Long-term impact of basin-wide wastewater management on faecal pollution levels along the entire Danube River.

The Danube River is, at 2857 km, the second longest river in Europe and the most international river in the world with 19 countries in its catchment. Along the entire river, faecal pollution levels are mainly influenced by point-source emissions from treated and untreated sewage of municipal origin under base-flow conditions. In the past 2 decades, large investments in wastewater collection and treatment infrastructure were made in the European Union (EU) Member States located in the Danube River Basin (DRB). Overall, the share of population equivalents with appropriately biologically treated wastewater (without disinfection) has increased from 69% to more than 85%. The proportion of tertiary treatment has risen from 46 to 73%. In contrast, no comparable improvements of wastewater infrastructure took place in non-EU Member States in the middle and lower DRB, where a substantial amount of untreated wastewater is still directly discharged into the Danube River. Faecal pollution levels along the whole Danube River and the confluence sites of the most important tributaries were monitored during four Danube River expeditions, the Joint Danube Surveys (JDS). During all four surveys, the longitudinal patterns of faecal pollution were highly consistent, with generally lower levels in the upper section and elevated levels and major hotspots in the middle and lower sections of the Danube River. From 2001 to 2019, a significant decrease in faecal pollution levels could be observed in all three sections with average reduction rates between 72 and 86%. Despite this general improvement in microbiological water quality, no such decreases were observed for the highly polluted stretch in Central Serbia. Further improvements in microbiological water quality can be expected for the next decades on the basis of further investments in wastewater infrastructure in the EU Member States, in the middle and lower DRB. In the upper DRB, and due to the high compliance level as regards collection and treatment, improvements can further be achieved by upgrading sewage treatment plants with quaternary treatment steps as well as by preventing combined sewer overflows. The accession of the Western Balkan countries to the EU would also significantly boost investments in wastewater infrastructure and water quality improvements in the middle section of the Danube. Continuing whole-river expeditions such as the Joint Danube Surveys is highly recommended to monitor the developments in water quality in the future.

Two-year Monitoring of Microbiological Water Quality in Small Water Supply Systems: Implications for Microbial Risk Management.

Small water supply systems (SWSSs) are often more vulnerable to waterborne disease outbreaks. In Japan, many SWSSs operate without regulation under the Waterworks Law, yet there is limited investigation into microbial contamination and the associated health risks. In this study, the microbiological water quality of four SWSSs that utilize mountain streams as water sources and do not install water treatment facilities were monitored for over 2 years. In investigated SWSSs, the mean heterotrophic plate counts were below 350 CFU/mL, and the total bacterial loads (16S rDNA concentration) ranged from 4.71 to 5.35 log10 copies/mL. The results also showed the consistent presence of fecal indicator bacteria (FIB), i.e., Escherichia coli and Clostridium perfringens, suggesting the potential of fecal pollution. E. coli was then utilized as an indicator to assess the health risk posed by E. coli O157:H7 and Campylobacter jejuni. The results indicated that the estimated mean annual risk of infection and disability-adjusted life years (DALYs) exceeded acceptable levels in all SWSSs for the two reference pathogens. To ensure microbial water safety, implementing appropriate water treatment facilities with an estimated mean required reduction of 5-6 log10 was necessary. This study highlighted the potential microbial contamination and health risk level in SWSSs that utilize mountain streams as water sources, even though the water sources were almost not affected by human activities. Furthermore, this study would also be helpful in supporting risk-based water management to ensure a safe water supply in SWSSs.

Seasonal Variations in Household Water Use, Microbiological Water Quality, and Challenges to the Provision of Adequate Drinking Water: A Case of Peri-urban and Informal Settlements of Hosanna Town, Southern Ethiopia.

