Modeling the Transport of Human Rotavirus and Norovirus in Standardized and in Natural Soil Matrix-Water Systems.

We modeled Group A Rotavirus (RVA) and Norovirus genogroup II (GII NoV) transport experiments in standardized (crystal quartz sand and deionized water with adjusted pH and ionic strength) and natural soil matrix-water systems (MWS). On the one hand, in the standardized MWS, Rotavirus and Norovirus showed very similar breakthrough curves (BTCs), showing a removal rate of 2 and 1.7 log10, respectively. From the numerical modeling of the experiment, transport parameters of the same order of magnitude were obtained for both viruses. On the other hand, in the natural MWS, the two viruses show very different BTCs. The Norovirus transport model showed significant changes; BTC showed a removal rate of 4 log10, while Rotavirus showed a removal rate of 2.6 log10 similar to the 2 log10 observed on the standardized MWS. One possible explanation for this differential behavior is the difference in the isoelectric point value of these two viruses and the increase of the ionic strength on the natural MWS.

Evaluation of Bacterial Contamination as an Indicator of Viral Contamination in a Sedimentary Aquifer in Uruguay.

In Uruguay, groundwater is frequently used for agricultural activities, as well as for human consumption in urban and rural areas. As in many countries worldwide, drinking water microbiological quality is evaluated only according to bacteriological standards and virological analyses are not mentioned in the legislation. In this work, the incidence of human viral (Rotavirus A, Norovirus GII, and human Adenovirus) and bacterial (total and thermotolerant coliform and Pseudomonas aeruginosa) contamination in groundwater in the Salto district, Uruguay, as well as the possible correlation between these groups of microorganisms, was studied. From a total of 134 groundwater samples, 42 (32.1%) were positive for Rotavirus, only 1 (0.7%) for both Rotavirus and Adenovirus, and 96 (72.6%) samples were positive for bacterial indicators. Results also show that Rotavirus presence was not associated with changes in chemical composition of the aquifer water. Bacteriological indicators were not adequate to predict the presence of viruses in individual groundwater samples (well scale), but a deeper spatial-temporal analysis showed that they are promising candidates to assess the viral contamination degree at aquifer scale, since from the number of wells with bacterial contamination the number of wells with viral contamination could be estimated.

Entero- and parechovirus distributions in surface water and probabilities of exposure to these viruses during water recreation.

Numerous studies have reported quantitative data on viruses in surface waters generated using different methodologies. In the current study, the impact of the use of either cell culture-based or molecular-based methods in quantitative microbial risk assessment was assessed. Previously and newly generated data on the presence of infectious human enteroviruses (HEV) and enterovirus and parechovirus RNA were used to estimate distributions of virus concentrations in surface waters. Because techniques for the detection of infectious human parechoviruses (HPeV) in surface waters were not available, a 'Parallelogram Approach' was used to estimate their concentrations based on the ratio infectious HEV/HEV RNA. The obtained virus concentrations were then used to estimate the probability of exposure for children during recreation in such virus contaminated surface waters. Human enterovirus cell culture/PCR ratios ranged from 2.3 × 10(-3) to 0.28. This broad range of ratios indicates that care should be taken in assuming a fixed ratio for assessing the risk with PCR based virus concentrations. The probabilities of exposure to both enteroviruses and parechoviruses were calculated, using our Parallelogram Approach for the calculation of infectious parechoviruses. For both viruses it was observed that the detection method significantly influenced the probability of exposure. Based on the calculated culture data, PCR data, and the ingestion volume, it was estimated that the mean probabilities of exposure, of recreating children, to surface water containing viruses were 0.087 (infectious enteroviruses), 0.71 (enterovirus particles), 0.28 (parechovirus particles) and 0.025 (calculated infectious parechoviruses) per recreation event. The mean probabilities of exposure of children recreating in surface water from which drinking water is produced to infectious enteroviruses were estimated for nine locations and varied between 1.5 × 10(-4) - 0.09 per recreation event. In this study, the use of the rotavirus dose response relationship as a surrogate was avoided. Instead, the probabilities of exposure were derived as a function of the distributions of the calculated doses. Our 'Parallelogram Approach' was used to estimate the unavailable infectious parechovirus concentrations using Monte Carlo simulations, and the exposure assessment carried out showed that virus concentrations present in surface waters could pose a health risk for children and other vulnerable populations.

Quantitative assessment of infection risk from exposure to waterborne pathogens in urban floodwater.

