Progress 10/01/10 to 09/30/13
Outputs
Target Audience: Our target audiences served by this project include shellfish industry through Spinney Creek Shellfish (SCS), Inc. (Eliot, ME-co-PI Howell, President) and regional oyster growers, shellfish programs and public health agencies. We have an ongoing collaboration with SCS in experimental design, data interpretation and use of their facilities. We have also conducted analysis for total and pathogenic V. parahaemolyticus, V. vulnificus and V. cholerae in oysters from industry partners in MA (Island Creek Oysters) and in CT that were part of the recalled oysters caused by V. parahaemolyticus outbreaks in those two states during 2013. During the summer and early fall of 2013, we also met with key personnel from the public health and shellfish program in ME (ME-DMR & ME-CDC), NH (NH-DES & NH-NHHS), MA (MA-DMF & MA-DPH) and CT (CT-Bureau of Ag.& CT-DPH) to share our knowledge about Vibrio ecology and pathogenesis and to start planning for collaborative work with these agencies to help prevent outbreaks during the summer of 2014. We have received and partially analyzed clinical V. parahaemolyticus isolates from patients from NH & MA, and will soon be receiving isolates from ME & CT to determine differences and similarities in characteristics between these clinical isolates and isolates from the environment that we have collected through the years in the Great Bay estuary. The project findings have helped these groups of colleagues to understand how best to manage harvesting during summertime to avoid vibrio-related public health issues. We have also begun to target citizen and non-industry related audiences to inform them of the potential for exposure to Vibrios through recreational and seafood (raw/undercooked) exposure. Efforts have included seminars given to high school and university students, academic colleagues, citizen volunteer groups, and local environmental groups to inform them of risks and strategies to ensure safe shellfish consumption and recreational activities in NH coastal environments. Citizen volunteers and students have also been educated through hands-on demonstrations of field and laboratory activities with shellfish. Several graduate and undergraduate students working on this project also represent ethnic minorities. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? The work has also provided training to four graduate students, eight undergraduate students, several non-student UNH technicians, and two summertime research fellows who have all gained improved skills and increased knowledge about Vibrios and field, laboratory, environmental and molecular-based research. How have the results been disseminated to communities of interest? The results of project results have been disseminated by a variety of mechanisms. We have made presentations to student, academic and citizen groups to help educate them in the potential risks of eating shellfish that may contain pathogenic Vibrio species. We have also made several talks and presented posters at national and international meetings where scientists, government officials, public health and natural resource professionals, and the shellfish industry have heard about our findings. The most significant new direction for us has been our presentations and discussions with state programs. We have met with and presented our research results on pathogenic Vibrios to the ME Center for Disease Control, MA Division of Marine Fisheries/Shellfish Program, NH Dept. of Health and Human Services and the Department of Environmental Services, the CT Bureau of Agriculture and CT Department of Public Health, and the NH oyster farmers at a NH Sea Grant sponsored meeting this summer. All of these groups are concerned about the risks of Vibrios in shellfish and have come to us because we have the most extensive data and information about these organisms in the Northeast. