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Physiological measures of animal welfare (2): 動物福利之生理學測定法 ( 二 ) Neuroendocrine 神經內分泌系統

Physiological measures of animal welfare (2): 動物福利之生理學測定法 ( 二 ) Neuroendocrine 神經內分泌系統. Module 4 模組 4. This module will enable you to 本模組的教學目標. Understand the relationship between welfare and physiology 了解動物福利與生理學的關係

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Physiological measures of animal welfare (2): 動物福利之生理學測定法 ( 二 ) Neuroendocrine 神經內分泌系統

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  1. Physiological measures of animal welfare (2):動物福利之生理學測定法(二)Neuroendocrine神經內分泌系統 Module 4 模組4

  2. This module will enable you to本模組的教學目標 • Understand the relationship between welfare and physiology了解動物福利與生理學的關係 • Examine how the neuroendocrine system is associated with welfare檢視神經內分泌系統與動物福利之關係 • Understand how neuroendocrine responses can be used to assess welfare了解如何利用神經內分泌反應測量動物福利 • Identify some pros/cons of using neuroendocrine measures to assess welfare動物福利測量方法的正反理由

  3. Change in welfare 動物福利改變װChange in physical & psychological states生理及心理狀態改變װPhysiological responses產生生理反應 Review複習模組3之內容

  4. Review複習模組3之內容 Change in welfare 當動物福利改變時Activation of CNS 中樞神經系統受刺激活化   Autonomic nervous responses 自主神經系統反應 Neuroendocrine responses 神經內分泌系統反應

  5. Neuroendocrine system關於神經內分泌系統 • A welfare change affects three sub-divisions of the neuroendocrine system: 動物福利變化影響下列三個神經內分泌子系統： • Hypothalamus 下視丘 • Pituitary gland 腦下垂體 • Adrenal gland 腎上腺

  6. Hypothalamic-Pituitary-Adrenal Axis (HPA)下視丘-腦下垂體-腎上腺（HPA軸） • Major mediator of endocrine responses內分泌反應的主要媒介 • HPA increases mobilisation of energy stores for physical response HPA加強動員儲存的能量，供生理反應所需 • HPA mediates responses to: HPA調節的反應包括： • Stress 壓力 ─Sexual activity 性行為 • Courtship 求偶 ─Food arrival / foraging 尋覓食物

  7. HPA cascadeHPA軸作用流程 Hypothalamus下視丘 腦下垂體前葉釋放激素 CRHcorticotrophin-releasing hormone Anterior pituitary gland 腦下垂體前葉 促腎上腺皮質激素 ACTH adrenocorticotrophic hormone Adrenal cortex 腎上腺皮質 糖皮質素 Glucocorticoids Liver 肝臟 Module 4

  8. HPA characteristicsHPA軸之特性 • Response not as immediate as sympathetic adreno-medullary system 反應比交感腎上腺系統慢 • Measure of acute welfare changes 可測量急性的動物福利變化 • For example, plasma glucocorticoids elevated between 2-10 minutes after stimulation 動物在受到刺激後2-10分鐘，糖皮質素的血中濃度提昇 • Can remain elevated for hours depending on severity of stimulus 依照刺激的強弱，其濃度可以在血中維持數小時

  9. Glucocorticoid measurement糖皮質素測量評估

  10. Glucocorticoid measurement糖皮質素測量評估 • Analytical methods 分析方法 • Enzyme-linked immunosorbent assay 酵素連結免疫吸附分析 • High performance liquid chromatography 高壓液相層析分析 • Competitive protein binding assay 蛋白質結合競爭分析 • Radioimmunoassay 同位素免疫分析

  11. Glucocorticoids糖皮質素 • Corticosterone: rodents, birds囓齒、鳥類 • Cortisol: humans, pigs, and monkeys人類、豬、猴 • Useful direct measure 有用的直接測量準則

  12. Glucocorticoid example 1糖皮質素作用例1 • Cortisol  in lambs in response to: • Tail-docking 小羊在實施斷尾 手術時糖皮質素會升高 • Castration小羊在實施閹割手 術時糖皮質素會升高 (Mellor & Murray, 1989)

