Microbial faecal pollution of river water in a watershed of tropical Ethiopian highlands is driven by diffuse pollution sources

Journal of Water and HealthTropical communities in the developing world depend heavily on riverine systems for their socioeconomic development. However, these resources are poorly protected from diffuse pollution, and there is a lack of quantitative information regarding the microbial pollution characteristics of riverine water, despite frequently reported gastrointestinal diseases. The aim of our study was to apply faecal taxation (i.e., faecal pellet counting in representative test areas to estimate the potential availability of diffuse pollution sources) in combination with a detailed microbiological faecal pollution analysis in a riverine environment to elucidate the importance of diffuse pollution. To realize this approach, ambient faecal pellets, a multiparametric data set for standard faecal indicator bacteria (SFIB), including Escherichia coli, Clostridium perfringens spores and enterococci from catchment soil and river water, and a number of riverine water physicochemical variables were analysed during a one-year cycle. We demonstrated that the abundance of ambient faecal pellets, which were consistently counted at reference sites in the catchment, was associated with faecal pollution in the river water. Water SFIB, dissolved oxygen, nutrients, conductivity and total suspended solids were strongly linked with the abundance of ambient faecal pellets in the river catchment, as demonstrated by principal component analysis (PCA). Elevated concentrations of SFIB in the riverine water in the absence of rainfall also suggested the direct input of faecal bacteria into the riverine water by livestock (e.g., during watering) and humans (e.g., during bathing). Statistical analyses further revealed that the microbiological water quality of the investigated riverine water was not influenced by SFIB potentially occurring in the soil. This study demonstrates the importance of diffuse faecal pollution sources as major drivers of the microbiological quality of riverine water in the Ethiopian highlands. In addition, the new successfully applied integrated approach could be very useful for developing predictive models, which would aid in forecasting riverine microbiological quality in tropical developing countriesinfo:eu-repo/semantics/publishedVersio

Spring Water of an Alpine Karst Aquifer Is Dominated by a Taxonomically Stable but Discharge-Responsive Bacterial Community

Alpine karst aquifers are important groundwater resources for the provision of drinking water all around the world. Yet, due to difficult accessibility and long-standing methodological limitations, the microbiology of these systems has long been understudied. The aim of the present study was to investigate the structure and dynamics of bacterial communities in spring water of an alpine limestone karst aquifer (LKAS2) under different hydrological conditions (base vs. event flow). The study was based on high-throughput 16S rRNA gene amplicon sequencing, study design and sample selection were guided by hydrology and pollution microbiology data. Spanning more than 27 months, our analyses revealed a taxonomically highly stable bacterial community, comprising high proportions of yet uncultivated bacteria in the suspended bacterial community fraction. Only the three candidate phyla Parcubacteria (OD1), Gracilibacteria (GN02), Doudnabacteria (SM2F11) together with Proteobacteria and Bacteroidetes contributed between 70.0 and 88.4% of total reads throughout the investigation period. A core-community of 300 OTUs consistently contributed between 37.6 and 56.3% of total reads, further supporting the hypothesis of a high temporal stability in the bacterial community in the spring water. Nonetheless, a detectable response in the bacterial community structure of the spring water was discernible during a high-discharge event. Sequence reads affiliated to the class Flavobacteriia clearly increased from a mean proportion of 2.3% during baseflow to a maximum of 12.7% during the early phase of the studied high-discharge event, suggesting direct impacts from changing hydrological conditions on the bacterial community structure in the spring water. This was further supported by an increase in species richness (Chao1) at higher discharge. The combination of these observations allowed the identification and characterization of three different discharge classes (Q1–Q3). In conclusion, we found a taxonomically stable bacterial community prevailing in spring waters from an alpine karst aquifer over the entire study period of more than 2 years. Clear response to changing discharge conditions could be detected for particular bacterial groups, whereas the most responsive group – bacteria affiliated to the class of Flavobacteriia – might harbor potential as a valuable natural indicator of “system disturbances” in karst aquifers

