scholarly journals Comparative microbial source tracking methods for identification of fecal contamination sources at Sunnyside Beach in the Toronto region area of concern

2016 ◽  
Vol 14 (5) ◽  
pp. 839-850 ◽  
Author(s):  
Zachery R. Staley ◽  
Thomas A. Edge
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Fecal Contamination ◽  
Bird Species ◽  
Microbial Source Tracking ◽  
Source Tracking ◽  
Pcr Marker ◽  
E Coli ◽  
Remediation Strategies ◽  
Humber River ◽  
Area Of Concern

Several beaches within the Toronto region area of concern have persistent issues with fecal contamination, causing a beach beneficial use impairment (BUI). In this study, Escherichia coli, including ampicillin-resistant strains, were enumerated via culturable and quantitative polymerase chain reaction (qPCR) methods. Microbial source tracking (MST) markers (for general Bacteroidales, human, ruminant/cow, gull, and dog) were detected and enumerated via PCR and qPCR to identify sources of fecal contamination at Sunnyside Beach and in the Humber River. Human, cow, and dog markers had good host-specificity, while gull markers sometimes amplified a few other bird species. The ruminant endpoint PCR marker amplified a variety of other animal species rendering it less useful. Both human and gull fecal contamination were prevalent in the Humber River, while Sunnyside Beach was predominantly impacted by gull fecal contamination. Human sewage impacts were more prevalent in the lower Humber River, particularly in Black Creek. However, to reduce Sunnyside beach postings, reducing bird fecal contamination in the river and at the beach would be necessary. When there are high levels of E. coli throughout a beachshed, an MST toolbox approach can add value to discriminate source(s) of E. coli contamination and guide decisions relating to public health risk and remediation strategies.

scholarly journals Microbial Source Tracking Using Quantitative and Digital PCR To Identify Sources of Fecal Contamination in Stormwater, River Water, and Beach Water in a Great Lakes Area of Concern

2018 ◽  
Vol 84 (20) ◽  
Author(s):  
Zachery R. Staley ◽  
Rachel J. Boyd ◽  
Phoenix Shum ◽  
Thomas A. Edge
Keyword(s):  
Great Lakes ◽  
Fecal Contamination ◽  
Digital Pcr ◽  
Microbial Source Tracking ◽  
Source Tracking ◽  
Content Type ◽  
Low Levels ◽  
Areas Of Concern ◽  
Area Of Concern ◽  
The Individual

ABSTRACTAreas of concern (AOCs) around the Great Lakes are characterized by historic and ongoing problems with microbial water quality, leading to beneficial use impairments (BUIs) such as beach postings and closures. In this study, we assessed river and beach sites within the Rouge River watershed, associated stormwater outfalls, and at Rouge Beach. The concentrations ofEscherichia colias well as human- and gull-specific qPCR microbial source tracking (MST) markers were assessed at all sites. A preliminary comparison of digital PCR (dPCR) methodologies for both MST markers was conducted regarding sensitivity and specificity. Within the watershed, the outfalls were found to be a prominent source of human fecal contamination, with two outfalls particularly affected by sewage cross-connections. However, the occurrence of human fecal contamination along Rouge Beach and in the lower portions of the watershed was largely dependent on rain events. Gull fecal contamination was the predominant source of contamination at the beach, particularly during dry weather. The multiplex human/gull dPCR methodology used in this study tended to be more sensitive than the individual quantitative PCR (qPCR) assays, with only a slight decrease in specificity. Both dPCR and qPCR methodologies identified the same predominance of human and gull markers in stormwater and beach locations, respectively; however, the dPCR multiplex assay was more sensitive and capable of detecting fecal contamination that was undetected by qPCR assays. These results demonstrate the dPCR assay used in this study could be a viable tool for MST studies to increase the ability to identify low levels of fecal contamination.IMPORTANCEFecal contamination of recreational water poses a persistent and ongoing problem, particularly in areas of concern around the Great Lakes. The identification of the source(s) of fecal contamination is essential for safeguarding public health as well as guiding remediation efforts; however, fecal contamination may frequently be present at low levels and remain undetectable by certain methodologies. In this study, we utilized microbial source tracking techniques using both quantitative and digital PCR assays to identify sources of contamination. Our results indicated high levels of human fecal contamination within stormwater outfalls, while lower levels were observed throughout the watershed. Additionally, high levels of gull fecal contamination were detected at Rouge Beach, particularly during drier sampling events. Furthermore, our results indicated an increased sensitivity of the digital PCR assay to detect both human and gull contamination, suggesting it could be a viable tool for future microbial source tracking studies.

