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Failure to Detect Equid Herpesvirus Type 1 DNA in Thoroughbred Placentae and Healthy New-Born Foals

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Failure to Detect Equid Herpesvirus Type 1 DNA in Thoroughbred Placentae and Healthy New-Born Foals Failure to detect equid herpesvirus type 1 DNA in Thoroughbred placentae and healthy new-born foals Principal researcher: Dr Lara Jean Brown Supervisor: Dr Geoff Brown, University of Pretoria, RSA Co-supervisor: Prof Martin Schulman, University of Pretoria, RSA Co-supervisor: Dr Julia Kydd, University of Nottingham, UK Co-supervisor: Prof Tom Stout, Utrecht University, Netherlands Declaration of originality I, Lara Jean Brown, do hereby declare that the research presented in this dissertation was conceived and executed by myself, and apart from the normal guidance of my supervisor, I have received no assistance. Neither the substance, nor any part of this dissertation, has been submitted in the past, nor is to be submitted for a degree at this university, nor any other university. This dissertation is presented in fulfilment of the requirements for the MSc (Production Animal Science) in the department of Production Animal Studies, Faculty of Veterinary Science, University of Pretoria. I hereby grant the University of Pretoria free licence to reproduce this dissertation, in part or as a whole, for the purpose of research or continuing education. 31/10/2017 Lara Jean Brown Date i Acknowledgements I would like to thank all who have assisted me with this project through giving their advice, time and expertise. I would particularly like to thank the following people: My supervisors, Dr Geoff Brown, Prof Martin Schulman, Dr Julia Kydd and Prof Tom Stout for their guidance during the sample collection period and for their support and guidance during the writing up of this thesis. Prof Alan Guthrie for his guidance and assistance in the setting up of our sampling protocol and during the data analysis period. Dr Bennie van der Merwe and Mrs Jacqui van der Merwe for their assistance during the data collection period and for allowing us the use of their horses in this study. I would also like to thank them for their hospitality and for being so welcoming and supportive. Furthermore, I would like to thank the Wits Health Consortium for providing the funding for this project. Without their support, this project would not have been possible. The staff of the Moutonshoek stud farm, in particular, Ms Romi Bettison and Mr Karel Skirmaans, for their invaluable assistance with the data collection and animal handling. The staff of the Veterinary Genetics Laboratory, in particular, Mr Mpho Monyai, for their assistance in performing the RT-PCR procedures. Ms Annemarie Human for her constant assistance in the administration of the budget and project as a whole. Mr Byron Snow for providing the illustrations included in this thesis and for his support throughout the running of this project. ii List of abbreviations AHS: African Horse Sickness CNS: Central nervous system DNA: deoxyribonucleic acid DOB: Date of birth eCG: equine chorionic gonadotropin EDTA: ethylenediaminetetraacetic acid EHM: Equid herpes myeloencephalopathy EHV-1,-2,-3,-4,-5: Equid alphaherpesvirus 1,2,3,4, and 5 ELISA: Enzyme-linked immunosorbent assay EI: Equine Influenza FSH: Follicle Stimulating Hormone IFNƴ+: Interferon gamma IgG: Immunoglobulin G LAT: Latency associated transcripts LH: Luteinising hormone PBMC: Peripheral blood mononuclear cell PCR: Polymerase chain reaction iii qPCR: Quantitative or Real-Time Polymerase Chain Reaction RNA: Ribonucleic acid RT-PCR: Reverse transcriptase- polymerase chain reaction SOP: Standard Operating Procedure ZP: Zona pellucida iv Table of contents Declaration of originality ................................................................................................................. i Acknowledgements ......................................................................................................................... ii List of abbreviations ....................................................................................................................... iii Table of contents ............................................................................................................................ v List of tables ................................................................................................................................... ix List of figures ................................................................................................................................... x Summary ......................................................................................................................................... xi 1 General introduction .............................................................................................................. 1 2 Literature review..................................................................................................................... 4 2.1 Equine pregnancy ............................................................................................................ 4 2.1.1 Overview ................................................................................................................... 4 2.1.2 Development of the equine conceptus and placenta ............................................ 4 2.2 Taxonomy of herpesviruses in domestic species ........................................................... 8 2.3 Herpesvirus structure ...................................................................................................... 8 2.3.1 Functions of the glycoproteins ................................................................................ 8 2.4 Virus-host interaction and immunology ....................................................................... 10 2.4.1 EHV-1 specific immunology in foals ...................................................................... 11 2.4.2 Latency .................................................................................................................... 13 v 2.5 Equid alphaherpesviruses 1 and 4 ................................................................................ 14 2.5.1 Pathogenesis of EHV-1 and -4 ............................................................................... 15 2.5.2 Clinical signs of EHV-1 and -4infection .................................................................. 18 2.5.3 Pathological findings in EHV-1 and -4 infections .................................................. 21 2.6 Other equine herpesviruses .......................................................................................... 23 2.6.1 Equid gammaherpesvirus type 2 and 5 ................................................................. 23 2.6.2 Equid alphaherpesvirus type 3 .............................................................................. 23 2.7 Diagnosis of herpesvirus infections .............................................................................. 24 2.7.1 Histopathology ....................................................................................................... 24 2.7.2 Virus isolation ......................................................................................................... 25 2.7.3 Immunostaining techniques .................................................................................. 25 2.7.4 Serology .................................................................................................................. 26 2.7.5 Molecular diagnostics ............................................................................................ 27 2.8 Differential diagnoses for EHV-1 ................................................................................... 28 2.9 Management of equid herpesviruses in breeding populations .................................. 30 2.9.1 Prevention of infection .......................................................................................... 30 2.9.2 Infection control following a suspected outbreak................................................ 32 2.10 Treatment of EHV-1 infection ....................................................................................... 34 2.10.1 Symptomatic treatment of fever, respiratory symptoms and abortion ............. 34 2.10.2 Symptomatic treatment of EHV-1 myeloencephalopathy ................................... 34 vi 2.10.3 Antiviral agents ....................................................................................................... 35 3 Research question ................................................................................................................ 36 3.1 Aim .................................................................................................................................. 36 3.2 Hypotheses ..................................................................................................................... 36 3.2.1 Null hypothesis ....................................................................................................... 36 3.2.2 Alternative hypothesis ........................................................................................... 36 4 Materials and Methods ........................................................................................................ 37 4.1 Study design and ethical approval ................................................................................ 37 4.2 Study location and population ...................................................................................... 37 4.3 Housing and feeding ...................................................................................................... 37 4.3.1 Foaling management ............................................................................................
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