Module 4 模組4 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 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 Review 複習模組3之內容 Change in welfare 動物福利改變 װ Change in physical & psychological states 生理及心理狀態改變 װ Physiological responses 產生生理反應 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 Neuroendocrine responses 自主神經系統反應 神經內分泌系統反應 Neuroendocrine system 關於神經內分泌系統 • A welfare change affects three subdivisions 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 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 HPA cascade HPA軸作用流程 Hypothalamus 下視丘 腦下垂體前葉釋放激素 CRH corticotrophin-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 Stimulus 刺激 肝臟 HAP vessels 下視丘前葉血管 Module 4 Circulation 血流 HPA characteristics HPA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 Glucocorticoid measurement 糖皮質素測量評估 Method Invasive Restrictive Disturbing 方法 侵入性 限制性 干擾性 Plasma or or 血漿 Saliva 唾液 Urine 尿液 Faeces 糞便 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 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 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) 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) 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 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 overcrowding => higher cortisol response than if less crowded 經長期 密飼後血中cortisol會升高 (Friend et al., 1977) Limitations to HPA measures HPA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 Limitations to HPA measures HPA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 Limitations to HPA measures HPA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 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 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) Temperature measurement 體溫測量 Method Site 方法 採樣位置 Core Temp 中心體溫 Direct, e.g. thermometer 直接法--體溫計 Telemetry 嵌入遙測裝置 Thermoimaging 體表溫度照影 Invasi Restrictive Disturbing ve 限制性 干擾性 侵入 性 Mouth,口 ear 耳 & rectal 肛 Internal 體內 +/- +/- Skin 皮膚 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 Opioid measurement 類鴉片類之測量 Method Invasive Restrictive Disturbing 方法 侵入性 干擾性 Blood 血液 Brain receptors 腦內受體 限制性 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) 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 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 & StephensLarson, 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) 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 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 Bulletin 50 (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 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. Endocrinology 118, 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 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 Behavior 2, 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 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