scholarly journals Comparison of Microbial and Chemical Source Tracking Markers To Identify Fecal Contamination Sources in the Humber River (Toronto, Ontario, Canada) and Associated Storm Water Outfalls

2016 ◽  
Vol 82 (21) ◽  
pp. 6357-6366 ◽  
Author(s):  
Zachery R. Staley ◽  
Josey Grabuski ◽  
Ed Sverko ◽  
Thomas A. Edge
Keyword(s):  
Ecosystem Health ◽  
Fecal Contamination ◽  
Storm Water ◽  
Source Tracking ◽  
Water Runoff ◽  
Content Type ◽  
E Coli ◽  
Storm Water Runoff ◽  
Sewage Contamination ◽  
Humber River

ABSTRACTStorm water runoff is a major source of pollution, and understanding the components of storm water discharge is essential to remediation efforts and proper assessment of risks to human and ecosystem health. In this study, culturableEscherichia coliand ampicillin-resistantE. colilevels were quantified and microbial source tracking (MST) markers (including markers for generalBacteroidalesspp., human, ruminant/cow, gull, and dog) were detected in storm water outfalls and sites along the Humber River in Toronto, Ontario, Canada, and enumerated via endpoint PCR and quantitative PCR (qPCR). Additionally, chemical source tracking (CST) markers specific for human wastewater (caffeine, carbamazepine, codeine, cotinine, acetaminophen, and acesulfame) were quantified. Human and gull fecal sources were detected at all sites, although concentrations of the human fecal marker were higher, particularly in outfalls (mean outfall concentrations of 4.22 log10copies, expressed as copy numbers [CN]/100 milliliters for human and 0.46 log10CN/100 milliliters for gull). Higher concentrations of caffeine, acetaminophen, acesulfame,E. coli, and the human fecal marker were indicative of greater raw sewage contamination at several sites (maximum concentrations of 34,800 ng/liter, 5,120 ng/liter, 9,720 ng/liter, 5.26 log10CFU/100 ml, and 7.65 log10CN/100 ml, respectively). These results indicate pervasive sewage contamination at storm water outfalls and throughout the Humber River, with multiple lines of evidence identifying Black Creek and two storm water outfalls with prominent sewage cross-connection problems requiring remediation. Limited data are available on specific sources of pollution in storm water, though our results indicate the value of using both MST and CST methodologies to more reliably assess sewage contamination in impacted watersheds.IMPORTANCEStorm water runoff is one of the most prominent non-point sources of biological and chemical contaminants which can potentially degrade water quality and pose risks to human and ecosystem health. Therefore, identifying fecal contamination in storm water runoff and outfalls is essential for remediation efforts to reduce risks to public health. This study employed multiple methods of identifying levels and sources of fecal contamination in both river and storm water outfall sites, evaluating the efficacy of using culture-based enumeration ofE. coli, molecular methods of determining the source(s) of contamination, and CST markers as indicators of fecal contamination. The results identified pervasive human sewage contamination in storm water outfalls and throughout an urban watershed and highlight the utility of using both MST and CST to identify raw sewage contamination.

