實(shí)驗(yàn)動(dòng)物學(xué)總論一實(shí)驗(yàn)動(dòng)物和實(shí)驗(yàn)用動(dòng)物

〔一〕實(shí)驗(yàn)動(dòng)物(Laboratoryanimals)是指經(jīng)人工培育或人工改造，對(duì)其攜帶的微生物實(shí)行控制；遺傳背景明確，來(lái)源清楚，用于科學(xué)實(shí)驗(yàn)、藥品、生物制品的生產(chǎn)和檢定及其他科學(xué)實(shí)驗(yàn)的動(dòng)物。〔二〕實(shí)驗(yàn)用動(dòng)物(Animalsforresearch；Experimentalanimals〕所有用于科學(xué)實(shí)驗(yàn)的動(dòng)物統(tǒng)稱為實(shí)驗(yàn)動(dòng)物。包括實(shí)驗(yàn)動(dòng)物，野生動(dòng)物，經(jīng)濟(jì)動(dòng)物和欣賞動(dòng)物。二實(shí)驗(yàn)動(dòng)物學(xué)的定義和范圍〔一〕定義

實(shí)驗(yàn)動(dòng)物學(xué)〔Laboratoryanimalscience〕是研究實(shí)驗(yàn)動(dòng)物和動(dòng)物實(shí)驗(yàn)的科學(xué)。〔二〕研究范圍1.實(shí)驗(yàn)動(dòng)物遺傳育種學(xué)〔Laboratoryanimalgeneticbreedingscience〕2.實(shí)驗(yàn)動(dòng)物微生物學(xué)和寄生蟲學(xué)〔Laboratoryanimalmicrobiologyandparasitology〕3.實(shí)驗(yàn)動(dòng)物環(huán)境生態(tài)學(xué)〔Laboratoryanimalenvironmentalecology〕4.實(shí)驗(yàn)動(dòng)物營(yíng)養(yǎng)學(xué)〔Laboratoryanimalnutriology〕

〔二〕研究范圍(續(xù))5.實(shí)驗(yàn)動(dòng)物飼養(yǎng)管理〔Laboratoryanimalhusbandry〕6.實(shí)驗(yàn)動(dòng)物醫(yī)學(xué)〔Laboratoryanimalmedicine〕7.比較醫(yī)學(xué)〔Comparativemedicine〕8.動(dòng)物實(shí)驗(yàn)〔Animalexperimentaltechniques〕三實(shí)驗(yàn)動(dòng)物的重要性〔一〕實(shí)驗(yàn)動(dòng)物是生命科學(xué)研究的支撐條件之一生命科學(xué)實(shí)驗(yàn)研究的四個(gè)支撐條件----AEIR要素A：Animal：實(shí)驗(yàn)動(dòng)物；E：Equipment：儀器設(shè)備；I：Information：信息；R：Reagent：試劑；〔二〕實(shí)驗(yàn)動(dòng)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(lái)源于實(shí)驗(yàn)動(dòng)物〔三〕實(shí)驗(yàn)動(dòng)物是人類的替身，起著“活的天秤〞和“活的化學(xué)試劑〞的作用〔四〕實(shí)驗(yàn)動(dòng)物工作實(shí)行法制化管理四實(shí)驗(yàn)動(dòng)物的分類〔一〕傳統(tǒng)的動(dòng)物學(xué)分類方法依據(jù)自然分類法，把整個(gè)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(jí)。以大家鼠為例，它屬于：脊椎動(dòng)物門(phylumvertebrata)哺乳動(dòng)物綱(classmammalia)嚙齒目(orderrodentea)鼠科(familymurinae)大家鼠屬(genusrattus)大家鼠種(speciesrattusnorvegicus)學(xué)名：褐家鼠〔大家鼠〕(Rattusnorvegicus)多數(shù)情況下，根據(jù)不同目的進(jìn)行種下分類，把實(shí)驗(yàn)動(dòng)物劃分為不同品系。〔二〕按實(shí)際用途分類1、實(shí)驗(yàn)動(dòng)物2、經(jīng)濟(jì)動(dòng)物〔Economicalanimals〕，或稱家畜、家禽3、野生動(dòng)物〔Wildanimals〕4、欣賞動(dòng)物〔Exihibitinganimals〕〔三〕按遺傳學(xué)控制分類

