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Serological and Molecular Biological Studies of Marine Predator-Prey Systems

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Serological and Molecular Biological Studies of Marine Predator-Prey Systems Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 1998 Serological and Molecular Biological Studies of Marine Predator-Prey Systems. Marilyn J. Hoyt Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Hoyt, Marilyn J., "Serological and Molecular Biological Studies of Marine Predator-Prey Systems." (1998). LSU Historical Dissertations and Theses. 6739. https://digitalcommons.lsu.edu/gradschool_disstheses/6739 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Historical Dissertations and Theses by an authorized administrator of LSU Digital Commons. For more information, please contact [email protected]. INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back o f the book. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6” x 9” black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. UMI A Bell & Howell Information Company 300 North Zed) Road, Ann Arbor MI 48106-1346 USA 313/761-4700 800/521-0600 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. SEROLOGICAL AND MOLECULAR BIOLOGICAL STUDIES OF MARINE PREDATOR-PREY SYSTEMS A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor o f Philosophy in The Department of Microbiology by Marilyn J. Hoyt B.S., Louisiana State University, 1986 D.V.M., Louisiana State University, 1991 August, 1998 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. UMI Number: 9902641 UMI Microform 9902641 Copyright 1998, by UMI Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. UMI 300 North Zeeb Road Ann Arbor, MI 48103 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. DEDICATION This work is dedicated to my husband, Phillip Hoyt and my two sons, Nathan and Brenten. Their patience and understanding have allowed this project to progress to completion. Thank you for being there when I needed you. ii Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ACKNOWLEDGMENT Thank you to Dr. R. J. Siebeling, for his support, encouragement and assistance with this project. He is an exceptional teacher and researcher, who provided guidance while encouraging self reliance. Thank you to all the members of my committee. Dr. Eric Achberger, provided assistance whenever necessary as did Dr. Randy Gayda. Dr. David Horohov and Dr. Greg Jarosik, were available for meetings and provided assistance when necessary. Thank you to Dr. David Baker as the appointed committee member for you assistance. Thank you, to Dr. John Fleeger, who provided much of the expertise of working with harpacticoids and grass shrimp and whose laboratory supplied most of the samples. He worked tirelessly on this project and was always available. Margaret Pace and Stacy Webb were responsible for the feeding trials and sample collections and contributed a lot to this project as it could not have been done without them. James Ditty, supplied and morphologically identified the anchovy specimens for this work. Thank you, Jim, as this project would not have progressed without your help Thanks are extended to Dr. V. R. Shrinivasan who always had a suggestion, especially when I need one and to Dr. Amy Zuppardo who generously shared the laboratory with me and was unfailing in her support and help, and to Cindy Hink for her suggestions and technical support with slides and presentations as well as being a friend. This research was funded by Sea Grant. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. TABLE OF CONTENTS DEDICATION................................................................................................... ii ACKNOWLEDGMENTS.................................................................................. iii LIST OF TABLES............................................................................................. v LIST OF FIGURES........................................................................................... vi ABSTRACT...................................................................................................... vii INTRODUCTION/LITERATURE REVIEW..................................................... 1 CHAPTER I. IDENTIFICATION OF ANCHOVY SPECIES BY RESTRICTION ENZYME FRAGMENT ANALYSIS OF PCR AMPLIFIED MITOCHONDRIAL NADH DEHYDROGENASE GENES 5 AND 6 ... 16 INTRODUCTION................................................................................... 17 MATERIALS AND METHODS............................................................ 19 RESULTS............................................................................................... 28 DISCUSSION......................................................................................... 43 CHAPTER n. SEROLOGICAL DETECTION OF A HARPACTICOID (Amphiscoides atopus) PROTEIN IN THE GUT CONTENTS OF GRASS SHRIMP (Palaemonetes pugio) ............................................................ 52 INTRODUCTION................................................................................... 53 MATERIALS AND METHODS............................................................ 55 RESULTS............................................................................................... 63 DISCUSSION......................................................................................... 70 SUMMARY/CONCLUSION............................................................................. 78 REFERENCES................................................................................................... 82 V ITA .................................................................................................................. 90 iv Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. LISTS OF TABLES 1. Serological analysis of rabbit anit-fish-RC serum hemagglutination with homologous and heterologous red cell suspensions ............... 30 2. Serological analysis of absorbed rabbit anti-fish-RBC serum hemagglutination with homologous and heterologous red cell suspensions ...................................................................... 30 3. Serological analysis of catfish anti-fish-RBC serum hemagglutination with homologous and heterologous red cell suspensions ............. 31 4. Comparison of Dpn II restriction enzyme fragment patterns of adult anchovy mtDNA to morphological identification ............................. 37 5. PCR amplification of mtDNA preserved larval anchovies .......................... 39 6. Restriction enzyme fragment analyses of anchovy larvae mtDNA collected on 8/26/96 (ethanol preserved) ......................................... 41 7. Restriction enzyme fragment analyses of anchovy larvae mtDNA collected on 7/24/94 and 5/22/97 (ethanol preserved) ................... 42 8. Restriction enzyme fragment analyses of anchovy larvae mtDNA undated (ethanol preserved) .......................................................... 42 9. Comparison of restriction fragments pattern analyses of mtDNA with morphological identification of larvae ................................... 44 10. Gut clearance in grass shrimp fed fluorescent bead-labeled food ...................67 11. Fluorescent bead-labeled food traced through digestive tract of grass shrimp by microscopy ..................................................... 68 v Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. LIST OF FIGURES 1. A diagram of generic fish mitochondrial DNA based on sequence from five species .................................................................. 9 2. Immunization schedules for anti-fish erythrocyte serum .......................... 21 3. PCR amplified product from four species of adult anchovies ...................
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