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The research to date has resulted in significant changes in our knowledge about the dynamics of Vibrios in oysters in Northeast US estuarine ecosystems. Our 2012-13 fieldwork has shown how V. parahaemolyticus and V. vulnificus in oysters, sediments and water are present in more areas and for longer periods of time compared to what was observed in the 1990's. The most likely cause is elevated temperatures in the Gulf of Maine Region. During the summer of 2012, which was the warmest on record for the Gulf of Maine, we observed elevated Vibrio levels during the late spring and summer, as expected with higher temperatures. In fact, the levels in oysters during August 2012 were the highest we have recorded from 2007 to 2013. In the fall of 2012, however, Vibrio levels dropped to undetectable levels, well before expected. At the oyster relay site, which we have monitored since 1991 and where V. parahaemolyticus levels have always been very low or undetectable, levels of V. parahaemolyticus were higher (93/g) than ever detected in 2012. This was not alarming, but the levels during the summer of 2013 were up to 2 orders of magnitude higher than in 2012 during July. This shift in ecosystem population levels, in the year following the warmest year on record for the areas, is a concern and focus of current research. The relay experiments showed relatively consistent and significant decreases in V. parahaemolyticus levels in relayed oysters after 10-14 days during 2011-12 and analysis of shellfish tissue samples for sequencing of the oyster microbiome are underway. The oyster relay research results also suggest Vibrio level dynamics and variability not reported elsewhere, and a biological factor is apparently required for effective removal of Vibrios. The relay experiments conducted during 2013, however, gave much different results because of elevated Vibrio levels in the relay waters, as previously (above) described. More in-depth research is now underway to better understand how oysters bioaccumulate Vibrios from the water column compared to the Vibrios actually colonizing the oyster tissue. Finally, we have expanded the array of genetic markers used to characterize differences and similarities between strains of V. parahaemolyticus and V. vulnificus beyond the routinely used genes for species identification and pathogenicity. Until this year (2013), we never detected what would be considered pathogenic strains of V. parahaemolyticus and V. vulnificus from oysters, water or sediments from the Great Bay estuary. In June-August 2013, we detected potentially pathogenic (containing tdh/trh genetic markers) strains of V. parahaemolyticus for the first time in local oysters. We have also detected pathogenic strains in oysters from MA and CT that were collected from areas closed to harvesting in 2013 because of V. parahaemolyticus infections. The detection of these putatively pathogenic strains is significant because these strains are rarely detected from environmental samples in New England waters. Partial sequencing analysis of clinical and environmental V. parahaemolyticus isolates shows some evidence of relatedness between these and strains from the environment that do not contain these genetic markers, suggesting that pathogenic strains may be evolving from within native V. parahaemolyticus populations. This is a significant outcome of the research.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2012
Citation:
Jones, S.H. 2012. Detection methods for pathogenic Vibrio species. Abstract. Invited presentation at the Northeast Aquaculture Conference and Exposition/Milford Aquaculture Seminar/International Conference on Shellfish Restoration Aquaculture Conference. December 12-14, 2012. Groton, CT.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
Taylor, M., J. Yu, C. Whistler, V. Cooper, T. Howell and S. Jones. 2013. Factors involved with effective relaying reduction of Vibrio parahaemolyticus from oysters (Crassostra virginica). Abstract. Presented at the 9th International Conference on Molluscan Shellfish Safety, Sydney, Australia. March 17-22, 2013.
- Type:
Conference Papers and Presentations
Status:
Under Review
Year Published:
2014
Citation:
Taylor, M., J. Yu, C. Whistler, V. Cooper, T. Howell and S. Jones. Under review. Treatment time and environmental factors affect the reduction of Vibrio parahaemolyticus concentrations in relayed oysters (Crassostrea virginica). Proceedings of the 9th International Conference on Molluscan Shellfish Safety, Sydney, Australia. March 17-22, 2013.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
Taylor, M., J. Yu and S. Jones. 2013. Fourteen?day natural relay to high salinity seawater decreases the presence of Vibrio parahaemolyticus in oysters (Crassostrea virginica). J. Food Protection 76 (Supplement A): 156.
