實驗動物學(xué)總論(NXPowerLite)

1、實驗動物學(xué)總論實驗動物學(xué)總論一一 實驗動物和實驗用動物實驗動物和實驗用動物（一）實驗動物（一）實驗動物(Laboratory animals)是指經(jīng)人工培育或人工改造，對其攜帶的微生物實行控制；遺傳背景明確，來源清楚，用于科學(xué)實驗、藥品、生物制品的生產(chǎn)和檢定及其他科學(xué)實驗的動物。（二）實驗用動物（二）實驗用動物(Animals for research ； Experimental animals）所有用于科學(xué)實驗的動物統(tǒng)稱為實驗動物。包括實驗動物，野生動物，經(jīng)濟動物和觀賞動物。二二 實驗動物學(xué)的定義和范圍實驗動物學(xué)的定義和范圍（一）定義（一）定義 實驗動物學(xué)（Laboratoryanimal

2、science）是研究實驗動物和動物實驗的科學(xué)。（二）研究范圍（二）研究范圍 1.實驗動物遺傳育種學(xué)（Laboratoryanimalgeneticbreedingscience）2.實驗動物微生物學(xué)和寄生蟲學(xué)（Laboratoryanimalmicrobiologyandparasitology）3.實驗動物環(huán)境生態(tài)學(xué)（Laboratoryanimalenvironmentalecology）4.實驗動物營養(yǎng)學(xué)（Laboratoryanimalnutriology）（二）研究范圍（二）研究范圍(續(xù)續(xù))5.實驗動物飼養(yǎng)管理（Laboratoryanimalhusbandry）6.實驗動物醫(yī)學(xué)（

3、Laboratoryanimalmedicine）7.比較醫(yī)學(xué)（Comparativemedicine）8.動物實驗（Animalexperimentaltechniques）三三 實驗動物的重要性實驗動物的重要性（一）實驗動物是生命科學(xué)研究的支撐條件之一（一）實驗動物是生命科學(xué)研究的支撐條件之一生命科學(xué)實驗研究的四個支撐條件-AEIR要素A：Animal：實驗動物；E：Equipment：儀器設(shè)備；I：Information：信息；R：Reagent：試劑； （二）實驗動物在生命科學(xué)研究中被廣泛應(yīng)用，很多重要（二）實驗動物在生命科學(xué)研究中被廣泛應(yīng)用，很多重要 的科研成果來源于實驗動物的科研成

4、果來源于實驗動物 （三）實驗動物是人類的替身，起著（三）實驗動物是人類的替身，起著“活活 的天秤的天秤”和和“活的化學(xué)試劑活的化學(xué)試劑”的作用的作用 （四）實驗動物工作實行法制化管理（四）實驗動物工作實行法制化管理 四四 實驗動物的分類實驗動物的分類 （一）傳統(tǒng)的動物學(xué)分類方法（一）傳統(tǒng)的動物學(xué)分類方法依據(jù)自然分類法，把整個生物，通常是用界(kingdom)、門(phylum)、綱(class)、目(order)、科(family)、屬(genus)、種(species)等劃分分類等級。以大家鼠為例，它屬于：脊椎動物門(phylumvertebrata)哺乳動物綱(classmammalia)

5、嚙齒目(orderrodentea)鼠科(familymurinae)大家鼠屬(genusrattus)大家鼠種(species rattus norvegicus)學(xué)名：褐家鼠（大家鼠）(Rattus norvegicus)多數(shù)情況下，根據(jù)不同目的進行種下分類，把實驗動物劃分為不同品系。（二）按實際用途分類（二）按實際用途分類1、實驗動物2、經(jīng)濟動物（Economicalanimals），或稱家畜、家禽3、野生動物（Wildanimals）4、觀賞動物（Exihibitinganimals）（三）按遺傳學(xué)控制分類（三）按遺傳學(xué)控制分類根據(jù)基因的純合程度，把實驗動物分成下列四類：1、近交系動物

6、（Inbredstrainanimals）2、突變系動物（Mutantstrainanimals）3、雜交群動物（Hybridcolonyanimals）（雜交一代，F(xiàn)1代動物）4、封閉群動物(Closedcolonyanimals)1、近交系動物（、近交系動物（Inbred strain animals）又叫純系動物。是采用同胞兄妹或親子交配，連續(xù)繁殖20代以上所培育出來的遺傳上達到高度一致的動物群?；蚣兒铣潭瓤蛇_99.8%。主要指嚙齒動物；可出現(xiàn)近親交配衰退。親子交配與兄妹交配不能混用。親子交配時必須采用年輕的雙親同其子女交配。較大動物純種培育很難獲得成功，因為世代間隔較長，費用較大，所

