Faecal indicator organism inputs to watercourses from streamside pastures grazed by cattle: Before and after implementation of streambank fencing.

Faecal indicator organisms (FIOs) are major pollutants in many catchments world-wide, with streamside pastures on livestock farms being potentially significant sources. Hitherto, few empirical studies have quantified FIO fluxes from such areas or investigated streambank fencing (SBF) and other possible mitigation measures. The aim of this two-phase (before/after intervention) study of the effectiveness of SBF was to generate an empirical evidence-base to enable regulatory authorities to make better-informed decisions concerning the implementation of this measure. It was undertaken during the summer bathing season along a 271 m stream reach in the River Tamar catchment, SW England. The study included: cattle distribution surveys; monitoring of changes in E. coli (EC) and intestinal enterococci (IE) concentrations and fluxes down the reach and of concentrations in ditch flow and surface runoff; phage tracer studies of surface runoff from pasture land; and experimental streambed trampling to investigate streambed FIO sources. The results show that cattle spend a disproportionately large amount of time in the watercourse/riparian zone along unfenced streams; identify direct defecation to the stream by wading livestock and the release/mobilisation of FIOs from cowpats by surface runoff from the adjacent pastures at times of high flow as key transmission routes; and demonstrate that FIOs become incorporated within streambed sediments, from which they may subsequently be released by trampling. Partial exclusion of cattle through SBF with a drinking bay greatly reduces the time cattle spend in streams. Total exclusion SBF, with provision of an alternative drinking supply, considerably reduces FIO load inputs to the stream reach, e.g. at times of high flow, which are critical in terms of pollutant fluxes to coastal waters, the mean EC and IE input loads to the reach fell by 0.842 and 2.206 log10, respectively.

Within-day variability in microbial concentrations at a UK designated bathing water: Implications for regulatory monitoring and the application of predictive modelling based on historical compliance data.

Prediction of bathing water quality is recommended by the World Health Organization (WHO), the European Union (EU) and the United States Environmental Protection Agency (USEPA) and is an established element in bathing water management designed to protect public health. Most commonly, historical regulatory compliance data are used for model calibration and provide the dependent variable for modelling. Independent (or predictor) variables (e.g. rainfall, river flow and received irradiance) measured over some antecedent period are used to deliver prediction of the faecal indicator concentration measured on the day of the regulatory sample collection. The implied linked assumptions of this approach are, therefore, that; (i) the independent variables accurately predict the bathing-day water quality; which is (ii) accurately characterized by the single regulatory sample. Assumption (ii) will not be the case where significant within-day variability in water quality is evident. This study built a detailed record of water quality change through 60 days at a UK coastal bathing water in 2011 using half-hourly samples each subjected to triplicate filtration designed to enhance enumeration precision. On average, the mean daily variation in FIO concentrations exceeded 1 log10 order, with the largest daily variations exceeding 2 log10 orders. Significant diurnality was observed at this bathing water, which would determine its EU Directive compliance category if the regulatory samples were collected at the same time each day. A sampling programme of this intensity has not been reported elsewhere to date and, if this pattern is proven to be characteristic of other bathing waters world-wide, it has significance for: (a) the design of regulatory sampling programmes; (b) the use of historical data to assess compliance, which often comprises a single sample taken at the compliance point on a regular, often weekly, basis; and (c) the use of regulatory compliance data to build predictive models of water quality.

Relationships between human adenoviruses and faecal indicator organisms in European recreational waters.

Human adenoviruses (HAdV) may be implicated in some disease outbreaks associated with recreational water exposures, typically in swimming pools. Modern molecular methods can be used to detect HAdV in environmental water samples. During the EU FP6 Project VIROBATHE a database of over 290 HAdV analyses with corresponding faecal indicator organism (FIO) determinations was gathered and used to explore statistical associations between HAdV and FIO results. The FIOs measured were Escherichia coli, intestinal enterococci and somatic coliphage. Statistically significant trends of increasing proportions of HAdV-positive results in categories of increasing FIO concentration were found in freshwater but not seawater samples. The analysis of these trends in freshwater samples was refined, the trends remaining statistically significant when using categories of 0.5 log(10) intervals of FIO concentration. Logistic regression models were then developed to predict the probability of a HAdV-positive outcome from FIO concentration. Potential applications of these models to predict the probability of HAdV-positive outcomes from routine FIO determinations used to describe recreational water quality exposures and to classify recreational water quality are discussed.

