Implementation and integration of microbial source tracking in a river watershed monitoring plan

Ballesté, Elisenda; Demeter, Katalin; Masterson, Bartholomew; Timoneda, Natàlia; Sala-Comorera, Laura; Meijer, Wim G.

doi:10.1016/j.scitotenv.2020.139573

Cited by 33 publications

(43 citation statements)

References 63 publications

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“…The amplification from the samples of non-targeted hosts was reported as a false-positive detection, and no amplification from the samples of the targeted host was reported as a false-negative result. The performance of the assays was evaluated by calculating sensitivity and specificity and accuracy with the following formulas 1–3 ( Boehm et al, 2013 ; Tiwari et al, 2018 ; Ballesté et al, 2020 ):…”

Section: Methodsmentioning

confidence: 99%

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The Use of Ribosomal RNA as a Microbial Source Tracking Target Highlights the Assay Host-Specificity Requirement in Water Quality Assessments

et al. 2021

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For microbial source tracking (MST), the 16S ribosomal RNA genes (rDNA) of host-specific bacteria and mitochondrial DNA (mtDNA) of animal species, known to cause fecal contamination of water, have been commonly used as molecular targets. However, low levels of contamination might remain undetected by using these DNA-based qPCR assays. The high copy numbers of ribosomal RNA (rRNA) could offer a solution for such applications of MST. This study compared the performance of eight MST assays: GenBac3 (general Bacteroidales), HF183 (human), BacCan (dog), Rum-2-Bac (ruminant), Pig-2-Bac (swine), Gull4 (gull), GFD, and Av4143 (birds) between rRNA-based and rDNA-based approaches. Three mtDNA-based approaches were tested: DogND5, SheepCytB, and HorseCytB. A total of 151 animal fecal samples and eight municipal sewage samples from four regions of Finland were collected for the marker evaluation. The usability of these markers was tested by using a total of 95 surface water samples with an unknown pollution load. Overall, the performance (specificity, sensitivity, and accuracy) of mtDNA-based assays was excellent (95–100%), but these markers were very seldom detected from the tested surface water samples. The rRNA template increased the sensitivity of assays in comparison to the rDNA template. All rRNA-based assays (except Av4143) had more than 80% sensitivity. In contrast, only half (HF183, Rum-2-Bac, Pig-2-Bac, and Gull4) of rDNA-based assays reached this value. For markers targeted to bird feces, the use of the rRNA-based assay increased or at least did not change the performance. Regarding specificity, all the assays had >95% specificity with a DNA template, except the BacCan assay (71%). While using the RNA template for the assays, HF183 and BacCan exhibited only a low level of specificity (54 and 55%, respectively). Further, the HF183 assay amplified from multiple non-targeted animal fecal samples with the RNA template and the marker showed cross-amplification with the DNA template as well. This study recommends using the rRNA-based approach for MST assays targeting bird fecal contamination. In the case of mammal-specific MST assays, the use of the rRNA template increases the sensitivity but may reduce the specificity and accuracy of the assay. The finding of increased sensitivity calls for a further need to develop better rRNA-based approaches to reach the required assay performance.

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Section: Methodsmentioning

confidence: 99%

“…Sensitivity, specificity, and accuracy of assays have been used for the characterization of the performance of different microbial methods ( Boehm et al, 2013 ; Tiwari et al, 2018 ; Ballesté et al, 2020 ). Among such criteria, specificity is the primarily important character for any given host-specific MST assay.…”

Section: Introductionmentioning

confidence: 99%

The Use of Ribosomal RNA as a Microbial Source Tracking Target Highlights the Assay Host-Specificity Requirement in Water Quality Assessments

et al. 2021

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“…Sewage may contain a variety of human viruses belonging to at least 41 viral families, including Adenoviridae, Polyomaviridae and Papillomaviridae, which may cause gastrointestinal and respiratory disease among other pathologies ( Cantalupo et al, 2011 ; Fernandez-Cassi et al, 2018 ; Martínez-Puchol et al, 2020 ). Pollution of coastal waters and rivers by sewage or by faecal contamination from non-point animal sources therefore poses a serious public health risk due to ingestion of contaminated water during recreational activities or contamination for of shellfish for human consumption ( Ballesté et al, 2020 ; Bellou et al, 2013 ; Soller et al, 2010 ; Yamahara et al, 2007 ). Human faecal contamination originating from sewerage misconnections, combined sewage overflows or discharges from wastewater treatment plants may occur frequently in urban areas ( Ahmed et al, 2019 ; Kay et al, 2008 ; Panasiuk et al, 2015 ; Reynolds et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Decay of infectious SARS-CoV-2 and surrogates in aquatic environments