Flooding and heavy rainfall have been associated with waterborne infectious disease outbreaks, however, it is unclear to which extent they pose a risk for public health. Here, risks of infection from exposure to urban floodwater were assessed using quantitative microbial risk assessment (QMRA). To that aim, urban floodwaters were sampled in the Netherlands during 23 events in 2011 and 2012. The water contained Campylobacter jejuni (prevalence 61%, range 14- >10(3) MPN/l), Giardia spp. (35%, 0.1-142 cysts/l), Cryptosporidium (30%, 0.1-9.8 oocysts/l), noroviruses (29%, 10(2)-10(4) pdu/l) and enteroviruses (35%, 10(3)-10(4) pdu/l). Exposure data collected by questionnaire, revealed that children swallowed 1.7 ml (mean, 95% Confidence Interval 0-4.6 ml) per exposure event and adults swallowed 0.016 ml (mean, 95% CI 0-0.068 ml) due to hand-mouth contact. The mean risk of infection per event for children, who were exposed to floodwater originating from combined sewers, storm sewers and rainfall generated surface runoff was 33%, 23% and 3.5%, respectively, and for adults it was 3.9%, 0.58% and 0.039%. The annual risk of infection was calculated to compare flooding from different urban drainage systems. An exposure frequency of once every 10 years to flooding originating from combined sewers resulted in an annual risk of infection of 8%, which was equal to the risk of infection of flooding originating from rainfall generated surface runoff 2.3 times per year. However, these annual infection risks will increase with a higher frequency of urban flooding due to heavy rainfall as foreseen in climate change projections.

Reduction of bacteriophage MS2 by filtration and irradiation determined by culture and quantitative real-time RT-PCR.

Molecular methods are increasingly applied for virus detection in environmental samples without rendering data on viral infectivity. Infectivity data are important for assessing public health risks from exposure to human pathogenic viruses in the environment. Here, treatment efficiencies of three (drinking) water treatment processes were estimated by quantification of the indicator virus bacteriophage MS2 with culture and real-time reverse transcription polymerase chain reaction (qRT-PCR). We studied the virus reduction by slow sand filtration at a pilot plant. No decay of MS2 RNA was observed, whereas infectious MS2 particles were inactivated at a rate of 0.1 day(-1). Removal of MS2 RNA and infectious MS2 particles was 1.2 and 1.6 log10-units, respectively. Virus reduction by UV and gamma irradiation was determined in laboratory-scale experiments. The reduction of MS2 RNA based on qRT-PCR data was negligible. Reduction of infectious MS2 particles was estimated at 3.0-3.6 log10-units (UV dose up to 400 or 800 J/m(2)) and 4.7-7 log10-units (gamma dose up to 200 Gray). As shown in this study, estimations of viral reduction, both inactivation and removal, obtained by molecular methods should be interpreted carefully when considering treatment options to provide virus-safe drinking water. Combining culture-based methods with molecular methods may provide supplementary information on mechanisms of virus reduction.

Effects of fluctuations in river water level on virus removal by bank filtration and aquifer passage--a scenario analysis.

Riverbank filtration is an effective process for removing pathogenic viruses from river water. Despite indications that changing hydraulic conditions during floods can affect the efficacy of riverbank filtration to remove viruses, the impact on advection and dispersion of viruses in the riverbank is not well understood. We investigated the effects of fluctuations in river water level on virus transport during riverbank filtration, considering 3-D transient groundwater flow and virus transport. Using constant removal rates from published field experiments with bacteriophages, removal of viruses with distance from the riverbank was simulated for coarse gravel, fine gravel and fine sandy gravel. Our simulations showed that, in comparison with steady flow conditions, fluctuations in river water level cause viruses to be transported further at higher concentrations into the riverbank. A 1-5 m increase in river water levels led to a 2- to 4-log (log10 reduction in concentration relative to the initial concentration in the river) increase in virus concentration and to up to 30% shorter travel times. For particular cases during the receding flood, changing groundwater flow conditions caused that pristine groundwater was carried from further inland and that simulated virus concentrations were more diluted in groundwater. Our study suggests that the adverse effect of water level fluctuations on virus transport should be considered in the simulation of safe setback distances for drinking water supplies.

Definition and quantification of initial anthropogenic pollutant release in swimming pools.