7、以成功率低。禽類和兔的血緣關(guān)系達到80%以上（相當于兄妹交配四代）時，即可稱為近交系。2、突變系動物（、突變系動物（Mutant strain animals）具有特殊突變基因的品系動物，正常染色體基因發(fā)生突變，并具有各種遺傳缺陷的動物。在長期繁殖過程中，動物的子代突然發(fā)生變異，變異的基因位點又可遺傳下去，或者即使沒有明確的基因位點，經(jīng)淘汰和選育后，仍能維持其穩(wěn)定的遺傳性狀。這種變異并能繼續(xù)保持遺傳基因特性的品系動物，稱為突變系動物。如無胸腺裸鼠、無K細胞、或無K、無B、無巨噬細胞等裸鼠。用于免疫研究、移植實驗等。3、雜交群動物（、雜交群動物（Hybrid colony animals）（雜交

8、一代，F(xiàn)1代動物）兩個近交品系動物之間進行有計劃交配所獲得的第一代動物。例如：C57BL/6JDBA/2B6D2F1（B6為C57BL/6J的縮寫，D2為DBA/2的縮寫。）4、封閉群動物、封閉群動物(Closed colony animals)以非近親交配方式進行繁殖生產(chǎn)的一個種群，在不從外部引入新的血緣條件，至少連續(xù)繁殖四代以上稱封閉群。封閉群又稱遠交群。（四）按微生物學(xué)控制分類四）按微生物學(xué)控制分類1、無菌動物（、無菌動物（Germ free animals，GF）體內(nèi)、外無任何可檢測出的活的微生物和寄生蟲的動物。來源于無菌手術(shù)剖腹取胎，飼養(yǎng)在無菌隔離器內(nèi)，人工喂乳或保姆代養(yǎng)培育而成。悉

9、生動物悉生動物(Gnotobiotes animals，GA)體內(nèi)攜帶有已知微生物的動物。這種動物來源于無菌動物，人為的投給已知的單菌、雙菌、三菌或多菌。這些均為已知菌，與無菌動物一樣，飼養(yǎng)在隔離器中。2、無特定病原體動物、無特定病原體動物(Specefic pathogen free animals，SPF）體內(nèi)不存在特定病原微生物和寄生蟲的的動物，簡稱SPF動物。是指無傳染病的健康動物。這種動物都是來自無菌動物或悉生動物，轉(zhuǎn)移到屏障系統(tǒng)中飼養(yǎng)。要在屏障系統(tǒng)環(huán)境設(shè)施中飼育繁殖和進行實驗，要進行嚴格消毒、檢疫、隔離并定期剖腹凈化。3、清潔動物、清潔動物(Clean animals，CL)又稱最

10、低限度疾病動物(Ginimaldiseaseanimals),體內(nèi)外不攜帶人畜共患的病原體或動物傳染病病原的動物，不能帶有體外寄生蟲和大部分體內(nèi)寄生蟲。4、普通動物（、普通動物（Conventional animals）未經(jīng)積極的微生物學(xué)控制，飼養(yǎng)在開放衛(wèi)生環(huán)境未經(jīng)積極的微生物學(xué)控制，飼養(yǎng)在開放衛(wèi)生環(huán)境 里的動物。墊料和飼料和飲水一般不消毒，飲用里的動物。墊料和飼料和飲水一般不消毒，飲用 普通自來水。普通自來水。 所謂普通動物也并不是對微生物沒有一定控制的所謂普通動物也并不是對微生物沒有一定控制的 一般動物，而是要求不帶能夠感染人的微生物和一般動物，而是要求不帶能夠感染人的微生物和 體外寄生蟲

11、。這種動物只能供教學(xué)和一般實驗用。體外寄生蟲。這種動物只能供教學(xué)和一般實驗用。 根據(jù)新修訂的國家實驗動物微生物、寄生蟲質(zhì)量根據(jù)新修訂的國家實驗動物微生物、寄生蟲質(zhì)量 標準，我國將實驗大、小鼠分為三類即清潔級、標準，我國將實驗大、小鼠分為三類即清潔級、 無特定病原體級、無菌級（包括悉生動物）。其無特定病原體級、無菌級（包括悉生動物）。其 它品種實驗動物仍然分為普通級、清潔級、無特它品種實驗動物仍然分為普通級、清潔級、無特 定病原體級、無菌級（包括悉生動物）四級。即定病原體級、無菌級（包括悉生動物）四級。即 從從2002年年5月月1日起取消了普通級大、小鼠標準。日起取消了普通級大、小鼠標準。The