High genetic diversity ofVibrio choleraein the European lake Neusiedler See is associated with intensive recombination in the reed habitat and the long-distance transfer of strains

Coastal marine Vibrio cholerae populations usually exhibit high genetic diversity. To assess the genetic diversity of abundant V. cholerae non-O1/non-O139 populations in the Central European lake Neusiedler See, we performed a phylogenetic analysis based on recA, toxR, gyrB and pyrH loci sequenced for 472 strains. The strains were isolated from three ecologically different habitats in a lake that is a hot-spot of migrating birds and an important bathing water. We also analyzed 76 environmental and human V. cholerae non-O1/non-O139 isolates from Austria and other European countries and added sequences of seven genome-sequenced strains. Phylogenetic analysis showed that the lake supports a unique endemic diversity of V. cholerae that is particularly rich in the reed stand. Phylogenetic trees revealed that many V. cholerae isolates from European countries were genetically related to the strains present in the lake belonging to statistically supported monophyletic clades. We hypothesize that the observed phenomena can be explained by the high degree of genetic recombination that is particularly intensive in the reed stand, acting along with the long distance transfer of strains most probably via birds and/or humans. Thus, the Neusiedler See may serve as a bioreactor for the appearance of new strains with new (pathogenic) properties.The study was financed by the Austrian Science Fund FWF, project nr P21625-B20. In addition, CP and ISD were partially supported by the Austrian Science Fund FWF, P25745-B20. CA thanks to the Ministerio de Educacion, Cultura y Deporte and FEDER funds for the grant AGL2014-58933-P. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.Peer Reviewe

Automated online monitoring of fecal pollution in water by enzymatic methods

RÉSUMÉ: To facilitate the prompt management of public health risks from water resources, the fluorescence-based detection of the enzymatic activity of β-d-glucuronidase (GLUC) has been suggested as a rapid method to monitor fecal pollution. New technological adaptations enable now its automated, near-real-time measurement in a robust and analytically precise manner. Large data sets of high temporal or spatial resolution have been reported from a variety of freshwater resources, demonstrating the great potential of this automated method. However, the fecal indication capacity of GLUC activity and the potential link to health risk is still unclear, presenting considerable limitations. This review provides a critical evaluation of automated, online GLUC-based methods (and alternatives) and defines open questions to be solved before the method can fully support water management

Enterococcus and Escherichia coli fecal source apportionment with microbial source tracking genetic markers - Is it feasible?

Fecal pollution is measured in surface waters using culture-based measurements of enterococci and Escherichia coli bacteria. Source apportionment of these two fecal indicator bacteria is an urgent need for prioritizing remediation efforts and quantifying health risks associated with source-specific pathogens. There are a number of quantitative real-time PCR (QPCR) assays that estimate concentrations of source-associated genetic markers; however, their concentrations are not necessarily amenable to source apportionment because the markers may differ in prevalence across sources. Here we mathematically derive and test, under ideal conditions, a method that utilizes the ratios of fecal source-associated genetic markers and culture and molecular measurements of general fecal indicators to apportion enterococci and E. coli. The source contribution is approximately equal to the ratio of the source-associated and the general fecal indicator concentrations in a water sample divided by their ratio in the source material, so long as cross-reactivity is negligible. We illustrate the utility of the ratio method using samples consisting of mixtures of various fecal pollution sources. The results from the ratio method correlated well with the actual source apportionment in artificial samples. However, aging of contamination can confound source allocation predictions. In particular, culturable enterococci and E. coli, the organisms presently regulated in the United States and much of the world, decay at different rates compared to source-associated markers and as a result cannot be apportioned using this method. However, limited data suggest a similar decay rate between source-associated and QPCR-measured Enterococcus and E. coli genetic markers, indicating that apportionment may be possible for these organisms; however further work is needed to confirm.Keywords: Water quality, Microbial source tracking, Fecal pollutio

Real-time monitoring of beta-d-glucuronidase activity in sediment laden streams: A comparison of prototypes

