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Aims This study aims to identify and address significant limitations in current culture-based regulatory methods used for monitoring microbiological water quality. Specifically, these methods' inability to distinguish between planktonic forms and aggregates containing higher bacterial loads and associated pathogens may lead to a severe underestimation of exposure risks, with critical public health implications. Methods and results We employed a novel methodology combining size fractionation with ALERT (Automatic Lab-in-a-vial E.coli Remote Tracking), an automated rapid method for comprehensive quantification of culturable fecal indicator bacteria (FIB). Our findings reveal a substantial and widespread presence of aggregate-bound indicator bacteria across various water matrices and geographical locations. Comprehensive bacterial counts consistently exceeded those obtained by traditional methods by significant multiples, such as an average of 3.4× at the Seine River 2024 Olympic venue, and occasionally up to 100× in irrigation canals and wastewater plant effluent. These results, supported by microscopic and molecular analyses, underscore a systematic bias in global water safety regulatory frameworks. Conclusions Our research demonstrates the inadequacy of traditional culture-based techniques in assessing microbiological risks posed by aggregate-bound FIB and associated pathogens, particularly in water matrices affected by FIB-rich fecal particles from recent sewer overflows or sediment, which can carry higher infectious risks. Incorporating comprehensive FIB analysis techniques, including molecular methods and rapid culture-based approaches as shown in this study, offers a promising and effective solution to these risk assessment limitations. [ABSTRACT FROM AUTHOR]

Marine sediments have been suggested as a reservoir for pathogenic bacteria, including Escherichia coli. The origins, and properties promoting survival of E. coli in marine sediments (including osmotolerance, biofilm formation capacity, and antibiotic resistance), have not been well-characterized. Phenotypes and genotypes of 37 E. coli isolates from coastal marine sediments were characterized. The isolates were diverse: 30 sequence types were identified that have been previously documented in humans, livestock, and other animals. Virulence genes were found in all isolates, with more virulence genes found in isolates sampled from sediment closer to the effluent discharge point of a wastewater treatment plant. Antibiotic resistance was demonstrated phenotypically for one isolate, which also carried tetracycline resistance genes on a plasmid. Biofilm formation capacity varied for the different isolates, with most biofilm formed by phylogroup B1 isolates. All isolates were halotolerant, growing at 3.5% NaCl. This suggests that the properties of some isolates may facilitate survival in marine environments and can explain in part how marine sediments can be a reservoir for pathogenic E. coli. As disturbance of sediment could resuspend bacteria, this should be considered as a potential contributor to compromised bathing water quality at nearby beaches. [ABSTRACT FROM AUTHOR]

The presence and level of faecal indicator bacteria are important factors in estimating the microbiological quality of surface water and the risk of human infection upon exposure to this water. Until 2014, ISO 9308-1:2000 was available and used to enumerate faecal indicator Escherichia coli in bathing water. In 2014, this ISO was technically revised and replaced by ISO 9308-1:2014. This ISO introduced a less selective method for enumeration of E. coli that allows non-specific growth from waters containing high levels of bacteria, such as surface waters. This implies that currently there is no suitable reference membrane filtration method for the compliance monitoring of official bathing sites for E. coli according to the European Bathing Water Directive. Here, the performance characteristics of three chromogenic culture media, namely Tryptone Bile X-glucuronide (TBX) agar, Chromogenic Coliform Agar (CCA), and CHROMagar E. coli/Coliform (ECC) were investigated at 44 °C for water with varying levels of bacteria according to ISO 13843:2017. Based on performance characteristics, colony counts, and practical usage, TBX appeared the most suitable culture medium for the enumeration of E. coli in bathing water and other waters with high levels of background bacteria, such as surface water in agricultural areas and wastewater discharge points. [ABSTRACT FROM AUTHOR]

Escherichia coli is a Gram-negative, facultative anaerobic, rod-shaped coliform bacterium which is widely used as an indicator of biological safety of water. Currently, 'gold' standard for the detection of E. coli (and other pathogens) are microbiological culture-based tests. Microbiological tests are very robust for the detection of live pathogens however it can take several days to get the results. A prospective option for the detection of E. coli in recreational waters in addition to the genetic PCR analyses can be an immunobiosensor, allowing to detect E. coli in 20 min and to issue timely warnings of the safety of water. To compare different methods – microbiological culturing, quantitative PCR analysis and antibody-based immunobiosensor for the analysis of bathing water samples from a popular urban beach Anne Canal in Tartu, Estonia were used. The median value of biosensor results for E. coliwas considerably higher than the results of microbiological cultivation and qPCR: for 40 times, and 4 times, respectively. In addition, the biosensor results were in significant correlation with the number of total coliforms. Considering the current requirements for the biological safety of bathing water, we propose that the indicative safety threshold for bathing water usingimmunosensor analyses is in the range 4 × 103–4 x 104E. coli cells/100 mL. [ABSTRACT FROM AUTHOR]

Water quality impairment is a substantial environmental hazard which impacts a wide variety of stakeholders and interests, particularly those who participate in outdoor water-based recreational activities. Recreational bathing water qualities are highly vulnerable to microbial pollution from municipal sewage, industrial effluents, agriculture run-off and river discharges. Fecal contamination impairs water quality and potentiates human health risks. The aim of this study was to see the 5-year trend of microbiological quality of recreational bathing waters in Albania. Every year we collected 1,071 samples taken 30 centimeters below the water's surface at least one meter deep. Assessment of bacterial load of the coastal waters was done nine times for every point, for the Escherichia coli and Intestinal enterococci, according to the methods ISO 7899-1 and ISO 9308-3. Bathing water assessment is to be classified according the categories indicated in the Directive 2006/7/EC and recommendations of WHO/UNEP-2010. Microbial Water Quality Assessment Category (cfu/100 ml water) done in 119 monitoring points were: During 2016, Excellent 53%, Sufficient 23%, Good 9% and Poor 15%. In 2017, Excellent 68%, Sufficient 15%, Good 6% and Poor 10%. During 2018, Excellent 82.4%, Sufficient 13%, Good 0.9% and Poor 3.7%. During 2019, Excellent 89%, Sufficient 2.5%, Good 6% and Poor 2.5%. During 2020 Excellent 89.9%, Sufficient 5%, Good 0.9%, and Poor 4.2%. Based on the above assessment, it is noticed a significant increase of microbial quality of recreational bathing waters in Albania due to investments in the sewerage system and better waste water treatment. [ABSTRACT FROM AUTHOR]