12、JacksonLaboratoryIn1929,Harvard-trainedgeneticistClarenceCookLittlefoundedTheJacksonLaboratory,basedonthethen-radicalideathatmicewerekeytounderstandingthegeneticbasisofhumandevelopment,diseasesanddisorders.Throughoutthe1900s,thatideawouldbecomeincreasinglycentraltotheprogressofbiomedicalresearch,rea

13、chingultimateconfirmationwhenthemouseandhumangenomesweresequencedattheturnofthecenturyandproveddefinitivelythatthetwospeciessharethevastmajorityoftheirgenes.【上海斯萊克實驗動物有限責任公司】【中國科學(xué)院上海實驗動物中心 】【國家嚙齒類實驗動物種子中心上海分中心】 Animal Care and UseLaboratoryanimalsareusedinbiomedicalresearchasmodelsforhumans.Observed

14、behavioralorphysiologicalchangesexhibitedbytheseanimals,whentheyareusedintheprotocol,areassumedtoberesponsestotheexperimentalprocedures.Ifnon-experimentalvariablescausedbyinadequatehousing,disease,orstressfromimproperhandlingareinadvertentlyintroducedintothestudytheycanelicitsimilarresponseswhichcou

15、ldskewthedataortotallyconfoundtheexperiment.Thesafeandeffectiveuseofanimalsinalaboratorysettingisanessentialelementofyourresearchactivities. The Animal Welfare Act (AWA), and PHS Policy require that all Animal Users complete training on the laws, regulations, and procedures associated with animal ca

16、re and use. This course is designed to fulfill that requirement, providing a quick overview of the relevant laws and regulations covering use of animals in a laboratory setting, and the recommended procedures for working with specific animals. The course includes practical and essential information

17、that will guide you in the everyday care and use of animals in your laboratory environment. Importance of Proper Animal CareYourprimaryresponsibilitywhenworkingwithlaboratoryanimalsistotreatthemhumanely.Withthatinmind,themajorchallengeistomanagetheanimalsinawaythatminimizestheirexposuretopainanddist

18、ressandmaintainstheintegrityoftheirphysiologicalfunctionstofacilitatethegenerationofreliableresearchdata.Thiscoursewillteachyouhowthepropertreatmentofanimalscarriestheaddedbenefitofminimizingtheintroductionofunwantedvariablesintotheresearchprocess.Importance of Proper Animal CareYourcareandtreatment

19、ofanimalsmustalsocomplywiththeappropriatelaws,regulations,andwithNIHpolicy.Manyoftheselaws,regulationsandpolicieswereenactedinresponsetopublicconcernaboutthetreatmentofanimalsintheresearchsetting.Consequently,yourobjectiveshouldbetotreatyouranimalsinamannerthatcomplieswiththeethicalandlegalimperativ

20、esprescribedforthehumanetreatmentoflaboratoryanimalsthatwillreflectpositivelyonNIHasaresearchinstitution.research institution.LawsandRegulations:The main regulations, policies and guidelines that apply to animal use at NIH are:TheAnimalWelfareAct(AWA)ThePublicHealthServicePolicyonHumaneCareandUseofL

21、aboratoryAnimals(PHSPolicy)U.S.GovernmentPrinciplesfortheUtilizationandCareofVertebrateAnimalsUsedinTesting,Research,andTraining(GovernmentPrinciples)GuidefortheCareandUseofLaboratoryAnimals(Guide)NIHManual3040-2:AnimalCareandUseintheIntramuralProgramStructure and AccreditationTheDDIRdelegatestotheD

22、irectoroftheOfficeofAnimalCareandUse(OACU)overallresponsibilityforensuringthattheACUprogramcomplieswithapplicablepoliciesandregulations.Pain and DistressThereisalsoapragmaticpointofscientificlogicthatsupportsminimizingtheexposureofresearchanimalstoexperimentalpainordistress.Animalsrespondtopainanddi

23、stresswithchangesintheirnormalphysiologywhichcanskewdatabeingcollected.Alleviating Pain and DistressAnimals,likehumans,canbeexpectedtogenerateastressresponsetonovelstimuli.Handlinganimalsveryearlyinlife,evenasnew-borns,togentlethemisaprocedurethathasbeenwidelyusedonavarietyofspeciestofamiliarizethea

