DIVISION OF AGRICULTURE
RESEARCH & EXTENSION
University of Arkansas System
Embr yoTransferinCattle
TomR.Troxel
ProfessorandAssociate
DepartmentHeadAnimal
Science
ArkansasIs
Thefirstcommercialembryo
OurCampus
theearly1970s.Initially,embryos
Visitourwebsiteat:
http://www.uaex.edu
UniversityofArkansas,UnitedStatesDepartmentofAgriculture,andCountyGovernmentsCooperating
AgricultureandNaturalResources
Introduction
Embryotransferisonestepinthe
processofremovingoneormore
embryosfromthereproductivetractof
adonorfemaleandtransferringthem
tooneormorerecipientfemales.
Embryosalsocanbeproducedinthe
laboratoryviatechniquessuchas
in vitro fertilizationorsomaticcell
cloning.Buttheactualtransferofan
embryoisonlyonestepinaseriesof
processesthatmayincludesomeorall
ofthefollowing:superovulationand
inseminationofdonors,collectionof
embryos,isolation,evaluationand
shorttermstorageofembryos,micro
manipulationandgenetictestingof
embryos,freezingofembryosand
embryotransfer.
Embryotransferincattlehas
recentlygainedconsiderablepopu
laritywithseedstockdairyandbeef
producers.Mostoftheapplicable
embryotransfertechnologywasdevel
opedinthe1970sand1980s;however,
thehistoryoftheconceptgoesback
muchfarther.Embryotransferwas
firstperformedandrecordedby
WalterHeapein1890.Hetransferred
twoAngorarabbitembryosintoa
gestatingBelgiandoe.TheBelgian
doeproducedamixedlitterofBelgian
andAngorabunnies.Embryotransfer
infoodanimalsbeganinthe1930s
withsheepandgoats,butitwasnot
untilthe1950sthatsuccessful
embryo transferswerereportedin
cattleandpigsbyJimRowsonat
Cambridge,England.
transfersinthiscountryweredonein
Thefirststepisselectingadonor
wererecoveredfromvaluabledonors
andtransferredtorecipientanimals
usingsurgicalprocedures.Itwasnot
FSA3119
untilnonsurgicalmethodswere
developedinthelate1970sthat
embryotransfergrewinpopularity.
PotentialValueofEmbryo
TransferinCattle
Thereproductivepotentialofeach
normalnewborncalfisenormous.
Thereareanestimated150,000
potentialeggsorovainthecowand
billionsofspermproducedbyeach
bull.Bynaturalbreeding,onlyafrac
tionofthereproductivepotentialofan
outstandingindividualisrealized.
Theaverageherdbullwillsire15to
50calvesperyear,andtheaverage
cowwillhaveonecalfperyear.With
artificialinsemination,itispossibleto
exploitthevastnumberofspermpro
ducedbyageneticallysuperiorbull;
however,thereproductivepotentialof
thefemalehasbeenlargelyunutilized.
Undernormalmanagementprograms,
acowproducesanaverageofeightto
tencalvesinherlifetime.Likeartifi
cialinseminationhasdoneforthe
bull,embryotransferisatechnique
thatcangreatlyincreasethenumber
ofoffspringageneticallyimportant
cowcanproduce.
StepsforEmbryoTransfer
inCattle
Virtuallyallcommercialembryo
transfersusenonsurgicalrecoveryof
theembryosratherthansurgicaltech
niques.Theprocessinvolvesseveral
stepsandconsiderabletimeaswellas
variableexpense.
1) SelectionoftheDonorCow
cow.Beefproducerswilldifferintheir
opinionsregardingthecriteriafor
selectingageneticallyoutstandingcow.
Whetherthecriteriaisperformancerecords,show
ringappealorboth,considerationmustbegivento
potentialdollarvalueofhercalves.Thereisconsider
ableexpenseincurredinachievingasuccessfully
transferredpregnancy.Therefore,thesalevalueof
thenewborncalfshouldbehighenoughtowarrant
theaddedexpenseoftheembryotransferprocedure.