Library-dependent and library-independent microbial source tracking to identify spatial variation in faecal contamination sources along a Lake Ontario beach (Ontario, Canada)

2010 ◽  
Vol 62 (3) ◽  
pp. 719-727 ◽  
Author(s):  
T. A. Edge ◽  
S. Hill ◽  
P. Seto ◽  
J. Marsalek
Keyword(s):  
Spatial Variation ◽  
Municipal Wastewater ◽  
Lake Ontario ◽  
Microbial Source Tracking ◽  
Pollution Sources ◽  
Source Tracking ◽  
Faecal Contamination ◽  
E Coli ◽  
Faecal Pollution ◽  
Humber River

Multiple microbial source tracking methods were applied to investigate spatial variation in faecal pollution sources impacting a 1.7 km freshwater beach on Lake Ontario (Canada). The highest E. coli concentrations measured in the study area were from interstitial sand pore water at Sunnyside Beach, reaching 2.6 × 106 CFU/100 ml. These E. coli concentrations exceeded those in the nearby Humber River and Black Creek, which are impacted by combined sewer overflows containing municipal wastewater and by stormwater conveying washoff from the urban area. Library-independent Bacteroidales HF183 analyses identified the more frequent occurrence of municipal wastewater contamination in the Humber River and at a Sunnyside Beach location closest to the mouth of the river. Library-dependent E. coli antibiotic resistance and rep-PCR DNA fingerprinting analyses identified the more frequent occurrence of bird faecal contamination at Sunnyside Beach locations away from the river mouth. These microbial source tracking results raise caution about managing beaches with multiple sources of contamination as a single entity without considering spatial variability in faecal pollution sources and the need for more localized beach management practices.

scholarly journals Microbial source tracking of private well water samples across at-risk regions in southern Ontario and analysis of traditional fecal indicator bacteria assays including culture and qPCR

2016 ◽  
Vol 14 (6) ◽  
pp. 1047-1058 ◽  
Author(s):  
Julia Krolik ◽  
Allison Maier ◽  
Shawna Thompson ◽  
Anna Majury
Keyword(s):  
At Risk ◽  
Water Samples ◽  
Rural Areas ◽  
Fecal Contamination ◽  
Microbial Source Tracking ◽  
Source Tracking ◽  
Well Water ◽  
Prevention Strategies ◽  
E Coli ◽  
Bacteriological Testing

Many people living in rural areas rely on privately owned wells as their primary source of drinking water. These water sources are at risk for fecal contamination of human, wildlife, and livestock origin. While traditional bacteriological testing involves culture-based methods, microbial source tracking (MST) assays present an opportunity to additionally determine the source of fecal contamination. This study investigated the main host sources of contamination in private well water samples with high levels of Escherichia coli (E. coli), using MST with human and multi-species specific markers. Fecal contamination of human origin was detected in approximately 50% of samples, indicating that current contamination prevention strategies require reconsideration. The relationship between cattle density and fecal contamination of bovine origin was investigated using a Bovine Bacteroidales specific MST assay. Regional variations of microbial sources were examined, and may inform local primary prevention strategies. Additionally, in order to assess MST and E. coli quantitative real time polymerase chain reaction (qPCR) assays as indicators of fecal contamination, these were compared to E. coli culture methods. Variation in results was observed across all assay methods investigated, suggesting the most appropriate routine bacteriological testing methodology cannot be determined without comparison to a method that directly detects the presence of fecal contamination.

Flood and rainfall mobilisation of E. coli and faecal source tracking markers from decomposing cowpats 

2021 ◽  
Author(s):  
Megan Devane ◽  
Brent Gilpin ◽  
Jennifer Webster-Brown ◽  
Louise Weaver ◽  
Pierre Dupont ◽  
...  
Keyword(s):  
Water Quality ◽  
Overland Flow ◽  
Quantitative Polymerase Chain Reaction ◽  
Linear Regression Analysis ◽  
Water Quality Monitoring ◽  
Quality Monitoring ◽  
Source Tracking ◽  
Faecal Contamination ◽  
E Coli ◽  
Faecal Indicators

