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519–524 Theriogenology71 (2009)

www.theriojournal.com

EffectsoftemporarycalfremovalandeCGonpregnancyrate
totimed-inseminationinprogesterone-treatedpostpartum
Nellorecows
#Pages
un [1] #Pages
un [1] #Pages
b [1] #Pages
un [1] #Pages
b [1]
V.G.Pinheiro, A.F.Souza, M.F.Pegorer, R.A.Satrapa, R.L.Ereno,
#Pages [1]
c #Pages
a,
* [1]
L.A.Trinca, C.M.Barros
un
DepartmentofPharmacology, UniversityofSaoPauloState (UNESP), 1868-000Botucatu, SaoPaulo, Brasil
b
DepartmentofBiostatistics-IB, UniversityofSaoPauloState (UNESP), 1868-000Botucatu, SaoPaulo, Brasil
c
DepartmentofAnimalReproduction-FMVZ, UniversityofSaoPauloState (UNESP), 1868-000Botucatu, SaoPaulo, Bras
Received12November2007; receivedinrevisedform18June2008; accepted14August2008

Resumen
Theobjectivewastoevaluatetheeffectsoftemporarycalfremoval (TCR), eCGadministration, orboth, basado e
Protocolo.SuckledNellorecows (40–80dpostpartum, n = 443) withbodyconditionscoresfrom2.0to3.5(5-pointscale) Co
farmswereallgivenasynchronizingprotocol(Pepe).Atthestart (designatedDay0), cowsweregivenanintravaginaldev
ofprogesterone) im and2.5mgofestradiolbenzoate (EB).OnDay8, thedevicewasremovedandcowsweregiv mg
2un(150
deD-cloprostenolim), followedin24hby1.0mgEBim, and30–36hthereafter, gira-timeAI.Thedesignwasa2 2factorial;
maineffectswereTCR (54–60h; fromdeviceremovaltoFTAI) andeCGtreatment (300IUim, 2unconcurrentw ).
TransrectalultrasonographywasdoneonDays 10and0todetectanestrus (absenceofaCLatbothexaminations) 30daftery
FTAI(pregnancydiagnosis).Datawereanalyzedbylogisticregression.Thefollowingvariablesdidnotsignificantlyaff
pregnancyrates:Farm,postpartuminterval,cyclicity,inseminators,andsemen(Sire).En general, el 77% ofthecowsweredee
anestro.Pregnancyratesweresimilar) P >0.05)amongtreatmentgroups:control(54/108=50.0%),TCR(44/106=41.5%
eCG(63/116=54.3%),andTCR+eCG(49/113=43.4%).Pregnancyratewashigherinmultiparousthanprimiparouscows
360,51.7%vs.24/83,28.9%, P <0.01), butwasnotsignificantlyaffectedbycyclicitystatusorbodyconditionscore.Inconclusio
temporarycalfremoval, eCG, orboth, didnotsignificantlyincreasepregnancyratetotimed-inseminationinaprogesterone-
synchronizationprotocolinpostpartumNellorecowswithacceptablebodycondition.
# 2009ElsevierInc.Allrightsreserved.

Palabras clave:
Fijo-timeartificialinsemination;Progesterona;Bovinos; eCG;Temporarycalfremoval

1 Introducción Theshortdurationofestrousbehavior (11 h), y

hace highincidenceofestrusatnight (30-50%)
Zebucattle) B o s t a u r u s)i n arepredominant
d i c u s difficulttodetectestrusinNellorefemales [1].
inBrazilandothertropicalandsubtropicalregions. Sin embargo, characterizationofbovinefollicular
dynamicsbyultrasonography [2] provideda
rationalbasisforpharmacologicalmanipulationof
* Correspondingauthor.Tel.:+551438116253; theestrouscycle, andsynchronizationofovulation
Fax: +551438153744. thatenabledfixed-timeartificialinsemination(FTAI)
E-mail: cmbarros@IBB.UNESP.br (C.M.Barros).withoutestrusdetection.

