實驗動物學總論課件

實驗動物學總論一實驗動物和實驗用動物

〔一〕實驗動物(Laboratoryanimals)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，用于科學實驗、藥品、生物制品的生產(chǎn)和檢定及其他科學實驗的動物。〔二〕實驗用動物(Animalsforresearch；Experimentalanimals〕所有用于科學實驗的動物統(tǒng)稱為實驗動物。包括實驗動物，野生動物，經(jīng)濟動物和欣賞動物。二實驗動物學的定義和范圍〔一〕定義

實驗動物學〔Laboratoryanimalscience〕是研究實驗動物和動物實驗的科學?！捕逞芯糠秶?.實驗動物遺傳育種學〔Laboratoryanimalgeneticbreedingscience〕2.實驗動物微生物學和寄生蟲學〔Laboratoryanimalmicrobiologyandparasitology〕3.實驗動物環(huán)境生態(tài)學〔Laboratoryanimalenvironmentalecology〕4.實驗動物營養(yǎng)學〔Laboratoryanimalnutriology〕

〔二〕研究范圍(續(xù))5.實驗動物飼養(yǎng)管理〔Laboratoryanimalhusbandry〕6.實驗動物醫(yī)學〔Laboratoryanimalmedicine〕7.比較醫(yī)學〔Comparativemedicine〕8.動物實驗〔Animalexperimentaltechniques〕三實驗動物的重要性〔一〕實驗動物是生命科學研究的支撐條件之一生命科學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〔三〕實驗動物是人類的替身，起著“活的天秤〞和“活的化學試劑〞的作用〔四〕實驗動物工作實行法制化管理四實驗動物的分類〔一〕傳統(tǒng)的動物學分類方法依據(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)嚙齒目(orderrodentea)鼠科(familymurinae)大家鼠屬(genusrattus)大家鼠種(speciesrattusnorvegicus)學名：褐家鼠〔大家鼠〕(Rattusnorvegicus)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?！捕嘲磳嶋H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、欣賞動物〔Exihibitinganimals〕〔三〕按遺傳學控制分類

根據(jù)基因的純合程度，把實驗動物分成以下四類：1、近交系動物〔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〔Inbredstrainanimals〕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到達高度一致的動物群。

基因純合程度可達99.8%。①

主要指嚙齒動物；可出現(xiàn)近親交配衰退。②

親子交配與兄妹交配不能混用。③

親子交配時必須采用年輕的雙親同其子女交配。④

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，所以成功率低。⑤

禽類和兔的血緣關(guān)系到達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〔Mutantstrainanimals〕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〔Hybridcolonyanimals〕〔雜交一代，F(xiàn)1代動物〕兩個近交品系動物之間進行有方案交配所獲得的第一代動物。例如：C57BL/6J×DBA/2→B6D2F1〔B6為C57BL/6J的縮寫，D2為DBA/2的縮寫。〕4、封閉群動物(Closedcolonyanimals)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。2、無特定病原體動物(Speceficpathogenfreeanimals，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(Cleananimals，CL)又稱最低限度疾病動物(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局部體內(nèi)寄生蟲。4、普通動物〔Conventionalanimals〕

未經(jīng)積極的微生物學控制，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。這種動物只能供教學和一般實驗用。

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。即從2002年5月1日起取消了普通級大、小鼠標準。TheJacksonLaboratory

In1929,Harvard-trainedgeneticistClarenceCookLittlefoundedTheJacksonLaboratory,basedonthethen-radicalideathatmicewerekeytounderstandingthegeneticbasisofhumandevelopment,diseasesanddisorders.Throughoutthe1900s,thatideawouldbecomeincreasinglycentraltotheprogressofbiomedicalresearch,reachingultimateconfirmationwhenthemouseand

humangenomesweresequencedattheturnofthecenturyandproveddefinitivelythatthetwospeciessharethevastmajorityoftheirgenes.【上海斯萊克實驗動物有限責任公司】【中國科學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】AnimalCareandUse

Laboratoryanimalsareusedinbiomedicalresearchasmodelsforhumans.Observedbehavioralorphysiologicalchangesexhibitedbytheseanimals,whentheyareusedintheprotocol,areassumedtoberesponsestotheexperimentalprocedures.Ifnon-experimentalvariablescausedbyinadequatehousing,disease,orstressfromimproperhandlingareinadvertentlyintroducedintothestudytheycanelicitsimilarresponseswhichcouldskewthedataortotallyconfoundtheexperiment.Thesafeandeffectiveuseofanimalsinalaboratorysettingisanessentialelementofyourresearchactivities.