Surveillance of adenoviruses and noroviruses in European recreational waters.

Exposure to human pathogenic viruses in recreational waters has been shown to cause disease outbreaks. In the context of Article 14 of the revised European Bathing Waters Directive 2006/7/EC (rBWD, CEU, 2006) a Europe-wide surveillance study was carried out to determine the frequency of occurrence of two human enteric viruses in recreational waters. Adenoviruses were selected based on their near-universal shedding and environmental survival, and noroviruses (NoV) selected as being the most prevalent gastroenteritis agent worldwide. Concentration of marine and freshwater samples was done by adsorption/elution followed by molecular detection by (RT)-PCR. Out of 1410 samples, 553 (39.2%) were positive for one or more of the target viruses. Adenoviruses, detected in 36.4% of samples, were more prevalent than noroviruses (9.4%), with 3.5% GI and 6.2% GII, some samples being positive for both GI and GII. Of 513 human adenovirus-positive samples, 63 (12.3%) were also norovirus-positive, whereas 69 (7.7%) norovirus-positive samples were adenovirus-negative. More freshwater samples than marine water samples were virus-positive. Out of a small selection of samples tested for adenovirus infectivity, approximately one-quarter were positive. Sixty percent of 132 nested-PCR adenovirus-positive samples analysed by quantitative PCR gave a mean value of over 3000 genome copies per L of water. The simultaneous detection of infectious adenovirus and of adenovirus and NoV by (RT)PCR suggests that the presence of infectious viruses in recreational waters may constitute a public health risk upon exposure. These studies support the case for considering adenoviruses as an indicator of bathing water quality.

Predicting microbial pollution concentrations in UK rivers in response to land use change.

The Water Framework Directive has caused a paradigm shift towards the integrated management of recreational water quality through the development of drainage basin-wide programmes of measures. This has increased the need for a cost-effective diagnostic tool capable of accurately predicting riverine faecal indicator organism (FIO) concentrations. This paper outlines the application of models developed to fulfil this need, which represent the first transferrable generic FIO models to be developed for the UK to incorporate direct measures of key FIO sources (namely human and livestock population data) as predictor variables. We apply a recently developed transfer methodology, which enables the quantification of geometric mean presumptive faecal coliforms and presumptive intestinal enterococci concentrations for base- and high-flow during the summer bathing season in unmonitored UK watercourses, to predict FIO concentrations in the Humber river basin district. Because the FIO models incorporate explanatory variables which allow the effects of policy measures which influence livestock stocking rates to be assessed, we carry out empirical analysis of the differential effects of seven land use management and policy instruments (fiscal constraint, production constraint, cost intervention, area intervention, demand-side constraint, input constraint, and micro-level land use management) all of which can be used to reduce riverine FIO concentrations. This research provides insights into FIO source apportionment, explores a selection of pollution remediation strategies and the spatial differentiation of land use policies which could be implemented to deliver river quality improvements. All of the policy tools we model reduce FIO concentrations in rivers but our research suggests that the installation of streamside fencing in intensive milk producing areas may be the single most effective land management strategy to reduce riverine microbial pollution.

Evaluating short-term changes in recreational water quality during a hydrograph event using a combination of microbial tracers, environmental microbiology, microbial source tracking and hydrological techniques: a case study in Southwest Wales, UK.

Quantitative assessment of multiple sources to short-term variations in recreational water quality, as indexed by faecal indicator organism (FIO) concentrations, is becoming increasingly important with adoption of modern water quality standards and catchment-based water quality management requirements (e.g. the EU Water Framework Directive, Article 11 'Programmes of Measures' and the US Clean Water Act, 'Total Maximum Daily Loads'). This paper describes a study combining microbial tracers, intensive FIO measurement, open channel hydrology and molecular microbial source tracking (MST) to enhance understanding of recreational water quality at Amroth in southwest Wales, UK. Microbial tracers were released from four stream inputs during a moderate hydrograph event. Tracers from two local streams impacted simultaneously with a period of maximum FIO concentrations at the near-shore compliance monitoring site. Connection between these inputs and this site were rapid (9-33 min). Water quality impairment from a more remote stream input followed, 12.85 h after tracer release, sustaining FIO concentrations above desired compliance levels. MST analysis showed dominance of ruminant Bacteroidales genetic markers, associated with agricultural pollution. This integration of tracers and MST offers additional information on the movement and individual sources causing water quality impairment.