0093-691 X / $–seefrontmatter
# 2009ElsevierInc.Allrightsreserved.
doi:10.1016/j.theriogenology.2008.08.018

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520 V.G.Pinheiroetal./Theriogenology71 (2009)–524
519

AmongthenumeroustreatmentstestedforFTAIin (Turvo n = 105, FarmB) andSantaCruzdoRioParto

beefcattle, protocolswithprogestinsweremost (n = 100, FarmC) inthecentral-westregionofthestate
efficacious, particularlyincattlewithpostpartum ofSaoPaulo, Brasil (latitude23 8 0 11 S, longitud
0
anestro [7].Theuseofprogesterone-releasingintrava 498 - 23 W), duringthe2004breedingseason.Nellore
inassociationwithadministrationof ginaldevices (P4) vacas) n = 443, 40–80dpostpartum, bodycondition
hasbeenextensivelyusedin estradiolbenzoate (EB) scoresfrom2.0to3.5 (5-pointscale [20]) fueron
FTAIprotocols [8].Theseprotocolstypicallyinclude maintainedonpasture) Brachiariadecumbenso
EBtreatmentconcurrentwithinsertionoftheprogester-Brachiariabrizantha ) conadlibitum mineralsalt
uno-releasingdevice (Day0), 2untreatmentatdevice
PGF suplementación.80-Fourcowswereprimiparous,
retiro (Days7or8), with1.0mgofEB24hlater, ywhereastheremainder) n = 360) weremultiparous.
FTAI30–36hafterEB.Sincethisprotocolinvolves
treatmentwithprogesterone, estrógenos, prostaglandinas,
2.2.Semenandinseminators
estrógeno, thisiscommonlydeemedthePEPEprotocol.
ThePEPEprotocolhasundergonemodificationsin OnFarmsA, BandC, semenfromthree (oneNellor
attemptstoimprovethecontrolofovarianfollicular andtwoRedAngus), dos (RedAngus), dos
growthandthesynchronizationofovulation.InrecentSireswereused (GuzeratandSimmental), respectivam
estudios, theadministrationofeCG (400 UI, im) Fijo-timeAIwasperformedbythesametwo
temprano
theapplicationofPGF 2uninthePEPEprotocoltendedto techniciansonthethreefarms.Semenmorphology
increasethepregnancyrateofcowswithpostpartum andmotility [21] wereevaluated, andsemenwas
anestro [10].Inthatregard, eCGstimulatesfollicular distributedhomogenouslyamongtheexperimental
growththroughitsFSHandLHaction, increasesfollicle grupos.
tamaño, ovulationrateandplasmaprogesteroneconcentra-
tionsduringdiestrussubsequenttoFTAI, así 2.3.Hormonaltreatments
enhancingembryonicdevelopmentandmaintenanceof
embarazo [12].AlthougheCGmayimprovepregnancy Withoutregardtothestageoftheestrouscycle (Inicio
ratesinsuckledbeefcowsinpoorbodycondition [13]oftreatmenty = Day0), cowsreceivedthePEPEprot
thosetreatedintheearlypostpartumperiodwhenthe withinsertionofaprogesteroneintravaginaldevice
1
incidenceofanestrusishigh [9], itmaynotbeefficacious (1.0 g, DIB, Syntex, Buenos Aires, Argentina)
incyclingcowsorthosewithgoodbodycondition [14]. administrationof2.5mgofestradiolbenzoate (EB,
1
Hasbeenutilized Temporarycalfremoval (TCR) Estrogin, Farmavet, SaoPaulo, SP, Brasil).Eightd
FTAIprotocolsinanattempttoimprovethenumberof más adelante (Day8), cowsweretreatedwithPGF
2un(150 mg D-
1
cowsovulatingandthesynchronyofovulation.Varioscloprostenolgivenim (Prolise, ARSAS.R.L.,Buenos
studieshaveshownthebeneficialeffectofTCRforthe Aires,Argentina),andtheDIBwasremoved.Twenty-
inductionofovulationincowsinpostpartumanestrus fourhoursafterDIBremoval, cowsreceivedEB
15 – [17].Meneghettietal. [18] reportedthatTCR(48h) (1.0 mg, im), and30–36hlater, allcowsreceivedFT
increasedthesizeofthedominantfollicleandthe withoutestrusdetection(Fig.1).
ovulationrateinducedbyexogenousGnRH.Further- Thedesignwasa2 2factorial; maineffectswere
más, TCR (48 h) increasedthepregnancyrateinTCR (54–60h; fromdeviceremovaltoFTAI)
Nellorecowsinpostpartumanestrousinprogesterone- tratamiento (300IUimconcurrentwithPGF 2un).Allí-
basedFTAIprotocols [19]. Fore, cowswereallocatedrandomlytofourGroups:
TheobjectiveofthepresentstudywastodetermineControl (PEPE), TCR, eCG, andTCR+eCG.I
whetheradministrationofeCG, TCR, orboth, sería TCRgroup, calveswereremovedtemporarilyfor54–
improvepregnancyratesinNellorecowsinwhich 60h (fromDIBremovaltoFTAI; Fig.1).Assoon
ovulationwassynchronizedwithexogenousprogester- calveswereremoved, thecowsweremovedtoanother
oneandAIwasdoneatafixedtime. pastureapproximately2.5kmaway.Cowswereallotted
totreatmentswithinafarm, takinginconsideration
2 Materialsandmethods paridad, postpartuminterval, andbodyconditionscore.
IntheeCGGroup, cowsreceivedPEPEtreatment
1
2.1.Locationandcattle pluseCG (300IUim;Novormon, Syntex, Buenos
unAdministración-
Aires, Argentina) concurrentwithPGF2
Theexperimentwasreplicatedonthreeprivate Tion.IntheTCR + eCGgroup, cowsweretreatedasi
granjas, (Piraju
n = 238, FarmA), EspiritoSantodo protocolTCR, plustheyweregiveneCGonDay8.

