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Numerous genetic markers for microbial source tracking (MST) have been evaluated by testing a panel of target and nontarget faecal samples. However, the performance of MST markers may vary between faecal and water samples, thereby resulting in inaccurate water quality assessment. In this study, a 30-day sampling study was conducted in an urban river impacted by human- and sewage-associated pollution to evaluate the performance of different human-associated markers in environmental water. Additionally, marker decay was assessed via a microcosms approach. Overall, Bacteroidales 16sRNA and crAssphage markers exhibited higher prevalence in the study area, and their detection frequencies exceeded 90%. In contrast, Bacteroidales protein markers exhibited poor detection frequencies compared to other markers, with the prevalence of Hum2 and Hum163 reaching only 63% and 84%, respectively. Regarding marker abundance, there was no significant difference in the detection concentrations between Bacteroidales 16sRNA and crAssphage markers (p > 0.05); however, the concentrations of Bacteroidales protein markers were nearly 1 order of magnitude lower than those of other MST markers. The microcosm experiments indicated that the decay rate of crAssphage markers was significantly lower than that of other bacterial target markers, which may improve their detectability when the pollution source is located far from the sampling site. Due to the observed differences in performance and decay patterns among Bacteroidales 16sRNA, crAssphage, and Bacteroidales protein markers, we recommend the simultaneous use of multiple markers from different target microorganisms to obtain a more comprehensive understanding of the pollution sources. This approach would also provide an accurate assessment of pollution levels and health risks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Pepper mild mottle virus (PMMoV), a plant pathogenic virus belonging to the family Virgoviridae, has been proposed as a potential viral indicator for human faecal pollution in aquatic environments. The present study investigated the occurrence, amount and diversity of PMMoV in water environments in Italy. A total of 254 water samples, collected between 2017 and 2019 from different types of water, were analysed. In detail, 92 raw sewage, 32 treated sewage, 16 river samples, 9 estuarine waters, 20 bathing waters, 67 groundwater samples and 18 drinking waters were tested. PMMoV was detected in 79% and 75% of untreated and treated sewage samples, respectively, 75% of river samples, 67% and 25% of estuarine and bathing waters and 13% of groundwater samples. No positive was detected in drinking water. The geometric mean of viral concentrations (genome copies/L) was ranked as follows: raw sewage (2.2 × 10 6 ) > treated sewage (2.9 × 10 5 ) > river waters (6.1 × 10 2 ) > estuarine waters (4.8 × 10 2 ) > bathing waters (8.5 × 10 1 ) > groundwater (5.9 × 10 1 ). A statistically significant variation of viral loads could be observed between raw and treated sewage and between these and all the other water matrices. PMMoV occurrence and viral loads did not display seasonal variation in raw sewage nor correlation with faecal indicator bacteria in marine waters and groundwater. This study represents the first report on the occurrence and quantification PMMoV in different water environments in Italy. Further studies are required to evaluate the suitability of PMMoV as a viral indicator for human faecal pollution and for viral pathogens in waters.

High-Throughput Sequencing technologies are providing unprecedented inventories of microbial communities in aquatic samples, offering an invaluable tool to estimate the impact of anthropogenic pressure on marine communities. In this case study, the Mediterranean touristic site of Aci Castello (Italy) was investigated by High-Throughput Sequencing of 16S and 18S rRNA genes. The sampling area falls within a Marine Protected Area and, notwithstanding, features an untreated urban wastewater discharge. Seawater samples were collected close to the wastewater output (COL) and at a second station about 400 m further off (PAN), before and after a summer increase in population. Prokaryotic communities clustered according to stations, rather than to seasons. While PAN showed a typical, not impacted, marine microbial composition, COL was consistently enriched in Epsilonproteobacteria and Firmicutes. Protist communities showed a peculiar clustering, as COL at springtime stood alone and was dominated by Ciliophora, while the other samples were enriched in Dinophyta. Analysis of alternative, detectable by High-Throughput Sequencing, microbial indicators, including both faecal- and sewage-associated, allowed uncovering the different sources of pollution in coastal and anthropogenically impacted marine ecosystems, underpinning the relevance of High-Throughput Sequencing-based screening as rapid and precise method for water quality management., (© FEMS 2019.)

This article describes a methodology for optimising predictive models for concentrations of faecal indicator organisms (FIOs) in coastal areas based on geographic and meteorological characteristics of upstream catchments. Concentrations of FIOs in mussels and water sampled from 50 sites in the south of Brazil from 2012 to 2013 were used to develop models to separately predict the spatial and temporal variations of FIOs. The geographical parameters used in predictive models for the spatial variation of FIOs were human population, urban area, percentage of impervious cover and total catchment area. The R 2 of models representing catchments located within 3.1 km from the monitoring points was up to 150% higher than that for the nearest catchment. The temporal variation of FIOs was modelled considering the combined effect of meteorological parameters and different time windows. The explained variance in models based on rainfall and solar radiation increased up to 155% and 160%, respectively., (Crown Copyright © 2018. Published by Elsevier Ltd. All rights reserved.)

