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Respiratory Pathogens in Thoroughbred Foals up to One Year of Age on a Stud Farm in South Africa

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Respiratory Pathogens in Thoroughbred Foals up to One Year of Age on a Stud Farm in South Africa University of Pretoria etd – Picard, J A (2005) RESPIRATORY PATHOGENS IN THOROUGHBRED FOALS UP TO ONE YEAR OF AGE ON A STUD FARM IN SOUTH AFRICA BY JACQUELINE ANITA PICARD i University of Pretoria etd – Picard, J A (2005) Submitted in partial fulfilment of the requirements for the degree of Master of Science (MSc) (Veterinary Science) in the Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, South Africa Date submitted: February 2005 ii University of Pretoria etd – Picard, J A (2005) ABSTRACT RESPIRATORY PATHOGENS IN THOROUGHBRED FOALS UP TO ONE YEAR OF AGE ON A STUD FARM IN SOUTH AFRICA by Jacqueline Anita Picard Supervisor : Professor J A W Coetzer Department of Veterinary Tropical Diseases Faculty of Veterinary Science University of Pretoria Pretoria South Africa Submitted in partial fulfilment of the requirements for the degree MSc(Veterinary Science) iii University of Pretoria etd – Picard, J A (2005) The project was undertaken to monitor a group of 30 foals on a farm both clinically and microbiologically from birth until one year of age, to determine the aetiology of upper respiratory tract infections and to establish immune profiles of some of the known respiratory viral pathogens. One to two months prior to the birth of their foals, blood for serology was collected from the mares. The same specimens were collected from the foals just after birth, prior to suckling and a day after suckling. Thereafter the foals were examined monthly for the presence of respiratory disease and specimens taken. The following specimens were collected from each foal: three nasopharyngeal swabs, (one for virus isolation, one for bacteria and fungus isolation, and one for mycoplasma isolation) and blood that was allowed to clot. Blood was collected in heparin from sick foals with elevated rectal temperatures. Virus isolation was done on roller tube cultures of equine embryonic lung (EEL), Vero cells and rabbit kidney 13 (RK13) cells. The bacteria (including mycoplasmas) and fungi were cultured from the swabs and identified using a variety of traditional methods. The serum neutralization test (SNT) was used to detect antibodies to equid herpesvirus 1 (EHV-1), equid herpesvirus 4 (EHV-4), equine rhinovirus 1 (ERV-1), equine rhinovirus 2 (ERV-2) and equine adenovirus 1 (EAdV-1). The complement fixation test (CFT) was used to detect antibodies to EHV-1 and EHV-4 and the haemagglutination inhibition test (HIT) antibodies to equine influenzavirus (EIV). Only EHV-4 was cultured from the nasopharyngeal swabs of nine foals when they were 5 to 6 months of age and from one foal two months later. A wide variety of bacteria and fungi were cultured and it was established that coagulase-negative staphylococci, viridans streptococci, Moraxella spp. and Flavobacterium spp. predominated in most of the samples. Several potential bacterial pathogens were isolated but the most common were Streptococcus equi subsp. zooepidemicus, Actinobacillus equuli and Staphylococcus aureus. Colostrum-derived antibodies were detected for all the viruses in all but two of the foals. It was found that the foals had similar or slightly higher titres than their mothers. The levels declined in direct proportion to what they initially were and were depleted by the time the foals were 2 to 7 months of age. Antibodies to natural infection was detected to EHV-4, ERV-2 and EAdV-1. A rise in antibody titres occurred when the foals were 5 to 6 months of age, two months later and when they were one year of age. Antibodies resulting from immunization was detected to EHV-1, EHV-4 and EIV. It was established that the most important virus causing upper respiratory tract disease of the foals from 5 to 12 months of age was EHV-1 with EAdV-1 playing a minor role. These viruses caused repeated bouts of infection with a two to five months interval. Streptococcus equi subsp. zooepidemicus was considered to be the most important secondary pathogen. Prior to this period most of the foals were healthy with only a few suffering from upper respiratory disease. The aetiology was not determined in these cases, iv University of Pretoria etd – Picard, J A (2005) but based on the bacteriology results, it was suspected that some of them were suffering from bacterial infections. KEY WORDS: foals, infectious respiratory tract disease, equid herpesvirus 1, equid herpesvirus 4, equine adenovirus, equine rhinovirus 1, equine rhinovirus 2, equine influenzavirus, South Africa v University of Pretoria etd – Picard, J A (2005) ACKNOWLEDGEMENTS I am deeply indebted to the following people for without their support, this project would not have been possible: Professor Koos Coetzer