Several studies have been conducted on household water use and microbial water quality globally. However, studies that considered seasonal variability of household water use and microbial water quality were limited. Therefore, this study investigated the seasonal variability of household water use, microbiological water quality, and challenges to the provision of adequate water in the peri-urban and informal settlements of Hosanna town, Southern Ethiopia. A longitudinal study was conducted on 288 households. The data was gathered using a pretested structured questionnaire, laboratory-analysis, interviews, storage-container inventories, focus group discussions, key-informant interviews, and an observational checklist. The data was analyzed using stepwise-multiple linear regression, bivariate and multivariable logistic regression, thematic-analysis, t-tests, and non-parametric-tests. Households were visited for 7 consecutive days during the dry and rainy seasons to account for changes in daily and seasonal variation of water use. 440 stored water and 12 source samples were analyzed for E. coli presence during dry and rainy seasons. The prevalence of stored water contamination with E. coli was 43.2% and 34.5% during the dry and rainy seasons, respectively. The per capita water consumption was 19.4 and 20.3 l during the dry and rainy seasons, respectively. Piped water on-premises, small family size, volume, and number of water storage containers were significant predictors of per capita water consumption in both seasons. Piped water off-premises, storing water for more than 3 days, uncovered, and wide-mouthed water storage containers were significantly associated with the presence of E. coli in water in both seasons. Seasonal variability of household water use and microbiological water quality was statistically significant, which is a significant public health concern and needs intervention to enhance water quantity and quality to mitigate the risk of waterborne diseases. Findings also suggest seasonal monitoring of the safety of drinking water to ensure that the water is safe and healthy.

Initiating guidance values for novel biological stability parameters in drinking water to control regrowth in the distribution system.

Nine novel biological stability parameters for drinking water have been developed recently. Here, we report data for these nine parameters in treated water from 34 treatment plants in the Netherlands to deduce guidance values for these parameters. Most parameters did not show a strong correlation with another biological stability parameter in the same sample, demonstrating that most parameters hold different information on the biological stability of drinking water. Furthermore, the novel biological stability parameters in treated water varied considerably between plants and five parameters in treated water were significantly lower for drinking water produced from groundwater than surface water. The maximum biomass concentration (MBC7), cumulative biomass potential (CBP14) from the biomass production potential test (BPP-W) and the total organic carbon concentration in treated water from groundwater were predictive parameters for HPC22 and Aeromonas regrowth in the distribution system. Guidance values of 8.6 ng ATP L-1, 110 d·ng ATP L-1 and 4.1 mg C L-1 were deduced for these parameters, under which the HPC22 and Aeromonas numbers remain at regulatory level. The maximum biomass growth (MBG7) from the BPP-W test, the particulate and/or high molecular organic carbon and the iron accumulation rate in treated water from surface water were predictive parameters for HPC22 and Aeromonas regrowth in the distribution system. Deduced guidance values for these biological stability parameters were 4.5 ng ATP L-1, 47 µg C L-1 and 0.34 mg Fe m-2 day-1, respectively. We conclude from our study that a multiple parameter assessment is required to reliable describe the biological stability of drinking water, that the biological stability of drinking water produced from groundwater is described with other parameters than the biological stability of drinking water produced from surface water, and that guidance values for predictive biological stability parameters were inferred under which HPC22 and Aeromonas regrowth is under control.

Microbiological quality of roof tank water in an urban village in southeastern China.