Discharge of inadequately treated human wastewater into surface waters used for recreation, drinking water, irrigation and shellfish cultivation may present a public health hazard due to the potential shedding of high concentrations of pathogenic viruses from the human gastrointestinal tract. Human adenovirus (HAdV) and human polyomavirus (HPyV) are ubiquitous in humans and have excellent survival characteristics in the environment, so are potential candidates for indicators of human sewage contamination. Using qPCR assays, the prevalence and quantity of HAdV and HPyV JC and BK were determined in influent and effluent wastewater and receiving waters (river, urban stream, estuarine), then compared with norovirus (NoV) presence, a significant human pathogen which is not necessarily ubiquitously excreted into the environment. HAdV and HPyV were frequently detected in high concentrations in wastewater and wastewater-contaminated waters confirming their use as potential indicators for the presence of human sewage. Overall, there was a correlation between the presence of HAdV and HPyV with NoV but there were some notable exceptions including the higher frequency of NoV compared to HAdV and HPyV in estuarine waters impacted by wastewater overflows. We found that HAdV and HPyV detection by qPCR was a suitable tool for evaluating water quality and that their detection can aid in determining pollution sources, thus providing useful information for health risk assessments., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Access to safe drinking water sources and appropriate sanitation facilities remains a dream in low and middle-income countries including South Africa. This study identified the origin of faecal pollution by using quantitative polymerase chain reaction (qPCR) targeting host-specific Bacteroidales genetic markers to track the distribution of human-specific (BacHum) and animal-specific (cattle—BacCow, chicken—Cytb, pig—Pig-2-Bac, dog—BacCan) markers in water sources used by rural communities of the Vhembe District Municipality (VDM). Results revealed the prevalence of BacHum, BacCow, and BacCan in all surface water sources in Thulamela Local Municipality (TLM) and Collins Chabane Local Municipality (CLM) during wet (100%) and dry seasons (50–75%). Cytb was not detected in untreated spring water in TLM and CLM, and Pig-2-Bac was not detected in untreated hand-dug well water in TLM during both seasons. Household-level analysis detected Cytb (28.8% wet, 17.5% dry), BacHum (34.4% wet, 25% dry for Pig-2-Bac) in stored untreated spring water in CLM, and Cytb (42.9% wet, 28.5% dry) in untreated hand-dug well water in TLM. Despite differences in detection frequencies of host-specific Bacteroidales, the study highlights the public health concern of faecal pollution in rural VDM households. [ABSTRACT FROM AUTHOR]

This study investigates the impact of water faecal contamination on highly threatened European brook lamprey larvae (Lampetra planeri). Water samples and the midgut contents of lampreys collected from a small lowland river upstream (site 1) and downstream from a wastewater treatment plant (WWTP) discharge (site 2) were analysed to check how the faecal microbial load of the habitat is reflected in the intestines of larval lampreys. The counts of viable mesophiles, psychrophiles, Escherichia coli and faecal streptococci as bacterial indicators of microbial (including faecal) water contamination were estimated. Microbial composition and abundance in larval midgut contents depended on the numbers of various microorganisms in the water environment. At site 2, the water was heavily microbiologically contaminated throughout the year by sewage inflow from the WWTP, and the amounts of studied bacteria were also high in the midgut of lampreys inhabiting site 2 regardless of the season. At site 1, water quality was better, and the levels of tested microbial indicators were lower in the intestines of the lampreys living there. The numbers of bacteria dependent on water temperature were growing in warmer seasons both in water and in intestines. Sewage pollution negatively influenced the condition of lampreys in site 2, where they exhibited lower body condition than in site 1. [ABSTRACT FROM AUTHOR]

A safe water supply is a necessity, but it remains one of the backlogs of services rendered in rural areas of developing countries. This leads to vulnerable communities using water from available sources that is unsafe as it is contaminated with faecal matter. Microbial source tracking (MST) methods are gold-standard techniques that detect the exact sources of faecal contamination. This study, therefore, tracked and identified the exact sources of faecal contamination from the catchment to the point of use in rural areas of Vhembe District Municipality. Collected water samples (n = 1048) were concentrated by membrane filtration for the enumeration and detection of E. coli, followed by DNA extraction. The extracted DNA was subjected to a quantitative polymerase chain reaction (qPCR) to track target host-specific Bacteroidales genetic markers from the water source to the point of use. Rivers and dams exhibited maximum E. coli counts of up to 90 CFU/100 mL during the wet season and up to 50 CFU/100 mL during the dry season. Due to the effective treatment of these water sources, no E. coli bacteria were detected in any of the sampled municipal drinking water treatment plants at the point of treatment, while this indicator bacterium was detected at the point of use (households), with a maximum of 4 CFU/100 mL recorded during both the wet and dry seasons. Overall, the most prevalent MST marker exhibited during the wet season was BacCan (dog-associated, 6.87%), followed by BacCow (cow-associated, 5.53%), while Pig-2-Bac (pig-associated, 2.48%) was the least prevalent. The most prevalent marker exhibited during the dry season was BacCan (5.34%), followed by BacCow, with Pig-2-Bac (1.72%) being the least prevalent. A positive correlation (r = 0.31, p = 0.001) was established between the presence of the MST markers and detected E. coli from water sources to the point of use. The knowledge of the faecal contamination attributes in both public and domestic domains will assist in developing prevention and control strategies. [ABSTRACT FROM AUTHOR]