et al. 2021

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The introduction of SARS-CoV-2 containing human stool and sewage into water bodies may raise public health concerns. However, assessment of public health risks by faecally contaminated water is limited by a lack of knowledge regarding the persistence of infectious SARS-CoV-2 in water. In the present study the decay rates of viable infectious SARS-CoV-2 and SARS-CoV-2 RNA were determined in river and seawater at 4 and 20°C. These decay rates were compared to S. typhimurium bacteriophage MS2 and pepper mild mottle virus (PMMoV). Persistence of viable SARS-CoV-2 was temperature dependent, remaining infectious for significantly longer periods of time in both freshwater and seawater at 4°C than at 20°C. T 90 for infectious SARS-CoV-2 in river water was 2.3 days and 3.8 days at 20°C and 4°C, respectively. The T 90 values were 1.1 days and 2.2 days in seawater at 20°C and 4°C, respectively. In contrast to the rapid inactivation of infectious SARS-CoV-2 in river and sea water, viral RNA was relatively stable. The RNA decay rates were increased in non-sterilised river and seawater, presumably due to the presence of microbiota. The decay rates of infectious MS2, MS2 RNA and PMMoV RNA differed significantly from the decay rate of SARS-CoV-2 RNA, suggesting that their use as surrogate markers for the persistence of SARS-CoV-2 in the environment is limited.

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“…If a specific site is classified as poor for 5 consecutive years, a permanent ban on bathing is required. For complex and unclear pollution sources, new tools are available, such as Microbial Source Tracking (28,29), which in the past decade has become indispensable for detecting the origin of microbial contamination in various aquatic systems. Genetic markers forming part of certain fecal microbes are closely related to a specific host (e.g., humans, livestock, dogs, and seagulls).…”

Section: Concentrations Of Microbiological Indicators In Lakesmentioning

confidence: 99%

Quality of Croatian Inland Bathing Areas: Reference to The Region and EU

Galić

Cenov²,

Lušić

et al. 2021

JHSCI

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Introduction: The Republic of Croatia (HR) has significantly invested in monitoring the quality of inland bathing areas during the past 10 years. The aim of this paper was to analyze the results obtained during the 2014-2019 period and to compare them with the Region (non-European Union [EU] countries neighboring Croatia) and EU. Moreover, bathing water quality data at two bathing areas of Zagreb (lakes Jarun and Bundek) were processed in more detail.Methods: The quality of inland bathing areas in the HR is monitored in five rivers and five lakes in nine Croatian counties; 19 of the 35 monitored sites are located in Zagreb County. Escherichia coli (EC) was determined by EN ISO 9308-3: 1998, while intestinal enterococci (ENT) were defined by EN ISO 7899-2: 2000. Statistical processing of the results was performed using the Microsoft Excel Statistical Package (Redmond, USA) and Statistica 13.5. (Stat.Sof.Inc., Tulsa, USA); the significance level was set to p < 0.05.Results: During the study period, the worst Croatian inland water quality was recorded in 2019 (excellent quality for only 25.9% of the bathing areas), while the best quality was recorded in 2015 (excellent quality in 57.1%). On average (2014-2019), the share of bathing areas with excellent quality in Croatia was 46.3% (i.e., under the EU average of 85.3%); with no poor locations (EU average was nearly 2%). A significant positive correlation was found between fecal indicators (EC&ENT) and the amount of precipitation, while a negative correlation with water and air temperature in lakes.Conclusions: The quality of Croatian inland bathing areas is significantly lower than the European average, while EU water quality is improving. Regarding the countries bordering Croatia, Italy, Slovenia, and Hungary have achieved better results, while Montenegro, Serbia, and Bosnia and Herzegovina are still in the process of developing the monitoring of the bathing water quality.

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Implementation and integration of microbial source tracking in a river watershed monitoring plan

Cited by 33 publications

References 63 publications

The Use of Ribosomal RNA as a Microbial Source Tracking Target Highlights the Assay Host-Specificity Requirement in Water Quality Assessments

The Use of Ribosomal RNA as a Microbial Source Tracking Target Highlights the Assay Host-Specificity Requirement in Water Quality Assessments

Decay of infectious SARS-CoV-2 and surrogates in aquatic environments

Quality of Croatian Inland Bathing Areas: Reference to The Region and EU

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