Contaminated marine bathing water has been reported to adversely affect human health. Our data demonstrated a correlation between total endotoxin (lipopolysaccharide; LPS) levels and degree of contamination of marine bathing waters. To assess the potential health implications of LPS present in marine bathing waters, the inflammation-inducing potency of water samples collected at different time points at multiple sampling sites were assessed using a cell culture-based assay. The numbers of fecal indicator bacteria (FIB) were also examined in the same samples. Water samples were used to stimulate two cell culture models: (1) a novel non-transformed continuously growing murine cell line Max Plank Institute (MPI) characteristic of alveolar macrophages and (2) human MonoMac 6 monocyte cell line. The inflammatory potential of the samples was assessed by measuring the release of inflammatory cytokines. The presence of high levels of LPS in contaminated bathing water led to induction of inflammatory response from our in vitro cell-based bioassays suggesting its potential health impact. This finding introduces an in vitro culture assay that reflects the level of LPS in water samples. These observations further promote previous finding that LPS is a reliable surrogate biomarker for fecal contamination of bathing water. [ABSTRACT FROM AUTHOR]

The Systems Approach Framework with an integrated Ecological-Social-Economic assessment was applied to address the issue of zebra mussel (Dreissena polymorpha) farming in the large Oder (Szczecin) Lagoon, southern Baltic Sea. Heavy eutrophication hampers the use of the lagoon and zebra mussel farming is considered as new use and potential measure to improve water quality. Three alternative scenarios were developed in interaction with local stakeholders: 1) the production of mussels as fresh feed and meal on a commercial basis seemed not profitable, because of a limited market for fresh mussels (zoos, aquaculture) and low prices for organic feed. 2) Mussel cultivation to improve transparency and attractiveness of bathing waters near beaches had only a limited potential (0.2 m improvement of Secchi depth). A higher mussel biomass would increase the risk of temporary hypoxia. 3) Mussels farms for improving the environmental status (according to EU Water Framework Directive) by supporting macrophyte restoration were considered as the most promising scenario. Our model simulations suggested that as soon as a compensation for nutrient removal is considered, all mussel farm scenarios could cover the costs. Experiments and literature confirm that the conditions for an environmental friendly farming approach in the lagoon are suitable. Steps towards and problems associated with an implementation, e.g. invasion of Dreissena bugensis (quagga mussel), are discussed. Each step of the Ecological-Social-Economic assessment and major lessons learnt are documented in detail. Altogether, the approach turned out to be very suitable for this issue. [ABSTRACT FROM AUTHOR]

To quantify the impact of fecal pollution on the microbiological bathing water quality, predictive modeling is being increasingly used in which the decay rate of the fecal indicators plays an important role. The decay of sewage-sourced enterococci and Escherichia coli culturable cells and their associated molecular markers (16SrRNA) quantified by Quantitative Reverse transcription PCR were measured in controlled microcosms as well in in situ conditions using different water types, from marine waters to fresh waters with intermediate salinity. All bacterial decays were fitted to a first order decay model. In the laboratory study, the light radiation was the most influent factor affecting E. coli and enterococci survival by culture methods although environmental conditions weakly impacted the decay of molecular markers. The results also indicated differential persistence of genetic markers and culturable organisms of fecal indicator bacteria in different water systems. For each bacteria indicator and analytical method, four equations were obtained to predict the time required to have a 90% reduction (T90) according irradiance, salinity and temperature parameters. The weighted model RMSE (Root Mean Square Error) calculated for all field experiments showed that quantification obtained with the equations defined by laboratory-based study compared reasonably well with in-situ observed quantification (0.4 and 0.2 log by standard culture methods for E. coli and Enterococcus spp. and 0.6 and 0.3 log by RT-qPCR for E. coli and Enterococcus spp. respectively). The modeling tool can be used to predict the presence of fecal pollution in marine and fresh waters in combination with either culture based- or rapid molecular methods. [ABSTRACT FROM AUTHOR]

This study developed, cross-validated and applied a regression-based model to predict concentrations of faecal indicator organisms (FIOs) under different environmental conditions in the North and South bays of Santa Catarina, South of Brazil. The model was developed using a database of FIO concentrations in seawater sampled at 50 sites and the validation was performed using a different database by comparing 288 pairs of measured and modelled results for 15 sites. The index of agreement between the model outputs and the FIO concentrations measured during the validation period was 66%; the mean average error was 0.43 log 10 and the root mean square error was 0.58 log 10 MPN.100 mL −1 . These validation results indicate that the model provides a fair representation of the FIO contamination in the bays for the meteorological conditions under which the model was trained. The simulation of different scenarios showed that under typical levels of resident human population in the catchments and median rainfall and solar radiation conditions, the median FIO concentration in the bays is 0.4 MPN.100 mL −1 . Under extreme meteorological conditions, the combined effect of high rainfall and low solar radiation increased FIO concentrations up to 5 log 10 MPN.100 mL −1 . The simulated scenarios also show that increases in resident population during the summer tourist season and average rainfall concentrations do not increase median FIO concentrations in the bays relative to periods of time with average population, possibly because of higher bacterial die-off in the waters. The models can be an effective tool for management of human health risks in bathing and shellfish waters impacted by sewage pollution. [ABSTRACT FROM AUTHOR]

