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Human fecal pollution of recreational waters remains a public health concern worldwide. As a result, there is a growing interest in the application of human-associated fecal source identification quantitative real-time PCR (qPCR) technologies for water quality research and management. However, there are currently no standardized approaches for field implementation and interpretation of qPCR data. In this study, a standardized HF183/BacR287 qPCR method was combined with a water sampling strategy and a novel Bayesian weighted average approach to establish a human fecal contamination score (HFS) that can be used to prioritize sampling sites for remediation based on measured human waste levels. The HFS was then used to investigate 975 study design scenarios utilizing different combinations of sites with varying sampling intensities (daily to once per week) and number of qPCR replicates per sample (2-14 replicates). Findings demonstrate that site prioritization with HFS is feasible and that both sampling intensity and number of qPCR replicates influence reliability of HFS estimates. The novel data analysis strategy presented here provides a prescribed approach for the implementation and interpretation of human-associated HF183/BacR287 qPCR data with the goal of site prioritization based on human fecal pollution levels. In addition, information is provided for future users to customize study designs for optimal HFS performance., (Published by Elsevier Ltd.)

Microbial source tracking (MST) methods were evaluated in the Source Identification Protocol Project (SIPP), in which 27 laboratories compared methods to identify host sources of fecal pollution from blinded water samples containing either one or two different fecal types collected from California. This paper details lessons learned from the SIPP study and makes recommendations to further advance the field of MST. Overall, results from the SIPP study demonstrated that methods are available that can correctly identify whether particular host sources including humans, cows and birds have contributed to contamination in a body of water. However, differences between laboratory protocols and data processing affected results and complicated interpretation of MST method performance in some cases. This was an issue particularly for samples that tested positive (non-zero Ct values) but below the limits of quantification or detection of a PCR assay. Although false positives were observed, such samples in the SIPP study often contained the fecal pollution source that was being targeted, i.e., the samples were true positives. Given these results, and the fact that MST often requires detection of targets present in low concentrations, we propose that such samples be reported and identified in a unique category to facilitate data analysis and method comparisons. Important data can be lost when such samples are simply reported as positive or negative. Actionable thresholds were not derived in the SIPP study due to limitations that included geographic scope, age of samples, and difficulties interpreting low concentrations of target in environmental samples. Nevertheless, the results of the study support the use of MST for water management, especially to prioritize impaired waters in need of remediation. Future integration of MST data into quantitative microbial risk assessments and other models could allow managers to more efficiently protect public health based on site conditions., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The last decade has seen development of numerous new microbial source tracking (MST) methodologies, but many of these have been tested in just a few laboratories with a limited number of fecal samples. This method evaluation study examined the specificity and sensitivity of 41 MST methodologies by analyzing data generated in 27 laboratories. MST methodologies that targeted human, cow, ruminant, dog, gull, pig, horse, and sheep were tested against sewage, septage, human, cow, dog, deer, pig, chicken, pigeon, gull, horse, and goose fecal samples. Each laboratory received 64 blind samples containing a single source (singletons) or two sources (doubletons), as well as diluted singleton samples to assess method sensitivity. Laboratories utilized their own protocols when performing the methods and data were deposited in a central database before samples were unblinded. Between one and seven laboratories tested each method. The most sensitive and specific assays, based on an analysis of presence/absence of each marker in target and non-target fecal samples, were HF183 endpoint and HF183SYBR (human), CF193 and Rum2Bac (ruminant), CowM2 and CowM3 (cow), BacCan (dog), Gull2SYBR and LeeSeaGull (gull), PF163 and pigmtDNA (pig), HoF597 (horse), PhyloChip (pig, horse, chicken, deer), Universal 16S TRFLP (deer), and Bacteroidales 16S TRFLP (pig, horse, chicken, deer); all had sensitivity and specificity higher than 80% in all or the majority of laboratories. When the abundance of MST markers in target and non-target fecal samples was examined, some assays that performed well in the binary analysis were found to not be sensitive enough as median concentrations fell below a minimum abundance criterion (set at 50 copies per colony forming units of enterococci) in target fecal samples. Similarly, some assays that cross-reacted with non-target fecal sources in the binary analysis were found to perform well in a quantitative analysis because the cross-reaction occurred at very low levels. Based on a quantitative analysis, the best performing methods were HF183Taqman and BacH (human), Rum2Bac and BacR (ruminant), LeeSeaGull (gull), and Pig2Bac (pig); no cow or dog-specific assay met the quantitative specificity and sensitivity criteria. Some of the best performing assays in the study were run by just one laboratory so further testing of assay portability is needed. While this study evaluated the marker performance in defined samples, further field testing as well as development of frameworks for fecal source allocation and risk assessment are needed., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1-3.3% and 1.9-7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1-2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

