Rapid QPCR-based assay for fecal Bacteroides spp. as a tool for assessing fecal contamination in recreational waters

Converse, Reagan R.; Blackwood, A. Denene; Kirs, Marek; Griffith, John F.; Noble, Rachel T.

doi:10.1016/j.watres.2009.06.036

Cited by 81 publications

(48 citation statements)

References 39 publications

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“…have been suggested as potential alternative indicators of human fecal material because human material is generally considered to be of greater risk to human health (Converse et al, 2009;Scott et al, 2005Scott et al, , 2007. In this study, positive PCR assays for general E. coli and ENT recovered strains, which were observed for all sediment samples, indicated the accumulation of FIB in the sediments deposited in Vidy Bay during the past several decades.…”

Section: Discussionmentioning

confidence: 65%

Characterization of fecal indicator bacteria in sediments cores from the largest freshwater lake of Western Europe (Lake Geneva, Switzerland)

Thevenon

Regier

Benagli

et al. 2012

Ecotoxicology and Environmental Safety

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a b s t r a c tThis study characterized the fecal indicator bacteria (FIB), including Escherichia coli (E. coli) and Enteroccocus (ENT), disseminated over time in the Bay of Vidy, which is the most contaminated area of Lake Geneva. Sediments were collected from a site located at $ 500 m from the present waste water treatment plant (WWTP) outlet pipe, in front of the former WWTP outlet pipe, which was located at only 300 m from the coastal recreational area (before 2001). E. coli and ENT were enumerated in sediment suspension using the membrane filter method. The FIB characterization was performed for human Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) and human specific bacteroides by PCR using specific primers and a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Bacterial cultures revealed that maximum values of 35.2 Â 10 8 and 6.6 Â 10 6 CFU g À 1 dry sediment for E. coli and ENT, respectively, were found in the sediments deposited following eutrophication of Lake Geneva in the 1970s, whereas the WWTP started operating in 1964. The same tendency was observed for the presence of human fecal pollution: the percentage of PCR amplification with primers ESP-1/ESP-2 for E. faecalis and E. faecium indicated that more than 90% of these bacteria were from human origin. Interestingly, the PCR assays for specifichuman bacteroides HF183/HF134 were positive for DNA extracted from all isolated strains of sediment surrounding WWPT outlet pipe discharge. The MALDI-TOF MS confirmed the presence of general E. coli and predominance E. faecium in isolated strains. Our results demonstrated that human fecal bacteria highly increased in the sediments contaminated with WWTP effluent following the eutrophication of Lake Geneva. Additionally, other FIB cultivable strains from animals or adapted environmental strains were detected in the sediment of the bay. The approaches used in this research are valuable to assess the temporal distribution and the source of the human fecal pollution in aquatic environments.

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Section: Discussionmentioning

confidence: 65%

Characterization of fecal indicator bacteria in sediments cores from the largest freshwater lake of Western Europe (Lake Geneva, Switzerland)

Thevenon

Regier

Benagli

et al. 2012

Ecotoxicology and Environmental Safety

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“…Bacteroides have long been considered a candidate indicator of fecal contamination, because they are among the dominant bacterial groups in the human gut (Wade et al 2006) and consequently their high occurrence in human feces can reach concentrations outnumbering E. coli concentrations by as much as 100-to 1000-fold (Health Canada 2012). Also, due to their inability to grow in the environment, they are expected to persist for much shorter periods than conventional FIB and their concentrations in animal scat are 2-5 orders of magnitude lower than human feces (Converse et al 2009). Bacteroides spp., are Gram-negative, obligate anaerobic bacteria, whose dominant species are B. fragilis, B. vulgatus, B. distasonis and B. thetaiotaomicron.…”

Section: Alternative Microbial Indicatorsmentioning

confidence: 99%

“…Bacteroides spp., are Gram-negative, obligate anaerobic bacteria, whose dominant species are B. fragilis, B. vulgatus, B. distasonis and B. thetaiotaomicron. Detection of Bacteroides genetic markers in RW is a relatively new area of research, but molecular markers have been developed for the genus that are generally more concentrated in human fecal material than animal (Converse et al 2009), and have been shown to be sensitive markers of human sewage in microbial sourcetracking efforts . The research undertaken to date suggests that Bacteroides markers may have a stronger role as a secondary indicator of fecal contamination, providing information on the potential sources of fecal material (Health Canada 2012).…”