Colistin is one of the antibiotics of last resort for human health. However, the dissemination of the plasmid-mediated colistin resistance gene mcr-1 is of great concern globally. In the One Health framework, the environment is an important component for managing antimicrobial resistance. However, little information is available concerning the prevalence of mcr-1 in water environments. We aimed to reveal the prevalence of mcr-1 in different water environments in Hanoi, Vietnam. Quantitative PCR was applied to detect mcr-1 in four urban drainages receiving untreated domestic wastewater, three rivers, five lakes and two groundwater samples. Urban drainages contained higher concentrations of mcr-1, suggesting that urban residents carry the gene. The class 1 integron-integrase gene was identified as a good surrogate of antibiotic resistance genes including mcr-1. A significant correlation was found between the levels of mcr-1 and the human-specific cross-assembly phage, which is an indicator of human faecal pollution. These results indicated that the primary source of mcr-1 in urban water environments is human faeces, which is consistent with the fact that most domestic wastewater is untreated in Hanoi. The control of untreated wastewater is critical for alleviating the spread of mcr-1 in water environments in Vietnam. [ABSTRACT FROM AUTHOR]

Recent studies have shown that crAssphage is abundant in human faecal samples worldwide. It has thus been postulated as a potential microbial source tracking (MST) marker to detect human faecal pollution in water. However, an effective implementation of crAssphage in water management strategies will depend on an understanding of its environmental dynamics. In this work, the abundance and temporal distribution of crAssphage was analysed in the effluent of wastewater treatment plants using different sewage treatments, and in two rivers (water and sediments) that differ in pollution impact and flow regime. Additionally, the influence of environmental conditions (temperature and rainfall) on the removal of the marker was studied along a river section, and natural inactivation was assessed by a mesocosms approach. Molecular and culture-based tools were used to compare crAssphage abundance and dynamics with those of bacteria and bacteriophages currently applied as global indicators (E. coli , somatic coliphages, Bacteroides GA17 bacteriophages, and the human-associated MST markers HF183 and HMBif). CrAssphage concentrations in sewage effluent and river samples were similar to those of HF183 and HMBif and higher than other general and/or culture-based indicators (by 2–3 orders of magnitude). Measurement of crAssphage abundance revealed no temporal variability in the effluent, although rainfall events affected the dynamics, possibly through the mobilisation of sediments, where the marker was detected in high concentrations, and an increase in diffuse and point pollution. Another factor affecting crAssphage inactivation was temperature. Its persistence was longer compared with other bacterial markers analysed by qPCR but lower than culturable markers. The results of this study support the use of crAssphage as a human source tracking marker of faecal pollution in water, since it has similar abundances to other molecular human MST markers, yet with a longer persistence in the environment. Nevertheless, its use in combination with infectious bacteriophages is probably advisable. Graphical abstract Image 1 Highlights • CrAssphage is highly abundant in sewage polluted environments. • CrAssphage inactivates faster at high than at low temperature. • CrAssphage persists more than molecular bacterial MST markers but less than culturable phages. • Sediments can become a reservoir of crAssphage. • CrAssphage is a good potential MST marker for water management in the studied context. [ABSTRACT FROM AUTHOR]

The aim of this study was to develop faecal pollution model structures with artificial neural networks (ANNs) for cost-effective lake water quality management studies. In this study 5 artificial neural networks model structures were applied to predict the Faecal coliform concentrations for 4 different coast areas "Göllüce, İnciraltı, Darka, Orhangazi" and all data of the coasts in Lake İznik- Turkey. The Levenberg-Marquardt and backpropagation algorithm was proposed for feed-forward neural networks training. According to performance functions root mean squared error (RMSE), neural network model structures provided acceptable results. Correlation values (R) were found between 0.590 and 0.999. Increasing the number of hidden layer in the model structures was not raised the model efficiency in each trial. Type and number of input parameters were more effective for some model efficiency. Increasing the number of hidden layer and inputs in the model structures did not raise the model efficiency in each trial. Because depending on the numbers and chemical compositions of the substrates in the lake water microorganism's metabolism and their growth rates could be influenced differently and the larger error values of the modeling results determined in Göllüce and Orhangazi Coasts which influenced by pollution sources. Water quality modeling studies and increasing of monitoring would provide more productive results for protection and management of coastal. [ABSTRACT FROM AUTHOR]

Aim: Ensuring the safety of potable water is a necessity of every human being. Taking into account long-term microbiological contamination of water supply network of Gorski Kotar, the results of 5-year County monitoring program of potable water safety in that region have been analysed. The aim of this study was to monitor the impact of amendment of legal standards as well as sanitary and technical requirements - the insertion of higher number of chlorinators on the number of examined samples deriving from the water supply system as well as the influence on the results of testing of the health parameters of water intended for human consumption. This study compared the contamination profile of water for human consumption in Gorski Kotar and other regions of the Primorje-Gorski Kotar County. Methods: This study presents the elaborated and interpreted results of the County monitoring program of potable water safety in the region of Gorski Kotar during the 5-year period (2011-2015; the number examined samples N = 7,047). Results: Application of the Regulations on the parameters of assessment and methods for the analysis of water for human consumption (Official Gazette 125/2013, 141/2013/, 128/2015) provides six times lesser number of samples for the monitoring of water supply system network in Gorski Kotar, but the number of analysed parameters was increased. Installation of automatic chlorinators (2012 and 2013) resulted with twice smaller number of non-compliant samples of drinking water. Conclusions: Application of the the Regulations (Official Gazette 125/2013, 141/2013, 128/2015) resulted with the significant reduction of samples for certain water supply systems making the objective assessment of their condition much harder. The installation of automatic chlorinators reduced the number of noncompliant water samples in Gorski Kotar by half. The main reason for non-complying samples of water for human consumption in the water supply systems of Gorski Kotar is faecal contamination, while for the other parts of the County major deviation from stipulated criteria are turbidity, increased number of colonies (Heterotrophic Plate Count - HPC) and chlorination by-products. [ABSTRACT FROM AUTHOR]