In recent years, global warming has led to a growing number of Vibrio cholerae infections in bathing water users in regions formerly unaffected by this pathogen. It is therefore of high importance to monitor V. cholerae in aquatic environments and to elucidate the main factors governing its prevalence and abundance. For this purpose, rapid and standardizable methods that can be performed by routine water laboratories are prerequisite. In this study, we applied a recently developed multiplex quantitative PCR (qPCR) strategy (i) to monitor the spatiotemporal variability of V. cholerae abundance in two small soda pools and a large lake that is intensively used for recreation and (ii) to elucidate the main factors driving V. cholerae dynamics in these environments. V. cholerae was detected with qPCR at high concentrations of up to 970,000 genomic units 100 ml-1 during the warm season, up to 2 orders of magnitude higher than values obtained by cultivation. An independent cytometric approach led to results comparable to qPCR data but with significantly more positive samples due to problems with DNA recovery for qPCR. Not a single sample was positive for toxigenic V. cholerae, indicating that only nontoxigenic V. cholerae (NTVC) was present. Temperature was the main predictor of NTVC abundance, but the quality and quantity of dissolved organic matter were also important environmental correlates. Based on this study, we recommend using the developed qPCR strategy for quantification of toxigenic and nontoxigenic V. cholerae in bathing waters with the need for improvements in DNA recovery. [ABSTRACT FROM AUTHOR]

Introduction: The twinning project entitled "Alignment in Bathing Water Monitoring" was implemented within the framework of EU Pre-Accession Financial Assistance IPA I-2010 Programming by the Ministry of Health, Public Health Institution of Turkey together with France-Italy consortium between the I. quarter of 2013 and II. quarter of 2015. The major aims of the project was to transpose the new bathing water Directive 2006/7/EC into the Turkish National Legislation and strengthening the bathing water quality monitoring system of Ministry of Health - Public Health Institution of Turkey within the framework of the new directive. Methods: The project was designed such as to ensure the alignment of the new bathing water Directive 2006/7/EC and preparation of the draft bylaw on bathing water quality management, transition from 76/160/EEC to 2006/7/EC Directive regarding the classification and quality assessment of bathing water starting gradually with the pilot applications and then disseminating to the whole bathing areas, the practice of establishing experimental bathing water profiles in the selected areas of the pilot provinces, compiling of sets of bathing water quality data, improvement of the bathing water quality monitoring system of the Ministry of Health in the direction of 2006/7/EC Directive, improvement of the technical capacity of the Public Health Laboratories to perform analysis according to the new bathing water directive. Results: The draft "By-law on bathing water quality management" has been prepared with the transposition of 2006/7/EC EU Directive by the Ministry of Health, Public Health Institution of Turkey with the contribution of the relevant institutions. Trainings and workshops were organized. Six guidance documents were prepared, published and distributed. Data quality assessment and classification practice were done at pilot provinces Antalya, Çanakkale and İstanbul. Conclusion: The all activities given in the project contract have been successfully completed and all project mandatory results were reached. The technical and institutional capacity for the full implementation of the 2006/7/EC Directive by the Ministry of Health was impoved successfully with the project. [ABSTRACT FROM AUTHOR]

This study investigated the public health risk from exposure to infectious microorganisms at Sandvika recreational beaches, Norway and dose–response relationships by combining hydrodynamic modelling with Quantitative Microbial Risk Assessment (QMRA). Meteorological and hydrological data were collected to produce a calibrated hydrodynamic model using Escherichia coli as an indicator of faecal contamination. Based on average concentrations of reference pathogens (norovirus, Campylobacter , Salmonella , Giardia and Cryptosporidium ) relative to E. coli in Norwegian sewage from previous studies, the hydrodynamic model was used for simulating the concentrations of pathogens at the local beaches during and after a heavy rainfall event, using three different decay rates. The simulated concentrations were used as input for QMRA and the public health risk was estimated as probability of infection from a single exposure of bathers during the three consecutive days after the rainfall event. The level of risk on the first day after the rainfall event was acceptable for the bacterial and parasitic reference pathogens, but high for the viral reference pathogen at all beaches, and severe at Kalvøya-small and Kalvøya-big beaches, supporting the advice of avoiding swimming in the day(s) after heavy rainfall. The study demonstrates the potential of combining discharge-based hydrodynamic modelling with QMRA in the context of bathing water as a tool to evaluate public health risk and support beach management decisions. [ABSTRACT FROM AUTHOR]

The purpose of this equivalence study was to compare an alternative method, Colilert-18 Quanti-Tray (ISO 9308-2) with the European bathing water directive (2006/7/EC) reference method, the miniaturised most probable number (MMPN) method (ISO 9308-3), for the analysis of Escherichia coli. Six laboratories analysed a total of 263 bathing water samples in Finland. The comparison was carried out according to ISO 17994:2004. The recovery of E. coli using the Colilert-18 method was 7.0% and 8.6% lower than that of the MMPN method after 48 hours and 72 hours of incubation, respectively. The confirmation rate of presumptive E. coli-positive wells in the Colilert-18 and MMPN methods was high (97.8% and 98.0%, respectively). However, the testing of presumptive E. colinegative but coliform bacteria-positive (yellow but not fluorescent) Colilert-18 wells revealed 7.3% false negative results. There were more false negatives in the naturally contaminated waters than in the samples spiked with waste water. The difference between the recovery of Colilert-18 and the MMPN method was considered not significant, and subsequently the methods are considered as equivalent for bathing water quality monitoring in Finland. Future bathing water method equivalence verification studies may use the data reported herein. The laboratories should make sure that any wells showing even minor fluorescence will be determined as positive for E. coli. [ABSTRACT FROM AUTHOR]