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Progress 10/01/11 to 09/30/12
Outputs
OUTPUTS: Activities included experiments and analyses to set the stage for continued in-depth research. Further testing of alternative isolation media continued with a new modified CHROMagar Vibrio culture medium for identifying Vibrio parahaemolyticus isolates from oyster tissue. The new modified CHROMagar used in conjunction with the existing CHROMagar provides more accurate identifications and is now included in our V. parahaemolyticus isolation and identification protocols. The main experiments involved relaying oysters monthly to determine removal rates and efficiencies for reducing levels of V. parahaemolyticus. Relaying involves moving oysters from harvest sites where Vibrios are present to areas where Vibrios are absent or present at low concentrations. Samples were analyzed from days 0, 7, 10 and 14 as V. parahaemolyticus levels tend to be removed between days 7 and 14. Starting in June, oysters were relayed from areas where vibrios were present in oysters to areas where they are absent. Samples were collected after 7, 10 and 14 days and levels of V. parahaemolyticus were determined using a qPCR-MPN (quantitative polymerase chain reaction-most probable number) approach. These monthly experiments continued through September and thus spanned environmental conditions of varying salinity and temperatures that dropped from 28 degrees C in July to 16 degrees C in September. Long-term monthly monitoring of V. parahaemolyticus levels in oysters, sediments and water at two other sites were continued to provide data from 2007 through 2012. Even though water temperatures were conducive for Vibrio presence, levels declined to very low levels by October in 2012, so no relaying was conducted after September. Isolates from 2010 were analyzed for phylogenetic relatedness and diversity using multilocus (2 housekeeping genes) sequence analysis. Isolates from paired freshly harvested and temperature abused oysters were different, suggesting temperature abuse changes the population and is not be a good strategy for studying relaying. Cold weather isolates were less diverse than those from warm weather. Events included presentation of the results of the research at six local and regional university and high school classes and meetings related to project related topics from climate change to shellfish safety. Dissemination involved frequent meetings with our shellfish industry partners, Spinney Creek Shellfish, to plan and carry out the oyster relaying experiments. We also engaged several researchers, outreach and education, extension and other interested shellfish industry partners from Virginia to Maine in discussions about project findings and collaborations. Project findings and data were shared with the NH Shellfish Program manager and the Piscataqua Region Estuaries Partnership to help frame critical regional research needs. An ongoing collaboration with the non-profit organization Clean Air/Cool Planet included working with a research fellow during the summer to help frame outreach materials on climate change and pathogenic vibrios for dissemination to the public, seafood industry and public health/resource managers, especially in the Northeast US. PARTICIPANTS: Individuals involved in the project include Dr. Steve Jones, who is the PI and supervises all other students and participants, and oversees experimental design, supply purchasing, augmenting funding from other sources, presentations and report writing. Mike Taylor is the PhD student who is using this research project as the basis for his PhD dissertation research. Mike is in charge of the day-to-day lab and fieldwork and supply purchasing, along with data analysis and direct supervision of student helpers. Jong Yu finished his Masters degree based on a previous AES project and has continued helping as a technician on this project. Drs. Cheryl Whistler, Vaughn Cooper and Kelley Thomas are UNH collaborators who take active roles in advising Mike Taylor in his efforts to optimize laboratory and data analysis approaches and to design experiments. Tom Howell, as president of our commercial shellfish depuration partner Spinney Creek Shellfish Inc., has helped by providing access to his facility, boat use, sampling equipment and expertise on shellfish handling and data analysis. Tom has been instrumental in making sure that the shellfish health aspects of the project experiments are not compromised. Besides the two graduate students, we have involved several citizen volunteers from a new volunteer group Dr. Jones is coordinating through the NH Sea Grant Program in sample collection, and we have included training of six undergraduate students in sample collection, processing and microbiological analyses. We also trained a summertime research fellow, Zoe O'Donoghue as part of a Clean Air/Cool Planet collaboration with the president, Adam Markham and Dr. Cooper. Collaborations with several local oyster farmers and Chris Nash, the NH Shellfish Program manager, have been established to help growers manage harvesting during summer to manage Vibrio levels in shellfish. TARGET AUDIENCES: Our target audiences served by this project include shellfish industry through Spinney Creek Shellfish, Inc. and the local NH oyster growers, along with the NH Shellfish Program manager. The project findings have helped them to understand how best to manage harvesting during summertime to avoid vibrio-related public health issues. We have also begun to target citizen audiences to inform them of the potential for exposure to vibrios through recreational and seafood (raw/undercooked) exposure. Several graduate and undergraduate students represent ethnic minorities. Efforts have included seminars given to high school and university students, academic colleagues, citizen volunteer groups, and local environmental groups to inform them of risks and strategies to ensure safe shellfish consumption and recreational activities in NH coastal environments. Citizen volunteers and students have also been educated through hands-on demonstrations of field and laboratory activities with shellfish. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The research to date has resulted in significant changes in our knowledge about the dynamics of Vibrios in oysters in Northeast US estuarine ecosystems. Our 2012 fieldwork showed how V. parahaemolyticus and V. vulnificus in oysters, sediments and water are present in more areas and for longer periods of time compared to what was observed in the 1990's. Our oyster relaying work has also shown unique dynamics and variability for Vibrio concentrations in the oysters not reported elsewhere, and suggests a biological factor is required for removal of Vibrios. The work has also provided training to two graduate students, several undergraduate students and a summertime research fellow who have all gained improved skills and increased knowledge about Vibrios and field, laboratory, environmental and molecular-based research. The results for the relaying experiments and ecological surveys are also under further analysis for publication and planned presentations in conferences.