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V.G.Pinheiroetal./Theriogenology71 (2009)–524
519 521

Inpostpartumsuckledcows Fig.1.Summaryoftreatmentsforaprogesterone-basedprotocolforfixed-timeAI (FTAI).Allcowsre

intravaginal, progesterona-releasingdevice (DIB) for8d, 2unatdeviceinsertionandremoval,
withEBandPGF respectivamente.Theexperimentaldesi
wasa2 2factorial; thefactorswereeCGtreatmentconcurrentwithPGF, fromdeviceremovaltoFTAI andtemporarycalfremoval (TC
2un
WasperformedtodetectthepresenceofaCL Ovarianultrasonography (Estados Unidos).

Cowswererandomlyallocatedtoreceiveeitheranew methoddescribedbyKnickerbockeretal. [22] y la

DIB) n = 157) oronethathadbeenpreviouslyusedfor samesourceofantiserumwasutilized.Antiserumto
( 8 nD= 286).Inthelattercase, priortoinsertion, el bovineprogesteronewasagiftfromDr.MagalyManzo
1
DIBwascleanedwithKilol (amixtureofascorbic, (FaculdadedeCienciasVeterinarias, Macaray, Vene-
citricandlacticacid (QUINABRA, SaoJosedos zuela).Theintra-andinter-assaycoefficientsof
Campos, SP, Brasil). variationwere4.8and10.6%,respectively,and la
sensitivityoftheassay (concentrationat90% cero
2.4.Ultrasonographyandradioimmunoassay 0.7PG / mL.
was50.2 vinculante) 100