24、nimalswiththehandlingprocess.Whenanimalsaretrainedthiswaytoassociatehandlingwithpleasantcircumstancestheyaremuchlesslikelytogenerateastressresponselaterinlifewhentheyarehandled.AlternativesThe Three Rs: Reduction:Reductionsinthenumbersofanimalsusedtoobtaininformationofacertainamountandprecision.Refi

25、nement:Decreaseintheincidenceorseverityofpainanddistressinthoseanimalsthatareused.Replacement:Substitutionofinsentientmaterialforanimalsorsubstitutionofalowerspecies,whichmightbelesssensitivetopainanddistress,forahigherspecies.Minimizing Exposure to Disease when Working with AnimalsStandard laborato

26、ry protective clothing includes:Uniforms-Uniformsareclothing,suchassurgicalscrubsuitsandcoveralls,dedicatedforwearduringworkintheanimalfacility.Auniformshouldnotbewornoutsideoftheanimalfacilityorresearchsetting.Forexample,uniformsshouldnotbeworninpubliccafeterias,lecturehalls,andmedicalpatientcarear

27、eas.Gloves-Lightweightvinylorlatexglovespreventcontaminationoftheskinwithpathogensthatmaybeonanimalsbodiesoronsurfacessoiledbytheirexcreta.Lab coats-Disposablelabcoatsandcoverallsprotectstreetclothesfromcontaminationwithanimalpathogens.Labcoatsshouldnotbewornoutsideoftheanimalfacilityorresearchsetti

28、ng.Forlongtermworkintheanimalfacility,auniformmaybesubstitutedforthesetypesofcoveringgarments.MucousMembraneProtection A device or a combination of devices such as face shields or surgical face masks combined with protective glasses or goggles, worn to protect the mouth, nose, and eyes from splash o

29、r droplet contamination. The degree of protection needed varies with the specific procedure being conducted and should be adjusted to the level of the anticipated risk. Full face shields provide splash protection for all of the facial mucus membranes. Partial mucous membrane protection is provided b

30、y using only a face mask and may be appropriate when the fecal oral route of contamination is the only concern. Shoe Covering-Stretchbooties,usuallymadeofpaperorplastic,arewornoverstreetshoestopreventthetransferofpathogenicorganisms.Bootiesshouldnotbewornoutsideoftheanimalfacility.Dedicatedfootwearc

31、anbesubstitutedforshoecoveringsduringlong-termactivities.Ifdedicatedshoesareworn,shoecoveringsmaybeusedtocovertheshoeswhenoutsidetheanimalfacility,buttheshoecoversmustberemovedonreturntothefacility.Cage SystemsCage SystemsCage systems that meet the general requirements for maintenance of normal body

32、 functions and provide moderate protection from exposure to pathogens are described as conventional cages. Conventional cages, however, do not adequately protect rats and mice from airborne diseases to which they are highly susceptible so cage systems that are specially designed to provide extra pro

33、tection from these pathogens are used.Rodent viruses and other airborne pathogens may be transported by clinging to dust particles and other particulate matter suspended in the air inside and above the animal cage. The risk of exposure to these organisms is reduced by increasing fresh air circulatin

34、g around the cages, or filtering the air.water water SanitationSanitationFiltered Air UnitFiltered Air Unit Observing and Reporting Problems with AnimalsWhileanimalhealthisthechargeoftheveterinarians,youwillhavearesponsibilitytoobserveandreportchangesinananimalshealthandtoinformtheveterinarianoranim

35、alfacilitymanagersothattheanimalcanbetreated.Immediateidentificationofsickorinjuredanimalsiscriticallyimportantinaresearchsettingbecausetheanimalsarecloselyconfined,andhousingspaceisfrequentlysharedbyseveralinvestigators.Infectiousdiseaseinonegroupofanimalscouldjeopardizealloftheanimalsintheroomorbu

36、ilding.Signs of Illness and Injury:Signs of Illness and Injury: Picking up mice.Whenpickingupmice,alwaysholdthemousebythebaseofthetail.Thiscanbedonewitheitherforcepsoryourfingers.DONOTpickmiceupbythetipofthetail.Ifyoupickamouseupbythetipofitstail,themousemayspinintightcircleswhenitsfeetlosecontactwi

37、ththecagesurface.Ifthishappens,thetipoftailcanbepulledoff.Additionally,amouseheldbythetipofitstailcanclimbupitstailandbiteyou!Separating mice/rats by gender.Separatingmice/ratsbygendercanbedifficult,especiallywhentheyareyoungandmalestesticleshavenotdescendedintothescrotum.Comparetherelativedistanceb