Most studies characterize microbial source tracking (MST) target performance using sensitivity and specificity metrics. However, it is important to also consider the temporal stability of MST targets in relation to regulated microbial pollutants. Differences among bacterial target stabilities may lead to erroneous conclusions about sources of contamination. The present study evaluates the relative stability of MST targets and fecal indicator organisms using the gull/pigeon-associated Catellicoccus marimammalium (CAT) marker and enterococci (ENT). The decay rates of CAT and ENT measured by culture (cENT) and QPCR (tENT) were compared in sand and seawater laboratory microcosms under environmentally relevant conditions (subject to tidal wetting versus no wetting in sand, and sunlit versus dark conditions in seawater). Bacterial targets were more persistent in beach sand than in seawater with decay rates on the order of 0.01–0.1 per day and 1 to 10 per day, respectively. Targets were more persistent in unwetted compared to wetted sand, and dark compared to sunlit seawater. During the first 8 days of the sand experiment, the decay rate k of CAT was greater than that of cENT. The decay rates of CAT, tENT, and cENT were similar in sand after day 8 and in dark seawater. In sunlit seawater, the decay rates were different between targets with k cENT > k CAT > k tENT . The decay rates presented here are useful for fate and transport models and also inform the use of MST marker concentrations to infer ENT sources in the environment. [ABSTRACT FROM AUTHOR]

Pollution of surface waters with pathogens from combined sewer overflows limits recreational use of surface waters. Large retention basins are a satisfactory solution but they are rarely sufficient for economic or space reasons. Fast disinfection during the overflow is an alternative, but few methods are known and each has problems. This work evaluated for the first time the full-scale disinfection using performic acid by the removal of the two currently regulated indicator bacteria for bathing water quality, Escherichia coli and Enterococcus . Experiments were performed at a sewage bypass through a sea-outfall pipe with a minimum hydraulic retention time of 24 min. The disinfection efficiency in the field was measured by analyzing samples taken before and after the treatment. Samples were also treated with performic acid in the laboratory to measure the disinfection effectiveness and kinetic of degradation of performic acid. Doses of 1–8 ppm of performic acid achieved 1.0–3.5 log removal of E. coli and 1.0–2.44 log removal of Enterococcus in the field, but were somewhat higher in laboratory conditions at 1.69–4.38 and 1.0–4.27 log units, respectively. Studies of the degradation of performic acid in collected real samples showed more than 50% was degraded in 20 min, and mostly degraded by 120 min. Comparison of field and laboratory dosed samples detected that performic acid synthesis did not start in one event and clogging of the sampler in another event. Overall the tests showed that the treatment was successful but it is indicated that online control could benefit treatments efficiency. [ABSTRACT FROM AUTHOR]

Waterborne enteric viruses may pose disease risks to bather health but occurrence of these viruses has been difficult to characterize at recreational beaches. The aim of this study was to evaluate water for human virus occurrence at two Southern California recreational beaches with a history of beach closures. Human enteric viruses (adenovirus and norovirus) and viral indicators (F+ and somatic coliphages) were measured in water samples over a 4-month period from Avalon Beach, Catalina Island (n = 324) and Doheny Beach, Orange County (n = 112). Human viruses were concentrated from 40 L samples and detected by nested reverse transcriptase polymerase chain reaction (PCR). Detection frequencies at Doheny Beach were 25.5% (adenovirus) and 22.3% (norovirus), and at Avalon Beach were 9.3% (adenovirus) and 0.7% (norovirus). Positive associations between adenoviruses and fecal coliforms were observed at Doheny (p = 0.02) and Avalon (p = 0.01) Beaches. Human viruses were present at both beaches at higher frequencies than previously detected in the region, suggesting that the virus detection methods presented here may better measure potential health risks to bathers. These virus recovery, concentration, and molecular detection methods are advancing practices so that analysis of enteric viruses can become more effective and routine for recreational water quality monitoring. [ABSTRACT FROM AUTHOR]

A total of 50 Finnish bathing water samples and 34 sewage effluent samples originating from 17 locations were studied in the summers of 2006 and 2007. Campylobacter were present in 58% and adenoviruses in 12% of all bathing water samples; 53% of all sewage effluent samples were positive for Campylobacter spp. and 59% for adenoviruses. C. jejuni was the most common Campylobacter species found and human adenovirus serotype 41 was the most common identified adenovirus type. Bathing water temperature displayed a significant negative relationship with the occurrence of Campylobacter. One location had identical pulsed-field gel electrophoresis patterns of C. coli isolates in the bathing water and in sewage effluent, suggesting that sewage effluent was the source of C. coli at this bathing site. The counts of faecal indicator bacteria were not able to predict the presence of Campylobacter spp. or adenoviruses in the bathing waters. Thus the observed common presence of these pathogens in Finnish sewage effluents and bathing waters may represent a public health risk. The low water temperature in Finland may enhance the prevalence of Campylobacter in bathing waters. More attention needs to be paid to minimizing the concentrations of intestinal pathogens in bathing waters. [ABSTRACT FROM AUTHOR]

In early autumn 2011, three dogs died after they had been exposed to a Microcystis aeruginosa bloom on Lake Amstelmeer, The Netherlands. The cyanobacterial scum from the lake contained up to 5.27 x 103 μg g-1 dry-weight microcystin, the vomit of one of the dogs contained on average 94 μg microcystin g-1 dry-weight. In both cases, microcystin-LR was the most abundant variant. This is the first report of dog deaths associated with a Microcystis bloom and microcystin poisoning in The Netherlands. [ABSTRACT FROM AUTHOR]