Microbiological contamination arising from faecal pollution is a widespread effect of human activity on aquatic systems, with management of the issue complicated because contamination sources are often unknown. Recent advances in molecular techniques have permitted the use of host-specific genetic markers to apportion sources of faecal pollution. In this study, we used a tiered approach employing faecal indicator bacteria and Bacteroidales markers to identify faecal sources in an urban setting. Canine sources were the most common source of faecal contamination, which led to a programme of education and targeted management. A single, but substantial, source of human faecal contamination was identified that was subject to corrective action and its effectiveness validated by supplementary monitoring. This study supports the use of a tiered approach for the identification of faecal contamination sources in New Zealand, including the use of faecal indicator bacteria and more complex source-tracking analysis using genetic markers. [ABSTRACT FROM PUBLISHER]

Laboratory-scale experiments on the survival of Escherichia coli in raw, undiluted, freshly collected, source-separated yellowwater were performed. Concentrations of E. coli and its survival at different temperature regimes and storage times were measured in yellowwater originally cross-contaminated with faeces and yellowwater purposely contaminated (deliberately spiked) with faecal material. The temperature regimes of cold (4°C), mild (10°C) and warm (22°C) were the limited factors, whereas the storage time of the contaminated yellowwater was unlimited and lasted until the E. coli concentrations reached the limit of detection of < 1 Most Probable Number (MPN)/100 mL. Temperature and pH played the main role in the inactivation and longevity of E. coli in source-separated yellowwater. The mild storage conditions were the most favourable for the persistence of E. coli, which reached 40 days with a concentration of 2.0 E+03 MPN in 100 mL of undiluted yellowwater. [ABSTRACT FROM AUTHOR]

The discharge of untreated or inadequately treated effluents has been identified among the activities responsible for the spread of a wide range of potentially infectious agents. The aim of this study was to determine whether inadequate treatment of wastewater and the faecal pollution load of effluents and receiving water bodies in Sedibeng District and Soshanguve peri-urban area of the Tshwane Metropolitan Municipality could be a potential threat to the health of the surrounding communities. Variations in the counts of faecal indicator bacteria and pathogenic microorganisms and compliance of the effluents and receiving water bodies with South African and World Health Organization standards were assessed between August 2011 and May 2012 using culture-based methods and molecular techniques. The overall quality of effluents did not comply with the South African special standard of no risk for unrestricted irrigation (zero Escherichia coli/100 ml). The quality of the receiving water bodies did not comply with South African regulatory limits set for domestic purposes (zero E. coli/100 ml, <30 faecal enterococci/100 ml and <1 somatic coliphages/100 ml), for full contact recreation (<20 somatic coliphages/100 ml) and aquaculture (<10 E. coli/100 ml) and WHO standards for full and intermediate contact recreational use (<1 E. coli/100 ml and <40 faecal enterococci/100 ml, respectively). The PCR results revealed the prevalence of pathogenic microorganisms; between 0 and 60 % of samples tested positive for Salmonella Typhimurium and Shigella dysenteriae, and between 20 and 60 % of samples tested positive for Vibrio cholerae. These findings demonstrated that potential health risks might be associated with the use of the target river waters for domestic, recreational and irrigation purposes. This study calls for a prompt intervention to improve wastewater management. [ABSTRACT FROM AUTHOR]

The aim of this study was to examine antibiotic resistance (AR) dissemination in coastal water, considering the contribution of different sources of faecal contamination. Samples were collected in Berlenga, an uninhabited island classified as Natural Reserve and visited by tourists for aquatic recreational activities. To achieve our aim, AR in Escherichia coli isolates from coastal water was compared to AR in isolates from two sources of faecal contamination: human-derived sewage and seagull faeces. Isolation of E. coli was done on Chromocult agar. Based on genetic typing 414 strains were established. Distribution of E. coli phylogenetic groups was similar among isolates of all sources. Resistances to streptomycin, tetracycline, cephalothin and amoxicillin were the most 2 frequent. Higher rates of AR were found among seawater and faeces isolates, except for last-line antibiotics used in human medicine. Multi-resistance rates in isolates from sewage and seagull faeces (29% and 32%) were lower than in isolates from seawater (39%). Seawater AR profiles were similar to those from seagull faeces and differed significantly from sewage AR profiles. Nucleotide sequences matching resistance genes blaTEM, sul1, sul2, tet(A) and tet(B), were present in isolates of all sources. Genes conferring resistance to 3rd generation cephalosporins were detected in seawater (blaCTX-M-1 and blaSHV-12) and seagull faeces (blaCMY-2). Plasmid-mediated determinants of resistance to quinolones were found: qnrS1 in all sources and qnrB19 in seawater and seagull faeces. Our results show that seawater is a relevant reservoir of AR and that seagulls are an efficient vehicle to spread human-associated bacteria and resistance genes. The E. coli resistome recaptured from Berlenga coastal water was mainly modulated by seagulls-derived faecal pollution. The repertoire of resistance genes covers antibiotics critically important for humans, a potential risk for human health. [ABSTRACT FROM AUTHOR]