Publications
- Ellis, C.N. B.M. Schuster, M.J. Striplin, S.H. Jones, C.A. Whistler and V.S. Cooper. 2012. Influence of seasonality on the genetic diversity of Vibrio parahaemolyticus in New Hampshire shellfish waters as determined by multi-locus sequence analysis. Appl. Environ. Microbiol.78: 3778-3782.
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Progress 10/01/10 to 09/30/11
Outputs
OUTPUTS: Activities included several key experiments to help set the stage for more in-depth research next year. An alternative isolation medium, CHROMagar, was tested for identifying Vibrio parahaemolyticus isolates from oyster tissue. CHROMagar was easier to use and gave more accurate identifications compared to the traditional TCBS agar, and is now our medium of choice for isolating V. parahaemolyticus. The main experiments involved relaying oysters monthly to determine removal rates and efficiencies for reducing levels of V. parahaemolyticus. Starting in July, oysters were relayed from areas where vibrios were present in oysters to areas where they are absent. Samples were collected after 2, 7 and 14 days and levels of V. parahaemolyticus were determined using a qPCR-MPN approach. These monthly experiments continued through November and thus spanned environmental conditions of varying salinity and temperatures that dropped from 29 degrees C in July to 7.5 degrees C in November. Long-term monthly monitoring of V. parahaemolyticus levels in oysters, sediments and water at two other sites were continued to provide data from 2007 through 2011. Events included presentation of the results of the research at six local, national and international meetings related to project related topics from climate change to shellfish safety. Dissemination involved frequent meetings with our shellfish industry partners, Spinney Creek Shellfish, to plan and carry out the oyster relaying experiments. We also reached out to other researchers, outreach and education, extension and other interested shellfish industry partners from Virginia to Maine to discuss wider participation in the project and potential collaborations. Meetings were also held at the International Conference on Molluscan Shellfish Safety in Charlottetown, PEI, Canada in June to share research findings and discuss future collaborations. PARTICIPANTS: Individuals involved in the project include Dr. Steve Jones, who is the PI and supervises all other students and participants, and oversees experimental design, supply purchasing, augmenting funding from other sources, presentations and report writing. Mike Taylor is the PhD student who is using this research project as the basis for his PhD dissertation research. Mike is in charge of the day-to-day lab and fieldwork and supply purchasing, along with data analysis and direct supervision of student helpers. Jong Yu is finishing up his Masters degree based on a previous AES project and has been helping as a technician on this project. Cheryl Whistler and Vaughn Cooper are both UNH collaborators and take active roles in advising Mike Taylor in his efforts to optimize laboratory and data analysis approaches and to design experiments. Tom Howell, as president of our commercial shellfish depuration partner Spinney Creek Shellfish Inc., has helped by providing access to his facility, boat use, sampling equipment and expertise on shellfish handling and data analysis. Tom has been instrumental in making sure that the shellfish health aspects of the project experiments are not compromised. Besides the two graduate students, we have involved several citizen volunteers from a new volunteer group Dr. Jones is coordinating through the NH Sea Grant Program in sample collection, and we have included training of three undergraduate students in sample collection, processing and microbiological analyses. TARGET AUDIENCES: Commercial shellfish growers and public health agency personnel PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The research to date has resulted in significant changes in our knowledge about the dynamics of Vibrios in oysters in our NH-Maine estuarine ecosystem, the Great Bay estuary, as well as during the process of oyster relay. Our 2011 fieldwork showed us vastly different results compared to past years for the levels of V. parahaemolyticus in oysters, sediments and water. We are still analyzing the results to enable publication of explanations. Our oyster relaying work has also revealed some results that differ from related work in other areas, like the Gulf of Mexico and Chesapeake Bay. The work has also provided training to two graduate students and several undergraduate students who have all gained improved skills and increased knowledge about Vibrios and both environmental and laboratory research. The results for the relaying experiments are also under further analysis for publication and planned presentations in conferences. Our project is in early stages and no changes in actions or conditions have yet occurred.