In many parts of the world, microbial contamination of surface waters used for drinking, recreation, and shellfishery remains a pervasive risk to human health, especially in Less Economically Developed Countries (LEDC). However, the capacity to provide effective management strategies to break the waterborne route to human infection is often thwarted by our inability to identify the source of microbial contamination. Microbial Source Tracking (MST) has potential to improve water quality management in complex river catchments that are either routinely, or intermittently contaminated by faecal material from one or more sources, by attributing faecal loads to their human or non-human sources, and thereby supporting more rational approaches to microbial risk assessment. The River Ouse catchment in southeast England (U.K.) was used as a model with which to investigate the integration and application of a novel and simple MST approach to monitor microbial water quality over one calendar year, thereby encompassing a range of meteorological conditions. A key objective of the work was to develop simple low-cost protocols that could be easily replicated. Bacteriophages (viruses) capable of infecting a human specific strain of Bacteroides GB-124, and their correlation with presumptive Escherichia coli, were used to distinguish sources of faecal pollution. The results reported here suggest that in this river catchment the principal source of faecal pollution in most instances was non-human in origin. During storm events, presumptive E. coli and presumptive intestinal enterococci levels were 1.1-1.2 logs higher than during dry weather conditions, and levels of the faecal indicator organisms (FIOs) were closely associated with increased turbidity levels (presumptive E. coli and turbidity, r = 0.43). Spatio-temporal variation in microbial water quality parameters was accounted for by three principal components (67.6%). Cluster Analysis, reduced the fourteen monitoring sites to six representative 'sentinel' sites. The correlation coefficient between presumptive E. coli and phages of Bacteroides GB-124 was very small (r = 0.05) whilst that between turbidity and suspended solids was high (r = 0.62). Variations in climate, animal and anthropogenic interferences were all, either directly or indirectly, related to faecal contamination. The findings show the importance of meteorological conditions, such as storm events, on microbial water quality, and suggest that any future increases in the frequency of storm events (associated with climate change) are likely to result in a greater incidence of FIO/pathogen loads. This low-cost approach could help to predict spatio-temporal 'hotspots' of elevated waterborne disease risk. The work also represents an important step towards integrating novel MST tools into river catchment modelling., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Knowledge of the variability in pathogen or indicator concentrations over time at a particular location (e.g. in drinking water sources) is essential in implementation of concentration-based regulations and in quantitative microbial risk assessment. Microbial enumeration methods, however, are known to yield highly variable counts (even among replicates) and some methods are prone to substantial losses (i.e. only a fraction of the target microorganisms in a sample are observed). Consequently, estimated microorganism concentrations may be biased and only a fraction of the variability that is observed in temporally distributed concentration estimates is due to variability in concentration itself. These issues have often been ignored in the past, and approaches to integrate knowledge about the measurement error associated with enumeration methods into decisions have not been standardized. Here, an existing model that describes variability in microorganism counts as a function of sample volume and the analytical recovery of the enumeration method is expanded to include temporal concentration variability and sample-specific recovery information. This model is used to demonstrate that microorganism counts and analytical recovery are not independent (as has often been assumed), even if the correlation is obscured by other sources of variability in the data. It is also used as an experimental design tool to evaluate strategies that may yield more accurate concentration estimates. Finally, the model is implemented in a Bayesian framework (with a Gibbs sampling algorithm) to quantify temporal concentration variability with appropriate consideration of measurement errors in the data and the analytical recovery of the enumeration method. We demonstrate by simulation that this statistical approach facilitates risk analyses that appropriately model variability in microorganism concentrations given the available data and that it enables decisions based on quantitative measures of uncertainty., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Urban part of Seine River serving as drinking water supply in Paris can be heavily contaminated by Cryptosporidium spp. and Giardia duodenalis. In the absence of agricultural practice in this highly urbanized area, we investigated herein the contribution of treated wastewater to the microbiological quality of this river focusing on these two parasites. Other microorganisms such as faecal bacterial indicators, enteroviruses and oocysts of Toxoplasma gondii were assessed concurrently. Raw wastewaters were heavily contaminated by Cryptosporidium and Giardia (oo)cysts, whereas concentrations of both protozoa in treated wastewater were lower. Treated wastewater, flowed into Seine River, had a parasite concentration closed to the one found along the river, in particular at the entry of a drinking water plant (DWP). Even if faecal bacteria were reliable indicators of a reduction in parasite concentrations during the wastewater treatment, they were not correlated to protozoal contamination of wastewater and river water. Oocysts of T. gondii were not found in both raw and treated wastewater, or in Seine River. Parasitic contamination was shown to be constant in the Seine River up to 40 km upstream Paris. Altogether, these results strongly suggest that treated wastewater does not contribute to the main parasitic contamination of the Seine River usually observed in this urbanized area., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Drinking water systems are vulnerable and subject to a wide range of risks. To avoid sub-optimisation of risk-reduction options, risk analyses need to include the entire drinking water system, from source to tap. Such an integrated approach demands tools that are able to model interactions between different events. Fault tree analysis is a risk estimation tool with the ability to model interactions between events. Using fault tree analysis on an integrated level, a probabilistic risk analysis of a large drinking water system in Sweden was carried out. The primary aims of the study were: (1) to develop a method for integrated and probabilistic risk analysis of entire drinking water systems; and (2) to evaluate the applicability of Customer Minutes Lost (CML) as a measure of risk. The analysis included situations where no water is delivered to the consumer (quantity failure) and situations where water is delivered but does not comply with water quality standards (quality failure). Hard data as well as expert judgements were used to estimate probabilities of events and uncertainties in the estimates. The calculations were performed using Monte Carlo simulations. CML is shown to be a useful measure of risks associated with drinking water systems. The method presented provides information on risk levels, probabilities of failure, failure rates and downtimes of the system. This information is available for the entire system as well as its different sub-systems. Furthermore, the method enables comparison of the results with performance targets and acceptable levels of risk. The method thus facilitates integrated risk analysis and consequently helps decision-makers to minimise sub-optimisation of risk-reduction options.