We hypothesized that the transport of Escherichia coli strains harvested from springs could be characterized by a similar set of cell characteristics and transport parameters. The hypothesis was tested by sampling springs throughout the Lubigi catchment in Kampala, Uganda. Chemo-physical parameters in addition to total coliform concentrations were determined. Furthermore, E. coli strains were harvested, and cell properties determined. Column experiments in saturated quartz columns of 7 cm height were conducted to determine transport parameters of selected E. coli strains. Using a two-site non-equilibrium sorption model, transport was modelled by fitting breakthrough data in HYDRUS 1D. Results indicated faecal contamination of the springs with high concentrations of total coliforms, chloride and nitrate. Furthermore, the maximum relative E. coli concentrations (C/C0)max in the column experiments were high. Compared with our previous work on E. coli strains, collected from a pasture and from zoo animals, attachment was low. Modelling revealed that both equilibrium and kinetic sorption were not important under conditions employed in the experiments. These observations are explained by the way in which the strains were harvested: from termination points of flow lines (springs). Such strains may possess characteristics that might have influenced their transport in the subsurface leading to their low attachment efficiency and possibly contributing to the lack of influence of equilibrium and kinetic sorption characteristics. There was no significant correlation between cell properties and transport parameters. Furthermore, 58% of the tested strains were of the O21:H7 serotype, and all definable serotypes identified were associated with diseases. We speculate that this serotype may possess characteristics that allow preferential transport through the aquifers of the area. We demonstrated that bacteria harvested from termination points of flow lines compared with those obtained from pollution sources, which have not undergone transport yet, present a good option for the assessment of bacteria transport characteristics in aquifers. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The Plankenburg and Eerste Rivers (Western Cape) have been reported to be contaminated with faecal coliforms. Water is drawn from both rivers for irrigation of fresh produce. The potential risk in the use of these rivers as irrigation sources was assessed by determining the fluctuations of 'indicator' and 'index' microbes over 3 years. Selected physico-chemical (water temperature, pH, COD, conductivity and alkalinity) and microbiological parameters, including coliforms, faecal coliforms, Escherichia coli and enterococci, as 'indicators' of faecal pollution, and Salmonella, Listeria and Staphylococcus, as 'index' of the presence of potential pathogens, were monitored. No correlation was found between water temperature and COD (r2 = 0.0003), whereas for temperature and pH a significant trend (p = 0.0004), but low correlation (r2 = 0.108), was observed. With the exception of the faecal coliforms (E. coli), no significant trends and no correlations between temperature and the dependent variables were found. For the faecal coliforms there was a significant trend (p = 0.0289) with temperature but not a good correlation (r2 = 0.0434), but the impact of temperature over time was significant (p = 0.0047). This is important, when the World Health Organisation (WHO) and South African Department of Water Affairs (DWA) guidelines for faecal coliforms are applied, as it indicates that temperature does impact the faecal coliform numbers. The presence of indicator organisms did not only indicate unsanitary conditions, but also the presence of potential pathogens such as Staphylococcus, Klebsiella, Listeria and Salmonella. Based on these results the microbial quality of these rivers was found to be unacceptable and does not meet the WHO and DWA guidelines for safe irrigation. There was also a high risk of exposure to human pathogens when water from these rivers is used to irrigate produce that is consumed without further processing. [ABSTRACT FROM AUTHOR]

Contamination of public water ways with sewage represents a serious environmental and health risk. We monitored pollution of the river Thames by enumerating the indicator organism Escherichia coli. Samples were taken from a site in central London near Waterloo Bridge in different seasons. E. coli were quantified using a membrane filtration method, and correlated with the tidal variations of the river and meteorological data on rainfall and temperature. More frequent and severe incidents of pollution occurred in the autumn. Heavy rainfall resulted in sharp peaks of E. coli contamination that implies a potential increase of numbers of pathogenic micro-organisms. Sixty percent of all samples were found to be in excess of the accepted upper limit of pollution set by European Union ( EU) legislation for bathing water. This study demonstrated that frequent sewage pollution of the Thames results in high numbers of E. coli and incidents of detectable levels of pathogenic bacteria demonstrating the need for regular monitoring of bacterial pollution. [ABSTRACT FROM AUTHOR]

Efficient management of deteriorating water bodies can be achieved by determining the sources of faecal pollution. Resourceful techniques for discrimination of the sources of Escherichia coli in surface water have recently been developed, including the use of river water to facilitate faecal indicator surveillance, identification of sources of faecal contamination and employing relevant management practices to maintain water quality. This study was conducted to employ microbial source tracking (MST) techniques for the determination of the sources of faecal pollution based on a water quality investigation of the physico-chemical characteristics and coliform count point of the Tirumanimuttar River. To accomplish this, an MST library-based antibiotic resistance analysis, serotyping and the genomic tool rep-PCR techniques were applied, and the obtained results were analysed statistically. Among 135 and 70 E. coli isolates present in the library and water samples collected from the river and nearby well water sources, respectively, most showed intrinsic, high or moderate resistance to antibiotics. Isolates from human and pig faecal sources were 92% homologous with the samples from the river, whereas isolates from sewage and dairy cattle showed 89% and 80% homology, respectively. These findings indicated that the Tirumanimuttar River is subjected to stress from anthropogenic activities and runoff contaminated with agricultural and human faecal contamination. The sources of faecal pollution identified in this study may facilitate the monitoring and management of the Tirumanimuttar River. [ABSTRACT FROM AUTHOR]