38、etweenanusandurinarypapilla.Thedistancebetweentheanusandtheurinarypapillaisgreaterinthemale.Thisphotographillustratesthatthemouseontherighthasthelesser(shorter)distancebetweentheanusandtheurinarypapilla,soitisthefemale.Restraining Rats.Ratscanbecaughtandliftedbythebaseoftheirtail,oryoucangraspthemar

39、oundtheirbodyasillustratedinthisphotograph.Ifyougrasptheratarounditsbodyyoucanpreventitfrombitingyoubyplacingyourindexfingersnuglyunderthejawagainstthepointoftheshoulderaheadoftherightfrontleg.Yourthumbisplacedbehindtheleftfrontlegwhichpushesitforwardagainsttheleftsideoftheneck.Thisgripsecuresthehea

40、d,andtheratcannotreacharoundtobite.Forlargerats,keepingagoodgrasponthetailwillgiveyouadditionalcontrol.Guinea Pigs. GuineaPigshavenotailandmustbecaughtbygraspingthemaroundtheirbody.Theyareveryinclinedtoavoidbeingcaughtbyscurryingaroundthecageandwhistlingloudly.Whentheyarecaughttheyrarelyattempttobit

41、ebutmayreachforwardwiththeirhindfeetandscratchwiththeirlongtoenails.Rabbits.Rabbitsarecharacterizedbytheirthinbones,fragileskeleton,andheavymusclesontheirbackandhindlegs.Whenrestrainingandliftingarabbit,themostimportantthingtorememberistosupporttherabbitsrumpasyoupickitup.Iftherabbitkicks,evenonetim

42、e,whenitsbackfeetleavethecageflooritcanbreakitsback,bepermanentlyparalyzedandhavetobeeuthanized.Thecorrecttechniqueistoapproachtherabbitwithyourhandheldhighovertheback,grasptheskinovertheshouldersandslideyourotherhandbetweenthebacklegstosupporttherumpbeforeyoulifttheanimaloffofthecagefloor.Intra-per

43、itoneal injection.1.Restrainthemousebygraspingtheskinalongitsbackwithyourlefthand(ifright-handed).2.Clampthetailbetweenyourringfingerandlittlefingerforadditionalcontrol.3.Positionthemousesothatitsheadisdown.(Intestines,stomach,etc.shouldfallforwardandcreateasmallspaceintheposteriorportionoftheperito

44、nealcavity.)4.Inserttheneedlealittleofftothesideofthecenterline(tomissthebladder).Collecting a blood sample.1.Givegeneralanesthesia.2.Usecapillarytubetreatedwithheparininsidesothattheblooddoesntclot.3.Introducethetubeintothemedialcanthusoftherodentseye,directedtowardthebackoftheorbit.4.Rotatethetube

45、tocutintothevenoussinusinthebackoftheorbit.Samplesmaybemoredifficulttocollectfromratsbecausetheyhaveavenousplexusratherthanavenoussinusatthebackoftheirorbitandthevesselscanbehardtolocate.5.Bloodflowsdownintothecapillarytube.6.Removetubeandapplygentlepressureagainsttheeyewithagauzespongetostoptheblee

46、ding.7.Watchtheanimalcloselytopreventcagematesfrominjuringitduringitsrecoveryfromtheanesthesia.Giving Injections/Taking SamplesRabbits:Rabbitsarefrequentlyusedtoproduceantibodiestoaninjectedantigenandlarge(50ml)bloodsamplesarecollectedforharvestingantibodies.Drawing a sample.1.Bloodisusuallycollecte

47、dfromtheauricularartery(darkline)thatliesalongthemiddleofthetopsurfaceoftheear.2.Alargebore(18guage)needleismostfrequentlyusedtodrawsamples.Giving Injections/Taking SamplesRabbits:Rabbitsarefrequentlyusedtoproduceantibodiestoaninjectedantigenandlarge(50ml)bloodsamplesarecollectedforharvestingantibod

48、ies.Intravenous injection.1.Usemarginalearvein,thedarkstructurealongthelowermarginofear.2.Shavethehairoverthevesseltobeused.3.Usesmallbore(23-27guage)needlewithasyringeorinsertasimilargaugescalpveinset(needlewithcatheter).Anesthesia/AnalgesiaPentobarbital Somnifer.Deliveredintravenously(IV)becauseit