根據(jù)基因的純合程度，把實(shí)驗(yàn)動(dòng)物分成以下四類：1、近交系動(dòng)物〔Inbredstrainanimals〕2、突變系動(dòng)物〔Mutantstrainanimals〕3、雜交群動(dòng)物〔Hybridcolonyanimals〕〔雜交一代，F(xiàn)1代動(dòng)物〕4、封閉群動(dòng)物(Closedcolonyanimals)1、近交系動(dòng)物〔Inbredstrainanimals〕又叫純系動(dòng)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(lái)的遺傳上到達(dá)高度一致的動(dòng)物群。

基因純合程度可達(dá)99.8%。①

主要指嚙齒動(dòng)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(shí)必須采用年輕的雙親同其子女交配。④

較大動(dòng)物純種培育很難獲得成功，因?yàn)槭来g隔較長(zhǎng)，費(fèi)用較大，所以成功率低。⑤

禽類和兔的血緣關(guān)系到達(dá)80%以上〔相當(dāng)于兄妹交配四代〕時(shí)，即可稱為近交系。

2、突變系動(dòng)物〔Mutantstrainanimals〕具有特殊突變基因的品系動(dòng)物，正常染色體基因發(fā)生突變，并具有各種遺傳缺陷的動(dòng)物。在長(zhǎng)期繁殖過(guò)程中，動(dòng)物的子代突然發(fā)生變異，變異的基因位點(diǎn)又可遺傳下去，或者即使沒(méi)有明確的基因位點(diǎn)，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(dòng)物，稱為突變系動(dòng)物。如無(wú)胸腺裸鼠、無(wú)K細(xì)胞、或無(wú)K、無(wú)B、無(wú)巨噬細(xì)胞等裸鼠。用于免疫研究、移植實(shí)驗(yàn)等。3、雜交群動(dòng)物〔Hybridcolonyanimals〕〔雜交一代，F(xiàn)1代動(dòng)物〕兩個(gè)近交品系動(dòng)物之間進(jìn)行有方案交配所獲得的第一代動(dòng)物。例如：C57BL/6J×DBA/2→B6D2F1〔B6為C57BL/6J的縮寫，D2為DBA/2的縮寫?！?、封閉群動(dòng)物(Closedcolonyanimals)以非近親交配方式進(jìn)行繁殖生產(chǎn)的一個(gè)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(yuǎn)交群。2、無(wú)特定病原體動(dòng)物(Speceficpathogenfreeanimals，SPF〕體內(nèi)不存在特定病原微生物和寄生蟲的的動(dòng)物，簡(jiǎn)稱SPF動(dòng)物。是指無(wú)傳染病的健康動(dòng)物。這種動(dòng)物都是來(lái)自無(wú)菌動(dòng)物或悉生動(dòng)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(jìn)行實(shí)驗(yàn)，要進(jìn)行嚴(yán)格消毒、檢疫、隔離并定期剖腹凈化。3、清潔動(dòng)物(Cleananimals，CL)又稱最低限度疾病動(dòng)物(Ginimaldiseaseanimals),體內(nèi)外不攜帶人畜共患的病原體或動(dòng)物傳染病病原的動(dòng)物，不能帶有體外寄生蟲和大局部體內(nèi)寄生蟲。4、普通動(dòng)物〔Conventionalanimals〕

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(dòng)物。墊料和飼料和飲水一般不消毒，飲用普通自來(lái)水。

所謂普通動(dòng)物也并不是對(duì)微生物沒(méi)有一定控制的一般動(dòng)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(dòng)物只能供教學(xué)和一般實(shí)驗(yàn)用。

根據(jù)新修訂的國(guó)家實(shí)驗(yàn)動(dòng)物微生物、寄生蟲質(zhì)量標(biāo)準(zhǔn)，我國(guó)將實(shí)驗(yàn)大、小鼠分為三類即清潔級(jí)、無(wú)特定病原體級(jí)、無(wú)菌級(jí)〔包括悉生動(dòng)物〕。其它品種實(shí)驗(yàn)動(dòng)物仍然分為普通級(jí)、清潔級(jí)、無(wú)特定病原體級(jí)、無(wú)菌級(jí)〔包括悉生動(dòng)物〕四級(jí)。即從2002年5月1日起取消了普通級(jí)大、小鼠標(biāo)準(zhǔn)。TheJacksonLaboratory

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

humangenomesweresequencedattheturnofthecenturyandproveddefinitivelythatthetwospeciessharethevastmajorityoftheirgenes.【上海斯萊克實(shí)驗(yàn)動(dòng)物有限責(zé)任公司】【中國(guó)科學(xué)院上海實(shí)驗(yàn)動(dòng)物中心】【國(guó)家嚙齒類實(shí)驗(yàn)動(dòng)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