TheAnimalWelfareAct(AWA),andPHSPolicy

requirethatallAnimalUserscompletetrainingonthelaws,regulations,andproceduresassociatedwithanimalcareanduse.Thiscourseisdesignedtofulfillthatrequirement,providingaquickoverviewoftherelevantlawsandregulationscoveringuseofanimalsinalaboratorysetting,andtherecommendedproceduresforworkingwithspecificanimals.Thecourseincludespracticalandessentialinformationthatwillguideyouintheeverydaycareanduseofanimalsinyourlaboratoryenvironment.ImportanceofProperAnimalCareYourprimaryresponsibilitywhenworkingwithlaboratoryanimalsistotreatthemhumanely.Withthatinmind,themajorchallengeistomanagetheanimalsinawaythatminimizestheirexposuretopainanddistressandmaintainstheintegrityoftheirphysiologicalfunctionstofacilitatethegenerationofreliableresearchdata.Thiscoursewillteachyouhowthepropertreatmentofanimalscarriestheaddedbenefitofminimizingtheintroductionofunwantedvariablesintotheresearchprocess.ImportanceofProperAnimalCareYourcareandtreatmentofanimalsmustalsocomplywiththeappropriatelaws,regulations,andwithNIHpolicy.Manyoftheselaws,regulationsandpolicieswereenactedinresponsetopublicconcernaboutthetreatmentofanimalsintheresearchsetting.Consequently,yourobjectiveshouldbetotreatyouranimalsinamannerthatcomplieswiththeethicalandlegalimperativesprescribedforthehumanetreatmentoflaboratoryanimalsthatwillreflectpositivelyonNIHasaresearchinstitution.researchinstitution.LawsandRegulations:Themainregulations,policiesandguidelinesthatapplytoanimaluseatNIHare:TheAnimalWelfareAct(AWA)

ThePublicHealthServicePolicyonHumaneCareandUseofLaboratoryAnimals(PHSPolicy)

U.S.GovernmentPrinciplesfortheUtilizationandCareofVertebrateAnimalsUsedinTesting,Research,andTraining(GovernmentPrinciples)

GuidefortheCareandUseofLaboratoryAnimals(Guide)

NIHManual3040-2:AnimalCareandUseintheIntramuralProgramStructureandAccreditationTheDDIRdelegatestotheDirectoroftheOfficeofAnimalCareandUse(OACU)overallresponsibilityforensuringthattheACUprogramcomplieswithapplicablepoliciesandregulations.PainandDistressThereisalsoapragmaticpointofscientificlogicthatsupportsminimizingtheexposureofresearchanimalstoexperimentalpainordistress.Animalsrespondtopainanddistresswithchangesintheirnormalphysiologywhichcanskewdatabeingcollected.AlleviatingPainandDistressAnimals,likehumans,canbeexpectedtogenerateastressresponsetonovelstimuli.Handlinganimalsveryearlyinlife,evenasnew-borns,to"gentlethem"isaprocedurethathasbeenwidelyusedonavarietyofspeciestofamiliarizetheanimalswiththehandlingprocess.Whenanimalsaretrainedthiswaytoassociatehandlingwithpleasantcircumstancestheyaremuchlesslikelytogenerateastressresponselaterinlifewhentheyarehandled.AlternativesTheThreeRs:

Reduction:Reductionsinthenumbersofanimalsusedtoobtaininformationofacertainamountandprecision.Refinement:Decreaseintheincidenceorseverityofpainanddistressinthoseanimalsthatareused.Replacement:Substitutionofinsentientmaterialforanimalsorsubstitutionofalowerspecies,whichmightbelesssensitivetopainanddistress,forahigherspecies.MinimizingExposuretoDiseasewhenWorkingwithAnimalsStandardlaboratoryprotectiveclothingincludes:

Uniforms-Uniformsareclothing,suchassurgicalscrubsuitsandcoveralls,dedicatedforwearduringworkintheanimalfacility.Auniformshouldnotbewornoutsideoftheanimalfacilityorresearchsetting.Forexample,uniformsshouldnotbeworninpubliccafeterias,lecturehalls,andmedicalpatientcareareas.Gloves-Lightweightvinylorlatexglovespreventcontaminationoftheskinwithpathogensthatmaybeonanimals'bodiesoronsurfacessoiledbytheirexcreta.Labcoats-Disposablelabcoatsandcoverallsprotectstreetclothesfromcontaminationwithanimalpathogens.Labcoatsshouldnotbewornoutsideoftheanimalfacilityorresearchsetting.Forlongtermworkintheanimalfacility,auniformmaybesubstitutedforthesetypesofcoveringgarments.MucousMembraneProtection–Adeviceoracombinationofdevicessuchasfaceshieldsorsurgicalfacemaskscombinedwithprotectiveglassesorgoggles,worntoprotectthemouth,nose,andeyesfromsplashordropletcontamination.Thedegreeofprotectionneededvarieswiththespecificprocedurebeingconductedandshouldbeadjustedtotheleveloftheanticipatedrisk.Fullfaceshieldsprovidesplashprotectionforallofthefacialmucusmembranes.Partialmucousmembraneprotectionisprovidedbyusingonlyafacemaskandmaybeappropriatewhenthefecaloralrouteofcontaminationistheonlyconcern.ShoeCovering-Stretchbooties,usuallymadeofpaperorplastic,arewornoverstreetshoestopreventthetransferofpathogenicorganisms.Bootiesshouldnotbewornoutsideoftheanimalfacility.Dedicatedfootwearcanbesubstitutedforshoecoveringsduringlong-termactivities.Ifdedicatedshoesareworn,shoecoveringsmaybeusedtocovertheshoeswhenoutsidetheanimalfacility,buttheshoecoversmustberemovedonreturntothefacility.CageSystems

Cagesystemsthatmeetthegeneralrequirementsformaintenanceofnormalbodyfunctionsandprovidemoderateprotectionfromexposuretopathogensaredescribedas"conventional"cages.Conventionalcages,however,donotadequatelyprotectratsandmicefromairbornediseasestowhichtheyarehighlysusceptiblesocagesystemsthatarespeciallydesignedtoprovideextraprotectionfromthesepathogensareused.Rodentvirusesandotherairbornepathogensmaybetransportedbyclingingtodustparticlesandotherparticulatemattersuspendedintheairinsideandabovetheanimalcage.Theriskofexposuretotheseorganismsisreducedbyincreasingfreshaircirculatingaroundthecages,orfilteringtheair.waterSanitationFilteredAirUnit

ObservingandReportingProblemswithAnimalsWhileanimalhealthisthechargeoftheveterinarians,youwillhavearesponsibilitytoobserveandreportchangesinananimal'shealthandtoinformtheveterinarianoranimalfacilitymanagersothattheanimalcanbetreated.Immediateidentificationofsickorinjuredanimalsiscriticallyimportantinaresearchsettingbecausetheanimalsarecloselyconfined,andhousingspaceisfrequentlysharedbyseveralinvestigators.Infectiousdiseaseinonegroupofanimalscouldjeopardizealloftheanimalsintheroomorbuilding.SignsofIllnessandInjury:

Pickingupmice.

Whenpickingupmice,alwaysholdthemousebythebaseofthetail.Thiscanbedonewitheitherforcepsoryourfingers.DONOTpickmiceupbythetipofthetail.Ifyoupickamouseupbythetipofitstail,themousemayspinintightcircleswhenitsfeetlosecontactwiththecagesurface.Ifthishappens,thetipoftailcanbepulledoff.Additionally,amouseheldbythetipofitstailcan"climbupitstail"andbiteyou!Separatingmice/ratsbygender.