Abstract: Microbial source tracking is an emerging tool developed to protect water sources from faecal pollution. In this study, we evaluated the suitability of real time-quantitative PCR (qPCR) Taqman assays developed for detection of host-associated Bacteroidales markers in a prairie watershed. The qPCR primers and probes used in this study exhibited high accuracy (88–96% sensitivity and ≥99% host specificity) in detecting Bacteroidales spp. that are associated with faeces from humans, ruminants, bovines, and horses. The ruminant- and human-associated markers were also found in high concentrations within individual faecal samples, ranging from 3.4 to 7.3log10 marker copy numberg−1 of individual host faeces. Following validation of host sensitivity and specificity, the host-associated Bacteroidales markers were detected in the Qu’Appelle Valley watershed of Saskatchewan, Canada which experiences a diversity of anthropogenic inputs. Concentrations of the ruminant marker were well-correlated with proximity to cattle operations and there was a correlation between the marker and Escherichia coli concentrations at these sites. Low concentrations of the human faecal marker were measured throughout the sampling sites, and may indicate a consistent influx of human faecal pollution into the watershed area. Persistence of each of the Bacteroidales host-associated marker was also studied in situ. The results indicated that the markers persist for shorter periods of time (99% decay in <8 days) compared with the conventional E. coli marker (99% decay in >15 days), suggesting they are effective at detecting recent faecal contamination events. The levels of Bacteroidales markers and E. coli counts did not correlate with the presence of the pathogenic bacteria, Salmonella spp. or Campylobacter spp. detected in the Qu’Appelle Valley. Collectively, the results obtained in this study demonstrated that the qPCR approach for detecting host-associated Bacteroidales spp. markers can be a useful tool in helping to determine host-specific impacts of faecal pollution into a prairie watershed. [Copyright &y& Elsevier]

In this study, faecal sterols were used to identify human faecal pollution in a nori-sewered catchment in Southeast Queensland, Australia. In all, 36 water samples were collected from six sites on six occasions and the concentration of sterols were determined using gas chromatography and mass spectrometry. The stanols concentration in water samples generally increased with increased catchment runoff. After moderate rainfall, high coprostanols levels found in water samples indicated human faecal pollution via defective septic systems. In contrast, it appears that during dry weather human faecal pollution is not occurring in the study catchment. Sterol profiles also pointed to a cattle farm polluting during modest catchment runoff. The method used in this study was able to identify the sources of faecal pollution to the catchment due to rainfall. [ABSTRACT FROM AUTHOR]

Water quality was assessed for five subregions of Lake Oconee and Lake Sinclair in Georgia, USA. The areas were chosen for their levels of human impact, including: (i) suburban development >30 years old; (ii) modern suburban development; (iii) industry; (iv) agriculture; and (v) an area of low human activity. The measured temperature, pH, dissolved oxygen concentration and turbidity values were normal for oligotrophic lakes. Faecal pollution in the lakes was determined using the membrane filtration method (Method 1600) on mEI plates for enterococci. There was a positive correlation between turbidity and the number of faecal enterococci in the lakes. The faecal pollution level was higher for the old suburban and agricultural areas. Faecal pollution in the agricultural region of Lake Oconee exceeded EPA regulatory standards. The faecal pollution source was identified using polymerase chain reaction detection, with Bifidobacterium adolescentis being a marker of human faecal pollution, and bovine-associated Bacteroides (BoBac) as a marker of cattle faecal pollution. Human faecal pollution was detected in the agricultural, old suburban and industrial areas of the lakes. In contrast, bovine faecal pollution was detected only in the agricultural area of Lake Oconee. Measurements of chlorophyll-a and relative algal community abundance indicated the least-impacted and modern suburban areas had significantly lower numbers of primary producers, being dominated by diatoms. [ABSTRACT FROM AUTHOR]

Recent studies have shown the increasing interest of F-specific RNA phage genotyping to identify major sources of faecal contamination in waters. This study, conducted in a river located in an urbanized watershed with recognized anthropogenic influences, was aimed at evaluating the relevance of direct phage genotyping by real-time RT-PCR. One hundred percent of positive results were obtained with a 5mL aliquot of river water (n =31). Phage distribution was modified after cultivation, since the ratio of the two most abundant genogroups (II and I) reached 1.51log10 by direct RT-PCR-based method versus 0.30log10 after cultivation (n =8). For the first time, positive correlations between the concentrations of genogroup II, bacterial indicators and human adenoviruses were observed, which may indicate a human faecal pollution. No correlation between genogroups II and I has been revealed. The concentration of genogroup I was only correlated with water turbidity, suggesting an animal pollution coming from upstream after rainfall events. Among the microbiological parameters studied, only genogroup II/genogroup I ratio shows variations occurring in the major sources of faecal pollution. [Copyright &y& Elsevier]

Multiple host-specific molecular markers were used to detect the sources of faecal pollution in a mixed land use non-sewered catchment in Southeast Queensland, Australia. These markers included human-specific Bacteroides (HF183 and HF134), cattle-specific Bacteroides (CF128), dog-specific Bacteroides (BacCan) and human-specific enterococci surface protein (esp) markers. The sensitivity and specificity of these markers were determined by testing 197 faecal samples from 13 host groups. The overall sensitivity and specificity of these markers was high (sensitivity ≥ 85% and specificity ≥ 93%) indicating their suitability for detecting the sources of faecal pollution. Of the 16 samples collected from the study area, 14 (87%) were positive for at least one of the molecular marker tested. Amongst all the markers, cattle-specific CF128 was more prevalent than others, followed by human-specific HF183 which was consistently detected in samples collected from sites within close proximity to urban development. Significant correlations were found between E. coli and enterococci concentrations with the positive/negative results of human-specific Bacteroides HF183 ( p < 0.001, p < 0.0001) and HF134 ( p < 0.001, p < 0.004) markers. No correlations were found between faecal indicators (E. coli or enterococci) with the CF128 or BacCan markers. A significant correlation was also found between enterococci concentrations and the presence/absence of the esp marker ( p < 0.02). Based on the results, it appears that the host-specific markers such as HF183 and esp are a sensitive measure of sources of human faecal pollution in surface waters in Southeast Queensland, Australia. [ABSTRACT FROM AUTHOR]