Abstract: In April and May 2011, three dogs died and one dog became ill after swimming in Lake IJmeer (The Netherlands). At the time, the lake was infested with the benthic cyanobacterial species Phormidium. A Eurasian Coot (Fulica atra) and a Black-headed Gull (Chroicocephalus ridibundus) also died near Lake IJmeer in the same period. One of the dogs and both birds were subjected to a pathological investigation. Furthermore, the Phormidium mat; algal samples from the dikes; contents of the animals’ digestive systems and organ tissues were analysed for the following cyanobacterial toxins: (homo)anatoxin-a; (7-deoxy-)cylindrospermopsin; saxitoxins and gonyautoxins by LC-MS/MS. Samples were also analysed for the nontoxic (homo)anatoxin-a metabolites dihydro(homo)anatoxin-a and epoxy(homo)anatoxin-a. The dog necropsy results indicated neurotoxicosis and its stomach contained Phormidium filaments. Anatoxin-a was detected in the Phormidium mat (272μgg−1 dry weight, stdev 65, n =3) and in the dog’s stomach contents (9.5μgg−1 dry weight, stdev 2.4, n =3). Both samples also contained the anatoxin-a metabolite dihydroanatoxin-a, and a trace of homoanatoxin-a was detected in the Phormidium mat. The birds were in bad nutritive condition at the time of necropsy and their stomachs and intestines did not contain any cyanobacterial material. Furthermore, no cyanobacterial toxins were detected in their stomachs, intestines and organs and they both had lesions that are not associated with cyanobacterial intoxication. This is the first report of anatoxin-a and homoanatoxin-a occurrence in The Netherlands, these toxins have likely caused the deaths of three dogs. The birds probably died of other causes. Dutch recreational waters are at this moment only screened for pelagic cyanobacterial species, the current bathing water protocol therefore does not protect humans and animals from negative effects of blooms of benthic cyanobacteria. [Copyright &y& Elsevier]

Abstract: For accompanying actions to pollution crisis management of recreational waters, rapid methods providing a quantified faecal indicator which can be completed in less than half a day, are currently needed. Two sensitive rRNA-targeted Reverse Transcription-quantitative PCR (RT-qPCR) methods were developed for quantifying Escherichia coli and Enterococcus spp. cells in marine bathing waters. Threshold cycle (Ct) values were found to be linear with a target quantity over a 4log dynamic range (from 105 to 100 cell equivalents per 100ml). In order to confirm the scope of application of such alternative methods, 80 and 85 seawater samples from the Atlantic Ocean and Mediterranean sea were analysed by both RT-qPCR and ISO culture-based methods for E. coli and Enterococcus spp. enumerations. This study demonstrates that by giving reliable results in 3h, the RT-qPCR method has high potential as a rapid test for recreational water quality monitoring. In natural waters, significant linear log–log relations between the RT-qPCR and culture method measurements for E. coli and Enterococcus spp. assays were shown (correlation coefficient (r) values of 0.814 and 0.715, p <0.0001, respectively). The sensitivities (defined as the probability of a sample testing positive if the criterion number is exceeded) of RT-qPCR for E. coli and Enterococcus spp. were respectively 94.4% and 83.9%. By contrast, specificities (defined as the probability of a test being negative if a contamination is truly absent) could be considered as lower (65.9% and 50.0% for E. coli and Enterococcus spp.). This reflects that RT-qPCR assays detect all viable cells (culturable and non-culturable) whereas culture methods only detect cells which are viable and culturable. [Copyright &y& Elsevier]

Abstract: In 2009, four bathing sites in The Netherlands were monitored for potentially human pathogenic Vibrio species to observe possible associations with environmental conditions and health complaints. Three slightly different enrichment procedures were used to isolate Vibrio species with different growth requirements. Waters were generally positive for Vibrio from May until October; median Vibrio concentrations ranged from 4 to 383 MPN per litre (maximum 105 MPN per litre). Isolated Vibrio species included V. alginolyticus (50.6%) and V. parahaemolyticus (8.5%) from bathing sites with salinities ranging between 2.8 and 3.5% and V. cholerae non-O1/O139 (6.9%) from sites with salinities ranging between 0.007 and 0.08%. Although more samples were positive for Vibrio at elevated water temperatures, a quantitative relation between Vibrio numbers in water samples and the water temperature was not observed which may be explained by maximum water temperatures of 21°C. Active surveillance yielded one case of a recreational water related Vibrio infection. V. cholerae non-O1/O139 was cultured from the patient''s wound and the implicated recreational water; PFGE profiles of the water and patient isolates were not identical. The number of patients that contract a Vibrio infection through exposure to Dutch recreational waters seems low, but may be underestimated. The common occurrence of Vibrio species in these waters stresses the need for providing information on Vibrio to risk groups to prevent infections. [Copyright &y& Elsevier]

Abstract: Bathing water compliant with bathing water legislation may nevertheless contain pathogens to such a level that they pose unacceptable health risks for swimmers. Quantitative Microbiological Risk Assessment (QMRA) can provide a proper basis for protective measures, but the required data on swimmer exposure are currently limited or lacking. The objective of this study was to collect exposure data for swimmers in fresh water, seawater and swimming pools, i.e. volume of water swallowed and frequency and duration of swimming events. The study related to swimming in 2007, but since the summer of 2007 was wet and this might have biased the results regarding surface water exposure, the study was repeated relating to swimming in 2009, which had a dry and sunny summer. Exposure data were collected through questionnaires administered to approximately 19 000 persons representing the general Dutch population. Questionnaires were completed by 8000 adults of whom 1924 additionally answered the questions for their eldest child (<15 years). The collected data did not differ significantly between 2007 and 2009. The frequency of swimming and the duration of swimming were different for men, women and children and between water types. Differences between men and women were small, but children behaved differently: they swam more often, stayed in the water longer, submerged their heads more often and swallowed more water. Swimming pools were visited most frequently (on average 13–24 times/year) with longest duration of swimming (on average 67–81 min). On average, fresh and seawater sites were visited 6–8 times/year and visits lasted 41–79 min. Dependent on the water type, men swallowed on average 27–34 ml per swimming event, women 18–23 ml, and children 31–51 ml. Data on exposure of swimmers to recreational waters could be obtained by using a questionnaire approach in combination with a test to measure mouthfuls of water for transformation of categorical data to numerical data of swallowed volumes of water. Previous assumptions on swimmer exposure were replaced with estimates of exposure parameters, thus reducing uncertainty in assessing the risk of infection with waterborne pathogens and enabling identification of risk groups. QMRA for Cryptosporidium and Giardia was demonstrated based on data from previous studies on the occurrence of these pathogens in recreational lakes and a swimming pool. [Copyright &y& Elsevier]

Abstract: 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, ) 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. [ABSTRACT FROM AUTHOR]

