computational fluid dynamic model under cruise conditions. Using the aerosol particle dispersion data, a quantitative microbial risk assessment was conducted for scenarios with an asymptomatic infectious person expelling aerosol particles by breathing and speaking. Effects of flight conditions were evaluated using generalized additive mixed models. RESULTS: Aerosol particle concentration decreased with increasing distance from the infectious person, and this decrease varied with direction. On a typical flight with an average shedder, estimated mean risk of contracting COVID-19 ranged from 1.3×10-3 to 9.0×10-2. Risk increased to 7.7×10-2 with a super shedder (<3% of cases) on a long flight. Risks increased with increasing flight duration: 2-23 cruise flights of typical duration and 2-10 flights of longer duration resulted in at least 1 case of COVID-19 due to onboard aerosol transmission by one average shedder, and in the case of one super shedder, at least 1 case in 1-3 flights of typical duration cruise and 1 flight of longer duration. DISCUSSION: Our findings indicate that the risk of contracting COVID-19 by aerosol transmission in an aircraft cabin is low, but it will not be zero. Testing before boarding may help reduce the chance of a (super)shedder boarding an aircraft and mask use further reduces aerosol transmission in the aircraft cabin. https://doi.org/10.1289/EHP11495.

Risk assessment of banknotes as a fomite of SARS-CoV-2 in cash payment transactions.

The COVID 19 pandemic has triggered concerns and assumptions globally about transmission of the SARS-CoV-2 virus via cash transactions. This paper assesses the risk of contracting COVID-19 through exposure to SARS-CoV-2 via cash acting as a fomite in payment transactions. A quantitative microbial risk assessment was conducted for a 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. Accounting for the likelihood of the scenario to occur by considering (1) a local prevalence of 100 COVID-19 cases/100,000 persons, (2) a maximum of about one-fifth 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). The depicted 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.

Removal of bacterial plant pathogens in columns filled with quartz and natural sediments under anoxic and oxygenated conditions.

Irrigation with surface water carrying plant pathogens poses a risk for agriculture. Managed aquifer recharge enhances fresh water availability while simultaneously it may reduce the risk of plant diseases by removal of pathogens during aquifer passage. We compared the transport of three plant pathogenic bacteria with Escherichia coli WR1 as reference strain in saturated laboratory column experiments filled with quartz sand, or sandy aquifer sediments. E. coli showed the highest removal, followed by Pectobacterium carotovorum, Dickeya solani and Ralstonia solanacearum. Bacterial and non-reactive tracer breakthrough curves were fitted with Hydrus-1D and compared with colloid filtration theory (CFT). Bacterial attachment to fine and medium aquifer sand under anoxic conditions was highest with attachment rates of max. katt1 = 765 day-1 and 355 day-1, respectively. Attachment was the least to quartz sand under oxic conditions (katt1 = 61 day-1). In CFT, sticking efficiencies were higher in aquifer than in quartz sand but there was no differentiation between fine and medium aquifer sand. Overall removal ranged between < 6.8 log10 m-1 in quartz and up to 40 log10 m-1 in fine aquifer sand. Oxygenation of the anoxic aquifer sediments for two weeks with oxic influent water decreased the removal. The results highlight the potential of natural sand filtration to sufficiently remove plant pathogenic bacteria during aquifer storage.

Dose-response relationship of Ralstonia solanacearum and potato in greenhouse and in vitro experiments.