  13. Glucocorticoid example 2糖皮質素作用—例二 • Urinary cortisol increases in domestic dogs if they are put in kennels 家犬隻若是被關入狗舍中，可在尿液中檢測到cortisol濃度上升(Hennessey et al 1997) • If the dog has not been in kennels before, the increase is greater than if the dog has experienced kennels before 而且沒有被關入狗舍經驗的犬隻反應更激烈，濃度上升越多(Hiby 2005)

  14. Chronic welfare assessment慢性福利反應評估 • Indirect assessment of HPA 間接HPA評估法 • Glucocorticoid receptors in the brain 腦中糖皮質素受體數目 • Number of receptors decreased in rats that were exposed to prolonged handling and disturbance 大鼠在經歷重複保定、驚擾的刺激後，發現腦中的受體數量減少 (Wadham, 1997) • ACTH challenge test 促腎上腺皮質素測試 • ACTH is administered, and resulting glucocorticoid levels measured 注射ACTH後測血液中糖皮質素量

  15. ACTH challenge test促腎上腺皮質素測試 • Example: Dairy cows 以乳牛飼養為例 • Long periods of over- crowding => higher cortisol response than if less crowded 經長期密飼後血中cortisol會升高(Friend et al., 1977)

  16. Limitations to HPA measuresHPA測量技術之限制 Individual differences: 個體差異 • Early experience 早年生活經驗 (Levine et al., 1967) • Sex 性別 (Livezey et al., 1985) • Metabolism 代謝狀態

  17. Limitations to HPA measuresHPA測量技術之限制（續） • Species 物種 • New World Monkeys 新世界猴(Klostermann et al., 1986) • Time when sampled 採樣時間 • Circadian rhythm 日夜規律

  18. Limitations to HPA measuresHPA測量技術之限制（續） • Disturbance due to measurement 測量之干擾 • Human presence, handling, restraint and sampling method 有人在動物旁邊、人觸摸動物、保定，甚至是採樣方式不當

  19. Other autonomic measures of animal welfare其他測量動物福利之項目 • Body temperature 體溫 • Opioid levels 類鴉片物質濃度 • Organ pathology 器官病理變化

  20. Body temperature體溫代表的意義 • Acute welfare measure 急性的福利測量 • Changes in other physiological systems (e.g. adrenal activity) => changes in core body temperature 其他生理系統產生變化（如腎上腺活性改變）造成中心體溫出現改變 • Rat body temperature  by storms and unfamiliar people 暴風與陌生人會使大鼠的體溫升高(Georgiev, 1978) •  body temperature in defeated tree shrews 社會地位挑釁落敗的樹鼩體溫反而下降(von Holst, 1986)

  21. Temperature measurement體溫測量

  22. Opioids類鴉片類物質 • Three types 3種型式 • Endorphins 腦內啡 • Enkephalins 腦啡肽 • Dynorphins 強啡肽 • Functions: 功能 • Stress-induced analgesia 調適緊迫的止痛劑 • Control hormone release控制激素之分泌

  23. Opioid measurement類鴉片類之測量

  24. Opioid examples類鴉片類作用之實例 • Increase in plasma ß-endorphin in lambs during castration, tail docking and mulesing 綿羊在受閹割、斷尾、切臀皮時，血中ß-腦內啡量會增加(Shutt et al.,1987) • Tethered sows had higher opioid receptor density than group-housed sows 繫頸鏈的豬比群飼豬有較高之類鴉片類受體密度(Zanella et al., 1996)

  25. Organ Pathology器官病變 • Chronic welfare changes => wide variety of pathologies 慢性動物福利變化可造成多處器官病變 • Most are measured post-mortem e.g., 大部分是藉由死後解剖來觀察，例如： • Adrenal hypertrophy 腎上腺肥大 • Kidney lesions 腎臟病變 • Myocardial lesions 心肌病變 • In vivo 活體觀察 • Stomach ulcers 胃潰瘍

  26. Pathology examples器官病變之例 • Social instability in female groups of rats = adrenal hypertrophy 置身於不安定社會之雌性大鼠出現腎上腺肥大的病變(Haller et al., 1999) • Subordinate mice show kidney lesions in the presence of dominant animals 在強勢小鼠之存在下，弱勢鼠產生腎臟的病變(Henry & Stephens-Larson, 1985) • Myocardial lesions associated with longer periods of restraint in pigs 豬受到長期保定與心肌病變有關(Wutzen et al., 1987)

