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Pandemic Vibrio parahaemolyticus

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Material Information

Title:
Pandemic Vibrio parahaemolyticus defining strains using molecular typing and a growth advantage at lower temperatures
Physical Description:
Book
Language:
English
Creator:
Davis, Carisa Renee
Publisher:
University of South Florida
Place of Publication:
Tampa, Fla
Publication Date:

Subjects

Subjects / Keywords:
PCR
Ribotyping
MLST
Proteomics
Growth curves
Dissertations, Academic -- Biochemestry & Molecular Biology -- Doctoral -- USF   ( lcsh )
Genre:
non-fiction   ( marcgt )

Notes

Summary:
ABSTRACT: Vibrio parahaemolyticus is a leading cause of seafood-borne illness with a newly emerged pandemic strain. Previous studies compared the pandemic and non-pandemic strains to understand the evolution of the pandemic strain but no definitive explanation for its emergence has been discovered. This study investigated the molecular characteristics of the pandemic strain and growth characteristics at different temperatures. The hypothesis tested was that pandemic strains of V. parahaemolyticus have modifications to their proteome that give a selective advantage over the other V. parahaemolyticus strains at temperatures normally encountered in the environment. Molecular typing techniques; automated ribotyping, pandemic specific PCR and multilocus sequence typing (MLST), were compared to determine the best method for pandemic strain determination. MLST was the best method because it was the most informative and accurate.Furthermore, nine Florida outbreak strains were identified as pandemic. Using representatives of both strains, growth curves were produced at four temperatures. The five pandemic strains had a significantly faster growth rate at 12°C than five non-pandemic strains. Temperature specific proteomic comparisons were completed using liquid chromatography followed by tandem mass spectroscopy. The proteome differences between these two groups at 12°C included three proteins (DnaA, DnaJ-related protein and DnaK-related protein) with functions related to cold stress. DnaA was expressed in the non-pandemic strain and not the pandemic strain, while the reverse was true for DnaJ-related and DnaK-related proteins. Western blot analysis and LC-MS/MS analysis on additional strains did not support the initial LC-MS/MS results. Growth studies using expression recombinants were employed to investigate these proteins on growth at 12°C.The overexpression of DnaA and DnaJ-related proteins did not significantly alter the growth rates compared to the control strain, but the overexpression recombinant strains DnaK-5 has a significantly slower growth rate than the control strain, the opposite direction as expected. The pandemic strain grows faster at lower temperatures, but the reason has not been determined. A theory is offered in which the pandemic growth advantage related to regulation of cold stress, leading to a shorter lag phase and faster growth rate after acclimation to the lower temperatures. Further experiments to investigate this theory are discussed.
Thesis:
Dissertation (Ph.D.)--University of South Florida, 2008.
Bibliography:
Includes bibliographical references.
System Details:
Mode of access: World Wide Web.
System Details:
System requirements: World Wide Web browser and PDF reader.
Statement of Responsibility:
by Carisa Renee Davis.
General Note:
Title from PDF of title page.
General Note:
Document formatted into pages; contains 120 pages.
General Note:
Includes vita.

Record Information

Source Institution:
University of South Florida Library
Holding Location:
University of South Florida
Rights Management:
All applicable rights reserved by the source institution and holding location.
Resource Identifier:
aleph - 002021189
oclc - 427639108
usfldc doi - E14-SFE0002531
usfldc handle - e14.2531
System ID:
SFS0026848:00001


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Full Text

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Davis, Carisa Renee.
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Pandemic Vibrio parahaemolyticus :
b defining strains using molecular typing and a growth advantage at lower temperatures
h [electronic resource] /
by Carisa Renee Davis.
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[Tampa, Fla] :
University of South Florida,
2008.
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Title from PDF of title page.
Document formatted into pages; contains 120 pages.
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Dissertation (Ph.D.)--University of South Florida, 2008.
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Includes bibliographical references.
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Text (Electronic dissertation) in PDF format.
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ABSTRACT: Vibrio parahaemolyticus is a leading cause of seafood-borne illness with a newly emerged pandemic strain. Previous studies compared the pandemic and non-pandemic strains to understand the evolution of the pandemic strain but no definitive explanation for its emergence has been discovered. This study investigated the molecular characteristics of the pandemic strain and growth characteristics at different temperatures. The hypothesis tested was that pandemic strains of V. parahaemolyticus have modifications to their proteome that give a selective advantage over the other V. parahaemolyticus strains at temperatures normally encountered in the environment. Molecular typing techniques; automated ribotyping, pandemic specific PCR and multilocus sequence typing (MLST), were compared to determine the best method for pandemic strain determination. MLST was the best method because it was the most informative and accurate.Furthermore, nine Florida outbreak strains were identified as pandemic. Using representatives of both strains, growth curves were produced at four temperatures. The five pandemic strains had a significantly faster growth rate at 12¨C than five non-pandemic strains. Temperature specific proteomic comparisons were completed using liquid chromatography followed by tandem mass spectroscopy. The proteome differences between these two groups at 12¨C included three proteins (DnaA, DnaJ-related protein and DnaK-related protein) with functions related to cold stress. DnaA was expressed in the non-pandemic strain and not the pandemic strain, while the reverse was true for DnaJ-related and DnaK-related proteins. Western blot analysis and LC-MS/MS analysis on additional strains did not support the initial LC-MS/MS results. Growth studies using expression recombinants were employed to investigate these proteins on growth at 12¨C.The overexpression of DnaA and DnaJ-related proteins did not significantly alter the growth rates compared to the control strain, but the overexpression recombinant strains DnaK-5 has a significantly slower growth rate than the control strain, the opposite direction as expected. The pandemic strain grows faster at lower temperatures, but the reason has not been determined. A theory is offered in which the pandemic growth advantage related to regulation of cold stress, leading to a shorter lag phase and faster growth rate after acclimation to the lower temperatures. Further experiments to investigate this theory are discussed.
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Mode of access: World Wide Web.
System requirements: World Wide Web browser and PDF reader.
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Co-advisor: Andrew Cannons, Ph.D.
Co-advisor: Huntington Potter, Ph.D.
653
PCR
Ribotyping
MLST
Proteomics
Growth curves
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Dissertations, Academic
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x Biochemestry & Molecular Biology
Doctoral.
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t USF Electronic Theses and Dissertations.
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u http://digital.lib.usf.edu/?e14.2531