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Pathogen Source Tracking–Library Independent Methods
Project Goal
To develop and evaluate novel molecular techniques capable of rapidly identifying human pathogens in the marine environment. Learn more about the background information for this project »
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Abstract
The HML OHH pathogen and source tracking project is developing tools to allow rapid pathogen detection from coastal waters and shellfish, and to allow sources of pollution to be identified. Recent accomplishments include microbial assessments of tidal creek systems and shellfish beds along the South Carolina coast. The tidal creek project has revealed a gradient of microbial concentrations in tidal creeks, with higher concentrations typically observed in the headwaters of the creeks. The shellfish project has demonstrated that shellfish from some sites harbored selected pathogens, and that pathogen abundances are not necessarily correlated with those of standard bacterial indicators. These field studies are also demonstrating the applicability of methods developed to date. Additional methods for pathogen detection, including Luminex xMap Ô technology that allows for rapid, simultaneous detection of numerous microbial species, are in early stages of development. The capacity to apply Luminex methodology for bacterial pathogens and real-time PCR methods for viral pathogens will provide NOAA the biotechnology and scientific information to integrate its resource assessments and research activities with those of public health institutions including NIH, NIEHS, NSF, CDC, State Health Departments and academic research centers. Together, these new tools can be used for coastal ecosystem assessments and protection of public health.
Objectives
- Test and evaluate the Luminex methodology for simultaneous detection of the pathogenic and source-specific bacteria; and
- Develop, refine and field test real-time PCR assays for detecting and quantifying viral pathogens.
Expected Results
The techniques developed through this project are expected to produce rapid, cost effective, and specific identifications and host source determinations of human pathogens in estuarine waters and marine organisms. These techniques will aid in understanding the fate, transport, and load of the selected microorganisms in coastal ecosystems, provide data to resource managers and ultimately provide tools that environmental managers can apply to make timely decisions about coastal water quality.
Accomplishments:
- Developed RT-PCR assays, including competitive internal positive controls, for direct detection of noroviruses, enteroviruses, and coliphage MS2 (viral indicator).
- Developed a multiplex Luminex assay to simultaneously detect Enterococcus spp. and Methanobrevibacter smithii as indicators of fecal pollution and human-source fecal pollution, respectively.
- Analyzed water and oyster samples from tidal creeks (in the Southeastern US) to determine the densities of fecal indicator organisms and pathogens; this study was conducted in collaboration with the Monitoring and Assessment Core. Preliminary results suggest that headwaters/sediments are reservoirs for fecal bacteria and that bacterial indicators do not adequately model enteric pathogens in these systems. Results from tidal creeks in North Carolina and Georgia demonstrate that these trends are present on a regional basis.
- Analyzed oyster samples from SCECAP/LTD sites along the South Carolina coast for fecal coliforms, enterococci, coliphages, noroviruses, Vibrio vulnificus, and Vibrio parahaemolyticus. The presence of human pathogens, i.e.,Vibrio vulnificus, V. parahaemolyticus, and enteric viral pathogens in addition to fecal indicator bacteria in these samples suggest that potential human health risks are present for people who consume shellfish from particular sites in the area.
Transfer of Results
The results will be delivered to researchers through technical journals and conferences, to shellfish and watershed managers through workshops at professional conferences such as the Interstate Shellfish Sanitation Conference and to public health wastewater treatment officials within regional areas. Currently, target audience identification and appropriate dissemination methods are being planned.
Publications/Presentations:
- Johnston, C., L. Webster, J. Stewart, and J. Gooch. Application of Luminex Technology for Detection of Marine Microbes of Public Health Significance. NOAA Oceans and Human Health All-PI Meeting. Charleston, SC Jan. 18-20, 2006.
- Gooch, J.A. and G.I. Scott (2005). Waterborne Microbial Agents as a Homeland Security Threat . ThinkTEC Meeting. Dec 1. Charleston , SC.
- Gooch, J.A., B.J. Robinson, B.C. Thompson, R.F. van Dolah, L.H. Coen, G. Riekerk, M.F. Fulton. (2006). Detection of enteric pathogens in South Carolina oysters. NOAA Oceans and Human Health Initiative All-PI Meeting. Jan. 18-20. Charleston, SC.
- Gooch, J.A., J.R. Stewart, L.F. Webster, B.J. Robinson, B.C. Thompson, and G.I. Scott. (2006). Microbial source tracking in South Carolina: Lessons learned and future directions. EPA Mobile Bay Water Quality Monitoring Workshop. Feb 2. Mobile, AL.
- Robinson, B.J. (2006). Efficiency of Detecting Viruses from Environmental Waters. Southern Section of AOAC International 20 th Annual Meeting. Atlanta, Georgia. April 25, 2006.
- Webster, L.F. (2006). Detection of E. coli Using the Luminex 100 System. Southern Section of AOAC International 20 th Annual Meeting. Atlanta, Georgia. April 25, 2006.
- Robinson, B.J., B.C. Thompson, L.F. Webster, G.T. DiDonato, J.R. Stewart, and J.A. Gooch. Microbial Water Quality of Tidal Creek Systems along the South CarolinaCoast. Poster presentation at The American Society for Microbiology General Meeting, Orlando, FL. May 21-25, 2006.
- Gooch, J.A., B.J. Robinson, B.C. Thompson, R.F. VanDolah, L. Coen, G.H.M. Riekerk, A. Hollis and M.H. Fulton. Determination of Human Enteric Pathogens in Oysters from Coastal South Carolina . Poster presentation at The American Society for Microbiology General Meeting, Orlando, FL. May 21-25, 2006.
- Webster, L.F., A.K. Leight, B.J. Robinson, S. Friedman, J. Macauley, G. Lauenstein, R.A. Warner, M.H. Fulton. Post Hurricane Katrina Assessment of Microbial Indicator Levels. Poster presentation at The American Society for Microbiology General Meeting, Orlando, FL. May 21-25, 2006.
- Gooch, J.A., B.J. Robinson, B.C. Thompson, L.D. Coen, D. Richardson, R.F. vanDolah. (2006). Joint SCECAP/oyster survey study: Results from summer and winter sampling. May 30. Brown Bag Seminar, HML. Charleston, SC.
- Gooch, J.A., C. Johnston, B.J. Robinson, J. Gregory, J.R. Stewart, B.C. Thompson, and L.F. Webster. (2006). Environmental microbiology program at CCEHBR. S1022 Annual Meeting. June 19. Charleston, SC.
(Additional publications/presentations are listed with the Pathogen Source Tracking Core)
Public Information and Outreach:
- Helped train Orange County, CA Sanitation District (OCSD) personnel in E. coli and Enterococcus qPCR techniques and procedures. Training was performed in part by Jason Gregory (NOAA) at a workshop sponsored by the Southern California Coastal Water Research Project (SCCWRP) and was conducted to allow OCSD employees to beta test rapid methods for use by local water quality labs.
- As demonstrated by the presentations cited above, this project is disseminating timely information on tools for detecting and tracking pathogens to a variety of targeted audiences and end-users.
For More Information
Contact: Jan Gooch, (843)762.8643
Email: Jan.Gooch@noaa.gov
Contact: Jill Stewart, (843)762.8609
Email: Jill.Stewart@noaa.gov