Publications
- C.N. Ellis, M.J. Striplin, C.A. Whistler, S.H. Jones, V.S. Cooper. 2010. Population genetic structure of Vibrio parahaemolyticus in New Hampshire's Great Bay Estuary. . In, Geomics Session Abstracts, Vibrios in the Environment 2010, November 7-12, 2010. Biloxi, MS. http://www.joss.ucar.edu/vibrios_2010/abstracts/index.html
- J. C. Mahoney, M. J. Gerding, S. H. Jones and C. A. Whistler. 2010. Temperature regulation of virulence-associated traits in environmental and clinical isolates of Vibrio parahaemolyticus. . In, Human Disease Session Abstracts,Vibrios in the Environment 2010, November 7-12, 2010. Biloxi, MS. http://www.joss.ucar.edu/vibrios_2010/abstracts/index.html
- Schuster, B.M., A. Tyzik, R. Donner, S.H. Jones, V.S. Cooper, C.A. Whistler. 2010. Phenotypic and Multi-locus sequence analysis of Vibrio cholera isolated from the Great Bay Estuary of New Hampshire reveals genetic recombination within this diverse endemic population. . In,Ecology of Vibrios Session Abstracts, Vibrios in the Environment 2010, November 7-12, 2010. Biloxi, MS. http://www.joss.ucar.edu/vibrios_2010/abstracts/index.html
- Jones, S, B. Schuster, J. Mahoney, J. Yu, C. Ellis, V. Cooper and C. Whistler. 2011. The Occurrence, Abundance, Phylogeny and Virulence Potential of Pathogenic Vibrio Species in New Hampshire Shellfish Waters, p. 79, In, National Shellfisheries Association Program and Abstracts of the 103rd Annual Meeting. March 27-31, 2011, Baltimore, MD.
- Mahoney J.C., M. J. Gerding, S. H. Jones & C.A. Whistler. 2010. Characterization of the pathogenic potential of environmental Vibrio parahaemolyticus compared to clinical strains indicates a role for temperature regulation in virulence. Appl. Environ. Microbiol. 76: 7459-7465.
- Jones, S., M. Striplin, J. Mahoney, V. Cooper and C. Whistler. 2010. Incidence and abundance of pathogenic Vibrio species in the Great Bay Estuary, New Hampshire, pp. 127-134, In, Proceedings of the Seventh International Conference on Molluscan Shellfish Safety. Lassus, P. (Ed.). Nantes, France, June 14-19, 2009. Quae Publishing, Versailles, France.
- B.M. Schuster, A. Tyzik, R. Donner, S.H. Jones, V.S. Cooper, C.A. Whistler. 2011. Population structure and ecological correlations of an endemic northern temperate population of Vibrio cholerae with close relatives to toxigenic isolates. Appl. Envrion. Microbiol.77: 7568-7575.
- Jones, S.H. 2011. Microbial Pathogens and Biotoxins: State of the Gulf of Maine Report. Gulf of Maine Council on the Marine Environment. http://www.gulfofmaine.org/stateofthegulf. 21 pp.
- Yu, JW, TL Howell, CA Whistler, VS Cooper, SH Jones. 2010. Oyster processing strategies for eliminating Vibrio parahaemolyticus and Vibrio vulnificus from New Hampshire oysters. In, Public Health and Related Factors Session Abstracts, Vibrios in the Environment 2010, November 7-12, 2010. Biloxi, MS. http://www.joss.ucar.edu/vibrios_2010/abstracts/index.html
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