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The Effect of Environmental Stressors on the Immune Response to Avian Infectious Bronchitis Virus (Ibv)

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The Effect of Environmental Stressors on the Immune Response to Avian Infectious Bronchitis Virus (Ibv) THE EFFECT OF ENVIRONMENTAL STRESSORS ON THE IMMUNE RESPONSE TO AVIAN INFECTIOUS BRONCHITIS VIRUS A thesis submitted in partial fulfilment of the requirement for the degree of Doctor of Philosophy at Lincoln University by Juan Carlos Lopez Lincoln University 2006 ii Abstract of a thesis submitted in partial fulfilment of the requirements for the Degree of Doctor of Philosophy THE EFFECT OF ENVIRONMENTAL STRESSORS ON THE IMMUNE RESPONSE TO AVIAN INFECTIOUS BRONCHITIS VIRUS (IBV) by Juan Carlos Lopez The first aim of this research was to determine the prevalence of IBV in broilers within the Canterbury province, New Zealand, in late winter and to search for associations with management or environmental factors. The second aim was to study how ambient stressors affect the immune system in birds, their adaptive capacity to respond, and the price that they have to pay in order to return to homeostasis. In a case control study, binary logistic regression analyses were used to seek associations between the presence of IBV in broilers and various risk factors that had been linked in other studies to the presence of different avian pathogens: ambient ammonia, oxygen, carbon dioxide, humidity and litter humidity. Pairs of sheds were selected from ten large broiler farms in Canterbury. One shed (case) from each pair contained poultry that had a production or health alteration that suggested the presence of IBV and the other was a control shed. Overall, IBV was detected by RTPCR in 50% of the farms. In 2 of the 5 positive farms (but none of the control sheds) where IBV was detected there were accompanying clinical signs that suggested infectious bronchitis (IB). Ambient humidity was the only risk factor that showed an association (inverse) with the prevalence of IBV (p = 0.05; OR = 0.92). It was concluded within the constraints of the totally enclosed management systems described, that humidity had an influence on the presence of IBV, but temperature, ammonia, carbon dioxide, oxygen or litter humidity had no effect. iii In another study environmental temperatures were changed in order to affect the biological function and adaptive capacity of chickens following infection with IBV. The ‘affective states’ of the animal were assessed by measuring levels of corticosterone (CORT) in plasma and tonic immobility (TI). It was found that low (10 +/- 2oC) and high (30 +/- 2oC) temperatures exacerbated the respiratory signs and lesions in birds infected with IBV as compared to those housed at moderate (20 +/- 2oC) temperatures. The chickens housed at high temperatures showed significantly decreased growth, a higher proportion of hepatic lesions (principally haemorrhages) and a longer tonic immobility period, but there was no significant alteration in the plasma levels of CORT. The birds housed at low temperatures developed a higher proportion of heart lesions (hydropericardium, ventricular hypertrophy) and had significantly higher levels of plasma CORT than birds housed under moderate and/or high temperatures. The specific antibody response to IBV decreased in birds housed under high temperatures. Interestingly the birds housed at high temperatures developed significantly higher levels of haemagglutinin antibodies to sheep red blood cells (SRBC) than those birds housed under low or moderated temperatures. Cell mediated immunity was not significantly affected by heat or cold stress in the first 13 days of treatment but at 20 days the levels of interferon gamma in the birds subjected to low temperatures were lower than in the high temperature group. In other trials, the exogenous administration of low physiological doses of oral CORT (as compared to high pharmacological doses typically used in such experiments) to birds resulted in suppression or enhancement of the immune response depending on duration of treatment and/or dose and nature of the antigen. To our knowledge, this is the first study to show that exogenous CORT can produce an enhancement in the immune response in chickens. iv In conclusion, environmental stressors such as high or low temperatures do affect the physiology of the fast-growing broiler. The adjustments the birds have to make to maintain homeostasis impacts on the course of common infectious diseases, such as IB, that normally is mild in the New Zealand poultry industry. The administration of exogenous CORT showed that this hormone may be part of the physiological stress