Ralstonia solanacearum is the causative agent of bacterial wilt of potato and other vegetable crops. Contaminated irrigation water contributes to the dissemination of this pathogen but the exact concentration or biological threshold to cause an infection is unknown. In two greenhouse experiments, potted potato plants (Solanum tuberosum) were exposed to a single irrigation with 50 mL water (non-invasive soil-soak inoculation) containing no or 102 - 108 CFU/mL R. solanacearum. The disease response of two cultivars, Kondor and HB, were compared. Disease development was monitored over a three-month period after which stems, roots and tubers of asymptomatic plants were analyzed for latent infections. First wilting symptoms were observed 15 days post inoculation in a plant inoculated with 5x109 CFU and a mean disease index was used to monitor disease development over time. An inoculum of 5x105 CFU per pot (1.3x102 CFU/g soil) was the minimum dose required to cause wilting symptoms, while one latent infection was detected at the lowest dose of 5x102 CFU per pot (0.13 CFU/g). In a second set of experiments, stem-inoculated potato plants grown in vitro were used to investigate the dose-response relationship under optimal conditions for pathogen growth and disease development. Plants were inoculated with doses between 0.5 and 5x105 CFU/plant which resulted in visible symptoms at all doses. The results led to a dose-response model describing the relationship between R. solanacearum exposure and probability of infection or illness of potato plants. Cultivar Kondor was more susceptible to brown-rot infections than HB in greenhouse experiments while there was no significant difference between the dose-response models of both cultivars in in vitro experiments. The ED50 for infection of cv Kondor was 1.1x107 CFU. Results can be used in management strategies aimed to reduce or eliminate the risk of bacterial wilt infection when using treated water in irrigation.

Die-off of plant pathogenic bacteria in tile drainage and anoxic water from a managed aquifer recharge site.

Managed aquifer recharge (MAR) can provide irrigation water and overcome water scarcity in agriculture. Removal of potentially present plant pathogens during MAR is essential to prevent crop diseases. We studied the die-off of three plant pathogenic bacteria in water microcosms with natural or filtered tile drainage water (TDW) at 10 and 25°C and with natural anoxic aquifer water (AW) at 10°C from a MAR site. These bacteria were: Ralstonia solanacearum (bacterial wilt), and the soft rot Pectobacteriaceae (SRP) Dickeya solani and Pectobacterium carotovorum sp. carotovorum (soft rot, blackleg). They are present in surface waters and cause destructive crop diseases worldwide which have been linked to contaminated irrigation water. Nevertheless, little is known about the survival of the SRP in aqueous environments and no study has investigated the persistence of R. solanacearum under natural anoxic conditions. We found that all bacteria were undetectable in 0.1 mL samples within 19 days under oxic conditions in natural TDW at 10°C, using viable cell counting, corresponding to 3-log10 reduction by die-off. The SRP were no longer detected within 6 days at 25°C, whereas R. solanacearum was detectable for 25 days. Whereas in anoxic natural aquifer water at 10°C, the bacterial concentrations declined slower and the detection limit was reached within 56 days. Finally, we modelled the inactivation curves with a modified Weibull model that can simulate different curve shapes such as shoulder phenomena in the beginning and long tails reflecting persistent bacterial populations. The non-linear model was shown to be a reliable tool to predict the die-off of the analysed plant pathogenic bacteria, suggesting its further application to other pathogenic microorganisms in the context of microbial risk assessment.

Evaluation of groundwater bacterial community composition to inform waterborne pathogen vulnerability assessments.

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.

Evaluation of water quality guidelines for public swimming ponds.

Swimming ponds are artificial ecosystems for bathing in which people imitate the conditions of natural waters. Swimming in natural water may pose health risks if the water quality is microbiologically poor. Swimming ponds are small water bodies that may be used by relatively large groups of people, moreover, the water is not disinfected, e.g. by using chlorine. The draft new swimming pool legislation in the Netherlands includes water quality requirements for swimming ponds. This study focused on the examination and evaluation of the new microbiological water quality requirements, including Escherichia coli, intestinal enterococci, Pseudomonas aeruginosa and Staphylococcus aureus, in thirteen public swimming pools. In eight of thirteen swimming ponds the water quality met the requirements for fecal indicators; 93-95% of the samples met the requirement for E. coli (≤100/100 ml) and intestinal enterococci (≤50/100 ml). The requirement for P. aeruginosa (≤10/100 ml) was met in eleven of thirteen swimming ponds (99% of the samples). In 68% of the samples the requirement for S. aureus (<1/100 ml) was met. A linear mixed effect analysis showed that E. coli and intestinal enterococci concentrations were significantly dependent on the log10 of turbidity. P. aeruginosa concentrations were significantly dependent on water temperature. 31-45% of the variation between swimming ponds was explained by considering 'pond' as a random effect in the analysis. The monitoring of microbiological parameters in swimming pond water needs selective analytical methods, such as those used in this study, due to large numbers of background bacteria. The draft new Dutch swimming pool legislation provides proper guidance to ensure the microbiological safety of swimming pond water; it would benefit from inclusion of turbidity as an extra parameter. S. aureus is a relevant parameter for non-fecal shedding, although scientific literature does not provide evidence for a norm value based on a dose-response relation for exposure to S. aureus in water.