Becausedairycattleareselectedmoreroutinelyon
onemajortrait(milkproduction),thedecisionscon
cerningdonorcowsaresomewhatlesscomplicated
thaninbeefcattle.However,theeconomicconsidera
tionsareequallyimportant.Embryotransferisnota
cureall.Itdoesnotmakeaveragecattlegoodor Thecorrectsiteforsemenplacementisinthebodyof
goodcattlebetter.Itissuitableforalimitednumber
ofseedstockproducerswithbeefordairycattlethat
canbebreedorspeciesimproversforoneormore
economicallyimportanttraits.
Thepotentialdonorcowshouldbereproductively
soundtoproducethedesiredresults.Thecowshould
haveanormalreproductivetractandpostpartum
history,especiallywithregardtoestrouslengthsof
18to24days.Bothbeefanddairycowsshouldbeat
least60dayspostpartumbeforethetransferproce
durebegins.Ithasbeensuggestedthatprospective
donorcowsinembryotransferprogramsbeselected
onthefollowingcriteria:
Regularestrouscyclescommencingata
young age
Ahistoryofnomorethantwobreedings
per conception
Previouscalveswithapproximately365day
intervals
Noparturitiondifficultiesorreproductive
irregularities
Noconformationalordetectablegeneticdefects
Thecowshouldbemaintainedatanutritionlevel
appropriateforhersizeandlevelofmilkproduction.
Boththeveryobesecowandthethincowwillhave
reducedfertility,soitisimportantthatthedonorbe
inanappropriatebodyconditionscoreatthetimeof
embryotransfer.
2) SuperovulationoftheDonorCow
Superovulationofthedonorcowisthenextstep
intheembryotransferprocess.Superovulationis
the releaseofmultipleeggsatasingleestrus.Cows
orheifersproperlytreatedcanreleaseasmanyas
ten or moreviableeggsatoneestrus.Approximately
85 percentofallnormalfertiledonorswillrespond
to superovulationtreatmentwithanaverageoffive
transferableembryos.Somecowsthatarerepeatedly
superovulatedat60dayintervalsmayproducefewer
number of eggs over time.The basic principle ofsuper
ovulationistostimulateextensivefolliculardevelop
mentthroughtheuseoffolliclestimulatinghormone
(FSH).Superovulationprotocolsmaydifferamong
embryotechnicians,butgenerally,FSHpreparations
areinjectedtwicedailyforfourdaysatthemiddle
or neartheendofanormalestrouscycle,whilea
functionalcorpusluteum(CL)isontheovary. A
prostaglandininjectiongivenonthefourthdayofthe
treatmentschedulewillcauseCLregressionand
estrustooccurapproximately48hours later.
3) InseminationoftheCow
Becauseofthereleaseofmanyovafrommultiple
follicles,thereisagreaterneedforviablespermcells
toreachtheoviductsofthesuperovulatedfemales.
Therefore,manyembryotransfertechnicianswill
choosetoinseminatethecowseveraltimesduring
andafterestrus.Oneschemeistoinseminatethe
superovulatedcowat12,24and36hoursafterthe
onsetofstandingestrus.Usinghighqualitysemen
withahighpercentageofnormal,motilecellsisa
verycriticalstepinanyembryotransferprogram.
theuterus.Thisisasmalltarget(1/2to1inch)just
infrontofthecervix.
4) FlushingtheEmbryos
Tocollecttheembryosnonsurgically,asmall
syntheticrubbercatheterisinsertedthroughthe
cervixofthedonorcow,andaspecialmediumis
flushedintoandoutoftheuterustocollectthe
embryossevendaysafterestrus(Figure1).
Figure1.Diagramofthe
embryoflushingand
recoveryprocedure.
