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Pathogenicity of Variant Field Isolates of Avian Reovirus And

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Pathogenicity of Variant Field Isolates of Avian Reovirus And PATHOGENICITY OF VARIANT FIELD ISOLATES OF AVIAN REOVIRUS AND MOLECULAR CHARACTERIZATION OF BRAZILIAN VARIANTS FROM COMMERCIAL BROILERS by RAFAEL AZAMBUJA BAMPI (Under the Direction of Holly S. Sellers) ABSTRACT Avian reoviruses are the causative agent of arthritis/tenosynovitis in broilers and turkeys. Since 2011, it has been observed an increased incidence of variant reoviruses inducing tenosynovitis in commercial poultry flocks in the U.S and worldwide, raising the question of cross species infection. Two turkey reoviruses isolates from clinical cases of tenosynovitis in commercial turkey breeders, 105057 and 105208, demonstrated to be pathogenic to commercial broilers, causing tenosynovitis with clinical signs of lameness (105057) and also, enteric disease (105208). Additionally, viral shedding was poor, demonstrating limited horizontal transmission. In commercial turkeys these isolates caused clinical disease and were efficiently shed. In addition, a chicken arthritis reovirus was inoculated in turkeys, but did not cause disease. Taken together, turkey arthritis reoviruses 105057 and 105208 represent a threat for the poultry industry. Additionally, it was determined for the first time the molecular characterization of arthritis reovirus variants from lame commercial broilers from Brazil. INDEX WORDS: Avian reovirus, chicken reovirus, turkey reovirus, tenosynovitis, arthritis, myocarditis, broilers, turkeys, FTA® cards PATHOGENICITY OF VARIANT FIELD ISOLATES OF AVIAN REOVIRUS AND MOLECULAR CHARACTERIZATION OF BRAZILIAN VARIANTS FROM COMMERCIAL BROILERS by RAFAEL AZAMBUJA BAMPI D.V.M, Universidade Federal de Santa Maria, Brazil, 2010 M.B.A, Fundação Getulio Vargas, Brazil, 2013 A Thesis Submitted to the Graduate Faculty of The University of Georgia in Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE ATHENS, GEORGIA 2016 © 2016 Rafael Azambuja Bampi All Rights Reserved PATHOGENICITY OF VARIANT FIELD ISOLATES OF AVIAN REOVIRUS AND MOLECULAR CHARACTERIZATION OF BRAZILIAN VARIANTS FROM COMMERCIAL BROILERS by RAFAEL AZAMBUJA BAMPI Major Professor: Holly S. Sellers Committee: Michael Day Monique França Electronic Version Approved: Suzanne Barbour Dean of the Graduate School The University of Georgia May 2016 DEDICATION I would like to dedicate this thesis to my parents that always encouraged me to study, and especially to my wife Angelica for all her love and support. iv ACKNOWLEDGEMENTS First, I would like to thank my major professor, Dr. Holly S. Sellers for taking me as a graduate student and providing the opportunity to work in her laboratory. I highly appreciate her excellent guidance and support throughout my MSc studies. I am sincerely grateful to Dr. Monique França and Dr. Michael Day for taking the time to monitor my progress, reviewing my work and providing constructive suggestions. I would also like to acknowledge Dr. Susan Williams for her help with the histological work and cooperation that was required during my studies. I extend my gratitude to Mr. Erich G. Linnemann and Dr. Durairaj for their great technical assistance during this period. I would take this opportunity to thank The University of Georgia, The Department of Population Health and Poultry Diagnostic and Research Center, for giving me the opportunity to work and learn during my Master’s program. Finally, a special thanks to my wife for her encouragement and moral support from the beginning until the end of this incredible journey. Without her, this dream would never come true. v TABLE OF CONTENTS Page ACKNOWLEDGEMENTS .................................................................................................v LIST OF TABLES ............................................................................................................. ix LIST OF FIGURES .......................................................................................................... xii CHAPTER 1 INTRODUCTION .............................................................................................1 References ....................................................................................................5 2 LITERATURE REVIEW ..................................................................................7 Introduction ..................................................................................................7 Literature review ..........................................................................................8 Viral classification .......................................................................................8 Physiochemical properties ...........................................................................8 Viral genome ................................................................................................9 Replication Cycle .......................................................................................16 Genetic diversity ........................................................................................19 Incidence and distribution ..........................................................................20 Transmission ..............................................................................................20 Pathogenesis and pathophysiology ............................................................21 Local immunity ..........................................................................................26 Cell mediated immunity .............................................................................27 vi Humoral immunity .....................................................................................28 Control .......................................................................................................29 Vaccination ................................................................................................29 Diagnostics .................................................................................................30 Virus Isolation ............................................................................................30 RT-PCR......................................................................................................31 Serology .....................................................................................................32 Virus Neutralization ...................................................................................32 References ..................................................................................................34 3 PATHOGENICITY OF TWO TURKEY ARTHRITIS REOVIRUS FIELD ISOLATES IN COMMERCIAL BROILERS .................................................49 Summary ....................................................................................................50 Key Words .................................................................................................51 Abbreviation ..............................................................................................51 Introduction ................................................................................................52 Materials and methods ...............................................................................54 Results ........................................................................................................60 Discussion ..................................................................................................64 References ..................................................................................................68 4 PATHOGENICITY OF TURKEY AND CHICKEN ARTHRITIS REOVIRUSES IN COMMERCIAL TURKEYS ............................................91 Summary ....................................................................................................92 Key Words .................................................................................................93 vii Abbreviation ..............................................................................................93 Introduction ................................................................................................94 Materials and methods ...............................................................................95 Results ......................................................................................................100 Discussion ................................................................................................103 References ................................................................................................109 5 DETECTION AND MOLECULAR CHARACTERZATION OF AVIAN REOVIRUSES FROM COMMERCIAL BROILERS IN BRAZIL .............127 Summary ..................................................................................................128 Key Words ...............................................................................................129 Abbreviation ............................................................................................129 Introduction ..............................................................................................130 Materials and Methods .............................................................................131 Results and discussion .............................................................................133 References ................................................................................................138 6 DISCUSSION ................................................................................................144 References ................................................................................................150
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