<p>The intensification of dairy farming on the agricultural landscape in New Zealand has raised concerns about pollution sources from dairy faecal runoff into waterways. The transport of faecal pollution from farms into waterways is facilitated by overland flow, which can result from rain and flood events, poorly designed irrigation practices and the washing down of milking sheds.</p><p>An important step for mitigation of pollution is the identification of the source(s) of faecal contamination. When elevated levels of faecal indicator bacteria (FIB) such as <em>Escherichia coli </em>are identified in a waterway, faecal source tracking (FST) tools such as microbial source tracking (MST) using quantitative polymerase chain reaction (qPCR), and faecal steroids (for example, cholesterol) provide information about the sources of faecal contamination. The understanding of the fate (degradation/persistence) and transport of these FST markers in the environment is recognised as an important requirement for the interpretation of water quality monitoring in aquatic environments.</p><p>This study investigated the effects of faecal decomposition on bovine faecal indicators (<em>E. coli </em>and FST markers: bovine-associated qPCR markers and ten faecal steroids) by monitoring the effect of flood and rainfall events on simulated cowpats over a five and a half month period under field conditions. Two separate spring/summer trials were conducted to evaluate: Trial 1) the mobilisation under simulated flood conditions of the faecal indicators from irrigated versus non-irrigated cowpats, Trial 2) the mobilisation of faecal indicators from non-irrigated cowpat flood runoff versus runoff after simulated rainfall onto non-irrigated cowpats.</p><p>The microbial community changes within the decomposing cowpat (as illustrated by amplicon-based metagenomic analysis) were expected to impact on the survival/persistence of the bacterial targets of the MST markers, and also alter the ratio between faecal sterols and their biodegradation products, the stanols. It was hypothesised, therefore, that there would be:</p><ul><li>Changes over time in the concentration of<em> E. coli </em>and the bovine-associated MST markers mobilised into the cowpat runoff</li> <li>Alterations in the FST ratio signature of the ten measured faecal steroids, resulting in a change from a bovine faecal steroid signature in fresh cowpat runoff to other animal faecal signatures in the runoff from decomposing cowpats</li> <li>A difference in the mobilisation decline rates of all FST and microbial markers within a treatment regime and between treatments.</li> </ul><p>Linear regression analysis was undertaken to establish mobilisation decline rates for each of the analytes in the mobilisable phase from the cowpat runoff treatments, with calculation of the time taken in days for reduction in 90% of the concentration (T<sub>90</sub>), and statistical comparison of the regression coefficients (slopes) of all analytes. The results will include a discussion of the impacts of the study’s observations on the interpretation of faecal indicator assessments for water quality monitoring in waterways influenced by sources of faecal contamination.</p>

The use of antibiotic resistance profiling as a means of tracing sources of fecal contamination in source waters

2010 ◽  
Vol 10 (2) ◽  
pp. 209-215
Author(s):  
M. S. Mthembu ◽  
P. T. Biyela ◽  
T. G. Djarova ◽  
A. K. Basson
Keyword(s):  
Antibiotic Resistance ◽  
Municipal Wastewater ◽  
Fecal Contamination ◽  
Source Tracking ◽  
Human Consumption ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
E Coli ◽  
Intestinal Pathogens ◽  
Source Waters

Fecal contamination of source waters and its associated intestinal pathogens continues to pose risks to public health although the extent and effect of microbial contamination of source waters gets very little attention in designing treatment plants in most developing countries. Coliform counts give an indication of the overall bacterial contamination of water and thus its safety for human consumption. However, their presence fails to provide information about the source of fecal contamination which is vital to managing fecal contamination problems in surface waters. This study explored the use of multiple antibiotic resistance (MAR) indexing as means of differentiating E. coli isolates from different sources. A total of 322 E. coli isolates were obtained from municipal wastewater and from fecal samples from domestic and wild animals. Conventional culture methods and standard chemical and biochemical tests were used for isolation and identification of E. coli. Isolates were assayed against 10 antibiotics using the micro-dilution technique. The results obtained generated antibiotic resistance profiles which were used to statistically group the isolates into different subsets. Correct source classification was obtained for 60% of human-derived and 95% non-human-derived E. coli respectively. These results indicate the validity of the usefulness of MAR indexing as a method of bacterial source tracking.

Multi-Year Microbial Source Tracking Study Characterizing Fecal Contamination in an Urban Watershed

2017 ◽  
Vol 89 (2) ◽  
pp. 127-143 ◽  
Author(s):  
Rebecca N. Bushon ◽  
Amie M.G. Brady ◽  
Eric D. Christensen ◽  
Erin A. Stelzer
Keyword(s):  
Fecal Contamination ◽  
Microbial Source Tracking ◽  
Source Tracking ◽  
Urban Watershed
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