Thiscollectionprocedureisrelativelysimpleandcan
becompletedin30minutesorlesswithoutharmto
thecow.Apresterilizedstyletisplacedinthelumen
ofthecathetertoofferrigidityforpassagethrough
thecervixintothebodyoftheuterus.Whenthetip
of thecatheterisinthebodyoftheuterus,thecuff
is slowlyfilledwithapproximately2mLofnormal
saline.Thecatheteristhengentlypulledsothatthe
cuffisseatedintotheinternalosofthecervix.Addi
tionalsalineisthenaddedtothecufftocompletely
sealtheinternalosofthecervix.AYconnectorwith
inflowandoutflowtubesisattachedtothecatheter.
Apairofforcepsisattachedtoeachtubetoregulate
theflowofflushingfluid.Thefluidissequentially
addedandremovedbygravity.Thefluidinthe
uterusisagitatedrectally,especiallyintheupper
onethirdoftheuterinehorn.Theuterusisfinally
filledwithmediumtoaboutthesizeofa40day
pregnancy.Oneliteroffluidisusedperdonor.Many
embryo transfer technicians useasmallervolumeand
flushoneuterinehornatatime.Eachuterinehornis
filledandemptiedfivetotentimeswith30to200mL
offluideachtime,accordingtosizeoftheuterus.The
embryosareflushedoutwiththisfluidandcollected
inafilterwiththefluid.Theporesinthefilterare
smallerthantheembryos,soexcessfluiddrainsout
ofthefilterwithoutlosingtheembryos.Embryosare
separatedfromtheflushmediaandexaminedunder
amicroscopetodeterminetheirqualityandstage
of development.

5) EvaluationoftheEmbryos
Astheindividualembryosarelocatedusinga
microscope,theyareevaluatedfortheirqualityand
classifiednumericallyastothepotentiallikelihoodof
successiftransferredtoarecipientfemale.Themajor
criteriaforevaluationinclude:
Regularityofshapeoftheembryo
Compactnessoftheblastomeres(thedividing
cellswithintheboundariesoftheembryo)
Variationincellsize
Colorandtextureofthecytoplasm(thefluid
withinthecellwall)
Overalldiameteroftheembryo
Presenceofextrudedcells Figure2.Morphologicallynormalbovineembryos
Regularityofthezonapellucida(theprotective
recoveredatvariousstagesofdevelopment.
layerofproteinandpolysaccharidesaroundthe
singlecelledembryo) 6) SelectionandPreparationof
Presenceofvesicles(smallbubblelike
RecipientFemales
structuresinthecytoplasm)

Properrecipientherdmanagementiscriticalto
Embryosareclassifiedaccordingtothese embryo transfer success.Cowsthatarereproductively
subjectivecriteriaas:
sound,thatexhibitcalvingeaseandthathavegood
milkingandmotheringabilityarerecipientprospects.
Grade1:Excellentorgood They must be ona properplane ofnutrition(body
Grade2:Fair
condition score 6 for beef cows and dairy bodycondition
Grade3:Poor score3to4).Thesecowsalsomustbeonasound
Grade4:Deadordegenerating herdhealthprogram.

Embryosalsoareevaluatedfortheirstageof
developmentwithoutregardtoquality.Thesestages
arealsonumbered:
Stage1:Unfertilized
Stage2:2to12cell
Stage3:Earlymorula
Stage4:Morula
Stage5:Earlyblastocyst
Stage6:Blastocyst
Stage7:Expandedblastocyst
Stage8:Hatchedblastocyst
Stage9:Expandinghatchedblastocyst
Thereisapparentlynodifferenceinpregnancy
ratesoffertilizedcellsindifferentstagesofdevelop
ment,assumingtheyaretransferredtotherecipient
femaleintheappropriatestageoftheestrouscycle.
Stages4,5and6embryosendurethefreezingand
thawingprocedureswiththegreatestviability.
