Occurrence of human-associated Bacteroidetes genetic source tracking markers in raw and treated wastewater of municipal and domestic origin and comparison to standard and alternative indicators of faecal pollution

AbstractRecent developments in water resource monitoring have increased the demand for the reliable identification of faecal pollution sources, also defined as microbial (faecal) source tracking (MST). Standardized faecal indicator bacteria (SFIB) enumeration does not directly support MST, as SFIB occur in animal and human sources. The aim of this study was to rigorously evaluate the applicability of host-associated faecal genetic MST markers detected by quantitative PCR (qPCR) at representative Austrian water resources (ground-, surface-, raw and treated wastewater, n = 196 samples) with high importance for the water management sector. Groundwater covered a gradient of non- (i.e., deep wells) to surface influenced resources (i.e., karst and shallow wells). In addition, single faecal excreta from humans as well as representative livestock and wildlife species were collected to evaluate the faecal source-specificity and -sensitivity of the MST assays. Genetic MST marker resistance against UV irradiation was evaluated in on-site ground and wastewater treatment installations. Bacteroides-based human- (HF183II, BacHum), ruminant- (BacR), and pig-associated (Pig2Bac) MST marker qPCR quantification was performed in concert with cultivation of E. coli, intestinal enterococci, and Clostridium perfringens (SFIB diagnostics). The selected MST makers revealed high faecal source identification capacity for the Austrian water compartments and quantitatively reflected the selected faecal pollution gradient. The study also demonstrated that SFIB data can efficiently be combined with MST data to solve previously unanswered questions in water safety monitoring and management (e.g., support pollution source-targeted catchment protection, hazard assessment, and health risk management). Further research and development needs are discussed to exploit the full power of MST technology. In conclusion, this study illustrates the capacity of molecular faecal pollution diagnostics to revolutionize water quality testing in the decades to come.

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Identification of source of faecal pollution of Tirumanimuttar River, Tamilnadu, India using microbial source tracking

Environmental Monitoring and Assessment ◽

10.1007/s10661-011-2398-7 ◽

2011 ◽

Vol 184 (10) ◽

pp. 6001-6012 ◽

Cited By ~ 12

Author(s):

Kasi Murugan ◽

Perumal Prabhakaran ◽

Saleh Al-Sohaibani ◽

Kuppusamy Sekar

Keyword(s):

Microbial Source Tracking ◽

Source Tracking ◽

Faecal Pollution

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Application of mussels as biosamplers for characterization of faecal pollution in coastal recreational waters

Water Science & Technology ◽

10.2166/wst.2010.910 ◽

2010 ◽

Vol 62 (3) ◽

pp. 586-593 ◽

Cited By ~ 12

Author(s):

P. Roslev ◽

A. S. Bukh ◽

L. Iversen ◽

H. Sønderbo ◽

N. Iversen

Keyword(s):

Molecular Markers ◽

Blue Mussel ◽

Unknown Origin ◽

Microbial Source Tracking ◽

Source Tracking ◽

Recreational Waters ◽

Human Waste ◽

Faecal Pollution ◽

Faecal Bacteria ◽

Seawater Samples

Sources of faecal pollution in coastal recreational waters may be identified by analysing different host associated microorganisms or molecular markers. However, the microbial targets are often present at low numbers in moderately impacted waters, and often exhibit significant temporal and spatial variability in waters with fluctuating faecal loads. This patchy occurrence can limit successful detection of relevant targets in microbial source tracking studies. In this study, we explored the possibility for using the blue mussel (Mytilus edulis) as a biosampler for accumulation of faecal bacteria relevant for microbial source tracking. Non-contaminated blue mussels were transferred to three coastal recreational waters affected by faecal pollution of unknown origin. Molecular markers associated with animal and human waste were targeted by PCR and compared in seawater and mussel samples. The results demonstrated that transplanted mussels in simple enclosures accumulated and retained elevated levels of molecular markers associated with different types of faecal pollution. The targets included a novel putative human associated E. coli subgroup B2 VIII clone, and animal and human associated markers in enterococci (esp, M19, M66, M90, and M91). Human (sewage) associated markers including esp and M66 were sometimes not detectable in seawater samples despite known wastewater contamination, whereas the markers were detectable in mussels. We suggest that transplanted mussels should be considered as potential biosamplers in studies focusing on identifying source of faecal pollution in low or moderately impacted recreational waters. Bioaccumulation of molecular markers in mussels for several days may represent the water quality better than traditional grab samples from the water column.