Urban villages are unique residential neighborhoods in urban areas in China. Roof tanks are their main form of water supply, and water quality deterioration might occur in this system because of poor hygienic conditions and maintenance. In this study, water samples were seasonally collected from an urban village to investigate the influence of roof tanks as an additional water storage device on the variation in the microbial community structure and pathogenic gene markers. Water stagnation in the roof tank induced significant decreases in chlorine (p < 0.05), residual chlorine was as low as 0.02 mg/L in spring. Propidium monoazide (PMA)-qPCR revealed a one-magnitude higher level of total viable bacterial concentration in roof tank water samples (2.14 ± 1.81 × 105 gene copies/mL) than that in input water samples (3.57 ± 2.90 × 104 gene copies/mL, p < 0.05), especially in spring and summer. In addition, pathogenic fungi, Mycobacterium spp., and Legionella spp. were frequently detected in the roof tanks. Terminal users might be exposed to higher microbial risk induced by high abundance of Legionella gene marker. Spearman's rank correlation and redundancy analysis showed that residual chlorine was the driving force that promoted bacterial colonization and shaped the microbial community. It is worth noted that the sediment in the pipe will be agitated when the water supply is restored after the water outages, which can trigger an increase in turbidity and bacterial biomass. Overall, the findings provide practical suggestions for controlling microbiological health risks in roof tanks in urban villages.

Direct evidence of microbiological water quality changes on bacterial quantity and community caused by plumbing system.

Drinking water quality deteriorates from treatment plant to customer taps, especially in the plumbing system. There is no direct evidence about what the differences are contributed by plumbing system. This study compared the water quality in the water main and at customer tap by preparing a sampling tap on the water main. The biomass was quantified by adenosine triphosphate (ATP) and the microbial community was profiled by 454 pyrosequencing. The results showed that in distribution pipes, biofilm contributed >94% of the total biomass, while loose deposits showed little contribution (< 2%) because of the low amount of loose deposits. The distribution of biological stable water had minor effects on the microbiocidal water quality regarding both quantity (ATP 1 ng/L vs. 1.7 ng/L) and community of the bacteria. Whereas the plumbing system has significant contribution to the increase of active biomass (1.7 ng/L vs. 2.9 ng/L) and the changes of bacterial community. The relative abundance of Sphingomonas spp. at tap (22%) was higher than that at water main (2%), while the relative abundance of Pseudomonas spp. in tap water (15%) was lower than that in the water from street water main (29%). Though only one location was prepared and studied, the present study showed that the protocol of making sampling tap on water main offered directly evidences about the impacts of plumbing system on tap water quality, which makes it possible to distinguish and study the processes in distribution system and plumbing system separately.

Diarrhoea among Children Aged 5 Years and Microbial Drinking Water Quality Compliance: Trends Analysis Study in South Africa (2008-2018).

In developing countries such as South Africa, diarrhoeal diseases are reported to be linked to inadequate drinking water quality, sanitation, and hygiene behaviours. The consumption of microbiologically contaminated drinking water has been reported to cause diarrhoea, mortality, and morbidity in children under the age of five years. This study evaluated the microbiological pathogens detected in municipal drinking water and diarrhoea trends for children under 5 years of age in South Africa between 2008 and 2018. A trends analysis study was conducted using secondary data on diarrhoea for children under the age of five years and microbial drinking water quality compliance. A negative correlation was found between the occurrence of microbial pathogens in water, morbidity, and mortality rates of children under the age of 5 years in South Africa. As compliance status improved, the mortality rate among children under 5 years old decreased by 31% over the study period. A conclusion can thus be drawn that the microbiological pathogens detected in drinking water at levels complying with SANS 241:2015 Edition 2 standards and diarrhoea incidences were not the primary cause of the mortality of children under 5 years old in South Africa between 2008 and 2018.

Is dialysis water a safe component for hemodialysis treatment in São Paulo State, Brazil?

Failure on the water treatment poses hemodialysis patients at risk of injury and death. Identifying if the patients are exposed to water quality related microbiological risks is an important objective to reduce the mortality for chronic renal patients and is the main issue of this study. We evaluated the microbiological water quality used by 205 dialysis services in São Paulo State, Brazil between 2010 to 2016. The study included heterotrophic bacteria count, total coliforms research, and bacterial endotoxin determination in 1366 dialysis water samples. The number of unsatisfactory clinics for at least one microbiological parameter decreased 16.0% between 2010 to 2015 but increased 57.2% in 2016. In 2010, the most frequent unsatisfactory parameter was related to heterotrophic bacteria count (54.8%) followed by endotoxin determination (45.2%). However, in 2013 an opposite situation was observed: endotoxin determination as the parameter of the higher incidence of nonconformities. Total coliform was verified at a lower frequency. We highlighted the importance of regular monitoring of dialysis water quality to prevent infections caused by dialytic procedures and to ensure that the water is a safe component of the treatment.