Ovarianultrasonography (US) wasperformed2.5.Statisticalanalysis

(AlokaSSD500with7.5MHzlinear-arraytransrectal
transductor;Aloka, Tokio, Japón), 10and0,
onDays Pregnancydatawereanalyzedbylogisticregression
inordertodetectthepresenceofaCL (10–15mm) (backwardsstepwiseregression),
y consideringthe
todesignatewhethereachcowwascyclingoranestrus followingfactorsinthemodel:Bull(Farm+insemina-
(absenceofCLinbothovaries).Bloodsampleswere Tor + Toro), paridad (primiparousvs.multiparous),
randomlycollected (BS) from22.7%(78/343) senceofaCL,de la bodyconditionscore (ascategorical
animalsthatdidnothaveaCLineitherofthetwo variables: <2.5;2.5 3.25and >3.25), días
ultrasonographicexaminations (días 10and0), a Postpartum(ascategoricalvariables: <45d, 45–70d,
confirmtheircyclicitystatus.Inthatregard, acowwith y >70dpostpartum), DIB (newvs.used), tem
aplasmaprogesteroneconcentration <1NG/mLatboth calfremoval (yesorno), eCG (yesorno) y
bloodsampleswasdeemedanestrus, whereasthosewith interactionsbetweencalfremovalandeCG, retiro
1NG/mLateitherorbothofthetwosampleswas andcyclicity,eCGandcyclicity.Theseanalyseswere
deemedcycling.Thesamples(10ml) wereobtainedperformedusingPROCGENMODfromtheStatistical
fromthejugularveinintoanevacuatedtubecontaining AnalysisSystem [23].
heparina, andimmediatelyplacedonice.Within4h
Posteriormente, plasmawasseparatedbycentrifugation 3 Resultados
(900 g andplasmawasseparatedand de for15min)
8
storedat Cuntiltheprogesteroneradioimmu-20 Pregnancyrateswerenotsignificantlydifferent
noassay.Theassaywasperformedaccordingtothe amongthefourgroups(table1).Pregnancyrateswere

Tabla1
Pregnancyrate (percentagewithinbrackets) tofixed-timeAIinNellorecows, 40–80dpostpartum, cyclingoranestrus (withorwithout
startoftreatment
CyclingControleCGTCReCG + TCR
Yes16/29(55.2)07/15(46.7)15/33(45.5)10/23(43.5)
No38/79(48.1)56/101(55.4)29/73(39.7)39/90(43.3)
Total54/108(50.0)63/116(54.3)44/106(41.5)49/113(43.4)
2factorial; maineffectsweretemporarycalfremoval (TCR;
Allcowsweretreatedwithaprogesterone-basedprotocol.Thedesignwasa2
Gira-timeAI) andeCGtreatment (concurrentwithPGF
).Pregnancyrateswerenotsignificantlydifferentamongthefourgroups, norbetween
2un
cyclingvs.anestruscows.

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522 V.G.Pinheiroetal./Theriogenology71(2009)519
–524