Evaluating the operational utility of a Bacteroidales quantitative PCR-based MST approach in determining the source of faecal indicator organisms at a UK bathing water.

Microbial source tracking techniques are used in the UK to provide an evidence-base to guide major expenditure decisions and/or regulatory action relating to sewage disposal. Consequently, it is imperative that the techniques used robustly index faecal indicator organisms (FIOs) that are the regulatory parameters for bathing and shellfish harvesting areas. This study reports a 'field-scale' test of microbial source tracking (MST) based on the quantitative PCR analyses of Bacteroidales 16S rRNA genetic marker sequences. The project acquired data to test the operational utility of quantitative Bacteroidales MST data, comparing it with FIO concentrations in streams, effluents and bathing waters. Overall, the data did not exhibit a consistent pattern of significant correlations between Bacteroidales MST parameters and FIOs within the different sample matrices (i.e. rivers, bathing waters and/or effluents). Consequently, there was little evidence from this study that reported concentrations and/or percentages of human and/or ruminant faecal loadings (that are based on Bacteroidales MST gene copy numbers) offer a credible evidence-base describing FIO contributions to receiving water 'non-compliance'. The study also showed (i) there was no significant attenuation of the Bacteroidales gene copy number 'signal' through the UV disinfection process; and (ii) single non-compliant samples submitted for Bacteroidales MST analysis, do not reliably characterise the balance of faecal loadings due to the high variability in the MST signal observed. At this stage in the development of the MST tool deployed, it would be imprudent to use the percentage human and/or ruminant contributions (i.e. as indicated by MST data acquired at a bathing water) as the sole or principal element in the evidence-base used to guide major expenditure decisions and/or regulatory action.

Results of field investigations into the impact of intermittent sewage discharges on the microbiological quality of wild mussels (Mytilus edulis) in a tidal estuary.

Field surveys were designed to examine the effects of sewage contamination from storm overflow effluent on faecal coliform and Escherichia coli concentrations in the flesh of wild mussels (Mytilus edulis). Bags containing 30 mussels each were fixed at known inter-tidal locations and retrieved at intervals following discharge from a nearby combined sewer overflow (CSO). Concentrations of faecal coliform bacteria and E. coli were measured in the shellfish flesh and in samples of overlying water prior to collection of the mussel samples. Faecal coliform and E. coli concentrations in shellfish increased rapidly after CSO discharge. E. coli concentrations exceeded the European shellfish hygiene class C limit of 46,000 100g(-1), and decayed during subsequent CSO discharge-free periods. The concentration and depuration response was independent of the magnitude of CSO spill volume. First-order exponential decay functions were fitted to the data. Decay rates were lower than those found in corresponding microcosm experiments. This relates to the repeated pattern of inundation and exposure associated with the tidal cycles in the estuary. Relationships between E. coli and faecal coliform concentrations in the shellfish and overlying water samples were relatively weak (r<0.60), a pattern often seen with data from uncontrolled environmental experiments.

Microbial source tracking: a forensic technique for microbial source identification?

As the requirements of the Water Framework Directive (WFD) and the US Clean Water Act (USCWA) for the maintenance of microbiological water quality in 'protected areas' highlight, there is a growing recognition that integrated management of point and diffuse sources of microbial pollution is essential. New information on catchment microbial dynamics and, in particular, the sources of faecal indicator bacteria found in bathing and shellfish harvesting waters is a pre-requisite for the design of any 'programme of measures' at the drainage basin scale to secure and maintain compliance with existing and new health-based microbiological standards. This paper reports on a catchment-scale microbial source tracking (MST) study in the Leven Estuary drainage basin, northwest England, an area for which quantitative faecal indicator source apportionment empirical data and land use information were also collected. Since previous MST studies have been based on laboratory trials using 'manufactured' samples or analyses of spot environmental samples without the contextual microbial flux data (under high and low flow conditions) and source information, such background data are needed to evaluate the utility of MST in USCWA total maximum daily load (TMDL) assessments or WFD 'Programmes of Measures'. Thus, the operational utility of MST remains in some doubt. The results of this investigation, using genotyping of Bacteroidetes using polymerase chain reaction (PCR) and male-specific ribonucleic acid coliphage (F + RNA coliphage) using hybridisation, suggest some discrimination is possible between livestock- and human-derived faecal indicator concentrations but, in inter-grade areas, the degree to which the tracer picture reflected the land use pattern and probable faecal indicator loading were less distinct. Interestingly, the MST data was more reliable on high flow samples when much of the faecal indicator flux from catchment systems occurs. Whilst a useful supplementary tool, the MST information did not provide quantitative source apportionment for the study catchment. Thus, it could not replace detailed empirical measurement of microbial flux at key catchment outlets to underpin faecal indicator source apportionment. Therefore, the MST techniques reported herein currently may not meet the standards required to be a useful forensic tool, although continued development of the methods and further catchment scale studies could increase confidence in such methods for future application.