A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Enterococcus spp. in water.

Faecal pollution of water and the resulting potential presence of human enteric pathogens is a predominant threat to public health. Microbiological water quality can be assessed by the detection of standard faecal indicator bacteria (SFIB) such as E. coli or certain Enterococcus species. In recent years, isothermal amplification methods have become a useful alternative to polymerase chain reaction (PCR), allowing molecular diagnostics with simple or no instrumentation. In this study, a novel screening method for the molecular detection of Enterococcus spp. by loop-mediated isothermal amplification (LAMP) is described. A set of six specific LAMP primers was designed to amplify a diagnostic fragment of the Enterococcus 23S rRNA gene, which is present in several enterococcal species targeted by quantitative PCR (qPCR), which is the standard technique recommended by the US Environmental Protection Agency. Sensitivity and specificity tests were performed using a set of 30 Enterococcus and non-target bacterial reference strains. It is shown that LAMP is equally sensitive and even more specific than the qPCR assay. A dilution series of Enterococcus faecalis DNA revealed that the LAMP method can reliably detect 130 DNA target copies per reaction within 45 min. Additionally, enterococci isolated from Austrian surface waterbodies, as well as a set of DNA extracts from environmental waters, were tested. Contingency analysis demonstrated a highly significant correlation between the results of the developed LAMP assay and the reference qPCR method. Furthermore, a simple staining procedure with a fluorescence dye demonstrated the identification of amplified products by eye. In conclusion, this method is an important component for the efficient screening and testing of water samples in low-resource settings lacking sophisticated laboratory equipment and highly trained personnel, requiring only a simple heating block.

Hydrological connectivity and Burkholderia pseudomallei prevalence in wetland environments: investigating rice-farming community's risk of exposure to melioidosis in North-East Thailand.

In our analysis of 136 water samples from wetland environments (rice paddies, natural wetland sites, man-made water bodies) in rural areas of North-East Thailand, Burkholderia pseudomallei was most prevalent in rice paddies (15 of the 30 positive sites). The high prevalence in the water of rice fields is indicative of the inherent vulnerability of farmers in rural agricultural areas in this area of Thailand and likely other locations in the tropics. Nearly all B. pseudomallei-positive sites were found within the vicinity of a large wetland associated with the Chi River, in the month of July 2014. Positive samples were found in water ranging in pH from 5.9 to 8.7, salinity ranging from 0.04 to 1.58 ppt, nitrate ranging from 0 to 10.8 ppm, and iron ranging from 0.003 to 1.519 ppm. Of these variables, only iron content was statistically higher in B. pseudomallei-positive versus B. pseudomallei-negative sites, suggesting that increasing concentrations of iron may encourage the growth of this bacterium, which is responsible for melioidosis. Our results, when combined with data from other published studies, support the notion that B. pseudomallei can exist in a wide range of environmental conditions. Thus, we argue that health safety education is a more appropriate means of addressing farmer vulnerability than chemical or physical alterations to fields at large scales. Further, it may be important to investigate melioidosis through transdisciplinary approaches that consider the complex social and ecological contexts in which the disease occurs.

Impact of Treated Sewage Effluent on the Microbiology of a Small Brook Using Flow Cytometry as a Diagnostic Tool.