Embryoqualityisalsoofutmostimportanceinthe
survivalofthefreezingandthawingstress.Grade 1
embryosgenerallyareconsideredtheonlyonesto
freeze.Grade2embryoscanbefrozenandthawed,
yetpregnancyratestypicallyarereduced.Ina
Louisianastudyinvolving1,116beefanddairycows
of15breeds,58percentofallembryosweretransfer
able,31percentwereunfertilizedand11percent
weredegenerated.
Itisdifficulttoknowhowmanyrecipientcows
areneededpereachdonorflushed.Toestablishan
averagefigureforthenumberofembryotransfer
calvesfromasingledonorcowinayearisdifficult.
Variationsinconditionsarewide,butifacowis
flushedevery90daysovera12monthperiodand
fivepregnanciesareobtainedpercollection,anaver
ageof20pregnanciesperyearcouldresult.Some
cowshaveproducedmorethan50pregnanciesper
yearbyembryotransferandprobablycouldhave
producedmoreifeconomicallyfeasible.Inthe
Louisianastudypreviouslymentioned,theaverage
numberofembryosfoundpercowwas7.4.Withonly
58percentofthesebeingtransferable,theaverage
was4.3transferableembryosperflush.
Tomaximizeembryosurvivalintherecipient
femalefollowingtransfer,conditionsintherecipient
reproductivetractshouldcloselyresemblethosein
thedonor.Thisrequiressynchronizationofthe
estrouscyclesbetweenthedonorandtherecipients,
optimallywithinonedayofeachother.Synchroniza
tionoftherecipientscanbedoneinasimilarmanner
andatthesameworkingtimeasthedonorcows.
Thereareanumberofdifferentestroussynchroniza
tionprotocolswithadvantagesanddisadvantagesfor
eachprotocol.Thecriticalpointregardingrecipient
cow estrous synchronizationisthetimingmust
matchthetimeofinseminationofthedonorcowso
thatthedonorandtherecipientshaveasimilar
uterineenvironmentsevendayslaterwhenthe
transfertakesplace.Synchronizingproductsare
more effective
onrecipientfemalesthatarealready
cycling.Anestrusornoncyclingcowsthataretoo
thinortoo shortindayspostpartumwillnotmake
usefulrecipients.
7) TransferoftheEmbryos
Thetransferoftheembryointotherecipientcow
firstrequiresloadingtheembryointoa1/4mL
inseminationstraw.Thisisdoneundermicroscopic
viewingwiththeaidofa1mLsyringeandrequires
considerablepractice,patienceanddexterity.Degen
eratedembryosorembryosofverylowgradeneednot
beloadedandcanbediscarded.Justpriortoembryo
transfer,theovariesoftherecipientarepalpatedrec
tallytodeterminewhichovaryhasovulated.Withthe
aidofanassistanttoholdopenthevulvaoftherecip
ientcow,thetransfergunorinseminationrodiscare
fullypassedthroughthecervix.Thetipoftherodis
thenallowedtoslideintothehornonthesameside
oftheovarywithanactivecorpusluteum.The
embryoisgentlyexpelledintheforwardtipofthat
uterinehorn.Greatcareistakentonotcausedamage
totheliningoftheuterus.Suchinflammationand
scarringwouldgreatlyreducetheprobabilityofthe
pregnancybeingestablished.Embryoflushingand
embryotransferarebothdoneafteranepiduralanes
thetichasbeengiventoblockcontractionsofthe
digestivetractandaidintheeaseofmanipulationof
thecervixandtheuterinehorns.Embryosshouldbe
transferredassoonaspossibleaftertheflush(within
8hoursatleast).
Thefreezingandthawingprocessisalsovery
intricateandusuallyresultsinapproximately10to
20percentreductioninpregnancyratesfromthose
observedwithfreshembryos.