Reply to comments on "Derivation of numerical values for the World Health Organization guidelines for recreational waters".

The contribution addressed reveals an optimistic design philosophy likely to systematically underestimate risk in epidemiologic studies into the health effects of bathing water exposures. The authors seem to recommend that data on the 'exposure' measure (i.e. water quality) in such studies should be acquired in a similar manner to that used for regulatory sampling. This approach may compromise the quality of the epidemiologic investigations undertaken. It may result in imprecise estimates of exposure because it ignores the fact that regulatory timescales and spatial resolution (even if artificially compressed to a bathing day) can mask large spatial and temporal variability in water quality. If this variability is ignored by taking some mean value and attributing that to all of those exposed in a period at a study location, many bathers may be misclassified and the studies may be biased to a 'no-effect' conclusion. A more appropriate approach is to maximise the precision of the epidemiologic investigations by measurement of individual exposure (or water quality) at the place and time of the exposure, as has been done in randomised volunteer studies in the UK and Germany. The precise epidemiologic relationships linking 'exposure' with 'illness' can then be related to the probability of exposure to particular water quality by a 'normal bather' using the known probability distribution of the exposure variable (i.e. faecal indicator concentration) in the regulated bathing waters. We suggest that any research protocol where poor sampling design for water quality assessment is justified because regulatory monitoring is equally imprecise may be fundamentally flawed. The rationale for this assessment is that the epidemiology is the starting point and evidence-base for 'standards'. If precision is not maximised at this stage in the process it compromises the credibility of the standards design process. The negative effects of the approach advocated in this 'comment' are illustrated using published research findings used to derive the figures illustrated in Wymer et al. [2005. Comment on derivation of numerical values for the World Health Organization guidelines for recreational waters. Water Research 39, 2774-2777].

Processes driving the episodic flux of faecal indicator organisms in streams impacting on recreational and shellfish harvesting waters.

Understanding the process controls on episodic fluxes of faecal indicator organisms (FIOs) is becoming increasingly important for the sustainable management and accurate modelling of water quality in both recreational and shellfish harvesting waters. Both environments exhibit transitory non-compliance with microbiological standards after rainfall episodes despite significant expenditures on control of sewage derived pollutant loadings in recent years. This paper demonstrates the role of wave propagation in the entrainment of FIOs from river channel beds as a contributor to episodes of poor microbial water quality. Previously reported data is reviewed in the light of relationships between wave and mean water travel velocities. High flows and rapid changes in river flow, driven by releases of bacterially pure reservoir water, resulted in elevated FIO concentrations and transient peaks in concentration. The new interpretation of these data suggest three modes of entrainment: (i) immediate wave-front disturbance, (ii) wave propagation lift and post-wave transport at mean flow velocity, and (iii) stochastic erosional mechanisms that maintain elevated bacterial concentrations under steady high flow conditions. This is a significant advance on the previously proposed mechanisms. Understanding these mechanisms provides an aid to managing streams intended for recreational use and emphasises the need to control the timing of high flow generation prior to use of the water body for e.g. canoeing events. In addition the processes highlighted have relevance for the protection of shellfish nurseries, drinking water supply intakes and episodes of poor bathing water quality, and associated health risks.

Predicting faecal indicator fluxes using digital land use data in the UK's sentinel Water Framework Directive catchment: the Ribble study.