Flow cytometry was applied to assess the microbiological impact of treated sewage effluent discharge into a small brook carrying surface runoff water. Increases in dissolved organic carbon and soluble reactive phosphorous were accompanied by increases in counts of intact bacteria by up to eightfold. Effluent ingress furthermore resulted in a pronounced shift of bacterial clusters. Whereas brook water upstream of the discharge point was characterised by a bacterial cluster with low nucleic acid (LNA) content, downstream water showed a shift to bacteria with high nucleic acid (HNA) content. Changes in the LNA/HNA ratio were largely maintained along the course of the brook. Results suggest that the LNA/HNA ratio can under certain conditions serve as an indicator of anthropogenic nutrient impact. Measuring impact on this low trophic level might be more sensitive and straightforward than measuring macroindicators. More evidence will however be required to assess the usefulness of LNA/HNA measurements to assess the ecological nutrient status of natural waters and the impact of nutrient pollution.

Evaluation of quantitative PCR combined with PMA treatment for molecular assessment of microbial water quality.

Microbial water quality assessment currently relies on cultivation-based methods. Nucleic acid-based techniques such as quantitative PCR (qPCR) enable more rapid and specific detection of target organisms and propidium monoazide (PMA) treatment facilitates the exclusion of false positive results caused by DNA from dead cells. Established molecular assays (qPCR and PMA-qPCR) for legally defined microbial quality parameters (Escherichia coli, Enterococcus spp. and Pseudomonas aeruginosa) and indicator organism group of coliforms (implemented on the molecular detection of Enterobacteriaceae) were comparatively evaluated to conventional microbiological methods. The evaluation of an extended set of drinking and process water samples showed that PMA-qPCR for E. coli, Enterococcus spp. and P. aeruginosa resulted in higher specificity because substantial or complete reduction of false positive signals in comparison to qPCR were obtained. Complete compliance to reference method was achieved for E. coli PMA-qPCR and 100% specificity for Enterococcus spp. and P. aeruginosa in the evaluation of process water samples. A major challenge remained in sensitivity of the assays, exhibited through false negative results (7-23%), which is presumably due to insufficient sample preparation (i.e. concentration of bacteria and DNA extraction), rather than the qPCR limit of detection. For the detection of the indicator group of coliforms, the evaluation study revealed that the utilization of alternative molecular assays based on the taxonomic group of Enterobacteriaceae was not adequate. Given the careful optimization of the sensitivity, the highly specific PMA-qPCR could be a valuable tool for rapid detection of hygienic parameters such as E. coli, Enterococcus spp. and P. aeruginosa.

Integrated modelling of faecal contamination in a densely populated river-sea continuum (Scheldt River and Estuary).

In order to simulate the long-term (months-years) median Escherichia coli distributions and variations in the tidal Scheldt River and Estuary, a dedicated module was developed for the Second-generation Louvain-la-Neuve Ice-ocean Model (SLIM, www.climate.be/slim). The resulting model (SLIM-EC2) presents two specific and new features compared to the older SLIM-EC model version. The first is that the E. coli concentrations in the river are split in three fractions: the free E. coli in the water column, the ones attached to suspended solids and those present in the bottom sediments, each with their own transport, decay and settling-resuspension dynamics. The bacteria attached to particles can settle and survive on the bottom, where they can be brought back in the water column during resuspension events. The second new feature of the model is that it is coupled to the catchment model SENEQUE-EC, which thus provides upstream boundary conditions to SLIM-EC2. The result is an integrated and multi-scale model of the whole Scheldt drainage network from its source down to the Belgian/Dutch coastal zone. This new model reproduces the long-term median E. coli concentration along the Scheldt River and Estuary. An extensive sensitivity study is performed demonstrating the relative robustness of the model with respect to the chosen parameterisations. In addition to reproducing the observed E. coli concentrations in 2007-2008 at various stations, two extreme wastewater management scenarios were considered. Overall, there is no doubt that the Scheldt Estuary acts as a cleaning filter of faecal contamination originating from large Belgian cities. As a result, at the mouth of the Scheldt Estuary E. coli concentration is negligible in all investigated conditions.