Frozenembryosareamarketablecommodityand
havebeenespeciallyusefulininternationalsalesof
UnitedStatesbeefanddairygenetics.Producerswho
believetheyowncattlewiththegeneticcapabilityto
bevaluableinothernationsmaywishtocontacttheir
stateDepartmentofAgricultureandaskaboutregu
lationsandmarketabilityoffrozenembryosfrom
theirherd.Differentnationshavedifferenthealth
requirementsofcattleproducingfrozenembryosfor
importintotheircountry.Therefore,individual
inquiriesarenecessarytolearnwhathealthandlegal
requirementsareexpected.
8) ExpectedEmbryoTransplantResults
Embryoproductionvariesgreatlyfromdonorto
donorandflushtoflush.Averageproductionis
approximatelysixfreezable(excellentandgood)and
eighttransferable(excellent,good,fairandpoor)
embryospersuperovulation.
PrintedbyUniversityofArkansasCooperativeExtensionServicePrintingServices.
DR.TOMR.TROXEL isprofessorandassociatedepartmenthead,
DepartmentofAnimalScience,UniversityofArkansasDivisionof
Agriculture,LittleRock.
FSA3119PD113RV
Pregnancyratesvaryfromflushtoflushwith
freshaverages60to70percentandfrozen50to
60 percent.Manyfactorsaffectpregnancyratessuch
asembryoquality,recipients,technicalabilityand
donor.Some donors consistently produceembryos
withhigherpregnancyratesthanotherswith
embryosofsimilargrade.Thislastfactorseemstobe
uncontrollableandunpredictable.
CostsofEmbryoTransfer
Thecostsofembryotransferareasvariableas
thecostsofbuyinganewtruck.Manydifferent
optionsandpackagesareofferedbyembryotransfer
technicians.Sometechniciansperformembryo
transferonlyontheranchwherethedonorcowsare
located.Othershavefacilitiestohouseandboard
donorandrecipientcowsandperformembryo
transferunderhospitallikeconditions.Manytechni
cianshavetheequipmentandexpertisetofreezeand
storeembryosforlatertransplantationorshipment
toothercountries.
Minimumcostsof$250perpregnancyhavebeen
reportedbyembryotransfertechnicians.Thesecosts
may notincludecostsforsuperovulationandcertainly
donotincludesemen,registration,embryotransfer
certificates,bloodtypingofdonorcowsandancestors
and,mostimportantly,thecostofmaintainingthe
donorcowuntilthecalfisweaned.Threetofive
strawsofvaluablesemencanbepricedfrom$45to
$300.Propernutrition,healthcareandsynchroni
zationofthedonorandtherecipientcanaddanother
$400to$500expensetoeachsuccessfulpregnancy.
Consequently,manypurebredoperationsconducting
embryotransferonaregularbasisconsiderthateach
ETcalfmusthaveamarketvalueof$1,500to
$2,000greaterthanothernaturallyconceivedand
rearedcalvesintheherdbeforeembryotransfer
is considered.
Beefanddairyproducersconsideringusing
embryotransfershouldfirstvisitwiththeirbreed
representativetodeterminethespecificrequirements
forcertificationandregistrationofembryotransfer
calvesinthatbreed.
AdaptedfromEmbryoTransferinCattle (F3158)by
G. Selk,OklahomaCooperativeExtensionService,
Stillwater,OK.
AdditionalSources
Davis,R.L.EmbryoTransferinBeefCattle.DavisRairdan
International.
Hasler,J.F.2004.FactorsInfluencingtheSuccessof
EmbryoTransferinCattle.23rdWorldBuiatrics
Congress,Qubec,Canada.
IssuedinfurtheranceofCooperativeExtensionwork,ActsofMay 8 and
June30,1914,incooperationwiththeU.S.DepartmentofAgriculture,
Director, Cooperative Extension Service, University of Arkansas. The
ArkansasCooperativeExtensionServiceoffersitsprogramstoalleligi
blepersonsregardlessofrace,color,nationalorigin,religion,gender,age,
disability,maritalorveteranstatus,oranyotherlegallyprotectedstatus,
andisanAffirmativeAction/EqualOpportunityEmployer.