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Examination and Comparison of Infectious Bronchitis Virus And EXAMINATION AND COMPARISON OF INFECTIOUS BRONCHITIS VIRUS AND COCCIDIA VACCINES AND VACCINATION METHODS by GRACE ASHBY ALBANESE (Under the Direction of Brian J. Jordan) ABSTRACT Of the myriad of diseases affecting commercial chickens, infectious bronchitis and coccidiosis cause the most significant economic losses. Vaccination for both of these pathogens, infectious bronchitis virus (IBV) and Eimeria spp., occurs at the hatchery using mass application strategies. IBV vaccines are live attenuated viruses, delivering serotype-specific immunity. The Arkansas IBV serotype is the most frequently detected serotype in the field, so vaccination to protect against disease from this type is commonplace. The current vaccine, ArkDPI, is not efficacious, and an alternative Ark-type vaccine is needed. Ark99, once a commercial IBV vaccine strain, was passaged once in embryonating chicken eggs to produce the ArkGA vaccine candidate. After 60 passages in embryos, the ArkGA vaccine was highly attenuated and provided good protection against Ark-type challenge when vaccinating broiler chickens, and is a suitable alternative to the ArkDPI vaccine. Like IBV, Eimeria spp. vaccines are often applied in the hatchery by mass vaccination methods. Vaccines may be applied using an aqueous spray, or a gel applicator bar may be used, although this technique is newer and not yet validated versus the traditional spray method. Vaccination using both a highly viscous and less viscous gel, applied by bar, was compared with liquid spray. Oocyst shedding differed slightly between vaccinated groups, although all groups were equally protected from E. maxima challenge, indicating that all the vaccine applications tested are effective. In addition to coccidiosis vaccine application, oocyst species used in vaccines are critical, because coccidia produces only species-specific immunity in chickens. An Eimeria type, E. mivati, has been contested as a species since its discovery, although it is included in a commercially licensed vaccine. Many claims suggest that E. mivati is a variant of E. mitis. Next generation Illumina sequencing was used to compare the mitochondrial cytochrome C oxidase subunit I gene sequences of samples containing E. mivati with known Eimeria sequences. None of the E. mivati samples contained sequence that matched with E. mivati in GenBank, although all matched with E. mitis. This data provides further evidence that E. mivati is likely a variant of E. mitis, although further genome examination is needed. INDEX WORDS: Infectious bronchitis virus, Broiler, Vaccination, Arkansas DPI, Arkansas GA, Coccidiosis, Eimeria mivati EXAMINATION AND COMPARISON OF INFECTIOUS BRONCHITIS VIRUS AND COCCIDIA VACCINES AND VACCINATION METHODS by GRACE ASHBY ALBANESE B.S., The University of Georgia, 2013 A Dissertation Submitted to the Graduate Faculty of The University of Georgia in Partial Fulfillment of the Requirements for the Degree DOCTOR OF PHILOSOPHY ATHENS, GEORGIA 2017 © 2017 Grace Ashby Albanese All Rights Reserved EXAMINATION AND COMPARISON OF INFECTIOUS BRONCHITIS VIRUS AND COCCIDIA VACCINES AND VACCINATION METHODS by Grace Ashby Albanese Major Professor: Brian J. Jordan Committee: Andrew P. Benson Richard W. Gerhold Mark W. Jackwood Holly S. Sellers Electronic Version Approved: Suzanne Barbour Dean of the Graduate School The University of Georgia December 2017 DEDICATION I would like to dedicate this dissertation to my husband, Jack. Thank you for your love, encouragement, and belief in me. I would also like to dedicate this work to my parents, Linden and Susan, and my sister, Savannah. I could not have achieved this work without the love and guidance of my wonderful husband and family. I am incredibly lucky to have you all as my support system and I am so thankful for it. iv ACKNOWLEDGEMENTS Firstly, I would like to acknowledge my advisor, Brian Jordan. I have truly appreciated your guidance and support over the past few years. I could not ask for a better mentor and I am so thankful that you took a chance on me as your first graduate student. I would also like to thank Mark Jackwood, Drew Benson, Rick Gerhold, and Holly Sellers for serving on my committee. I so appreciate your advice, suggestions, and feedback to guide my research. I truly believe that exposure to your respective research focuses has enriched me greatly as a scientist and I am forever appreciative of the time you have taken to serve on my committee. To Dr. Mike Lacy, thank you for your guidance during my undergraduate program and your mentorship. Having you in my corner opened so many doors for me, and I will forever be grateful for your help. I know that I would not be where I am today without your gentle push for me to go to graduate school, and I so value having you to help me along for so many years. Thank you. To the members of the Jackwood and Jordan labs: Debbie, Sunny, Christina, Emily, Johnny, Julia, and Laura, I could not have completed this research without your help and support. I am so grateful to have been part of a lab that works as a team and I am thankful for all of your help over the years. To the undergraduates and students that have spent time working in the lab, thank you for all of the assistance with the many projects that we worked on. To my family and friends, I love you, and I am so thankful for your support through this experience. Finally, to Merck Animal Health, thank you for sponsoring my doctoral research. I am so grateful to have had the opportunity to work with such a prestigious and impactful company, and exposure to poultry vaccine research and development projects has truly enriched me as a scientist and helped me to realize my passion for poultry vaccine research. v TABLE OF CONTENTS Page ACKNOWLEDGEMENTS .............................................................................................................v LIST OF TABLES ......................................................................................................................... ix LIST OF FIGURES ....................................................................................................................... xi CHAPTER 1 INTRODUCTION .........................................................................................................1 Specific Aims ...........................................................................................................4 References ................................................................................................................9 2 LITERATURE REVIEW ............................................................................................13 1 Review of the United States poultry industry ....................................................13 2 Vaccination of commercial poultry ...................................................................17 3 Infectious bronchitis virus..................................................................................20 4 Infectious bronchitis virus: Arkansas serotype ..................................................27 5 Coccidiosis .........................................................................................................30 6 Coccidiosis: Eimeria mivati ...............................................................................35 References ..............................................................................................................38 3 ATTENUATION CHARACTERISTICS OF A NOVEL ARKANSAS SEROTYPE INFECTIOUS BRONCHITIS VACCINE THAT IS PROTECTIVE AGAINST CHALLENGE..............................................................................................................70 Abstract ..................................................................................................................71 vi Introduction ............................................................................................................73 Materials and methods ...........................................................................................75 Results ....................................................................................................................82 Discussion ..............................................................................................................88 References ..............................................................................................................91 Tables .....................................................................................................................94 Figures....................................................................................................................99 Supplemental materials ........................................................................................107 4 EVALUATION OF A COCCIDIA VACCINE USING SPRAY AND GEL APPLICATIONS .......................................................................................................112 Abstract ................................................................................................................113 Introduction ..........................................................................................................115 Materials and methods .........................................................................................117 Results ..................................................................................................................123 Discussion ............................................................................................................127 References
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