Abstract: This paper reports on a successful application of statistical and inductive learning methods to determine optimal discriminating parameters and develop predictive models for the determination of faecal sources in waters, recently and heavily polluted with wastewaters (microbial source tracking). The data comes from an international study in which various microbial and chemical parameters were determined in heavily polluted waters from diverse geographical areas. A total of 38 variables derived from the microbial and chemical parameters were defined to characterise the available 103 observations. Four methods were evaluated: Euclidean k-nearest-neighbour, linear Bayesian classifier, quadratic Bayesian classifier and a support vector machine. The main aim was the obtention of highly accurate predictive models using the lowest number of variables possible. After a strong feature selection process, the obtained results show that predictive models using only two variables emerge with 100% correct classification. The obtained solutions make use of a linear combination of a discriminating tracer (the enumeration of phages infecting Bacteroides thetaiotaomicron) and a universal non-discriminant faecal indicator. Other models not using the discriminant tracer were developed, though a higher number of variables was needed to achieve a high rate of correct classification. [Copyright &y& Elsevier]

A direct, microscopic fluorescent antibody method was developed to detect the occurrence of Enterococcus faecium in coastal aquatic environments and was compared with the conventional membrane filtering method. The “in situ” application of the antibody-based protocol in the analysis of water samples collected from coastal polyhaline habitats demonstrated good sensitivity and ease of implementation. Data obtained with the microscopic technique were in agreement with those obtained from culture counts. The fluorescent antibody method proved to be a rapid and reliable technique for the detection of E. faecium. The advantages and limitations intrinsic to the method are discussed, highlighting the potential of this new technique for monitoring coastal aquatic environments. [Copyright &y& Elsevier]

Abstract: In this study water quality (major and minor inorganic components) of the Khaudum National Park in Namibia has been assessed for consumption by game. Although almost every modern country has drinking water guidelines for human consumption, for game such guidelines are rare. Therefore, as well as providing a database on water quality of the Khaudum National Park, existing recommendations for the quality of water supplied to game are revised in this study. Two out of 13 sources used for game water supply in the Khaudam Park are, in fact, unsuitable for this purpose. In general, water quality in the park is improved by supply from abstracted groundwater. However, a comparison of the abstracted groundwater with the supplied water in waterholes shows that the quality of the provided water is often lowered due to wash-in of faeces. It is suggested that reasonable guidelines for game water quality be established, and that water monitoring in game reserves and national parks should become obligatory. [Copyright &y& Elsevier]

Current measures for controlling the public health risks associated with bivalve molluscan shellfish consumption rely on the use of Escherichia coli to indicate the sanitary quality of shellfish harvesting areas. However, it has been demonstrated that E. coli is an inadequate indicator of the viral risk associated with shellfish. An alternative indicator, male-specific B+ coliphages, have been investigated as viral indicators of faecal contamination that may provide source-specific information for impacted environmental waters. This study compared the distribution of E. coli and F+ RNA bacteriophages in shellfish grown in harvesting areas of Greece and also examined the presence and proportions of the different subgroups of F+ RNA coliphages in shellfish. F+ RNA bacteriophages were present in shellfish at higher concentrations than E. coli. Elevated numbers of F+ RNA bacteriophages observed in the winter concur with the known increased viral risk associated with shellfish harvested at that time of year in Greece. The majority of F+ RNA coliphages detected in shellfish samples belonged to group IV which indicated the possible presence of animal faecal material in sample harvesting areas. Phages of groups II and III (human waste and human faecal material, respectively) were present at low levels. Finally, 8% of the phages hybridised were found to belong to group I. The presence of group IV showed seasonal distribution (more in winter, less in summer) whereas the other groups did not show any difference. Monitoring of F+ coliphage subgroups may indicate the presence and major sources of microbial inputs to surface waters; however, environmental effects on the relative occurrence of different groups need to be considered. [ABSTRACT FROM AUTHOR]

A model describing the dynamics of faecal coliforms (FC) in the Seine estuary has been developed and coupled with a three-dimensional hydrodynamic model. As input to this model three types of FC sources were considered: (1) FC transported by the Seine river flow at the estuary entrance at Poses dam; (2) FC brought in through the tributaries of the Seine estuary; and (3) the outfalls of the treated effluents of the WWTPs located along the estuary. As previous studies in the Seine estuary showed that a large proportion of FC was attached to SM, two pools of FC were considered separately in the model: free FC and FC attached to SM. Two main processes controlling the fate of FC in the estuary were considered: mortality and settling. The model calculates for a given discharge of the Seine river at Poses the longitudinal distribution of FC along the estuary. The model was validated by comparison of model calculations with experimental data in various hydrological conditions. The model also correctly reproduced the impact of the main river flow rate on the level of estuarine water's contamination. Finally, the model was used to test different scenarios of sanitation, suggesting priorities for sanitation efforts. [ABSTRACT FROM AUTHOR]