Transport of bacteriophage MS2 and PRD1 in saturated dune sand under suboxic conditions.

Soil passage of (pretreated) surface water to remove pathogenic microorganisms is a highly efficient process under oxic conditions, reducing microorganism concentrations about 8 log10 within tens of meters. However, under anoxic conditions, it has been shown that removal of microorganisms can be limited very much. Setback distances for adequate protection of natural groundwater may, therefore, be too short if anoxic conditions apply. Because removal of microorganisms under suboxic conditions is unknown, this research investigated removal of bacteriophage MS2 and PRD1 by soil passage under suboxic conditions at field scale. At the field location (dune area), one injection well and six monitoring wells were installed at different depths along three suboxic flow lines, where oxygen concentrations ranged from 0.4 to 1.7 mg/l and nitrate concentrations ranged from 13 to 16 mg/L. PRD1 and MS2 were injected directly at the corresponding depths and their removal in each flow line was determined. The highest bacteriophage removal was observed in the top layer, with about 9 log removal of MS2, and 7 log removal of PRD1 after 16 meters of aquifer transport. Less removal was observed at 12 m below surface, probably due to a higher groundwater velocity in this coarser grained layer. MS2 was removed more effectively than PRD1 under all conditions. Due to short travel times, inactivation of the phages was limited and the reported log removal was mainly associated with attachment of phages to the aquifer matrix. This study shows that attachment of MS2 and PRD1 is similar for oxic and suboxic sandy aquifers, and, therefore, setback distances used for sandy aquifers under oxic and suboxic conditions provide a similar level of safety. Sticking efficiency and the attachment rate coefficient, as measures for virus attachment, were evaluated as a function of the physico-chemical conditions.

Climate change impact on infection risks during bathing downstream of sewage emissions from CSOs or WWTPs.

Climate change is expected to influence infection risks while bathing downstream of sewage emissions from combined sewage overflows (CSOs) or waste water treatment plants (WWTPs) due to changes in pathogen influx, rising temperatures and changing flow rates of the receiving waters. In this study, climate change impacts on the surface water concentrations of Campylobacter, Cryptosporidium and norovirus originating from sewage were modelled. Quantitative microbial risk assessment (QMRA) was used to assess changes in risks of infection. In general, infection risks downstream of WWTPs are higher than downstream CSOs. Even though model outputs show an increase in CSO influxes, in combination with changes in pathogen survival, dilution within the sewage system and bathing behaviour, the effects on the infection risks are limited. However, a decrease in dilution capacity of surface waters could have significant impact on the infection risks of relatively stable pathogens like Cryptosporidium and norovirus. Overall, average risks are found to be higher downstream WWTPs compared to CSOs. Especially with regard to decreased flow rates, adaptation measures on treatment at WWTPs may be more beneficial for human health than decreasing CSO events.

Long-term inactivation of bacteriophage PRD1 as a function of temperature, pH, sodium and calcium concentration.

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.

Fate of Extended-Spectrum ß-Lactamase-Producing Escherichia coli from Faecal Sources in Surface Water and Probability of Human Exposure through Swimming.

The goal of this study was to determine the fate of ESBL-producing Escherichia coli (ESBL-EC) emitted from faecal sources in surface water, and the probability of human exposure through swimming. Concentrations of ESBL-EC were measured in recreational waters and in source waters, being water in ditches surrounding poultry farms and municipal wastewater. Additionally, the potential of ESBL-EC in source waters to reach recreational waters located downstream of these sources was modeled. Modeled ESBL-EC concentrations in recreational waters appeared to be mostly determined by the concentrations in the source waters and by subsequent dilution in surface water, and less by inactivation. The mean (95%) risk of human exposure to ESBL-EC per person per swimming event, as assessed from measured ESBL-EC concentrations in recreational waters, was 0.16 (0.89) for men, 0.13 (0.72) for women and 0.20 (0.95) for children. Similar exposure risks were estimated for hypothetical recreational waters containing 100- or 1000-times diluted source water, located 10 days water travel time downstream of the sources. Human exposure to ESBL-EC through swimming is likely, if recreational waters are located downstream of poultry farms and municipal wastewater discharge points.