Abstract: There has been an ongoing dilemma for agencies that set criteria for safe recreational waters in how to provide for a seasonal assessment of a beach site versus guidance for day-to-day management. Typically an overall ‘safe’ criterion level is derived from epidemiologic studies of sewage-impacted beaches. The decision criterion is based on a percentile value for a single sample or a moving median of a limited number (e.g. five per month) of routine samples, which are reported at least the day after recreator exposure has occurred. The focus of this paper is how to better undertake day-to-day recreational site monitoring and management. Internationally, good examples exist where predictive empirical regression models (based on rainfall, wind speed/direction, etc.) may provide an estimate of the target faecal indicator density for the day of exposure. However, at recreational swimming sites largely impacted by non-sewage sources of faecal indicators, there is concern that the indicator-illness associations derived from studies at sewage-impacted beaches may be inappropriate. Furthermore, some recent epidemiologic evidence supports the relationship to gastrointestinal (GI) illness with qPCR-derived measures of Bacteroidales/Bacteroides spp. as well as more traditional faecal indicators, but we understand less about the environmental fate of these molecular targets and their relationship to bather risk. Modelling pathogens and indicators within a quantitative microbial risk assessment framework is suggested as a way to explore the large diversity of scenarios for faecal contamination and hydrologic events, such as from waterfowl, agricultural animals, resuspended sediments and from the bathers themselves. Examples are provided that suggest that more site-specific targets derived by QMRA could provide insight, directly translatable to management actions. [ABSTRACT FROM AUTHOR]

The article provides insights into issues concerning to Marine Strategy Directive (MSD) in Europe. According to the author, MSD is framework legislation which more than double the geographic scope of European Commission environmental law. The author adds that the MSD comprises all marine waters, including sea-bed and subsoil, as measured from the coastal baseline to the extent of Member State jurisdiction.

Abstract: A simple model predicting bathing water concentrations of Escherichia coli from livestock in the Irvine catchment in SW Scotland has been adapted for intestinal enterococci (IE). This has been used to predict risk of bather illness by extrapolation of published data on bather IE exposure vs incidence of gastro-enteritis. Simulated reduction in the risk of illness by reduced faecal loading was multiplied by a willingness to pay for risk reduction to estimate the annual benefits of mitigation. Health benefits of reducing loading by 75% at Irvine Beach were estimated by a willingness to pay method to be about £276kpa. Estimated annualised costs of diffuse pollution mitigation measures across the catchment were higher (>£1m), and it is very unlikely that 75% mitigation is achievable with current stocking rates. Further work should explore the influence of uncertainty of model parameters, and use emerging epidemiological information on specific zoonotic pathogens such as E. coli O157 and Cryptosporidium. Other components of the value of clean water should also be included to obtain a complete estimate of the cost:benefit of mitigation. [Copyright &y& Elsevier]

The objectives of this study were (i) to apportion sources of faecal coliforms (FC) and faecal streptococci (FS) loads from a dairy farm to a stream in the Irvine catchment; (ii) to assess efficacy of pollution mitigation measures installed on this farm; (iii) to assess frequency with which observed FC loads contribute to high risk of failure of coastal bathing water to meet microbial standards. FC and FS loads in a primary stream running through the farm were estimated at up to four stations (above the farm, above the steading, below the farm and below a pre-existing 0.6ha area of open water/wetland), in summer 2004 and 2005. During this period, steading and field mitigation measures were being installed. We estimated that farm FC loads likely to cause bathing water failure were 8.9×108 colony-forming units (cfu) ha−1d−1 (guideline standards) and 1.7×1010 cfu ha−1d−1 (mandatory standards). In 2005, the guideline exceedance risk (42%) was associated about equally with field and steading sources, whereas the mandatory exceedance risk (11%) was mainly due to steading sources. The pond/wetland below the farm reduced these exceedance risks to 20% and <1%, respectively, but was a source of FS at high discharge. The exceedence curves below the farm were similar in 2004 and 2005, despite lower flows and installation of pollution mitigation measures. The results suggest that further mitigation efforts on this farm should be focused on improving management of the steading FC sources and that wetlands to receive steading runoff can be cost-effective mitigation tools for faecal indicator pollution. [Copyright &y& Elsevier]

The occurrence and pathogenicity of vibrios in bathing water were investigated along the Conero Riviera (Adriatic Sea, Central Italy). Vibrio spp. enumeration was performed on thiosulfate-citrate-bile-salts-sucrose-agar by the membrane filter method, and identification was done through a biochemical protocol. All isolates were tested for the presence of cytotoxicity, protease, lipase, elastase, gelatinase, urease, haemolytic activity, ctx, tdh and trh genes by conventional methods. In all, 200 vibrios were isolated from 132 samples that were analysed. Vibrio harveyi and Vibrio alginolyticus were the species most frequently recovered. All strains were cytotoxic and some of them showed protease, gelatinase, lipase, elastase, urease and haemolytic activity. One isolate of V. alginolyticus and one of V. harveyi had the trh gene, while another strain of V. harveyi and one of Vibrio parahaemolyticus had the ctx gene. These results demonstrate the presence of potentially pathogenic vibrios in the Conero Riviera and the risk of infection due to bathing water exposure. [Copyright &y& Elsevier]

Abstract: 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]. [Copyright &y& Elsevier]

Abstract: 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. [Copyright &y& Elsevier]

Abstract: ‘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. [Copyright &y& Elsevier]

This paper primarily addresses underlying laws and regulations related to protection of the coastal environment and examines their implementation. In this context, Directive 76/160/EEC which is the leading directive on the quality of bathing water and its revision, i.e. Commission Proposal COM (2000) 860 Final, is investigated particularly and criticized on technical and scientific manner; and similar and dissimilar issues between the Community Directives and Turkish Laws are evaluated. The conducted study reveals that although the Turkish legislation in the field of water quality is largely in line with the acquis, further efforts are also needed with respect to implementation in order to achieve full harmonization. [ABSTRACT FROM AUTHOR]