The Ribble drainage basin is the single UK sentinel study area chosen for examining the implementation of the EU Water Framework Directive (WFD 20/60/EC). The study which has generated the data for this paper was initiated to quantify 'catchment-derived' fluxes of faecal indicators originating from both point and diffuse sources to inform the competent authorities on the potential for, and prioritization of, further options for reducing the faecal indicator loadings to this crucial coastal environment. It represents the first UK drainage basin-scale 'profile' of faecal indicator sources as recommended by WHO [1999. Health Based Monitoring of Recreational Waters: The Feasibility of a New Approach; the "Annapolis Protocol". World Health Organisation Geneva, Switzerland; 2003. Guidelines for Safe Recreational-Water Environments Volume 1: Coastal and Fresh-Waters. World Health Organisation Geneva, Switzerland] and incorporated into current drafts of the revised Bathing Water Directive [Anon, 2004. Council of the European Communities Amended proposal for a Directive of the European Parliament and of the Council concerning the management of bathing water quality. Brussels 23rd June]. This paper focuses on the relationships between land use and faecal indicator organism concentrations in surface waters within this very large drainage basin (1583 km2) containing some extensive urban areas. A geographical information system comprising readily available digital elevation, remotely sensed land cover and digital map data was used to generate the land use variables for subcatchments draining to 41 locations across the study area. Presumptive concentrations of coliforms, Escherichia coli and enterococci (colony forming unit (cfu) 100 ml(-1)) were measured at each location on at least 20 occasions over a 44-day period within the 2002 bathing season. The sampling programme targeted hydrograph events. Hydrometric records were used to allocate results as either base flow or high flow. At each site, geometric mean faecal indicator organism concentrations were significantly elevated at high flow compared to base flow. Stepwise regression modelling produced statistically significant models predicting geometric mean base and high-flow faecal indicator organism concentrations from land use variables (r2: 49.5-68.1%). The dominant predictor variable in each case was the proportion of built-up land in subcatchments, suggesting that this land use type, with associated sewage-related inputs, is a critical source of faecal indicator organisms in this drainage basin.

Sustainable reduction in the flux of microbial compliance parameters from urban and arable land use to coastal bathing waters by a wetland ecosystem produced by a marine flood defence structure.

'Natural' treatment systems such as wetlands and reed beds have been proposed as sustainable means of reducing fluxes of faecal indicator organisms (FIOs) to recreational and shellfish harvesting waters. This is because FIO fluxes to coastal waters from both point (effluent) and diffuse (catchment) sources can cause non-compliance with microbiological standards for bathing and shellfish harvesting waters. The Water Framework Directive requires competent authorities in the member states to manage both point and diffuse sources of FIOs in an integrated manner to achieve compliance with 'good' water quality as defined in a series of daughter Directives. This study was undertaken to investigate the relative sources of FIOs to the popular bathing waters around Clacton, UK. In this predominantly arable (mainly cereal cropping) farming area, the principal land use predictor, explaining 76% of the variance in geometric mean presumptive Escherichia coli concentration at sub-catchment outlets during the bathing season, was the proportion of built-up (i.e. urbanised) land in each sub-catchment. This new finding contrasts with earlier studies in livestock farming regions where the proportion of improved grassland has proven to be the strongest predictor of microbial concentration. Also novel in this investigation, a flood defence wall has been built creating a wetland area which discharges every tidal cycle. The wetland produces over 97% reduction in the flux and concentrations of FIOs to the marine recreational waters. Also, FIO concentrations in water draining through the wetland to the sea were similar to concentrations measured in six UK sewage treatment plant effluents subject to secondary (biological) treatment followed by UV disinfection.

Derivation of numerical values for the World Health Organization guidelines for recreational waters.

In April 2001, draft 'Guidelines' for safe recreational water environments were developed at a World Health Organization (WHO) expert consultation. Later the same month, these were presented and discussed at the 'Green Week' in Brussels alongside the on-going revision of the European Union Bathing Water Directive 76/160/EEC. The WHO Guidelines cover general aspects of recreational water management as well as define water quality criteria for various hazards. For faecal pollution, these include faecal indicator organism concentrations and an assessment of vulnerability to faecal contamination. Central to the approach set out in the WHO Guidelines are: (i) the concept of beach profiling to produce a 'sanitary inspection category' which implies a priori hazard assessment as a core management tool and (ii) the prediction of poor water quality to assist in real time risk assessment and public health protection. These management approaches reflect a harmonized approach towards the assessment and management of risk for water-related infectious disease being applied by WHO. Numerical microbiological criteria for intestinal enterococci are proposed in the new Guidelines. These were developed using a novel approach to disease burden assessment, which has been applied to both recreational waters and urban air quality. This paper explains the scientific rationale and mathematical basis of the new approach, which is not presented in the WHO Guidelines for recreational waters.

Faecal-indicator concentrations in waters draining lowland pastoral catchments in the UK: relationships with land use and farming practices.

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.