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Morphologic and Molecular Pathogenesis Study of Condemned Kidneys in Swine From UNIVERSITY OF CALGARY Morphologic and molecular pathogenesis study of condemned kidneys in swine from Alberta by Claudia Benavente A THESIS SUBMITTED TO THE FACULTY OF GRADUATE STUDIES IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE DEPARTMENT OF MICROBIOLOGY AND INFECTIOUS DISEASES CALGARY, ALBERTA DECEMBER, 2010 © Claudia Benavente 2010 Library and Archives Bibliothèque et Canada Archives Canada Published Heritage Direction du Branch Patrimoine de l'édition 395 Wellington Street 395, rue Wellington Ottawa ON K1A 0N4 Ottawa ON K1A 0N4 Canada Canada Your file Votre référence ISBN: 978-0-494-75207-4 Our file Notre référence ISBN: 978-0-494-75207-4 NOTICE: AVIS: The author has granted a non- L'auteur a accordé une licence non exclusive exclusive license allowing Library and permettant à la Bibliothèque et Archives Archives Canada to reproduce, Canada de reproduire, publier, archiver, publish, archive, preserve, conserve, sauvegarder, conserver, transmettre au public communicate to the public by par télécommunication ou par l'Internet, prêter, telecommunication or on the Internet, distribuer et vendre des thèses partout dans le loan, distrbute and sell theses monde, à des fins commerciales ou autres, sur worldwide, for commercial or non- support microforme, papier, électronique et/ou commercial purposes, in microform, autres formats. paper, electronic and/or any other formats. The author retains copyright L'auteur conserve la propriété du droit d'auteur ownership and moral rights in this et des droits moraux qui protege cette thèse. Ni thesis. Neither the thesis nor la thèse ni des extraits substantiels de celle-ci substantial extracts from it may be ne doivent être imprimés ou autrement printed or otherwise reproduced reproduits sans son autorisation. without the author's permission. In compliance with the Canadian Conformément à la loi canadienne sur la Privacy Act some supporting forms protection de la vie privée, quelques may have been removed from this formulaires secondaires ont été enlevés de thesis. cette thèse. While these forms may be included Bien que ces formulaires aient inclus dans in the document page count, their la pagination, il n'y aura aucun contenu removal does not represent any loss manquant. of content from the thesis. Abstract Fifty condemned and 10 normal kidneys were collected from slaughtered pigs in southern Alberta and tested for pathogens as etiological agents of interstitial nephritis. A 75% of the samples were positive for bacterial isolation; bacteria caused an unspecific immune response in the host tested by western blot. In 50% of the tissues, porcine parvovirus was detected by PCR; 17.5% of samples were porcine circovirus type 2 positive; and there was no evidence of porcine reproductive and respiratory syndrome virus infection. Fungi and leptospires were not detected by histochemistry and/or immunohistochemistry. The majority of the condemned kidneys (76%) showed subacute to chronic interstitial nephritis; a 16% glomerulonephritis; and 8% had minimal histological changes. In contrast, normal kidneys (70%) were histologically normal. Taken together, these data suggest that the chronicity of the lesions did not determine the etiological agent, and manipulation of these kidneys may not represent a high human health risk. ii ACKNOWLEDGEMENTS First and foremost, my sincere thanks are due to Dr. Carmen Fuentealba. It is difficult to find words to express my deep gratitude to her. A deep appreciation to Dr. Glen Armstrong, he has contributed in many ways toward the completion of this study, through his support, encouragement and advice. I am grateful for the guidance of Dr. Rebakah De Vinney, who provided helpful suggestions during the course of this project. I thank Professor Oscar Illanes for his valuable discussions and providing helpful critiques in the review of this manuscript. My thanks also go to Dr. Markus Czub and the virology laboratory people for the training, use of equipment, and scientific discussions which were an integral part of my graduate experience. Gratitude is expressed for assistance provided by Dr. Anne Muckle and Dr. Carmencita Yason from the Atlantic Veterinary College, UPEI. The technical assistance needs special mention: Maureen Bukhari, Ramona Taylor, Barbara Waddell, Jan Giles, and Lorraine Lund is greatly appreciated, they helped me in many ways, for which I am thankful to them. I wish to thank Wendy Hutchins for the skills learned in the Biotechnology Training Centre at the University of Calgary. I also express my gratitude to Lisa Ashton of Canadian Food Inspection Agency for her special contribution in the collection of samples. I gratefully acknowledge the financial support provided by UCVM Studentship- Fellowship Award, UCVM Research Fellowship Department of Ecosystem and Public Health, and UCVM C. Fuentealba Start-up Grant from the University of Calgary. Finally, I am indebted to my family for always providing patience, love and words of encouragement during this challenging process. iii DEDICATION To my family, my husband Eduardo and children Sebastian, Macarena and Rodrigo. iv TABLE OF CONTENTS Abstract ............................................................................................................................... ii Acknowledgements ............................................................................................................ iii Dedication .......................................................................................................................... iv Table of Contents .................................................................................................................v List of Tables .................................................................................................................... vii List of Figures and Illustrations ....................................................................................... viii List of Symbols, Abbreviations and Nomenclature .............................................................x CHAPTER ONE: INTRODUCTION ..................................................................................1 1.1 Nephritis in swine ......................................................................................................2 1.1.1 Classification of nephritis ..................................................................................2 1.1.2 Causes of nephritis ............................................................................................3 1.2 Characteristics of emergent/re-emergent pathogens Actinobacillus suis and Actinobacillus equuli ...............................................................................................4 CHAPTER TWO: MATERIALS AND METHODS ........................................................12 2.1 Study design .............................................................................................................12 2.2 Sample collection .....................................................................................................12 2.3 Classification of gross lesions in the kidneys ..........................................................13 2.4 Histology ..................................................................................................................13 2.4.1 Classification of microscopic lesions in the kidneys .......................................14 2.5 Histochemistry .........................................................................................................14 2.6 Bacteriology .............................................................................................................16 2.7 Virology [Polymerase Chain Reaction (PCR)] ........................................................17 2.8 Western blot .............................................................................................................21 2.9 Statistical Analyses ..................................................................................................23 CHAPTER THREE: RESULTS ........................................................................................25 3.1 Classification of gross lesions in the kidneys ..........................................................25 3.2 Classification of microscopic lesions in the kidneys ...............................................25 3.2.1 Tubulointerstitial nephritis ..............................................................................29 3.2.2 Glomerulonephritis ..........................................................................................30 3.3 Histochemistry .........................................................................................................31 3.4 Bacteriology .............................................................................................................40 3.5 Polymerase Chain Reaction for PCV-2, PPV, and PRRS viruses ...........................41 3.6 Western blot .............................................................................................................42 CHAPTER FOUR: DISCUSSION ....................................................................................55 Conclusions ........................................................................................................................69 REFERENCES ..................................................................................................................72 1. APPENDIX 1 ............................................................................................................86
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