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A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters
Published on: March 9, 2016
Potential use of a host associated molecular marker in Enterococcus faecium as an index of human fecal pollution
Troy M Scott1, Tracie M Jenkins, Jerzy Lukasik
1Biological Consulting Services of N Florida, Inc, Gainesville, Florida 32609, USA. tscott@gator.net
Abstract:
Several genotypic and phenotypic microbial source tracking (MST) methods have been proposed and utilized to differentiate groups of microorganisms, usually indicator organisms, for the purpose of tracking sources of fecal pollution. Targeting of host-specific microorganisms is one of the approaches currently being tested. These methods are useful as they circumvent the need to isolate individual microorganisms and do not require the establishment of reference databases. Several studies have demonstrated that the presence and distribution of Enterococcus spp. in feces seems to be influenced by the host species. Here, we present a method for detection of genetic sequences in culturable enterococci capable of identifying human sources of fecal pollution in the environment. The human fecal pollution marker designed in this study targets a putative virulence factor, the enterococcal surface protein (esp), in Enterococcus faecium. This gene was detected in 97% of sewage and septic samples but was not detected in any livestock waste lagoons or in bird or animal fecal samples. Epidemiological studies in recreational and groundwaters have shown enterococci to be useful indicators of public health risk for gastroenteritis. By identifying the presence of human fecal pollution, and therefore the possible presence of human enteric pathogens, this marker allows for further resolution of the source of this risk.
Insights
A new method detects human fecal pollution by identifying the enterococcal surface protein (esp) gene in Enterococcus faecium. This marker accurately distinguishes human waste from animal sources, aiding public health risk assessment.
Area of Science:
- Environmental microbiology
- Molecular biology
- Public health
Background:
- Microbial source tracking (MST) methods identify fecal pollution sources.
- Host-specific microbial markers offer an alternative to traditional MST.
- Enterococcus species distribution varies by host, suggesting potential as an indicator.
Purpose of the Study:
- To develop a molecular method for detecting human fecal pollution.
- To identify a specific genetic marker in Enterococcus faecium indicative of human origin.
Main Methods:
- Targeted detection of the enterococcal surface protein (esp) gene in Enterococcus faecium.
- Analysis of fecal samples from various sources including sewage, septic systems, livestock, and wildlife.
Main Results:
- The esp gene was detected in 97% of human sewage and septic samples.
- The esp gene was absent in all tested livestock waste and animal/bird fecal samples.
- Enterococci are established indicators of public health risks like gastroenteritis.
Conclusions:
- The esp gene in Enterococcus faecium serves as a reliable marker for human fecal pollution.
- This method enhances the resolution of fecal pollution sources in environmental waters.
- Identifying human fecal pollution aids in assessing and mitigating public health risks associated with enteric pathogens.
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