Section: Alternative Microbial Indicatorsmentioning

confidence: 99%

Assessment of the microbiological quality of recreational waters: indicators and methods

Rodrigues

Cunha

2017

Euro-Mediterr J Environ Integr

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The microbiological quality of recreational waters (RW) has been assessed through the use of microbial indicators. Usually, three typical fecal bacteria, namely fecal coliforms, Escherichia coli and Enterococcus spp., are deemed by legislation and regulatory agencies for routine monitoring of the fecal contamination in RW. However, these traditional indicators have some limitations, as they underestimate health risks and do not determine the sources of contamination. Currently, the increasing concern with the minimization of human health risks associated with recreational water use and the protection of water resources outweigh the actual normative perspective. The potentials and limitations of conventional and alternative fecal contamination detection systems must be carefully evaluated, taking into account the specificities of each recreational water body and the natural or anthropogenic sources of environmental variability. The careful choice of analysis for detection and quantification of indicators in the light of their potential and shortcomings is of major importance for the timely evaluation of risks and implementation of strategies that effectively protect users and preserve water quality. This paper presents a critical literature review of the traditional and innovative methodologies for the analysis of fecal indicators in recreational water and intends to discuss their contribution for the management of the microbiological quality and safety of leisure surface waters.

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“…However, none of these species-specific markers were completely specific to sewage. The performance of the forward primer HF183 along with a previously designed reverse primer (BFDrev) was superior to all six newly developed species-specific qPCR assays [46,47].…”

Section: Introductionmentioning

confidence: 98%

Current Status of Marker Genes of Bacteroides and Related Taxa for Identifying Sewage Pollution in Environmental Waters

Ahmed

Hughes

Harwood

2016

Water

112

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Abstract:Microbial source tracking (MST) endeavors to determine sources of fecal pollution in environmental waters by capitalizing on the association of certain microorganisms with the gastrointestinal tract and feces of specific animal groups. Several decades of research have shown that bacteria belonging to the gut-associated order Bacteroidales, and particularly the genus Bacteroides, tend to co-evolve with the host, and are, therefore, particularly suitable candidates for MST applications. This review summarizes the current research on MST methods that employ genes belonging to Bacteroidales/Bacteroides as tracers or "markers" of sewage pollution, including known advantages and deficiencies of the many polymerase chain reaction (PCR)-based methods that have been published since 2000. Host specificity is a paramount criterion for confidence that detection of a marker is a true indicator of the target host. Host sensitivity, or the prevalence of the marker in feces/waste from the target host, is necessary for confidence that absence of the marker is indicative of the absence of the pollution source. Each of these parameters can vary widely depending on the type of waste assessed and the geographic location. Differential decay characteristics of bacterial targets and their associated DNA contribute to challenges in interpreting MST results in the context of human health risks. The HF183 marker, derived from the 16S rRNA gene of Bacteroides dorei and closely related taxa, has been used for almost two decades in MST studies, and is well characterized regarding host sensitivity and specificity, and in prevalence and concentration in sewage in many countries. Other markers such as HumM2 and HumM3 show promise, but require further performance testing to demonstrate their widespread utility. An important limitation of the one-marker-one-assay approach commonly used for MST is that given the complexities of microbial persistence in environmental waters, and the methodological challenges of quantitative PCR (qPCR) in such samples, the absence of a given marker does not ensure the absence of fecal pollution in the source water. Approaches under development, such as microarray and community analysis, have the potential to improve MST practices, thereby increasing our ability to protect human and ecosystem health.

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Rapid QPCR-based assay for fecal Bacteroides spp. as a tool for assessing fecal contamination in recreational waters

Cited by 81 publications

References 39 publications

Characterization of fecal indicator bacteria in sediments cores from the largest freshwater lake of Western Europe (Lake Geneva, Switzerland)

Characterization of fecal indicator bacteria in sediments cores from the largest freshwater lake of Western Europe (Lake Geneva, Switzerland)

Assessment of the microbiological quality of recreational waters: indicators and methods

Current Status of Marker Genes of Bacteroides and Related Taxa for Identifying Sewage Pollution in Environmental Waters

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