vibrio.pdf


Preview of PDF document vibrio.pdf

Page 1 2 34587

Text preview


TABLE OF CONTENTS
ACKNOWLEDGMENTS………………………………………………………………………..ii
LIST OF TABLES………………………………………………………………………………..v
LIST OF FIGURES……………………………………………………………………………..vii
ABSTRACT……………………………………………………………………………………...ix
CHAPTER 1: GENERAL INTRODUCTION…………………………………………………...1
1.1 Vibrio Background………………………………………………………………........1
1.2 Pathogenicity Factors.....................................................................................................3
1.3 Deepwater Horizon Oil Spill.........................................................................................5
1.4 Vibrio parahaemolyticus and Oil...................................................................................5
CHAPTER 2: IMPACT OF THE DEEPWATER HORIZON OIL SPILL ON THE VIBRIO
PARAHAEMOLYTICUS TYPE THREE SECRETION SYSTEM..........................8
2.1 Purpose and Hypotheses................................................................................................8
2.2 Materials and Methods..................................................................................................8
2.2.1 Sample Collection...........................................................................................8
2.2.1.1 Sample Locations.............................................................................9
2.2.2 Sample Preparation and Plating....................................................................10
2.2.2.1 Oyster Preparation and Plating......................................................10
2.2.2.2 Water Preparation and Plating.......................................................11
2.2.2.3 Sediment Preparation and Plating..................................................12
2.2.3 Colony Lifting and Hybridization.................................................................13
2.2.3.1 Proteinase-K (Pro-K).....................................................................14
2.2.3.2 Hybridization.................................................................................14
2.2.4 Pathogenic Vp Isolation................................................................................16
2.2.5 DNA Extraction............................................................................................17
2.2.6 Multiplex Polymerase Chain Reaction (PCR) Screening.............................18
2.2.7 Agarose Gel Electrophoresis.........................................................................19
2.2.8 Statistical Analysis........................................................................................20
2.3 Results..........................................................................................................................20
2.3.1 EID Data.......................................................................................................20
2.3.2 Presence/Absence TTSS Chi-Square Analysis.............................................22
2.4 Discussion and Future Research..................................................................................23
2.4.1 Discussion.....................................................................................................23
2.4.2 Future Research............................................................................................28
CHAPTER 3: IMPACT OF THE DEEPWATER HORIZON OIL SPILL ON THE VIBRIO
COMMUNITY......................................................................................................30
3.1 Purpose and Hypotheses..............................................................................................30
3.2 Materials and Methods................................................................................................30
3.2.1 Water Collection...........................................................................................30
iii