notsignificantlyaffectedbyeCGtreatment(48.9%vs. increaseinprogesteroneconcentrationinanimals
45.8%respectively,forcowswithvs.withouteCG),treatedwithPEPE/eCGhasbeenreportedinother
TCR(37.9%vs.52.2%forcowswithvs.withoutTCR), experiments[12],andhigherP4concentrationhasbeen
orcyclicitystatus(48.0%vs.47.2%forcowsthatwere associatedwithembryodevelopmentandthecapacity
cyclingvs.anestrus).Therewerenosignificant oftheconceptustosecreteinterferon-tau [26],and,
interactionsbetween:TCRandeCG;eCGandcyclicity; consequently,enhancedconceptionrates [27].
TCRandcyclicity;andTCR,eCGandcyclicity.CowsInthepresentstudy,TCRhadnosignificanteffecto
1
treatedwithaneworusedDIB hadsimilarpregnancy pregnancyrate.Penteadoetal. [28] determinedthe
rates(51.6%and45.1%).Similarly,whencowswere effectsoftreatmentwitheCG(400IU),TCR,orbo
groupedaccordingtopostpartumintervalorbody onpregnancyrateinanorgestomet-basedprotocol.In
condition,therewerenosignificantdifferencesamong thatstudy,eithereCG,TCRorbothimprovedthe
thesegroupsforpregnancyrate.However,pregnancy pregnancyratecomparedtothecontrolgroup(52.7,
ratewashigherinmultiparousthanprimiparouscows 47.4,58.4,and37.3%,respectively).Inthepresent
(186/360,51.7%vs.24/83,28.9%, P <0.01). studytherewasnosignificantdifferencebetweenthe
Plasmaprogesteroneconcentrationsweredeter- protocolsthatusedTCR(GroupsTCRand
minedin78cowsdesignatedasanestrus;17%had eCG+TCR)ornot(GroupsControlandeCG).
concentrations>1ng/mL,indicatingthepresenceof Accordingtosomeauthors,thereproductiveresponses
functionalCL. obtainedwithTCRcanvarydependingoncyclicityof
thecow [19],ageofthecalf [16],nutritionalstatus,or
4.Discussion eventhegenotypeoftheherd [29].Ithasbeen
demonstratedthatTCR(48h)ofanestrouscows
Inthepresentstudy,administrationofeCG,TCR,or increasedpulsatilityofLHsecretion [30,31].Further
both,didnotsignificantlyimprovepregnancyratesin more,Meneghettietal. [18] reportedthatremovalof
Nellorecowsinwhichovulationwassynchronizedwithcalvesfor48h,increasedthesizeofthedominant
exogenousprogesteroneandinseminationwascon- follicleandtheovulationrateinducedbyexogenous
ductedatafixedtime. GnRH,whencomparedtocowswhosecalveshadnot
Pregnancyrateswerenotsignificantlyaffectedby beenremoved(85.4%vs.51.0%,respectively).Eren
eCGtreatment.ThedoseofeCGusedinthisstudy etal. [17] reportedthatTCRhadinconsistenteffectson
(300IU)wasless [10,12,24] orthesame [25]asthat pregnancyratesincowstreatedwiththePEPE
reportedbyothers.Thatmostcows(92.1%)inthe protocol.Ononefarm,TCRsignificantlyimproved
presentexperimentwereingoodbodycondition(score pregnancyrates(48/71,67.6%vs.35/77,45.6%,
2.5)atthebeginningoftreatmentmayaccountfortheP <0.05),howeveronanotherfarm,therewasno
a p p a r e n t d i f f e r e n t r e s u l t s o b t a i n e d b y o t h[esignificantdifference.Conversely,Vasconcelosetal.
1r0s] , i n
whichanimalswithlowerbodycondition,comparedto [19] utilizedTCRinlactatingNellorecowsbeforethe
thecowsusedinthisstudy,wereused.Inthatregard, insertionofanintravaginaldevicecontainingproges-
eCGincreasedthepercentageofanimalsovulatingand teroneandadministrationofGnRH(Day0)andagain
pregnancyratesincowsexperiencingnutritionalstress utilizedTCRfromremovaloftheintravaginaldevice
[12].However,thepresentresultswereinagreement (Day7)toFTAIandaseconddoseofGnRH(Day
withCutaiaetal. [11];theyreportednosignificant Theydetectedaninteraction(P <0.05)between
increaseinpregnancyrateinresponseto400IUofeCG cyclicityandTCR;incyclingcows,TCRdecreased
inpostpartumcowstreatedwithaPEPEprotocol.Itwastheconceptionrate(29.9%vs.61.0%),whereasincows
noteworthythatmostcowsinthatstudywerecycling,as inpostpartumanestrous,TCRimprovedtheconception
indicatedbythepresenceofaCL. rate(53.8%vs.35.3%).
ThereareindicationsthatincludingeCGinaPEPE Inthepresentstudy,therewasnosignificanteffectof
protocolcanincreasepregnancyratesinzebu[10,12] or cyclicityonpregnancyrates,indicatingthatthe
cross-bredcattle [12],particularlyifthetheyarein protocolsusedwereequallyeffectiveinanestrusand
postpartumanestrousandhavealowbodycondition cycliccows,corroboratingresultsreportedpreviously
score( 2,onascalefrom0to5) [12].Barusellietal. [10,12,32].Therefore,theprogesterone(DIB)treatme
[24] reportedthateCGinducedsignificantincreasesinwasveryeffectiveininducingovulationinpostpartum
folliculardiameter,ovulationrateandplasmaconcen-anestrousNellorecows.However,primiparouscows
trationofprogesterone(12dafterFTAI)inlactatinghadasignificantlylowerpregnancyratethanmulti-
primiparouscross-breedcowstreatedwithPEPE.This parouscows,aspreviouslyreported [16,33].
 V.G.Pinheiroetal./Theriogenology71(2009)519
–524 523

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