While millions of people drink arsenic-contaminated tube well water across Bangladesh, there is no recent scientific explanation which is able to either comprehensively explain arsenic mobilization or to predict the spatial distribution of affected wells. Rather, mitigation strategies have focused on the sinking of deep tube wells into the currently arsenic-free Pleistocene aquifer. In this study, Bangladesh shallow tube wells identified as contaminated and uncontaminated, as well as deep tube wells, were analyzed for geochemical and in situ microbiological composition. Whereas arsenic was detected in all Holocene aquifer wells, no arsenic was found in wells accessing the Pleistocene aquifer. Bacterial genera, including Comamonadaceae, Acidovorax, Acinetobacter, and Hydrogenophaga, associated with tolerance of high arsenic concentrations, rather than dissimilatory Fe(III) or As(V) reduction, were identified in shallow tube wells, indicating that mobilization may not occur at depth, but is rather due to drawdown of surface contaminated water. Deep tube wells contained microbes indicative of aerobic conditions, including the genera Aquabacterium, Limnobacter, and Roseomonas. It is concluded that through drawdown of arsenic or organic matter, further utilization of the Pleistocene aquifer could result in contamination similar to that observed in the Holocene aquifer.

Norovirus (NoV) is a leading cause of acute gastroenteritis and is often spread via wastewater contamination. Little is known about how the wastewater treatment process affects norovirus, and which factors influence virus concentrations. To investigate this, we collected wastewater samples monthly during one year at eight different key sites at the municipal wastewater treatment plant in Gothenburg, Sweden. Virus particles were concentrated using ultracentrifugation, viral RNA was subsequently extracted, and transformed into cDNA by reverse transcription. The quantification was performed with real-time PCR assays for NoV genogroups I (GGI) and II (GGII), respectively. We found seasonal changes of NoV genogroups, with the highest concentration of NoV GGII during the winter months, and the highest concentration of NoV GGI during the summer months. Virus transmission in wastewater was more stable for NoV GGI, with NoV GGII demonstrating larger seasonal peaks. Virus reduction took place at similar rates in the primary settling, and in the activated sludge in combination with the secondary settling. Different physicochemical parameters and incoming virus concentrations were correlated to reduction of NoV between different treatment sites. This study gives new information about NoV transmission and virus reduction in a wastewater treatment plant.

Unlabelled: Each year the National Resource Defense Council addresses the quality of US beaches by routine bacterial indicators. In the Great Lakes region the indicator used is Escherichia coli and for 2007 more beaches were closed and impacted than ever before. In this study, water quality was addressed at two Lake Michigan Beaches over the 2004 swimming season by monitoring infectious enteric viruses by cell culture and integrated PCR and for a human sewage marker based on the Enterococcal Surface Protein (esp). Our goals for this study were to 1) examine the occurrence and variety of human enteric viruses present during peak usage of the beaches 2) determine key variables for development of predictive models for viruses; and 3) use quantitative risk assessment to estimate the potential health impact. Our results demonstrate that for both beaches predictive models of virus pollution were best described utilizing physical parameters like wind speed, wind direction and water temperature. The esp marker was not predictive of human viruses. The daily risk of acquiring a viral infection at either of the beaches ranged from 0.2 to 2.4/1000 swimmers using a quantitative microbial risk assessment model, with three swims during a day at the beach for children and over the season, the risk was 9-15/1000 swimmers using adenovirus as the model., Conclusions: Lake Michigan recreational beaches are being adversely impacted by human fecal pollution. Monitoring for the traditional indicators of water quality does not address viral risks and models can be developed and potentially used as real-time water quality forecasting tools.

