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RHODOCOCCUS EQUI</Em> in the FOAL – IMPROVING DIAGNOSTIC University of Kentucky UKnowledge Theses and Dissertations--Veterinary Science Veterinary Science 2018 RHODOCOCCUS EQUI IN THE FOAL – IMPROVING DIAGNOSTIC AND PREVENTION MEASURES Fernanda Bicudo Cesar University of Kentucky, [email protected] Digital Object Identifier: https://doi.org/10.13023/etd.2018.315 Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Bicudo Cesar, Fernanda, "RHODOCOCCUS EQUI IN THE FOAL – IMPROVING DIAGNOSTIC AND PREVENTION MEASURES" (2018). Theses and Dissertations--Veterinary Science. 36. https://uknowledge.uky.edu/gluck_etds/36 This Doctoral Dissertation is brought to you for free and open access by the Veterinary Science at UKnowledge. It has been accepted for inclusion in Theses and Dissertations--Veterinary Science by an authorized administrator of UKnowledge. For more information, please contact [email protected]. STUDENT AGREEMENT: I represent that my thesis or dissertation and abstract are my original work. Proper attribution has been given to all outside sources. I understand that I am solely responsible for obtaining any needed copyright permissions. I have obtained needed written permission statement(s) from the owner(s) of each third-party copyrighted matter to be included in my work, allowing electronic distribution (if such use is not permitted by the fair use doctrine) which will be submitted to UKnowledge as Additional File. I hereby grant to The University of Kentucky and its agents the irrevocable, non-exclusive, and royalty-free license to archive and make accessible my work in whole or in part in all forms of media, now or hereafter known. I agree that the document mentioned above may be made available immediately for worldwide access unless an embargo applies. I retain all other ownership rights to the copyright of my work. I also retain the right to use in future works (such as articles or books) all or part of my work. I understand that I am free to register the copyright to my work. REVIEW, APPROVAL AND ACCEPTANCE The document mentioned above has been reviewed and accepted by the student’s advisor, on behalf of the advisory committee, and by the Director of Graduate Studies (DGS), on behalf of the program; we verify that this is the final, approved version of the student’s thesis including all changes required by the advisory committee. The undersigned agree to abide by the statements above. Fernanda Bicudo Cesar, Student Dr. David W. Horohov, Major Professor Dr. Daniel Howe, Director of Graduate Studies RHODOCOCCUS EQUI IN THE FOAL – IMPROVING DIAGNOSTIC AND PREVENTION MEASURES ________________________________________ DISSERTATION _________________________________________ A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the College of Agriculture, Food and Environment at the University of Kentucky By Fernanda Bicudo Cesar Lexington, Kentucky Director: Dr. David W. Horohov, Professor of Immunology Lexington, KY 2018 Copyright © Fernanda Bicudo Cesar 2018 ABSTRACT OF DISSERTATION RHODOCOCCUS EQUI IN THE FOAL – IMPROVING DIAGNOSTIC AND PREVENTION MEASURES Although Rhodococcus equi (R. equi), previously known as Corynebacterium equi, was first isolated from pneumonic foals almost a century ago, it remains the most common cause of subacute or chronic granulomatous bronchopneumonia in foals. While the majority of foals exposed to R. equi develop a protective immune response (regressors), others exhibit a unique susceptibility to infection (progressors). The determinants for either outcome are not completely understood. Therefore, current diagnostic and preventive measures are suboptimal and require betterment. In light of this current need, we hypothesized that immunoglobulin G subisotype T [IgG(T)] against the virulence-associated protein A (VapA) of R. equi, and whole blood cytokine expression profile of foals predict the outcome of infection and can be used as diagnostic markers of clinical disease. Further, we hypothesized that the use of R. equi hyperimmune plasma (HIP) decreases severity of disease in naturally infected foals, playing an important role in disease prevention in the field. Lastly, we hypothesized that specific anti-Rhodococcus equi pili antibodies passively acquired by foals via colostrum after immunization of pregnant mares with a Rhodococcus equi pili-based candidate vaccine will confer protection against induced disease, and therefore have an immediate impact on R. equi pneumonia prophylaxis. The objectives of this study were: (1) to describe the humoral immune response of progressor and regressor foals to R. equi following experimental challenge and natural infection, (2) to compare the cytokine and cell-marker expression profile in whole blood of progressor