AbstractDetection of enzymatic activities has been proposed as a rapid surrogate for the culture-based microbiological pollution monitoring of water resources. This paper presents the results of tests on four fully automated prototype instruments for the on-site monitoring of beta-d-glucuronidase (GLUC) activity. The tests were performed on sediment-laden stream water in the Hydrological Open Air Laboratory (HOAL) during the period of March 2014 to March 2015. The dominant source of faecal pollution in the stream was swine manure applied to the fields within the catchment. The experiments indicated that instrument pairs with the same construction design yielded highly consistent results (R2 = 0.96 and R2 = 0.94), whereas the results between different designs were less consistent (R2 = 0.71). Correlations between the GLUC activity measured on-site and culture-based Escherichia coli analyses over the entire study period yielded R2 = 0.52 and R2 = 0.47 for the two designs, respectively. The correlations tended to be higher at the event scale. The GLUC activity was less correlated with suspended sediment concentrations than with E. coli, which is interpreted in terms of indicator applicability and the time since manure application. The study shows that this rapid assay can yield consistent results over a long period of on-site operation in technically challenging habitats. Although the use of GLUC activity as a proxy for culture-based assays could not be proven for the observed habitat, the study results suggest that this biochemical indicator has high potential for implementation in early warning systems

Assessment of the faecal contamination along the Sava River and identification of pollution sources

The contamination of water by faecal pollution leads to exposure to pathogens via drinking water production, recreation or irrigation. However, monitoring of microbiological quality of surface waters is quite neglected despite its importance for human health. In the case of Sava River Basin, many of the settlements situated on the river banks discharge high quantities of untreated or improperly treated wastewaters directly into surface waters. Due to usage of water for irrigation, the evaluation of microbiological quality of the Sava River becomes essential for further river management. Water samples were collected during September 2014 on 17 sites and during September 2015 on 15 sites situated along the Sava River. In 2015, additional samples were collected from 4 wastewater outlets detected onsite. Microbiological analyses comprised monitoring the standard indicators of faecal pollution within the surveys and long term monitoring data (obtained within 5 years of routine monitoring at 4 stations). For detection of total coliforms, Escherichia coli and enterococci, Defined Substrate Technology (DST) was used with quantification performed by Colilert Quanti-Tray 2000 system, which provides a Most Probable Number result. Detection of presumptive Clostridium perfringens was performed by membrane filtration method according to ISO 14189:2013. To identify the origin of pollution, microbial source tracking (MST) analyses were employed based on the human-associated BacHum and HF183II, the ruminant-associated BacR and the pigassociated Pig2Bac genetic Bacteroidetes faecal markers. Microbiological indicators showed the existence of hotsposts of faecal pollution in the Sava River. MST confirmed that the pollution is human associated. Long term data at selected sites indicated persistent faecal contamination which leads to conclusion that the sites are under the impact of continuous discharge of wastewaters

Have genetic targets for faecal pollution diagnostics and source tracking revolutionised water quality analysis yet?

The impacts on faecal pollution analysis using nucleic acid-based methods, such as PCR and sequencing, in health-related water quality research were assessed by rigorous literature analysis. A wide range of application areas and study designs has been identified since the first application more than 30 years ago (>1,100 publications). Given the consistency of methods and assessment types, we suggest defining this emerging part of science as a new discipline: genetic faecal pollution diagnostics (GFPD) in health-related microbial water quality analysis. Undoubtedly, GFPD has already revolutionised faecal pollution detection and microbial source tracking, the current core applications. GFPD is also expanding to many other research areas, including infection and health risk assessment, evaluation of microbial water treatment, and support of wastewater surveillance. In addition, storage of DNA extracts allows for biobanking, which opens up new perspectives. The tools of GFPD can be combined with cultivation-based standardised faecal indicator enumeration, pathogen detection, and various environmental data types, in an integrated data analysis approach. This comprehensive meta-analysis provides the scientific status quo of this field, including trend analyses and literature statistics, outlining identified application areas, and discussing the benefits and challenges of nucleic acid-based analysis in GFPD