Calidad microbiológica del agua de unidades odontológicas de una clínica universitaria de Bogotá / Microbiological quality of water in dental units of a university clinic in Bogotá

Este estudio tiene como objetivo evaluar la calidad microbiológica del agua destinada al uso en unidades odontológicas de una clínica universitaria de Bogotá, mediante el recuento de E. coli, Coliformes Totales, Enterococcus y Pseudomonas, con el fin de mejorar la calidad del agua, optimizar la prestación del servicio y proporcionar una mayor seguridad y confiabilidad a los pacientes y odontólogos. En este trabajo se determina que el agua destinada al uso de las unidades odontológicas no cumple, en lo referente a las características microbiológicas, con lo establecido por la Resolución 2115 del 2007, y la Norma Técnica Colombiana 813 (NTC 813) porque excede los límites aceptables para coliformes totales y Enterococcus, además presenta un importante recuento de Pseudomonas, las cuales al considerar factores como el estado inmunológico del paciente pueden llegar a causar enfermedad.

This study aims to evaluate the microbiological quality of water intended for use in dental units of a university clinic of Bogotá, using the E. coli count, total coliforms, Enterococcus, Pseudomonas, in order to improve the quality of the water, optimize service quality, and provide greater safety and reliability to the patients and dentists. In this work, it was determined that the water intended for the use of the dental units does not meet, in regard to the microbiological characteristics, with the provisions of Resolution 2115 of 2007, and the Colombian Technical Standard 813 (NTC 813) because it exceeds the acceptable limits for total coliforms and Enterococcus, it also presents an important count of Pseudomonas which can lead to disease when considering factors such as the immune status of the patient.

Calidad microbiológica del agua de un área agrícola-ganadera del centro sur de Chile y su posible implicancia en la salud humana / Microbiological quality of water in livestock area of southern Chile and its possible implications on human health

Introduction: A significant proportion of water for human consumption has an underground origin becoming 80% in Chilean rural areas where profound wells represent the only search of water for human and animal beverage. Aim: To study the microbiological quality of water from agricultural land for livestock production in the province of Valdivia (40°S), Chile and its potential impact on human health. Material and Method: Water samples were collected monthly (2008 - 2009), at the entrance and exit of a water stream running through the field and in well water used for human and animal consumption. The total coliform (Tc) and Escherichia coli were determined by the confirmatory method Quanti-Tray together with other physicochemical assessments in the water and climatic variables. Results: In samples from the stream water and wells, the Most Probable Number of Tc and E. coli exceeded the standard Chilean Norm of Water Quality (NCh 409/1) for human consumption. Conclusion: These results show the need to regulate the environmental impact of farming and cattle production and to monitor the drinking water to meet the minimum standards of health protection.

Introducción: Parte importante del agua para consumo humano corresponde a agua subterránea, alcanzando un 80% en áreas rurales de Chile, donde los pozos profundos son la única fuente de agua de bebida para consumo humano y animal. Objetivo: Estudiar la calidad microbiológica del agua de un predio agrícola-ganadero en la provincia de Valdivia (40° S), Chile, y su posible impacto en la salud humana. Material y Método: Muestras de agua fueron recolectadas mensualmente (2008-2009), a la entrada y salida de un estero que atraviesa el predio y en agua de pozos usadas para consumo humano y animal. Las especies coliformes totales (Ct) y Escherichia coli se determinaron por el método confirmativo Quanti-Tray, se evaluaron variables físico-químicas del agua y climáticas del sitio. Resultados: En las muestras de agua del estero y pozos el número más probable de Ct y E. coli sobrepasaron la norma chilena de calidad del agua (NCh 409/1) para consumo humano. Conclusión: Estos resultados muestran la necesidad de regular el impacto ambiental de la actividad agrícola-ganadera y monitorear el agua de bebida para cumplir con los estándares mínimos de protección de la salud.