Removal or inactivation of viruses in drinking water treatment processes can be quantified by measuring the concentrations of viruses or virus indicators in water before and after treatment. Virus reduction is then calculated from the ratio of these concentrations. Most often only the average reduction is reported. That is not sufficient when treatment efficiency must be characterized in quantitative risk assessment. We present three simple models allowing statistical analysis of series of counts before and after treatment: distribution of the ratio of concentrations, and distribution of the probability of passage for unpaired and paired water samples. Performance of these models is demonstrated for several processes (long and short term storage, coagulation/filtration, coagulation/sedimentation, slow sand filtration, membrane filtration, and ozone disinfection) using microbial indicator data from full-scale treatment processes. All three models allow estimation of the variation in (log) reduction as well as its uncertainty; the results can be easily used in risk assessment. Although they have different characteristics and are present in vastly different concentrations, different viruses and/or bacteriophages appear to show similar reductions in a particular treatment process, allowing generalization of the reduction for each process type across virus groups. The processes characterized in this paper may be used as reference for waterborne virus risk assessment, to check against location specific data, and in case no such data are available, to use as defaults.

In an effort to validate previous research suggesting remarkable resistance of norovirus to free chlorine disinfection, we characterized the disinfection response of purified and dispersed Norwalk virus (NV) by bench-scale free chlorine disinfection using RT-PCR for virus assays. The inactivation of NV by two doses of free chlorine (1 and 5mg/L) at pH 6 and 5 degrees C based on two RT-PCR assays was similar to that of coliphage MS2, but much faster than that of poliovirus 1. Despite the underestimation of virus inactivation by RT-PCR assays, the predicted CT values for NV based on RT-PCR assays are lower than the ones for most other important waterborne viruses and the CT guidelines for chlorine disinfection of viruses under the Surface Water Treatment Rule by the United States Environmental Protection Agency. Overall, the results of this study indicate that NV is not highly resistant to free chlorine disinfection as suggested by previous research and it is likely that NV contamination of drinking water can be controlled by adequate free chlorine disinfection practices with provision of proper pre-treatment processes before chlorination.

Potassium peroxymonosulfate activation leads to the formation of highly reactive species, mainly the sulfate radicals. Activated potassium peroxymonosulfate (from now on peroxymonosulfate) was tested against specific pollutants such as ammonium ion, creatinine, chlorinated creatinine products, arginine and Escherichia coli (E. coli), all constituents or derivatives of human discharges. The objective was to assess whether activated peroxymonosulfate can be a viable treatment reagent in recreational water applications. It was found that organic molecules such as creatinine, chlorinated creatinine products and arginine could be effectively treated with activated peroxymonosulfate. Ammonium ion was oxidized only by chlorine species and only in de-ionized water. Chlorine species were formed from the reaction of sulfate radicals with chloride ions. In pool water, the reaction of sulfate radicals with chloride ions and the subsequent ammonium ion oxidation were scavenged by the presence of bicarbonate ions. The Co/Peroxymonosulfate system was also shown to be an effective disinfection reagent, since 99.99% (4-log) kill of E. coli was achieved in 60 min of treatment. At the concentrations tested here, however, it is still not efficacious enough to qualify as an EPA-registered sanitizer for swimming pools (requires 6-log kill of E. coli, ATCC 11229, and Enterococcus faecium, ATCC 6569, in 30s).

Pulsed UV (PUV) is a novel UV irradiation system that is a non-mercury lamp-based alternative to currently used continuous-wave systems for water disinfection. PUV polychromatic irradiation disinfection efficiency was compared to that from continuous-wave monochromatic low-pressure (LP) and polychromatic medium-pressure (MP) UV systems, using two types of actinometry (ferrioxalate and iodide-iodate) and an absolute spectral emission method for fluence measurement. All three methods were in good agreement. Once accurate and reliable methods for fluence measurement were established, the inactivation of Escherichia coli and pathogen surrogates phage T4 and T7 were investigated under each technology. Inactivation was significantly faster using PUV irradiation compared to LP or MP UV lamps at equivalent fluence levels. A significant fraction of the enhanced PUV inactivation efficiency was due to wavelengths greater than 295 nm.

Advances in microbial source tracking (MST) have largely been driven by the need to comply with water quality standards based on traditional indicator bacteria. Recently, a number of culture-independent, and library-independent methods based on polymerase chain reaction (PCR) have been gaining popularity among source trackers. However, only a limited number of these methods have been successfully used in field applications, primarily due to the fact that many of them are still being developed. In this critical outlook, we examine different viewpoints associated with the practical use of MST to identify critical research gaps, propose a priority-based timeline to address them, and outline emerging technologies that will likely impact the future of source tracking. We propose that it is necessary to consider each of these aspects in order to advance towards a unifying framework in source identification, so that fecal pollution monitoring can be reliably used for comprehensive environmental microbial monitoring, to develop risk assessment models, and to implement and validate adequate management practices.