  27. Conclusion總結與討論 • Neuroendocrine system => acute and chronic measures of welfare 神經內分泌系統可作為動物福利之急性與慢性測量 • Direct and indirect ANS measures中樞神經系統反應有直接與間接之測量 • Limitations of measures神經內分泌系統之測量有其限制，故評估時要小心 • Other physiological responses can also be used to assess welfare e.g. temperature, organ pathology, and opioid levels 其他生理學之反應亦可用來評估動物福利，包括體溫、器官之病理變化、與血中類鴉片類濃度

  28. Further reading進階閱讀 • FRIEND, T.H., POLAN, C.E., GWAZDAUSKAS, F.C. & HEALD, C.W., 1977: Adrenal glucocorticoid response to exogenous adrenocorticotrophin mediated by density and social disruption in dairy cows. J. Diary Sci.60, 1958-1963 • GEORGIEV J. 1978 Influence of environmental conditions and handling on temperature rhythm of the rat. Biotelemetry Patient Monit. 5, 229-234 • HALLER J, FUCHS E, HALASZ J & MAKARA GB. 1999 Defeat is a major stressor in males while social instability is stressful mainly in females: Towards the development of a social stress model in female rats. Brain Research Bulletin50 (1): 33-39 • HENNESSEY MB, DAVIS HN, WILLIAMS MT, MELLOTT C & DOUGLAS CW 1997 Plasma cortisol levels of dogs at a county animal shelter. Physiol Behav 62: 485-490

  29. Further reading進階閱讀 • HENRYJP & STEPHENS-LARSON P. 1985 Specific aspects of stress on disease processes. In: MOBERG, G. P. (Ed.) Animal Stress. American Physiological Association, Maryland, USA • HIBY EF 2005 The Welfare of Kennelled Domestic Dogs. PhD Thesis, University of Bristol • KALIN NH, CARNES M, BARKSDALE CM, SHELTON SE, STEWART RD & RISCH SC. 1985 Effects of acute behavioural stress on plasma and cerebrospinal fluid, ACTH and -endorphin in rhesus monkeys. Neuroendocrinology 40, 97-101 • KLOSTERMANN LL, MURAI JT & SIITERI PK. 1986 Cortisol levels, binding and properties of corticosteroid-binding globulin in the serum of primates. Endocrinology118, 424-434 • LANG RE, HEIL JWE, GANTEN D, HERMANN K & UNGER T. 1983 Oxytocin, unlike vasopressin, is a stress hormone in the rat. Neuroendocrinology 37, 314-316

  30. Further reading進階閱讀 • LANE J 2006 Can non-invasive glucocorticoid measures be used as reliable indicators of stress in animals? Animal Welfare 15: 331-342 • LEVINE S, HALTMEYER GC, KARAS GG & DENENBERG VH. 1969 Physiological and behavioural effects of infantile stimulation. Physiology and Behavior2, 55-59 • LIVEZEY GT, MILLER JM & VOGEL WH. 1985 Plasma norepinephrine, epinephrine and corticosterone stress responses to restraint in individual male and female rats and their correlations. Neurosci. Lett. 62, 51-56 • MELLOR DJ & MURRAY L. 1989 Effects of tail docking and castration on behaviour and plasma cortisol concentrations in young lambs. Res. Vet. Sci.46, 387-391

  31. Further reading進階閱讀 • SHUTT, D.A., FELL, L.R., CONNELL, R., BELL, A.K., WALLACE, C.A. & SMITH, A.I., 1987: Stress-induced changes in plasma concentrations of immunosuppressive ß-endorphin and cortisol in response to routine surgical procedures in lambs.Aust. J. Biol. Sci. 40, 97-103. • VON HOLST D. 1986 Vegetative and somatic components of tree shrews’ behaviour. J. Auton. Nerv. Syst. Suppl., 657-670 • WADHAM JJB. 1997 Recognition and reduction of adverse effects in research on rodents. PhD Thesis, Faculty of Medicine and Dentistry, The University of Birmingham • WUTZEN J, LEWICKI Z, ROZYCKA Z, CEREMUZYNSKI L & BARCIKOWSKI R. 1987 Effects of immobilisation stress on histological, histochemical and electron-microscopic changes in porcine myocardium. Mat. Med. Pol.19, 78-84. • ZANELLA AJ, BROOM DM, HUNTER JC, MENDL MT. 1996 Brain opioid receptors in relation to stereotypies, inactivity, and housing in sows. Physiol Behav 59: 769-775

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