Pollutants, brought into a swimming pool by bathers, will react with chlorine to form disinfection by-products (DBPs). Some of these DBPs are found to be respiratory and ocular irritant and might be associated with asthma, or might even be carcinogenic. As DBPs in swimming pools are formed from bather-shed-pollutants, a reduction of these pollutants will lead to a reduction of DBPs. Until now, however, the release of pollutants by bathers has not been studied in detail. The study described in this paper focuses on the release of these pollutants, further called anthropogenic pollutants. The objective was to define and quantify the initial anthropogenic pollutants, by using a standardised shower cabin and a standardised showering protocol in laboratory time-series experiments and on-site experiments in swimming pools. The time-series experiments resulted in a definition of the initial anthropogenic pollutant release: the amount of pollutants released from a person in a standardised shower cabin during the first 60 s of showering. The data from the time-series experiments were used to create a model of pollutant release. The model can be used to predict the initial anthropogenic pollutant release as well as the effects of showering. On-site experiments were performed at four different swimming pools, including one outdoor pool. Results of these on-site showering experiments correspond with the time-series and model outcomes. Anthropogenic pollutant release (both chemical and microbiological) in swimming pool water can be reduced by pre-swim showering, very likely resulting in decreased DBPs formation and chlorine demand.

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

Temperature is considered as the major factor determining virus inactivation in the environment. Food industries, therefore, widely apply temperature as virus inactivating parameter. This review encompasses an overview of viral inactivation and virus genome degradation data from published literature as well as a statistical analysis and the development of empirical formulae to predict virus inactivation. A total of 658 data (time to obtain a first log(10) reduction) were collected from 76 published studies with 563 data on virus infectivity and 95 data on genome degradation. Linear model fitting was applied to analyse the effects of temperature, virus species, detection method (cell culture or molecular methods), matrix (simple or complex) and temperature category (<50 and ≥50°C). As expected, virus inactivation was found to be faster at temperatures ≥50°C than at temperatures <50°C, but there was also a significant temperature-matrix effect. Virus inactivation appeared to occur faster in complex than in simple matrices. In general, bacteriophages PRD1 and PhiX174 appeared to be highly persistent whatever the matrix or the temperature, which makes them useful indicators for virus inactivation studies. The virus genome was shown to be more resistant than infectious virus. Simple empirical formulas were developed that can be used to predict virus inactivation and genome degradation for untested temperatures, time points or even virus strains.

Vulnerability of unconfined aquifers to virus contamination.

An empirical formula was developed for determining the vulnerability of unconfined sandy aquifers to virus contamination, expressed as a dimensionless setback distance r(s)(*). The formula can be used to calculate the setback distance required for the protection of drinking water production wells against virus contamination. This empirical formula takes into account the intrinsic properties of the virus and the unconfined sandy aquifer. Virus removal is described by a rate coefficient that accounts for virus inactivation and attachment to sand grains. The formula also includes pumping rate, saturated thickness of the aquifer, depth of the screen of the pumping well, and anisotropy of the aquifer. This means that it accounts also for dilution effects as well as horizontal and vertical virus transport. Because the empirical model includes virus source concentration it can be used as an integral part of a quantitative viral risk assessment.

Long-term inactivation study of three enteroviruses in artificial surface and groundwaters, using PCR and cell culture.

Since the transmission of pathogenic viruses via water is indistinguishable from the transmission via other routes and since the levels in drinking water, although significant for health, may be too low for detection, quantitative viral risk assessment is a useful tool for assessing disease risk due to consumption of drinking water. Quantitative viral risk assessment requires information concerning the ability of viruses detected in drinking water to infect their host. To obtain insight into the infectivity of viruses in relation to the presence of virus genomes, inactivation of three different enteroviruses in artificial ground and surface waters under different controlled pH, temperature, and salt conditions was studied by using both PCR and cell culture over time. In salt-peptone medium, the estimated ratio of RNA genomes to infectious poliovirus 1 in freshly prepared suspensions was about 10(0). At 4 degrees C this ratio was 10(3) after 600 days, and at 22 degrees C it was 10(4) after 200 days. For poliovirus 1 and 2 the RNA/infectious virus ratio was higher in artificial groundwater than in artificial surface water, but this was not the case for coxsackievirus B4. When molecular detection is used for virus enumeration, it is important that the fraction of infectious virus (based on all virus genomes detected) decays with time, especially at temperatures near 22 degrees C.

Monitoring of waterborne pathogens in surface waters in amsterdam, the Netherlands, and the potential health risk associated with exposure to cryptosporidium and giardia in these waters.

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.