While humans require water for life, one-sixth of our species lives without access to safe water. In Africa, the situation is particularly acute because of global warming, the progression of the Sahara desert, civil unrest and poor governance, population growth, migration and poverty. In rural areas, the lack of adequate safe water and sanitary infrastructures leaves millions with doubtful water quality, increasing the harshness of daily life. In this paper, a pilot study was conducted during the wet season on Bolama Island (Guinea-Bissau, West Africa), a UNESCO Man and the Biosphere Reserve. Twenty-eight shallow wells, supplying water to most of the population, were sampled for microbiological, physical and chemical water quality characteristics. A ten-parameter water quality index (WQI) adapted to tropical conditions was applied to compare the different wells. About 79% of the wells showed moderate to heavy fecal contamination. From the surveyed parameters, it was found that chemical contamination was less important, although all samples were acidic, with the pH averaging 5.12+/-0.08. The WQI was 43+/-4% (0%-worst; 100%-best quality), showing that the water from the majority of wells was polluted but should be suitable for domestic use after appropriate treatment. At the onset of the wet season, diarrhea represented 11.5% of all medical cases, 92.5% of which were children aged <15. This paper suggests inexpensive steps to reduce the fecal contamination and control the pH in order to increase the potability of the well water and, concomitantly, to raise the living standards of the population in one of the poorest countries of the world.

Veterinary antibiotics are widely used at concentrated animal feeding operations (CAFOs) to prevent disease and promote growth of livestock. However, the majority of antibiotics are excreted from animals in urine, feces, and manure. Consequently, the lagoons used to store these wastes can act as reservoirs of antibiotics and antibiotic-resistant bacteria. There is currently no regulation or control of these systems to prevent the spread of these bacteria and their genes for antibiotic resistance into other environments. This study was conducted to determine the disinfection potential of chlorine, ultraviolet light and ozone against swine lagoon bacteria. Results indicate that a chlorine dose of 30 mg/L could achieve a 2.2-3.4 log bacteria reduction in lagoon samples. However, increasing the dose of chlorine did not significantly enhance the disinfection activity due to the presence of chlorine-resistant bacteria. The chlorine resistant bacteria were identified to be closely related to Bacillus subtilis and Bacillus licheniformis. A significant percentage of lagoon bacteria were not susceptible to the four selected antibiotics: chlortetracycline, lincomycin, sulfamethazine and tetracycline (TET). However, the presence of both chlorine and TET could inactivate all bacteria in one lagoon sample. The disinfection potential of UV irradiation and ozone was also examined. Ultraviolet light was an effective bacterial disinfectant, but was unlikely to be economically viable due to its high energy requirements. At an ozone dose of 100 mg/L, the bacteria inactivation efficiency could reach 3.3-3.9 log.

Amino acid-like fluorescence has been used as an indicator of biological activity in wastewater effluent and in natural waters, and can be detected using fluorescence spectroscopy. Little or no work has been able to state conclusively that these so called 'amino acid-like' fluorophores are associated with proteins present as a result of bacterial activity. This work aims to ascertain whether bacteria are one possible cause of these 'amino acid-like' peaks observed in natural waters. In addition, fluorescence derived solely from one bacterial source was determined as a function of the growth time and temperature. The bacterium Pseudomonas aeruginosa was isolated from the urban River Tame, Birmingham, UK, and planktonic bacteria were grown in sterile, sealed glass jars, in 100 ml growth media. Bacteria were grown at 11, 25 and 37 degrees C, over a maximum of 10 days. A 3D Excitation-Emission Matrix (EEM) plot was generated from fluorescence analysis of the samples. Both tryptophan and tyrosine-like fluorescence, resembling that observed in natural and waste waters, was observed in these samples, indicating that observed fluorescence signals from aquatic systems in the literature were of biotic origin. Significant differences in fluorescence signals were obtained from planktonic cells grown at different temperatures. At 25 and 37 degrees C, cells were found to produce predominantly tryptophan-like fluorescence, with some tyrosine-like fluorescence also detected. A further unknown fluorophore was also detected (emission wavelength of approximately 460 nm, with three excitation centres at 225, 260 and 390 nm), likely to be a bacterially produced metabolite. At 11 degrees C, a more environmentally realistic temperature in temperate environments, quantitative and qualitative differences were observed in fluorescence signals when compared with the higher temperatures, indicating that laboratory observations conducted at higher temperatures may not be easily used to interpret environmental processes.