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Microbial source tracking to identify human and ruminant sources of faecal pollution in an ephemeral Florida river

Journal of Applied Microbiology ◽

10.1111/jam.12007 ◽

2012 ◽

Vol 113 (6) ◽

pp. 1396-1406 ◽

Cited By ~ 27

Author(s):

E. Chase ◽

J. Hunting ◽

C. Staley ◽

V.J. Harwood

Keyword(s):

Microbial Source Tracking ◽

Source Tracking ◽

Faecal Pollution

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Genetic source tracking of an anthrax outbreak in Shaanxi province, China

Infectious Diseases of Poverty ◽

10.1186/s40249-016-0218-6 ◽

2017 ◽

Vol 6 (1) ◽

Cited By ~ 5

Author(s):

Dong-Li Liu ◽

Jian-Chun Wei ◽

Qiu-Lan Chen ◽

Xue-Jun Guo ◽

En-Min Zhang ◽

...

Keyword(s):

Shaanxi Province ◽

Source Tracking ◽

Genetic Source

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Automated Sampling Procedures Supported by High Persistence of Bacterial Fecal Indicators and Bacteroidetes Genetic Microbial Source Tracking Markers in Municipal Wastewater during Short-Term Storage at 5°C

Applied and Environmental Microbiology ◽

10.1128/aem.00998-15 ◽

2015 ◽

Vol 81 (15) ◽

pp. 5134-5143 ◽

Cited By ~ 13

Author(s):

R. E. Mayer ◽

J. Vierheilig ◽

L. Egle ◽

G. H. Reischer ◽

E. Saracevic ◽

...

Keyword(s):

Municipal Wastewater ◽

Storage Conditions ◽

Microbial Source Tracking ◽

Basic Knowledge ◽

Treated Wastewater ◽

Source Tracking ◽

Fecal Indicators ◽

Content Type ◽

Automated Sampling ◽

Sampling Procedures

ABSTRACTBecause of high diurnal water quality fluctuations in raw municipal wastewater, the use of proportional autosampling over a period of 24 h at municipal wastewater treatment plants (WWTPs) to evaluate carbon, nitrogen, and phosphorus removal has become a standard in many countries. Microbial removal or load estimation at municipal WWTPs, however, is still based on manually recovered grab samples. The goal of this study was to establish basic knowledge regarding the persistence of standard bacterial fecal indicators andBacteroidetesgenetic microbial source tracking markers in municipal wastewater in order to evaluate their suitability for automated sampling, as the potential lack of persistence is the main argument against such procedures. Raw and secondary treated wastewater of municipal origin from representative and well-characterized biological WWTPs without disinfection (organic carbon and nutrient removal) was investigated in microcosm experiments at 5 and 21°C with a total storage time of 32 h (including a 24-h autosampling component and an 8-h postsampling phase). VegetativeEscherichia coliand enterococci, as well asClostridium perfringensspores, were selected as indicators for cultivation-based standard enumeration. Molecular analysis focused on total (AllBac) and human-associated geneticBacteroidetes(BacHum-UCD, HF183 TaqMan) markers by using quantitative PCR, as well as 16S rRNA gene-based next-generation sequencing. The microbial parameters showed high persistence in both raw and treated wastewater at 5°C under the storage conditions used. Surprisingly, and in contrast to results obtained with treated wastewater, persistence of the microbial markers in raw wastewater was also high at 21°C. On the basis of our results, 24-h autosampling procedures with 5°C storage conditions can be recommended for the investigation of fecal indicators orBacteroidetesgenetic markers at municipal WWTPs. Such autosampling procedures will contribute to better understanding and monitoring of municipal WWTPs as sources of fecal pollution in water resources.

