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Edwardsiella Ictaluri: Pathogenic Mechanisms and Antigens Recognized by Infected Channel Catfish

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Edwardsiella Ictaluri: Pathogenic Mechanisms and Antigens Recognized by Infected Channel Catfish Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 1992 Edwardsiella Ictaluri: Pathogenic Mechanisms and Antigens Recognized by Infected Channel Catfish. Thomas James Baldwin Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Baldwin, Thomas James, "Edwardsiella Ictaluri: Pathogenic Mechanisms and Antigens Recognized by Infected Channel Catfish." (1992). LSU Historical Dissertations and Theses. 5421. https://digitalcommons.lsu.edu/gradschool_disstheses/5421 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 corner 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 of 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. University Microfilms International A Bell & Howell Information Company 300 North Zeeb Road. Ann Arbor, Ml 48106-1346 USA 313/761-4700 800/521-0600 Order Number 9316952 Edwardsie.Ua ictalurix Pathogenic mechanisms and antigens recognized by infected channel catfish Baldwin, Thomas James, Ph.D. The Louisiana State University and Agricultural and Mechanical Col., 1992 UMI 300 N. ZeebRd. Ann Arbor, MI 48106 EDWARDSIELLA ICTALURI: PATHOGENIC MECHANISMS AND ANTIGENS RECOGNIZED BY INFECTED CHANNEL CATFISH 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 of Philosophy in The School of Veterinary Medicine by Thomas J. Baldwin B.S., Washington State University, 1985 D.V.M., Washington State University, 1988 December 1992 MANUSCRIPT THESES Unpublished theses submitted for the Master’s and Doctor's Degrees and deposited in the Louisiana State University Libraries are available for inspection. Use of any thesis is limited by the rights of the author. Bibliographical references may be noted, but passages may not be copied unless the author has given permission. Credit must be given in subsequent written or published work. A library which borrows this thesis for use by its clientele is expected to make sure that the borrower is aware of the above restrictions. LOUISIANA STATE UNIVERSITY LIBRARIES TABLE OF CONTENTS LIST OF T ABLES .......................................... V LIST OF FIGURES ............................................ vi ABSTRACT .................................................viii INTRODUCTION ............................................ 1 CHAPTER I. LITERATURE REVIEW .......................... 5 Enteric septicemia of catfish . ............. 5 Geographic distribution ...................... 5 Species affected ............................ 5 Bacterial morphology and biochemical characteristics ........................ 7 Isolate homogeneity .......................... 7 Bacterial components ........................ 13 C a p s u l e ....................................13 F i m b r i a ....................................13 F l a g e l l a ....................................14 Lipopolysaccharides .................... 15 Outer membrane proteins ............... 17 P l a s m i d s ....................................19 Occurrence and transmission ................. 22 Clinical signs ............................... 2 3 Gross lesions ................................... 24 Histopathology ............................... 25 Pathogenesis .......................... 27 D i a g n o s i s ..................... 30 T r e a t m e n t ........................................ 31 Vaccination ..... ........................ 33 S a l m o n e l l a ............................................ 34 Mucosal invasion ............................ 35 Cell culture pa s s ag e ...........................36 Chemotaxis and motility ...................... 37 A t t a c h m e n t ......................................38 Penetration ......................................41 Transcytosis ................................. 43 T o x i n s .......................................... 44 Serum and phagocyte resistance ................46 Iron siderophores............................... 48 Genetic control of virulence ............... 48 S h i g e l l a .............................................. 50 A t t a c h m e n t ......................................52 Penetration ......................................54 Intracellular multiplication ............... 55 T o x i n s .......................................... 56 Iron siderophores ............... ...... 58 Genetic control of virulence ............... 59 Regulation of virulence genes ............... 60 ii Y e r s i n i a .............................................. 61 Mucosal invasion ............................ 62 Chemotaxis and motility ........... ..... 64 Attachment and penetration ................. 64 Transcellular passage ........................ 69 Serum and phagocyte resistance ................ 69 T o x i n s .......................................... 70 Iron siderophores............................... 70 Genetic control of virulence ............... 71 Study objectives ......................................73 CHAPTER II. EARLY EVENTS IN THE PATHOGENESIS OF ENTERIC SEPTICEMIA OF CHANNEL CATFISH, CAUSED BY EDWARDSIELLA ICTALURI: LIGHT MICROSCOPIC AND BACTERIOLOGIC FINDINGS ............................ 76 Introduction ....................................... 76 Materials and methods ............................ 77 B a c t e r i a ........................................ 77 Maintenance of channel catfish fingerlings . 78 Experimental infection ...................... 79 Tissue collection ............................ 79 Tissue processing ............................ 80 Quantitative cultures ........................ 81 Rabbit anti-E. ictaluri sera ............. 81 Immunohistochemical detection of E. i c t a l u r i ................... 82 Statistical evaluation . 83 R e s u l t s .............................................. 84 Bacterial cultures .......................... 84 Gross lesions ................................... 86 Histologic findings .......................... 86 I n t e s t i n e ............. 86 L i v e r ......................................90 Spleen, trunk kidney, and head kidney . 90 D i s c u s s i o n ............................................ 91 CHAPTER III. EARLY EVENTS IN THE PATHOGENESIS OF ENTERIC SEPTICEMIA OF CATFISH: ELECTRON MICROSCOPIC FINDINGS ............................... 97 Introduction ....................................... 97 Materials and methods ............................ 98 B a c t e r i a ........................................ 98 Rabbit anti-E. ictaluri sera ............... 98 Maintenance and experimental infection of c a t f i s h ................................... 99 Tissue collection ............................ 99 Bacterial culture ............................ 100 Tissue processing and immunogold staining . 100 Results ....................................... 101 Bacterial cultures .......................... 101 Electron microscopic findings ......... 102 I n t e s t i n e ................................ 102 iii L i v e r ..................................... 106 Spleen, trunk kidney, and head kidney . 106 D i s c u s s i o n ............................................ 109 CHAPTER IV. ANTIGENS OF EDWARDSIELLA ICTALURI RECOGNIZED IN NATURALLY INFECTED CHANNEL CATFISH................................................ 113 Introduction ........................................ 113 Materials and methods ............................. 114 B a c t e r i a ........................................114 Enriched bacterial fractions ................ 115 Outer membrane proteins ................ 115 F l a g e l l a ................................... 116 Lipopolysaccharide ...................... 117 Whole cell lysates ............................ 118 Quantification of LPS and p r o t e i n s ...........119 Gel electrophoresis............................ 119 Immunoblots ............. 120 Production of rabbit anti-catfish IgM . .122 R e s u l t s .............................................
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