The paper describes a modelling study of near-shore coastal waters, undertaken to assess the impact of various bacterial input loads on the receiving waters in a coastal basin in the UK. Total and faecal coliforms, used as the indicators for bathing water quality under the European Union (EU) Bathing Water Directive, were numerically modelled using a 2D depth integrated hydro-environmental model. Details are given of the governing equations and solution methods used in the numerical model, together with a discussion of the recent development in faecal bacterial indicator modelling. Details are also given of a field data collection exercise, which involved initially collecting existing information on effluent input loads and followed by an intensive field survey. Using the water quality model, the mortality rate of the pathogen bacteria was investigated. Three methods were used to represent the relationship between the decay rate and the level of solar radiation including: a constant decay rate, day- and night-time decay rates and a solar radiation related time varying decay rate. Relatively close agreement between model predicted and measured total and faecal coliform concentration distributions were obtained for different day- and night-time decay rates and time varying decay rates. No significant differences were found in the optimum decay rates for total and faecal coliform levels. Finally, the impact of the individual inputs on the bathing water quality of the basin was also statistically and numerically investigated. Results showed that the River Irvine was the most significant input during high river flows, and that under these conditions the bathing waters were likely to fail to comply with the European Union Bathing Water Directive. For base river flow conditions the Meadowhead effluent input was found to be critical for both total and faecal coliform level predictions.

To evaluate the success of almost 20 years of pollution abatement in the Bilbao estuary watershed in northern Spain, we analyzed temporal trends in pollution discharges and water quality from 1993 to 2003. Over that period a great portion of the raw wastewater discharge was intercepted and treated, leading to a significant reduction in the pollution load to the estuary (51.8% in biochemical oxygen demand, 70.9% in ammonia nitrogen and 81.9% in faecal coliforms). Temporal trends of mean annual levels of water quality variables showed statistically significant increases in dissolved oxygen saturation (between 2.04 and 4.11%/year) and decreases in ammonia nitrogen (between -4.15 and -175.75 microM NH3/year) and faecal coliforms concentrations (from 2.55 x 10(5) to 2.13 x 10(4) CFU/100ml). The improvement of the Bilbao estuary water quality reported in this paper is primarily attributed to the pollution abatement measures accomplished by the local water authority. Finally, as a result of these pollution control efforts, European bathing water quality standards were met at local beaches.

The microbiological quality of drinking water from 144 private water supplies in the Netherlands was tested and additionally the occurrence of Escherichia coli O157 was examined. Faecal indicators were enumerated by using standard membrane filtration methods. The presence of E. coli O157 was determined using a specific enrichment method. Eleven percent of the samples contained faecal indicators whereas E. coli O157:H7 was isolated from 2.7% of the samples that otherwise met the drinking water standards. The E. coli O157 positive water supplies were located on camp-sites in agricultural areas with large grazer densities. Pulsed field gel electrophoresis (PFGE) analysis suggested that cattle might have been the cause of contamination. Our results indicate that compliance with microbiological quality standards obtained in routine monitoring does not always guarantee the absence of pathogens. The presence of pathogens such as E. coli O157 may suggest possible health consequences; however, a risk assessment process should be performed as the monitoring of both faecal indicator parameters and pathogens do not predict the effect of microbial contamination of drinking water on a population.

Analyses of Irish bathing water quality data sets are reported to investigate whether the parametric calculations proposed in the draft Bathing Water Directive are valid. Faecal coliforms (assumed to be Escherichia coli) and faecal streptococci (assumed to be intestinal enterococci) have been analysed separately. It was noted that classifications based on the parametric 95th percentile calculations disagreed with those based on percentage compliance with the standards on 13.8% of occasions. When these disagreements were studied, it was found that the datasets frequently contained many censored data points (Result < 1). Also, the datasets were not log normally distributed on at least 85% of occasions. Both these findings fatally undermine the validity of using a parametric method for calculating 95th percentiles to classify bathing water quality. By contrast the non-parametric Hazen method is a better estimate of true population 95th percentiles, but essentially gives very similar classifications to the percentage compliance approach, fully agreeing on over 95% of occasions. The same is also true when considering 90th percentiles. A series of Monte Carlo studies were also conducted to determine the impact of small numbers. It was ascertained that small sample sizes are very undependable in determining bathing water classification and the parametric method in particular is particularly unreliable. In conclusion, the parametric method for calculating bathing water compliance is so severely and fatally flawed statistically that it should not form the basis of any legislation. The Hazen method gives a better estimate of true 90th or 95th percentiles, though as the resultant classifications agree with percentage compliance so closely it is doubtful that there is any statistical value in using a percentile approach over the long established and well understood percentage compliance approach.