Achieving compliance with the mandatory standards of the 1976 Bathing Water Directive (76/160/EEC) is required at all UK identified bathing waters. In recent years, the Fylde coast has been an area of significant investments in 'point source' control, which have not proven, in isolation, to satisfactorily achieve compliance with the mandatory, let alone the guide, levels of water quality in the Directive. The potential impact of riverine sources of pollution was first confirmed after a study in 1997. The completion of sewerage system enhancements offered the potential for the study of faecal indicator delivery from upstream sources comprising both point sources and diffuse agricultural sources. A research project to define these elements commenced in 2001. Initially, a desk study reported here, estimated the principal infrastructure contributions within the Ribble catchment. A second phase of this investigation has involved acquisition of empirical water quality and hydrological data from the catchment during the 2002 bathing season. These data have been used further to calibrate the 'budgets' and 'delivery' modelling and these data are still being analysed. This paper reports the initial desk study approach to faecal indicator budget estimation using available data from the sewerage infrastructure and catchment sources of faecal indicators. [ABSTRACT FROM AUTHOR]

Abstract: Intestinal enterococci are the principal ‘health-evidence-based’ parameter recommended by WHO for the assessment of marine recreational water compliance. Understanding the survival characteristics of these organisms in nearshore waters is central to public health protection using robust modelling to effect real-time prediction of water quality at recreation sites as recently suggested by WHO and the Commission of the European Communities Previous models have more often focused on the coliform parameters and assumed two static day-time and night-time T 90 values to characterise the decay process. The principal driver for enterococci survival is the received dose of irradiance from sunlight. In the water column, transmission of irradiance is determined by turbidity produced by suspended material. This paper reports the results of irradiated microcosm experiments using simulated sunlight to investigate the decay of intestinal enterococci in relatively turbid estuarine and coastal waters collected from the Severn Estuary and Bristol Channel, UK. High-turbidity estuarine waters produced a T 90 value of 39.5h. Low-turbidity coastal waters produced a much shorter T 90 value of 6.6h. In experiments receiving no irradiation, high-turbidity estuarine waters also produced a longer T 90 of 65.1h compared with corresponding low-turbidity coastal waters, T 90 24.8h. Irradiated T 90 values were correlated with salinity, turbidity and suspended solids (, ). The results suggest that enterococci decay in irradiated experiments with turbidity >200 NTU is similar to decay observed under dark conditions. Most significantly, these results suggest that modelling turbidity and or suspended solids offers a potential means of predicting T 90 values in ‘real-time’ for discrete cells of a hydrodynamic model. [Copyright &y& Elsevier]

Three alternative approaches to predicting delivery of faecal indicators from livestock sources to surface water in the catchment of the River Irvine, Ayrshire, Scotland, are described. These are a soil transport model which assumes all E. coli are transported through the soil, a regression model using observed E. coli concentrations in surface waters, and a distributed catchment model (PAMIMO). Each of these is linked to a simple group of equations describing inputs of E. coli from livestock to land, transport and inactivation in the river Irvine and mixing and inactivation in the sea. The models predict E. coli content of bathing water for Irvine beach. The regression model gives the best predictions of bathing water quality. The low values predicted by the soil transport model suggests that preventing surface runoff of faecal indicators from livestock would provide an adequate solution to the problem of bathing water contamination. [Copyright &y& Elsevier]

Somatic coliphages, F-specific RNA bacteriophages, bacteriophages infecting Bacteroides fragilis, Escherichia coli and enterococci were counted in bathing waters in the late spring and summer. We tested fresh and marine bathing waters from North, South, East and West Europe expected to contain between 100 and 500 E. coli per 100 ml, although wider ranges were sometimes found. Bacteriophages were counted after concentration, since a preliminary study proved that this step was necessary to obtain positive counts. During monitoring, a first-line quality control with reference materials for bacteria and bacteriophages was performed by all the laboratories participating in the study. The same microbes were also counted in raw sewage samples from various areas in Europe, where the bacterial indicators and the three groups of bacteriophages were detected in roughly the same numbers. All groups of bacteriophages were detected in both fresh and marine bathing waters throughout Europe. Reliable and complete results from 147 samples showed that for log-transformed values, E. coli and bacteriophages were slightly correlated. However, the slope of the regression line changed according to E. coli concentration and the correlation diminished when this concentration was close to zero per 100 ml. The ratios between E. coli and phages in bathing waters differed significantly from those in sewage. The relative amounts of bacteriophages, mainly somatic coliphages and phages infecting Bact. fragilis RYC2056, increased in bathing waters with low E. coli concentration, especially in seawater samples containing <100 E. coli per 100 ml. The relationship of bacteriophages with respect to enterococci paralleled that of bacteriophages with respect to E. coli. Somatic coliphages and bacteriophages infecting Bact. fragilis are useful to predict the presence of some pathogens with the same origin as present bacterial indicators but with higher survival rates. [Copyright &y& Elsevier]

During the European project ‘Bacteriophages in bathing waters’ (January 1996–June 1999), research was carried out to optimise the method for detection and enumeration of F-specific (RNA) phages in water. It was evaluated whether further optimisation would be possible/needed for the procedure as described in the standard method of the International Organisation for Standardisation (ISO) 10705-1. The research focused mainly on optimisation of the different steps for culturing the host strain WG49 Salmonella Typhimurium. It was concluded that all steps described in ISO 10705-1 are necessary and, if followed carefully, using a culture of host strain WG49 Salmonella Typhimurium of good quality, reliable results could be obtained for the enumeration of F-specific RNA phages. [Copyright &y& Elsevier]