Transport of Escherichia coli and solutes during waste water infiltration in an urban alluvial aquifer.

Recharge of waste water in an unconsolidated poorly sorted alluvial aquifer is a complex process, both physically and hydrochemically. The aim of this paper is to analyse and conceptualise vertical transport mechanisms taking place in an urban area of extensive wastewater infiltration by analysing and combining the water balance, the microbial (Escherichia coli) mass balance, and the mass balance for dissolved solutes. For this, data on sediment characteristics (grain size, organic carbon, reactive iron, and calcite), groundwater levels, and concentrations of E. coli in groundwater and waste water were collected. In the laboratory, data on E. coli decay rate coefficients, and on bacteria retention characteristics of the sediment were collected via column experiments. The results indicated that shallow groundwater, at depths of 50 m below the surface, was contaminated with E. coli concentrations as high as 10(6) CFU/100 mL. In general, E. coli concentrations decreased only 3 log units from the point of infiltration to shallow groundwater. Concentrations were lower at greater depths in the aquifer. In laboratory columns of disturbed sediments, bacteria removal was 2-5 log units/0.5 cm column sediment. Because of the relatively high E. coli concentrations in the shallow aquifer, transport had likely taken place via a connected network of pores with a diameter large enough to allow bacterial transport instead of via the sediment matrix, which was inaccessible for bacteria, as was clear from the column experiments. The decay rate coefficient was determined from laboratory microcosms to be 0.15 d(-1). Assuming that decay in the aquifer was similar to decay in the laboratory, then the pore water flow velocity between the point of infiltration and shallow groundwater, coinciding with a concentration decrease of 3 log units, was 0.38 m/d, and therefore, transport in this connected network of pores was fast. According to the water balance of the alluvial aquifer, determined from transient groundwater modelling, groundwater flow in the aquifer was mainly in vertical downward direction, and therefore, the mass balance for dissolved solutes was simulated using a 1D transport model of a 200 m column of the Quaternary Alluvium aquifer. The model, constructed with PHREEQC, included dual porosity, and was able to adequately simulate removal of E. coli, cation-exchange, and nitrification. The added value of the use of E. coli in this study was the recognition of relatively fast transport velocities occurring in the aquifer, and the necessity to use the dual porosity concept to investigate vertical transport mechanisms. Therefore, in general and if possible, microbial mass balances should be considered more systematically as an integral part of transport studies.

Determination of protection zones for Dutch groundwater wells against virus contamination--uncertainty and sensitivity analysis.

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 x 10(8) 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.

Effect of goethite coating and humic acid on the transport of bacteriophage PRD1 in columns of saturated sand.

The transport of bacteriophage PRD1, a model virus, was studied in columns containing sediment mixtures of quartz sand with goethite-coated sand and using various solutions consisting of monovalent and divalent salts and humic acid (HA). Without HA and in the absence of sand, the inactivation rate of PRD1 was found to be as low as 0.014 day(-1) (at 5+/-3 degrees C), but in the presence of HA it was much lower (0.0009 day(-1)), indicating that HA helps PRD1 to survive. When the fraction of goethite in the sediment was increased, the removal of PRD1 also increased. However, in the presence of HA, C/C0 values of PRD1 increased by as much as 5 log units, thereby almost completely eliminating the effect of addition of goethite. The sticking efficiency was not linearly dependent on the amount of goethite added to the quartz sand; this is apparently due to surface charge heterogeneity of PRD1. Our results imply that, in the presence of dissolved organic matter (DOM), viruses can be transported for long distances thanks to two effects: attachment is poor because DOM has occupied favourable sites for attachment and inactivation of virus may have decreased. This conclusion justifies making conservative assumptions about the attachment of viruses when calculating protection zones for groundwater wells.

Evaluation of data from the literature on the transport and survival of Escherichia coli and thermotolerant coliforms in aquifers under saturated conditions.

