Studies on Equine Placentitis
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University of Kentucky UKnowledge Theses and Dissertations--Veterinary Science Veterinary Science 2014 Studies on Equine Placentitis Igor Frederico Canisso University of Kentucky, [email protected] Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Frederico Canisso, Igor, "Studies on Equine Placentitis" (2014). Theses and Dissertations--Veterinary Science. 21. https://uknowledge.uky.edu/gluck_etds/21 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. Igor Frederico Canisso, Student Dr. Barry A. Ball, Major Professor Dr. Daniel Howe, Director of Graduate Studies STUDIES ON EQUINE PLACENTITIS 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 Igor Frederico Canisso Lexington, Kentucky Director: Dr. Barry A. Ball, Professor of Equine Reproduction Copyright © Igor Frederico Canisso 2014 iii ABSTRACT OF DISSERTATION Two types of placentitis were studied: ascending and nocardioform placentitis. Although the first diagnosis of nocardioform placentitis was made three decades ago, little is known about the disease, due to the lack of an experimental model. In attempt to develop a research model, Crossiella equi was inoculated through intrauterine, intravenous, intrapharyngeal, and oral routes, but none of the routes resulted in nocardioform placentitis. This may indicate that unidentified factors may play a role in disease pathogenesis and that simple presence of bacteria is not sufficient to induce nocardioform placentitis. The second and major component of this dissertation involved the identification of diagnostic markers for placentitis. Because ascending bacterial placentitis is readily and predictably induced using existing experimental models, this model was used to identify diagnostic markers for placentitis in maternal plasma and fetal fluids. Three potential biomarkers were examined: acute phase inflammatory proteins, steroid hormones produced by the fetoplacental unit, and protein composition of the fetal fluids. Of the three acute phase proteins investigated, serum amyloid A and haptoglobin but not fibrinogen increased in association with experimentally induced ascending placentitis. Androgens and progestins appear to be poor markers for placentitis. Serum estradiol 17β concentrations were reduced in mares with experimentally induced placentitis and appear to be a good marker for placentitis in mares. Different methods were used to study the protein composition of the fetal fluids. Alpha- fetoprotein was characterized as a major protein present in the equine fetal fluids, and this protein was elevated in plasma of mares with placentitis. In another study, using a high- throughput proteomic technique several new proteins were characterized in the amniotic and allantoic fluids of mares carrying normal pregnancies, and several previously uncharacterized proteins were detected in the allantoic fluid of mares with placentitis. Three secreting proteins were elevated in allantoic fluid of mares with experimentally induced ascending placentitis. Key-words: ascending placentitis, nocardioform placentitis, pregnancy loss, premature delivery, abortion iv Igor Frederico Canisso Student’s Siganture December, 17, 2014 v STUDIES ON EQUINE PLACENTITIS By Igor Frederico Canisso __________________________________ Dr. Barry A. Ball Director of Thesis __________________________________ Dr. Daniel Howe Director of Graduate Studies December 17, 2014 vi Et in te! Credo in somni! Labor omnia vincit! Igor Canisso This dissertation is dedicated to my wife Erotides Capistrano da Silva and to my daughter Pietra Capistrano Canisso. The completion of this work has stolen several valuable hours of their personal time. In memoriam of the beloved ones that did not have the chance to make through the end of the journey that started 16 years ago: Alexandre Canisso Antonio Frederico Maria M. Canisso vii ACKNOWLEDGMENTS I would like to thank the graduate program of the University of Kentucky for allowing me to join the Veterinary Science program in 2011. The Geoffrey Hughes Foundation is particularly thanked for providing the fellowship that supported my graduate studies in this program. The Kentucky Thoroughbred Association and Kentucky Thoroughbred Owners and Breeders are thanked for supporting all the studies in this dissertation. My adviser Professor Barry A Ball is acknowledged for his mentorship. I feel that I belong to a privileged group of people that had opportunity to work under his guidance. Professor Mats Troedsson is thanked for serving on my committee and for his expertise with equine placentitis. Professor Thomas Curry is thanked for participating on my graduate committee and for opening the doors of his laboratory, and letting us use the facilities. His assistant, Carole Bryant is thanked for her help during the measurements of estradiol and alpha-fetoprotein. Dr. Neil Williams, who served on my graduate committee for about 2 years, is thanked for his services and for contributing with the pathological examinations performed in this dissertation. Professor Edward Squires is thanked for stepping in and serving on my committee in the last 5 months of my graduate studies. Dr. Phillip Bridges is thanked for participating as an external examiner during my defense. All the trainees in the reproduction laboratory and other laboratories at the Maxwell H. Gluck Equine Research Center are thanked for direct or indirect interactions during my graduate studies. Dr. Thomas Scwerczek is thanked for his help with several studies being carried out at the animal pathology building. Faculty members at Department of Veterinary Science are thanked for several positive interactions that in one way or another helped the iii progression of my professional development while I was a PhD student at the Maxwell H. Gluck Equine Research Center. Drs. Karen Wolfsdorf and Alan Dorton are thanked for helping with sample collections for mares in chapter II, by thanking them I would like to extend my gratitude to their colleagues for assistance with these samplings. I would like to thank Mr. Lynn Ennis and Mr. Kevin Gallacher for helping with the animal work and its associated arrangements; by acknowledging them, I would like to extend my gratitude to all the workers at the Maine Chance farm that contributed to this thesis. Drs. Kirsten Scoggin and Alejandro Esteller-Vico are thanked for their providential laboratorial assistance and collaboration during this dissertation. Deb Mollett, Roy Leach, Gail Waltkins, Diane Furry and Edith Eberhart are all thanked for their help during my stay at the Maxwell Gluck Equine Research Center. Drs. Carolyn Cray (University of Miami) and Dr. Erdal Erol (University of Kentucky) are thanked for collaborating with our studies. My parents Antonio Canisso and Rosa Canisso and my parents in law Mauricio Capistrano and Dionice Capistrano are also thanked for always being presented when we needed them. The journey leading to the achievement of this degree has been a bumpy road, full with detours, fulfilled with enjoyable moments and less pleasant moments. As I am entering the professorship atmosphere at the College of Veterinary Medicine of the University of Illinois, I am looking forward to have a calmer journey with fewer detours. iv TABLE OF CONTENTS LIST OF TABLES ......................................................................................................................... vii LIST OF FIGURES ...................................................................................................................... viii Chapter 1 .........................................................................................................................................