and regressor foals after challenge, (3) to evaluate the Vap-A specific IgG profile of a commercially available HIP product and its value as a prophylactic tool on an endemic farm, and (4) to evaluate the efficacy of a vaccine based on the Rhodococcus equi pili (Rpl). Although the IgG(T) response of progressor foals after challenge or following natural infection tended to be more pronounced than that observed in regressor foals, its performance as a diagnostic test for predicting disease outcome was poor. Likewise, whole blood cell-marker and cytokine expression profiles of progressor and regressor foals were not significantly different, undermining its reliability as a diagnostic tool. Evaluation of the association of HIP VapA specific IgG profile and rhodococcal disease outcome in the field resulted in the conclusion that progressor foals received significantly less VapA specific IgG, suggesting that HIP may have provided some protection to regressor foals. Although HIP appeared to have provided some protection against clinical pneumonia, Rpl maternally-derived IgG failed to confer any advantage to foals born from vaccinated mares. The Rpl candidate vaccine failed to confer protection to foals after challenge, and did not decrease disease severity in comparison to a control group. In summary, the results of this study do not support the use of VapA specific IgG(T) or whole blood cytokine expression profile as predictors of disease outcome. Further, our results suggest a positive effect of HIP on disease outcome. Lastly, the presence of systemic and local Rpl antibodies was not protective in foals. KEY WORDS: Rhodococcus equi, foal, VapA, IgG(T), hyperimmune plasma, vaccine. Fernanda Cesar August 1, 2018 RHODOCOCCUS EQUI IN THE FOAL – IMPROVING DIAGNOSTIC AND PREVENTION MEASURES By Fernanda Bicudo Cesar Dr. David W. Horohov Director of Dissertation Dr. Daniel Howe Director of Graduate Studies June 2018 To my husband Patrick and our families, for their endless support. To my son Benjamin, hopefully as a future example of perseverance. ACKNOWLEDGEMENTS I would like to acknowledge and thank the following important people who have been instrumental in the completion of this doctorate. Firstly, I would like to express my gratitude to my advisor, Dr. David Horohov, for his unwavering support, trust and guidance throughout this research project. I would also like to thank the members of my committee for their continuous insight. Special thanks to Dr. John Timoney who carefully reviewed this dissertation, providing very detailed feedback and improvement. I would like to express my gratitude to all the hands-on help I received from the UK farm personnel, local veterinarians and farms managers. Their contribution and eagerness to aid in answering our research questions drove a significant part of our work. Finally, I would like to thank my mother-in-law Carolyn Godfrey who has given up so much of her personal time to care for my son Benjamin since he was 7 weeks old. Her generosity has stamped us and definitely made this possible. iii TABLE OF CONTENTS ACKNOWLEDGEMENTS ............................................................................................................ iii LIST OF TABLES .......................................................................................................................... vi LIST OF FIGURES ....................................................................................................................... vii LIST OF ABBREVIATIONS .......................................................................................................... x CHAPTER 1: LITERATURE REVIEW AND RESEARCH HYPOTHESES ............................... 1 Introduction .................................................................................................................................. 1 Innate immunity of the healthy foal ............................................................................................. 2 Phagocyte function ................................................................................................................... 3 Expression of cytokines, activation markers, and cell receptors.............................................. 4 Adaptive immunity of the healthy foal ........................................................................................ 6 Cell-mediated immunity .......................................................................................................... 6 Humoral immunity ................................................................................................................... 9 Foal’s innate immune response to R. equi ................................................................................
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