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Bifidobacterial survival in surface water and implications for microbial source tracking

Canadian Journal of Microbiology ◽

10.1139/w09-007 ◽

2009 ◽

Vol 55 (6) ◽

pp. 642-647 ◽

Cited By ~ 7

Author(s):

Jakob R. Ottoson

Keyword(s):

Surface Water ◽

Screening Tool ◽

Poultry Manure ◽

Source Tracking ◽

Animal Origin ◽

Faecal Coliforms ◽

Sewage Pollution ◽

Faecal Pollution ◽

Log Linear ◽

Linear Manner

Bifidobacteria are common in the intestines of humans and other animals. Sorbitol-fermenting Bifidobacteria (SFB) are associated with human faecal pollution. The ratio of total Bifidobacteria (TB) to SFB has been proposed to be a means for determining the source (human or animal) of faecal pollution. In this study, SFB were detected in all sewage samples (n = 39) and in 42% of samples of animal origin (n = 24): pig (5/6 positive), poultry (4/8), horse (1/4), and cow (0/6). The TB:SFB ratio was significantly higher in animal samples than in sewage samples (p < 0.01), with the exception of stored poultry manure, which had ratios (1.4–11) similar to the sewage samples (1–17). The lowest ratio in the other animal samples was 40 (pig). The TB:SFB ratio in sewage remained stable over time in surface water, and it may be a useful screening tool to determine the source of faecal pollution in surface water. Bifidobacteria died off in a log linear manner at 4 and 22 °C, with rates of disappearance of 0.16 and 0.58 log(days–1), respectively. Faecal coliforms (FC) disappeared more slowly (0.09 and 0.43 log(days–1), respectively). The FC:SFB ratio proved to be an inadequate tool for determining the age of sewage pollution.

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Genetic source tracking of human plague cases in Inner Mongolia-Beijing, 2019

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0009558 ◽

2021 ◽

Vol 15 (8) ◽

pp. e0009558

Author(s):

Jianyun Li ◽

Yumeng Wang ◽

Fang Liu ◽

Xiaona Shen ◽

Yiting Wang ◽

...

Keyword(s):

Inner Mongolia ◽

Meriones Unguiculatus ◽

Source Tracking ◽

Snp Analysis ◽

Nucleotide Polymorphism ◽

Single Nucleotide ◽

Genome Wide ◽

Genetic Source ◽

Inner Mongolia Autonomous Region ◽

Relationship Of

On 12 November 2019, one couple from the Sonid Left Qi (County) in the Inner Mongolia Autonomous Region was diagnosed with pneumonic plague in Beijing. The wife acquired the infection from her husband. Thereafter, two bubonic plague cases were identified in Inner Mongolia on November 16th and 24th. In this study, genome-wide single nucleotide polymorphism (SNP) analysis was used to identify the phylogenetic relationship of Yersinia pestis strains isolated in Inner Mongolia. Strains isolated from reservoirs in 2018 and 2019 in Inner Mongolia, together with the strain isolated from Patient C, were further clustered into 2.MED3m, and two novel lineages (2.MED3q, 2.MED3r) in the 2.MED3 population. According to the analysis of PCR-based molecular subtyping methods, such as the MLVA 14 scheme and seven SNP allele sequencing, Patients A/B and D were classified as 2.MED3m. In addition, strains from rodents living near the patients’ residences were clustered into the same lineage as patients. Such observations indicated that human plague cases originated from local reservoirs. Corresponding phylogenetic analysis also indicated that rodent plague strains in different areas in Inner Mongolia belong to different epizootics rather than being caused by spreading from the same epizootic in Meriones unguiculatus in 2019.

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