Abstract: A 3 year study was conducted to evaluate mussels as bioindicators of faecal contamination in coastal ecosystems of California. Haemolymph samples from 4680 mussels (Mytilus spp.) were tested for Cryptosporidium genotypes using PCR amplification and DNA sequence analysis. Our hypotheses were that mussels collected from sites near livestock runoff or human sewage outflow would be more likely to contain the faecal pathogen Cryptosporidium than mussels collected distant to these sites, and that the prevalence would be greatest during the wet season when runoff into the nearshore marine environment was highest. To test these hypotheses, 156 batches of sentinel mussels were collected quarterly at nearshore marine sites considered at higher risk for exposure to livestock runoff, higher risk for exposure to human sewage, or lower risk for exposure to both faecal sources. Cryptosporidium genotypes detected in Haemolymph samples from individual mussels included Cryptosporidium parvum, Cryptosporidium felis, Cryptosporidium andersoni, and two novel Cryptosporidium spp. Factors significantly associated with detection of Cryptosporidium spp. in mussel batches were exposure to freshwater outflow and mussel collection within a week following a precipitation event. Detection of Cryptosporidium spp. was not associated with higher or lower risk status for exposure to livestock faeces or human sewage sources. This study showed that mussels can be used to monitor water quality in California and suggests that humans and animals ingesting faecal-contaminated water and shellfish may be exposed to both host-specific and anthropozoonotic Cryptosporidium genotypes of public health significance. [Copyright &y& Elsevier]

Guanabara Bay, located adjacent to Rio de Janeiro in Brazil, is highly contaminated by substances derived from domestic and industrial effluents as well as from agricultural runoff. In the present work, sedimentary coprostanol and other faecal sterols were used to investigate historical trends in sewage contamination. Sediment cores were collected from eight different (and characterized) locations in the bay and were sectioned into dated segments. Organic carbon was determined by dry combustion and sterols were separated and quantified by GC/MS. The space-distribution of organic carbon and faecal sterol concentrations generally coincided with the presence of known pollution sources. Concentrations of coprostanol as high as 40 μg g-1 were found, indicating areas of severe sewage contamination. Faecal sterol ratios, commonly used as tools to investigate contamination in temperate environments, however, were found to have limited applicability to this tropical estuarine system, probably due to the influence of intensive primary production and microbial processes. [Copyright &y& Elsevier]

We assessed antimicrobial resistance (AMR) potential to seven major classes of antibiotics in Central Thailand's coastal aquaculture region using high-throughput qPCR targeting 295 antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs). In parallel, we used MinION next generation sequencing (NGS) of 16S rRNA gene amplicons, qPCR of faecal pollution makers, and conventional plate count methods for the comprehensive characterization of environmental microbiomes. We tested the hypothesis that aquaculture increases environmental AMR by comparing ARGs and MGEs in water and sediment samples from five aquaculture sites and their associated canals. There was no evidence from the ARG and MGE data that aquaculture is a major driver of environmental AMR in Central Thailand. Instead, the highest relative prevalence of resistance traits was found in Hua Krabue canal water influenced by urban pollution from Bangkok at the inland edge of the coastal aquaculture region. The sum of ARGs and MGEs, relative to 16S rRNA genes used as markers for overall bacterial abundance, was between 0.495 ± 0.011 and 0.498 ± 0.013 in Hua Krabue canal water, compared with at most 0.132 ± 0.005 in all the other environmental samples. Corresponding patterns were observed for most faecal pollution markers, which were also elevated in Hua Krabue canal water. [Display omitted] • Canal and aquaculture water and sediment were investigated in Central Thailand. • Antimicrobial resistance (AMR) was studied with HT-qPCR and NGS. • High AMR prevalence was associated with high fecal indicator bacteria prevalence. • AMR prevalence was highest in peri-urban canal water. • Urban water pollution appears to drive environmental AMR. [ABSTRACT FROM AUTHOR]

The last decades have seen the development of several source tracking (ST) markers to determine the source of pollution in water, but none of them show 100% specificity and sensitivity. Thus, a combination of several markers might provide a more accurate classification. In this study Ichnaea® software was improved to generate predictive models, taking into account ST marker decay rates and dilution factors to reflect the complexity of ecosystems. A total of 106 samples from 4 sources were collected in 5 European regions and 30 faecal indicators and ST markers were evaluated, including E. coli , enterococci, clostridia, bifidobacteria, somatic coliphages, host-specific bacteria, human viruses, host mitochondrial DNA, host-specific bacteriophages and artificial sweeteners. Models based on linear discriminant analysis (LDA) able to distinguish between human and non-human faecal pollution and identify faecal pollution of several origins were developed and tested with 36 additional laboratory-made samples. Almost all the ST markers showed the potential to correctly target their host in the 5 areas, although some were equivalent and redundant. The LDA-based models developed with fresh faecal samples were able to differentiate between human and non-human pollution with 98.1% accuracy in leave-one-out cross-validation (LOOCV) when using 2 molecular human ST markers (HF183 and HMBif), whereas 3 variables resulted in 100% correct classification. With 5 variables the model correctly classified all the fresh faecal samples from 4 different sources. Ichnaea® is a machine-learning software developed to improve the classification of the faecal pollution source in water, including in complex samples. In this project the models were developed using samples from a broad geographical area, but they can be tailored to determine the source of faecal pollution for any user. Image 1 • Samples from 5 geographical sources were analysed with 30 faecal markers and indicators. • A machine learning software was used to develop faecal source discriminant models. • An in-silico matrix was generated using faecal samples, adding dilution and inactivation. • LDA models' output was a combination of markers able to improve the accuracy of classification. • Models using between 2 and 5 source tracking markers can achieve LOOCV accuracies of over 95%. [ABSTRACT FROM AUTHOR]