response and acts as a messenger to prepare the immune system for potential challenges (e.g., infection). v Acknowledgements I would like to acknowledge the advice, guidance, patience and friendship of Dr Robin McFarlane. I doubt that I will ever be able to express my appreciation fully, but I owe him my eternal gratitude. I would like to thank my cosupervisor Dr Graham Barrell for his valuable suggestions. Special thanks go to Martin Wellby, Omega Amoafo, Janette Busch and Karl Gately - without whose knowledge and assistance this study would not have been successful. Thanks to NZAID for the scholarship and Tegel Foods Ltd for their collaboration and support. I would like to thank Alison Lister for assistance with the statistical analysis. Finally, I would like to thank my family, especially my wife Adriana, for support and help with my experiments. Without her help and encouragement, I would not have finished my PhD. vi List of Contents Abstract ii Acknowledgments v List of contents vi List of tables xi List of figures xiii Abbreviations xvi Chapter 1 Introduction 1 Chapter 2 Literature review 5 2.1 Seasonal cycles of infectious diseases 5 2.2 Welfare and stress in poultry 5 2.2.1 Mechanisms of the general adaptation syndrome (GAS) 7 2.2.2 Hypothalamopituitary-adrenal axis (HPA) 8 2.3 Body temperature and avian thermoregulation 10 2.3.1 Physiological changes due to effect of heat or cold 11 2.3.2 Effect of temperature on the immune system in birds 12 2.4 Infectious bronchitis 13 2.4.1 IB in New Zealand 14 2.4.2 Aetiology 15 2.4.3 Strain classification 15 2.4.4 Transmission and carriers 17 2.4.5 Clinical signs 17 2.4.6 Pathogenesis 18 2.4.7 Lesions 19 2.4.8 Diagnosis 20 vii 2.4.9 Treatment 22 2.5 Poultry immune system 22 2.5.1 Immune response against IBV 23 2.5.1.1 Innate immunity 23 2.5.1.2 Humoral immunity 23 2.5.1.3 Cellular immunity 25 2.5.1.4 Passive immunity 26 2.5.2 Vaccination against IBV 26 2.5.2.1 Types of vaccines 26 2.5.2.2 Vaccination programs 27 Chapter 3 The influence of environmental factors on the prevalence of infectious bronchitis 30 3.1 Introduction 30 3.2 Materials and methods 31 3.2.1 Farms 31 3.2.2 Clinical signs and detection of IBV 32 3.2.3 Ambient measurements 34 3.2.4 Questionnaire and data analysis 34 3.3 Results 34 3.4 Discussion 36 viii Chapter 4 Effect of thermal change on immune responses and physiological function in chickens 40 4.1 Introduction 40 4.2 Materials and methods 42 4.2.1 Animals and housing conditions 42 4.2.2 Immunization 43 4.2.3 Blood sampling 45 4.2.4 Clinical signs 45 4.2.5 Lesions and mortality 46 4.2.6 Body weights, haematocrit and body temperature 47 4.2.7 Tonic immobility (TI) 47 4.2.8 Corticosterone 47 4.2.9 Antibody 47 4.2.10 Interferon gamma 48 4.2.11 Statistical analysis 49 4.3 Results 49 4.3.1 Clinical signs 49 4.3.2 Lesions and mortality 51 4.3.3 Weight gain, haematocrit and body temperature 53 4.3.4 Tonic immobility and plasma corticosterone levels 53 4.3.5 Humoral immune response 55 4.3.6 Cell mediated immune response 57 4.4. Discussion 58 ix Chapter 5 Effect of corticosterone on the immune response in broiler chickens 64 5.1 Introduction 64 5.2 Materials and methods 66 5.2.1 Titration of oral corticosterone treatment 66 5.2.2 Animals, housing conditions and corticosterone treatment 67 5.2.3 Infection and immunization 68 5.2.4 Clinical signs 69 5.2.5 Lesions 69 5.2.6 Relative spleen weights 69 5.2.7 Tonic immobility (TI) 69 5.2.8 Plasma corticosterone 70 5.2.9 Determination of antibody titres 70 5.2.10 Interferon gamma 70 5.2.11 Intradermal hypersensitivity 70 5.2.12 Statistical analysis 71 5.3 Results 71 5.3.1 Dose effect of corticosterone (pilot trial) 71 5.3.2 Clinical signs 72 5.3.3 Lesions 73 5.3.4 Body weight and relative spleen weights 74 5.3.5 Tonic immobility and plasma CORT levels 75 5.3.6 Humoral immune response 76 5.3.7 Cell mediated immune response 77 5.4 Discussion 79 x Chapter 6 Effects of corticosterone on the immune response in chickens depend on duration of action 84 6.1 Introduction 84 6.2 Material and methods 85 6.2.1 Animal and housing conditions 85 6.2.2 Experimental infection, immunization and blood sampling 86 6.2.3 Clinical signs and lesions 87 6.2.4 Tonic immobility (TI) 88 6.2.5 Corticosterone levels in drinking water and plasma 88 6.2.6 Determination of antibody titres 88 6.2.7 Interferon gamma (IFN γ) 88 6.2.8 Statistical analysis 88 6.3 Results 89 6.3.1 Clinical signs and lesions 89 6.3.2 Body and spleen weights 90 6.3.3 Tonic immobility and CORT levels 92 6.3.4 Humoral immune response 93 6.3.5 Cell mediated immune response 94 6.4 Discussion 97 Chapter 7 General discussion 102 References 110 Appendices 136 xi List of Tables Table 3.1.
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