Enteric viruses have been detected in many drinking water supplies all over the world. A meaningful number of these supplies were treated and disinfected according to internationally acceptable methods. In addition, counts of bacterial indicators (coliform bacteria and heterotrophic plate count organisms) in these water supplies were within limits generally recommended for treated drinking water and these findings have been supported by epidemiological data on infections associated with drinking water. The shortcomings of conventional treatment methods and indicator organisms to confirm the absence of enteric viruses from drinking water, was generally ascribed to the exceptional resistance of these viruses. In this study, the prevalence of enteroviruses detected from July 2000 to June 2002 in sewage, river-, borehole-, spring- and dam water as well as drinking water supplies treated and disinfected according to international specifications for the production of safe drinking water was analysed. A glass wool adsorption-elution technique was used to recover viruses from 10--20 l of sewage as well as environmental water samples, in the case of drinking water from more than 100 l. Recovered enteroviruses were inoculated onto two cell culture types (BGM and PLC/PRF/5 cells) for amplification of viral RNA with nested-PCR being used to detect the amplified viral RNA. Results from the study demonstrated the presence of enteroviruses in 42.5% of sewage and in 18.7% of treated drinking water samples. Furthermore, enteroviruses were detected in 28.5% of river water, in 26.7% of dam/spring water and in 25.3% of borehole water samples. The high prevalence of coxsackie B viruses found in this study suggested, that a potential health risk and a burden of disease constituted by these viruses might be meaningful. These findings indicated that strategies, other than end-point analysis of treated and disinfected drinking water supplies, may be required to ensure the production of drinking water that does not exceed acceptable health risks. More reliable approaches to ensure acceptable safety of drinking water supplies may be based on control by multiple-barrier principles from catchment to tap using hazard assessment and critical control point (HACCP) principles.

In recent years, considerable investment has been committed to sewerage infrastructure and new sewage treatment plants in the catchment surrounding an estuarine basin along the north-west coast of England. Although this capital investment has resulted in a marked reduction in the input of bacterial loads, relatively high counts of faecal indicator organisms are still being encountered in the coastal receiving waters, and the local bathing waters continue to fail on occasions to comply with the European Community (EC) Bathing Water Directive (1976) mandatory standards. Details are given herein of a comprehensive modelling study aimed at quantifying the impact of various bacterial inputs into the estuary and surrounding coastal waters on the bathing water quality. The model domain includes the coastal area and the entire estuary (namely the Ribble) up to the tidal limits of its tributaries. Faecal coliforms have been used as the main water quality indicator organisms. The numerical model developed for this study combines a depth integrated two-dimensional coastal model and a cross-sectionally integrated one-dimensional river model, and is capable of predicting water surface elevations, velocity fields and faecal coliform concentration distributions across the entire model domain. The hydrodynamic model was calibrated using water level and velocity measurements from three surveys and then validated against measured data from three other surveys. In order to predict the faecal coliform concentration distributions, variable faecal coliform decay rates were used, i.e. different values of decay rates were applied to the coastal and riverine waters, for day- and nighttime, and for wet and dry weather conditions. The maximum and minimum decay rates used were 2.32/day and 0.71/day for the dry and wet weather surveys, respectively. The model was then applied to (i) assess the impact of previous discharge strategies and investigate the effectiveness of future capital investment works and (ii) predict the impact of a range of strategic options, including: the effects of adding UV treatment, constructing storm water storage tanks and incorporating various combined sewer overflow (CSO) discharge scenarios for different weather conditions.

Faecal-indicator budget studies have shown marine bathing water quality at two small UK coastal resorts, Staithes and Newport, to be adversely affected by riverine inputs from lowland pastoral catchments (J. Chartered Inst. Water Environ. Mangt. 12 (1998) 414). The present paper reports on presumptive coliform (PC), presumptive Escherichia coli (PE) and presumptive streptococci (PS) concentrations at 43 sampling points on watercourses within these catchments, and on their relationship with land use and livestock-related management practices, such as grazing and slurry/manure applications. The results show > 10-fold elevations in geometric mean faecal-indicator concentrations under high-flow conditions, compared with low flow, with maximum high-flow geometric mean PC, PE and PS concentrations of 2.6 x 10(6), 1.8 x 10(6) and 4.4 x 10(5) cfu/100 ml, respectively. High-flow geometric mean concentrations exhibit highly significant positive correlations with land use/management variables associated with intensive livestock farming, both within the individual catchments and in the two combined. Additional factors, such as antecedent weather conditions and topography, contribute to inter-catchment variability in water quality. Although inputs from diffuse and point sources of pollution were not quantified, point sources (e.g. runoff from farm yards) seem likely to be significant. The findings suggest that it may be possible to develop generic statistical models to predict microbial water quality from land use and farm management data. They also provide indirect evidence that channel bed sediment 'stores' closely reflect land use within their catchments and that there is little die-off of organisms along watercourses.