Effect of Climate Change on the Concentration and Associated Risks of Vibrio Spp. in Dutch Recreational Waters.

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.

Bacteriophage PRD1 batch experiments to study attachment, detachment and inactivation processes.

Knowledge of virus removal in subsurface environments is pivotal for assessing the risk of viral contamination of water resources and developing appropriate protection measures. Columns packed with sand are frequently used to quantify attachment, detachment and inactivation rates of viruses. Since column transport experiments are very laborious, a common alternative is to perform batch experiments where usually one or two measurements are done assuming equilibrium is reached. It is also possible to perform kinetic batch experiments. In that case, however, it is necessary to monitor changes in the concentration with time. This means that kinetic batch experiments will be almost as laborious as column experiments. Moreover, attachment and detachment rate coefficients derived from batch experiments may differ from those determined using column experiments. The aim of this study was to determine the utility of kinetic batch experiments and investigate the effects of different designs of the batch experiments on estimated attachment, detachment and inactivation rate coefficients. The experiments involved various combinations of container size, sand-water ratio, and mixing method (i.e., rolling or tumbling by pivoting the tubes around their horizontal or vertical axes, respectively). Batch experiments were conducted with clean quartz sand, water at pH 7 and ionic strength of 20 mM, and using the bacteriophage PRD1 as a model virus. Values of attachment, detachment and inactivation rate coefficients were found by fitting an analytical solution of the kinetic model equations to the data. Attachment rate coefficients were found to be systematically higher under tumbling than under rolling conditions because of better mixing and more efficient contact of phages with the surfaces of the sand grains. In both mixing methods, more sand in the container yielded higher attachment rate coefficients. A linear increase in the detachment rate coefficient was observed with increased solid-water ratio using tumbling method. Given the differences in the attachment rate coefficients, and assuming the same sticking efficiencies since chemical conditions of the batch and column experiments were the same, our results show that collision efficiencies of batch experiments are not the same as those of column experiments. Upscaling of the attachment rate from batch to column experiments hence requires proper understanding of the mixing conditions. Because batch experiments, in which the kinetics are monitored, are as laborious as column experiments, there seems to be no major advantage in performing batch instead of column experiments.

A mathematical model for removal of human pathogenic viruses and bacteria by slow sand filtration under variable operational conditions.

Slow sand filtration (SSF) in drinking water production removes pathogenic microorganisms, but detection limits and variable operational conditions complicate assessment of removal efficiency. Therefore, a model was developed to predict removal of human pathogenic viruses and bacteria as a function of the operational conditions. Pilot plant experiments were conducted, in which bacteriophage MS2 and Escherichia coli WR1 were seeded as model microorganisms for pathogenic viruses and bacteria onto the filters under various temperatures, flow rates, grain sizes and ages of the Schmutzdecke. Removal of MS2 was 0.082-3.3 log10 and that of E. coli WR1 0.94-4.5 log10 by attachment to the sand grains and additionally by processes in the Schmutzdecke. The contribution of the Schmutzdecke to the removal of MS2 and E. coli WR1 increased with its ageing, with sticking efficiency and temperature, decreased with grain size, and was modelled as a logistic growth function with scale factor f0 and rate coefficient f1. Sticking efficiencies were found to be microorganism and filter specific, but the values of f0 and f1 were independent of microorganism and filter. Cross-validation showed that the model can be used to predict log removal of MS2 and ECWR1 within ±0.6 log. Within the range of operational conditions, the model shows that removal of microorganisms is most sensitive to changes in temperature and age of the Schmutzdecke.

Effect of dissolved calcium on the removal of bacteriophage PRD1 during soil passage: the role of double-layer interactions.

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 10mM, 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 risk estimation for a Legionella pneumophila infection due to whirlpool use.

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 <100 cfu Legionella/L water. This study suggests that stricter provisions might be required to assure adequate public health protection.

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.

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.