Separatingmice/ratsbygendercanbedifficult,especiallywhentheyareyoungandmales'testicleshavenotdescendedintothescrotum.Comparetherelativedistancebetweenanusandurinarypapilla.Thedistancebetweentheanusandtheurinarypapillaisgreaterinthemale.Thisphotographillustratesthatthemouseontherighthasthelesser(shorter)distancebetweentheanusandtheurinarypapilla,soitisthefemale.RestrainingRats.

Ratscanbecaughtandliftedbythebaseoftheirtail,oryoucangraspthemaroundtheirbodyasillustratedinthisphotograph.Ifyougrasptheratarounditsbodyyoucanpreventitfrombitingyoubyplacingyourindexfingersnuglyunderthejawagainstthepointoftheshoulderaheadoftherightfrontleg.Yourthumbisplacedbehindtheleftfrontlegwhichpushesitforwardagainsttheleftsideoftheneck.Thisgripsecuresthehead,andtheratcannotreacharoundtobite.Forlargerats,keepingagoodgrasponthetailwillgiveyouadditionalcontrol.GuineaPigs.GuineaPigshavenotailandmustbecaughtbygraspingthemaroundtheirbody.Theyareveryinclinedtoavoidbeingcaughtbyscurryingaroundthecageandwhistlingloudly.Whentheyarecaughttheyrarelyattempttobitebutmayreachforwardwiththeirhindfeetandscratchwiththeirlongtoenails.

Rabbits.

Rabbitsarecharacterizedbytheirthinbones,fragileskeleton,andheavymusclesontheirbackandhindlegs.Whenrestrainingandliftingarabbit,themostimportantthingtorememberistosupporttherabbitsrumpasyoupickitup.Iftherabbitkicks,evenonetime,whenitsbackfeetleavethecageflooritcanbreakitsback,bepermanentlyparalyzedandhavetobeeuthanized.Thecorrecttechniqueistoapproachtherabbitwithyourhandheldhighovertheback,grasptheskinovertheshouldersandslideyourotherhandbetweenthebacklegstosupporttherumpbeforeyoulifttheanimaloffofthecagefloor.Intra-peritonealinjection.

1.Restrainthemousebygraspingtheskinalongitsbackwithyourlefthand(ifright-handed).

2.Clampthetailbetweenyourringfingerandlittlefingerforadditionalcontrol.

3.Positionthemousesothatit'sheadisdown.(Intestines,stomach,etc.shouldfallforwardandcreateasmallspaceintheposteriorportionoftheperitonealcavity.)

4.Inserttheneedlealittleofftothesideofthecenterline(tomissthebladder).Collectingabloodsample.

1.Givegeneralanesthesia.

2.Usecapillarytubetreatedwithheparininsidesothattheblooddoesn'tclot.

3.Introducethetubeintothemedialcanthusoftherodent'seye,directedtowardthebackoftheorbit.

4.Rotatethetubetocutintothevenoussinusinthebackoftheorbit.Samplesmaybemoredifficulttocollectfromratsbecausetheyhaveavenousplexusratherthanavenoussinusatthebackoftheirorbitandthevesselscanbehardtolocate.5.Bloodflowsdownintothecapillarytube.

6.Removetubeandapplygentlepressureagainsttheeyewithagauzespongetostopthebleeding.

7.Watchtheanimalcloselytopreventcagematesfrominjuringitduringitsrecoveryfromtheanesthesia.GivingInjections/TakingSamplesRabbits:Rabbitsarefrequentlyusedtoproduceantibodiestoaninjectedantigenandlarge(50ml)bloodsamplesarecollectedforharvestingantibodies.Drawingasample.

1.Bloodisusuallycollectedfromtheauricularartery(darkline)thatliesalongthemiddleofthetopsurfaceoftheear.

2.Alargebore(18guage)needleismostfrequentlyusedtodrawsamples.GivingInjections/Taki