Thc inactivation behaviour of the bacteriophages PHI X 174 (ssDNA virus). MS2 (ssRNA virus) and B40-8 (dsDNA) toward non-ionizing (UV-253.7 nm) as well as to ionizing radiation (gamma radiation) was studied in order to evaluate their potential as viral indicators for water disinfection by irradiation. Previous findings of the high UV-253.7 nm resistance of MS2 were confirmed whereas an unexpected high sensitivity to gamma radiation compared to the two other phages was found. On the other hand, PHI X 174 revealed an enhanced UV sensitivity but a high resistance to ionizing radiation. B40-8 had an intermediate position between the other two bacteriophages relative to both types of radiation. As expected, the data of E. coli reconfirmed the unreliability of fecal indicator bacteria for the purpose of predicting responses of viruses to water treatment. In UV disinfection the influence of water matrix may be adequately controlled by considering the UV (253.7 nm) absorption of the water whereas so far no such parameter has existed for the influence of the water quality on ionizing irradiation with respect to the scavenger concentration.

The occurrence of Escherichia coli O157:H7 in selected water samples in South Africa was investigated. The chromogenic Rainbow agar O157 medium designed for the rapid identification of E. coli O157:H7 was used for the detection of these organisms in various river-water samples in the Vaal Barrage Reservoir drainage basin of South Africa. A total of 204 samples were obtained from 15 sites where water was used for direct and indirect human consumption. Samples were filtered through Gelman filter-units and incubated on Rainbow agar O157 which produced different colours according to the bacterial chromogenic properties. Six hundred and sixty-three suspected E. coli O157:H7 colonies, with colours ranging between dark blue, grey and black, were subcultured onto sorbitol-MacConkey agar and screened for different virulence factors specific for E. coli O157:H7 and agglutination with anti-E. coli O157 antiserum. The results indicated that none of the suspected colonies contained all of the virulence factors necessary to classify them as E. coli O157:H7. None of these organisms agglutinated with antisera against E. coli O157. The probability of being infected with E. coli O157:H7 from direct or indirect consumption of these river water sources is therefore low. Some samples did, however, contain enterohaemorrhagic E. coli virulence properties, such as Stx1, Stx2 and enterohaemolysin, which might impose a health risk if ingested.

A set of ecological indicators including structural, functional, and system-level aspects were proposed for a lake ecosystem health assessment, according to the structural, functional, and system-level responses of lake ecosystems to chemical stresses including acidification, eutrophication and copper, oil and pesticide contamination. The structural indicators included phytoplankton cell size and biomass, zooplankton body size and biomass, species diversity, macro- and micro-zooplankton biomass, the zooplankton phytoplankton ratio, and the macrozooplankton microzooplankton ratio. The functional indicators encompassed the algal C assimilation ratio, resource use efficiency, community production, gross production/respiration (i.e. P/R) ratio, gross production standing crop biomass (i.e. P/B) ratio, and standing crop biomass unit energy flow (i.e. B/E) ratio. The ecosystem-level indicators conisisted of ecological buffer capacities, energy, and structural energy. Based on these indicators, a direct measurement method (DMM) and an ecological modeling method (EMM) for lake ecosystem health assessment were developed. The DMM procedures were designed to: (1) identify key indicators; (2) measure directly or calculate indirectly the selected indicators; and, (3) assess ecosystem health on the basis of the indicator values. The EMM procedures were designed to: (1) determine the structure and complexity of the ecological model according to the lake's ecosystem structure; (2) establish an ecological model by designing a conceptual diagram, establishing model equations, and estimating model pararmeters; (3) compare the simulated values of important state variables and process rates with actual observations; (4) calculate ecosystem health indicators using the ecological model; and, (5) assess lake ecosystem health according to the values of the ecological indicators. The results of a case study demonstrated that both methods provided similar results which corresponded with the lake's actual trophic state.

Water samples and pipe coupons were collected from the Brussel's drinking water distribution system (DS). A treated surface water and various groundwaters feed this DS. Parameters related to bacterial regrowth have been measured on these samples: temperature, concentrations of free residual chlorine, concentration of biodegradable dissolved organic carbon (BDOC), abundance of suspended bacteria, densities of fixed bacteria and levels of bacterial activity. Results showed that groundwaters were less susceptible to favor bacterial regrowth in the DS pipes. Treated surface water and mixed waters had the highest potential of bacterial regrowth in the DS dead ends. Results also showed that the potential regrowth induced by the distribution of a treated surface water could be reduced if: (1) the BDOC levels were below 0.25 mg C/l at the outlet of the surface water treatment plant; (2) a significant free chlorine residual was present within the whole DS. Second-stage biological filtration using granular activated carbon is now under construction at the surface water treatment plant feeding a part of this DS. This treatment implementation should reduce BDOC levels and chlorine demand of the treated surface water and will further reduce the slight regrowth phenomena observed in this DS.

Colonies and single cells of Microcystis aeruginosa and the hepatotoxin microcystin were retained by salad lettuce after growth with spray irrigation water containing the microcystin-producing cyanobacteria. These findings are discussed in terms of crop spray irrigation with water containing cyanobacteria and potential human exposure to cyanobacterial toxins via plant foods grown in such circumstances.