• Europe-wide analysis of organisms of public health concern in bathing waters. • Critical lack of surveillance of pathogenic organisms in bathing water is evident. • Need to integrate current monitoring methods with more inclusive parameters. • Highlight's health risk from organisms undetected by current regulatory methods. • Greater understanding of health risk associated with bathing waters is needed. Globally, water-based bathing pastimes are important for both mental and physical health. However, exposure to waterborne organisms could present a substantial public health issue. Bathing waters are shown to contribute to the transmission of illness and disease and represent a reservoir and pathway for the dissemination of antimicrobial resistant (AMR) organisms. Current bathing water quality regulations focus on enumeration of faecal indicator organisms and are not designed for detection of specific waterborne organisms of public health concern (WOPHC), such as antimicrobial resistant (AMR)/pathogenic bacteria, or viruses. This investigation presents the first scoping review of the occurrence of waterborne organisms of public health concern (WOPHC) in identified natural bathing waters across the European Union (EU), which aimed to critically evaluate the potential risk of human exposure and to assess the appropriateness of the current EU bathing water regulations for the protection of public health. Accordingly, this review sought to identify and synthesise all literature pertaining to a selection of bacterial (C ampylobacter spp., Escherichia coli, Salmonella spp. , Shigella spp. , Vibrio spp. , Pseudomonas spp., AMR bacteria), viral (Hepatitis spp. , enteroviruses , rotavirus, adenovirus, norovirus), and protozoan (Giardia spp., and Cryptosporidium spp.) contaminants in EU bathing waters. Sixty investigations were identified as eligible for inclusion and data was extracted. Peer-reviewed investigations included were from 18 countries across the EU, totalling 87 investigations across a period of 35 years, with 30% published between 2011 and 2015. A variety of water bodies were identified, with 27 investigations exclusively assessing coastal waters. Waterborne organisms were classified into three categories; bacteria, viruses, and protozoa; amounting to 58%, 36% and 17% of the total investigations, respectively. The total number of samples across all investigations was 8,118, with detection of one or more organisms in 2,449 (30%) of these. Viruses were detected in 1281 (52%) of all samples where WOPHC were found, followed by bacteria (865(35%)) and protozoa (303(12%)). Where assessed (442 samples), AMR bacteria had a 47% detection rate, emphasising their widespread occurrence in bathing waters. Results of this scoping review highlight the potential public health risk of exposure to WOPHC in bathing waters that normally remain undetected within the current monitoring parameters. [Display omitted] [ABSTRACT FROM AUTHOR]

As part of a prospective cohort study among triathletes to determine a relationship between the microbiological quality of fresh bathing water and the risk of acquiring an intestinal infection, the exposure of the triathletes to microbiological contaminants was assessed. Waters were collected at seven triathlons (swimming course 1-1.5km) held in the summer of 1993 and 1994 to have a range of water qualities. All were influenced by sewage effluents, most also by agricultural run-off. Samples were collected several weeks before the event to establish a sampling programme (1993) and during the actual exposure of the triathletes (1993 and 1994) and examined for thermotolerant coliforms alone (samples preceding the event) and for E. coli, faecal enterococci, Staphylococcus aureus, F-specific RNAphages, enteroviruses (1993 and 1994) and for thermophilic Campylobacter. Salmonella, Aeromonas, Plesiomonas shigelloides and Pseudomonas aeruginosa (1993). The samples taken in the weeks before the exposure showed significant differences in thermotolerant coliform concentration between locations, depths and times. Also during swimmer exposure, significant differences occurred in microorganism levels at the different sampling points over the swimming course. As the triathletes swam as a group, they were exposed to approximately the same water at the same time. The geometric mean concentration was used to characterise each site. In the epidemiological study, the risk of an intestinal infection correlated with the concentration of thermotolerant coliforms and E. coli but not with the other parameters. The geometric mean concentration of thermotolerant coliforms at the triathlons ranged from 11-330/100mL and 54-1,200/100mL E. coil Ranking of the seven sites by faecal pollution level, based on the geometric mean concentration of a faecal indicator, resulted in a different ranking for each indicator. At the fresh water sites studied, only the ratio between the geometric mean density of E. coli and thermotolerant coliforms was constant. The ratio between the other parameters related to faecal pollution (faecal enterococci, F-specific RNA phages, enteroviruses) varied considerably. Water quality standards relating to faecal pollution can only be based on parameters that show a significant correlation with risk of intestinal illness. [ABSTRACT FROM AUTHOR]

Antibiotic resistance represents the greatest challenge to healthcare systems around the world. As antibiotic resistance genes (ARGs) are shed in faeces, many studies have focused on how wastewater effluent contributes to ARG pollution in rivers. However, small urban streams and bathing waters not impacted by treated wastewater have received little attention though they may be important reservoirs of ARGs. The main objective of this study was to assess the extent to which ARG and faecal pollution impact small urban streams and bathing waters and to determine if there is a relationship between these contaminants. For one year, bi-monthly water samples were collected from two urban streams and Dublin city's three designated bathing waters. The Liffey Estuary, that receives treated wastewater, was also sampled. The sul1 , tet (O), qnrS , bla TEM , bla SHV and bla CTX-M ARGs were quantified. E. coli and intestinal enterococci levels were determined and the source of faecal pollution (human, dog, gull) quantified by microbial source tracking. Our results show that the Liffey Estuary, the urban streams and the bathing waters are highly impacted by ARGs and human faeces. There were clear correlations between all of the studied faecal indicators and ARGs in the Liffey Estuary. In the urban streams relationships were observed for only some of the ARGs and faecal indicators, which is likely a result of non-continuous sewage leaks and overflows to the streams. Similarly, only some ARGs correlated with faecal indicators in the urban bathing waters. The source of ARGs in the bathing waters is likely to be multifaceted as we detected sporadic dog and gull faecal markers. This study demonstrates that small urban streams and bathing waters are reservoirs of ARGs and that they may pose a previously unrecognised public health risk as they have the potential to transmit enteric pathogens and antibiotic resistance determinants. Unlabelled Image • All faecal indicators and ARGs correlated in the wastewater impacted Liffey Estuary • Urban streams were intermittently impacted by high levels of faecal indicators. • ARGs and faecal indicators sporadically correlate in small urban streams. • Faecal indicators and ARGs sporadically correlated in urban bathing waters. • Human, gull and dog source tracking markers were quantified in urban bathing waters. [ABSTRACT FROM AUTHOR]