who made me keep going and supervised the dissertation. Professors Joop Boomker and Roy Tustin who assisted greatly in editing this dissertation. Dr Neil and Mrs Elizabeth Orford for allowing their farm to be used for this project and their unfailing support and hospitality. Professor Alan Guthrie and the Equine Research Centre, Faculty of Veterinary Science, South Africa who provided me with the funds to successfully complete this project. Mrs Trudie Goosen who provided me with the training and lots of advice in the culture and identification of viruses and also the virus neutralization test. She also provided me with endless numbers of cells and culture media. Mr Johann Carstens and Mr Johan Gouws who taught me how to culture and identify bacteria, especially mycoplasmas. Mrs Eleanor Stylianides who taught me how to perform the complement fixation test and provided me with reagents. Mr Martinus Bezuidenhout and Dr Nicky Altschul who assisted me in the serology. Professor Peter Howell who allowed me to tap into his deep experience in virology and viral diseases. Mr Hercules Els who did the electron microscopy. The FDA that supplied me with a student grant for Dr Altschul so that she could assist me in setting up and performing the haemagglutination test for equine influenza virus. All the members of the Department of Veterinary Tropical Diseases who both provided me with assistance and moral support. vi University of Pretoria etd – Picard, J A (2005) ABBREVIATIONS ASPV Acid-stable picornavirus AHS African horse sickness AHSV African horse sickness virus AHV-1 Asinine herpesvirus 1 AHV-3 Asinine herpesvirus 3 ATCC American Type Culture Collection ATV Trypsin-versene solution BAL Broncho-alveolar lavage CF Complement fixing CFT Complement fixation test CID Combined immunodeficiency disease CO2 Carbon dioxide CPE Cytopathic effect °C Degrees Celsius DMSO Dimethylsulphoxide DVTD Department of Veterinary Tropical Diseases EAdV-1 Equine adenovirus 1 EAV Equine arteritis virus EEL Equine embryonic lung EEV Equine encephalosis virus EHV-1 Equid herpesvirus 1 EHV-2 Equid herpesvirus 2 EHV-4 Equid herpesvirus 4 EHV-5 Equid herpesvirus 5 EI Equine influenza EIV Equine influenzavirus EMEM Eagles-minimum essential medium ERV-1 Equine rhinovirus 1 ERV-2 Equine rhinovirus 2 ERV-3 Equine rhinovirus 3 EVA Equine viral arteritis FCS Foetal calf serum G Gauge hypodermic needle G Gravitational force GM Gram-negative GP Gram-positive H Haemagglutinin HI Haemagglutination inhibiting antibodies HIT Haemagglutination inhibition test IFAT Immunofluorescent antibody test kg Kilogram MBA Microbiotic agar mg Milligram MHC The minimum haemolytic dose of complement MHD Minimum haemolytic dose ml Millilitres MP Matrix protein N Neuraminidase NP Nucleoprotein PBS Phosphate buffered saline PBS- Calcium and magnesium free phosphate buffered saline PI-3 Parainfluenza-3 virus SDA Sabouraud’s dextrose agar vii University of Pretoria etd – Picard, J A (2005) SEM Scanning electron microscopy SN Serum neutralizing SNT Serum neutralization test SRBC Sheep red blood cells TCID50 Tissue Culture Infective Dose50 TEM Transmission electron microscope TSI Triple sugar iron agar TTA Transtracheal aspirate/wash VBS Veronal buffered saline µl Microlitres XLD Lysine dextrose viii University of Pretoria etd – Picard, J A (2005) TABLE OF CONTENTS ABSTRACT ............................................................................................................................. iii ACKNOWLEDGEMENTS ....................................................................................................... vi ABBREVIATIONS .................................................................................................................. vii TABLE OF CONTENTS .......................................................................................................... ix List of tables ............................................................................................................................ xi List of figures.......................................................................................................................... xii List of figures.......................................................................................................................... xii CHAPTER ONE .......................................................................................................................1 1.1 BACKGROUND .........................................................................................................1 1.2 STUDY OBJECTIVES ...............................................................................................1 1.3 LITERATURE REVIEW .............................................................................................2 1.3.1 Aetiology.............................................................................................................2
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