Escherichia coli and thermotolerant coliforms are of major importance as indicators of fecal contamination of water. Due to its negative surface charge and relatively low die-off or inactivation rate coefficient, E. coli is able to travel long distances underground and is therefore also a useful indicator of fecal contamination of groundwater. In this review, the major processes known to determine the underground transport of E. coli (attachment, straining and inactivation) are evaluated. The single collector contact efficiency (SCCE), eta0, one of two parameters commonly used to assess the importance of attachment, can be quantified for E. coli using classical colloid filtration theory. The sticking efficiency, alpha, the second parameter frequently used in determining attachment, varies widely (from 0.003 to almost 1) and mainly depends on charge differences between the surface of the collector and E. coli. Straining can be quantified from geometrical considerations; it is proposed to employ a so-called straining correction parameter, alpha(str). Sticking efficiencies determined from field experiments were lower than those determined under laboratory conditions. We hypothesize that this is due to preferential flow mechanisms, E. coli population heterogeneity, and/or the presence of organic and inorganic compounds in wastewater possibly affecting bacterial attachment characteristics. Of equal importance is the inactivation or die-off of E. coli that is affected by factors like type of bacterial strain, temperature, predation, antagonism, light, soil type, pH, toxic substances, and dissolved oxygen. Modeling transport of E. coli can be separated into three steps: (1) attachment rate coefficients and straining rate coefficients can be calculated from Darcy flow velocity fields or pore water flow velocity fields, calculated SCCE fields, realistic sticking efficiency values and straining correction parameters, (2) together with the inactivation rate coefficient, total rate coefficient fields can be generated, and (3) used as input for modeling the transport of E. coli in existing contaminant transport codes. Areas of future research are manifold and include the effects of typical wastewater characteristics, including high concentrations of organic compounds, on the transport of E. coli and thermotolerant coliforms, and the upscaling of experiments to represent typical field conditions, possibly including preferential flow mechanisms and the aspect of population heterogeneity of E. coli.

Transport of E. coli in columns of geochemically heterogeneous sediment.

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 0.18-0.50mm quartz sand, grains coated with goethite, calcite grains or grains of activated carbon (AC), in varying fractions (lambda=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 (alpha(total)) showed that upon increasing the fraction of favourable mineral grains (lambda) 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.

Effect of climate changes on waterborne disease in The Netherlands.

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.

Determining straining of Escherichia coli from breakthrough curves.

Though coliform bacteria are used world wide as an indication of faecal pollution, the parameters determining the transport of Escherichia coli in aquifers are relatively unknown, especially for the period after the clean bed collision phase brought about by prolonged infiltration of waste water. In this research, the breakthrough curves of E. coli after total flushing of 50-200 pore volumes were studied for various influent concentrations in various sediments at different pore water flow velocities. The results indicated that straining in Dead End Pores (DEPs) was an important process that dominated bacteria breakthrough in fine-grained sediment (0.06-0.2 mm). The filling of the DEP space with bacteria took 5-65 pore volumes and was dependent on concentration. Column breakthrough curves were modelled and from this the DEP volumes were determined. These volumes (0.21-0.35% of total column volume) corresponded well with values calculated with a formula based on purely geometrical considerations and also with values calculated with a pore size density function. For this function the so-called Van Genuchten parameters of the sediments used in the experiments were determined. The results indicate that straining might be a dominant process affecting colloid transport in the natural environment and therefore it is concluded that proper knowledge of the pore size distribution is crucial to an understanding of the retention of bacteria.

Elimination of viruses, bacteria and protozoan oocysts by slow sand filtration.

The decimal elimination capacity (DEC) of slow sand filters (SSF) for viruses, bacteria and oocysts of Cryptosporidium has been assessed from full-scale data and pilot plant and laboratory experiments. DEC for viruses calculated from experimental data with MS2-bacteriophages in the pilot plant filters was 1.5-2 log10. E. coli and thermotolerant coliforms (Coli44) were removed at full-scale and in the pilot plant with 2-3 log10. At full-scale, Campylobacter bacteria removal was 1 log10 more than removal of Coli44, which indicated that Coli44 was a conservative surrogate for these pathogenic bacteria. Laboratory experiments with sand columns showed 2-3 and >5-6 log10 removal of spiked spores of sulphite-reducing clostridia (SSRC; C. perfringens) and oocysts of Cryptosporidium respectively. Consequently, SSRC was not a good surrogate to quantify oocyst removal by SSF. Removal of indigenous SSRC by full-scale filters was less efficient than observed in the laboratory columns, probably due to continuous loading of these filter beds with spores, accumulation and retarded transport. It remains to be investigated if this also applies to oocyst removal by SSF. The results additionally showed that the schmutzdecke and accumulation of (in)organic charged compounds in the sand increased the elimination of microorganisms. Removal of the schmutzdecke reduced DEC for bacteria by +/-2 log10, but did not affect removal of phages. This clearly indicated that, besides biological activity, both straining and adsorption were important removal mechanisms in the filter bed for microorganisms larger than viruses.

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

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