Animal Reproduction Science 136 (2013) 170 177

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Animal Reproduction Science

journa l h omepa g e: www.elsev ier .com/ locate /an i reprosc i

Embryo transfer in domestic South American

Julio B. SFaculty of Vete

a r t i c l

Article history:Available onlin

Keywords:AlpacaLlamaEmbryo transf

embryed tecw rapi

presertion abis provce of sinces in

future

1. Introduction

The embryo transfer technique in South Americancamelids involves embryo collection from a female donorby uterine lavage 78 days after breeding, and transfer toa recipient donor. Embthe potentiity of donorto facilitatewild camelifor identifydefects becgenetics an

Throughten offsprinreported a transfer pro(2008) repotransfer pro

This papeence on Camelby Gregg P. Ad

E-mail add

been done and studies are on-going to develop of a super-ovulation protocol (Bourke et al., 1992; Correa et al., 1997;Gomez et al., 2002; Vaughan and Tibary, 2006; Huancaet al., 2009). Using a superovulation protocol, Vaugham(2012) reported an average of 2.53 embryos per uter-

0378-4320/$ http://dx.doi.ofemale that has been synchronized with theryo transfer in South American camelids offersal to take advantage of the reproductive capac-s with high genetic merit in domestic herds, and

preservation and repopulation of endangeredd species. Embryo transfer is also a valuable tooling and eliminating genetic aws or hereditaryause it involves rapid proliferation of knownd a detailed data-base of parentage.

embryo transfer, a donor can produce eight org per year instead of just one. Taylor et al. (2000)66% pregnancy rate in a commercial embryogram in llamas in the United States, and Sumarrted a pregnancy rate of 40% in and embryogram in alpacas in Peru. Many studies have

r is part of the special issue entitled: International Confer-id Genetics and Reproductive Biotechnologies, Guest Editedams and Ahmed Tibary.

ress: jbsumar@gmail.com

ine ush; hence a single donor can then produce up to 21embryos per year.

Finally, the embryo transfer technique enables thepreservation of endangered species such as vicuna, guana-cos and some breeds of alpacas and llamas with speciccharacteristics (Del Campo et al., 1995). One interestingmethod used to preserve valuable species or breeds of lla-mas and alpacas is to perform interspecies embryo transfer(Taylor et al., 2001; Von Baer et al., 2003; Sumar, 2008).Another alternative to achieve preservation of endangeredspecies of South American camelids is embryo freezing, atechnique that is actively being studied (Aller et al., 2002;Lattanzi et al., 2002; Palasz and Adams, 2000; Vasquez et al.,2007, 2011).

2. History of embryo transfer in domestic SouthAmerican camelids

The rst surgical embryo collection and transfer inalpacas was reported 42 years ago in Peru. Novoa andSumar (1968) reported in alpacas the rst collection of

see front matter 2012 Published by Elsevier B.V.rg/10.1016/j.anireprosci.2012.10.029umarrinary Medicine, San Marcos University, Lima, Peru

e i n f o

e 1 November 2012

er

a b s t r a c t

Intraspecic and interspecic developing into a well-establishductive biotechnologies to allomerit (e.g., high ber quality,to provide up-to-date informacamelids. Specic information ent synchronization, the practiand transfer techniques, advaninter-specic transfer, and thecamelids. camelids

o transfer in domestic South American camelids ishnique. Reports reveal many benets of using repro-d propagation of alpacas and llamas of high geneticve color variation). The objective of this review isout embryo transfer in domestic South American

ided on criteria for male selection, donor and recipi-gle- vs. super-ovulation protocols, embryo recovery

cryopreservation of embryos, results of intra- andof the embryo transfer in domestic South American

2012 Published by Elsevier B.V.

J.B. Sumar / Animal Reproduction Science 136 (2013) 170 177 171

llection

zygotes fro(Day 1 = daRinger soluto the uterprocedure tight sphincsible to ucamelids, eovulating fein the uteriwith forcepstages were1974), supe750 IU of eCwere collecafter matinoviducts of nancies and

The rstransfer tec(1985) in ttion and ovand collectdone 7 dayafter transfeBourke et alation can collected nocessful pregChile. Excein domesticpreviously (Tibary et aTibary, 200

3. Selectio

Breederfree of lesidetected byembryo traspring unlemales of thembryo traaverage in gtion and frereproductiv

fore, freferalpatiotial co

enitalio to

rally rwithored foot yetn, betwroducrs shomic hesirableicolor opost-p), the ting (4ient atterine, and a

me of

earlyg resures to as slauctivelyt the m

matins at 10acas aday) r2, 48mpactFig. 1. Method of oviductal ushing at laparotomy for embryo co

m the oviduct after fertile mating. At Day 3y of mating), the embryos were ushed withtion from the mbriated end of the oviductus (ante grade surgical ush), similar to theused by Smith and Murphy (1987) in ewes. Ater at the uterotubal junction makes it impos-sh liquids from the uterus to the oviducts inmbryos were recovered from 80% of single-males by placing a pipette through an incisionne wall, and clamping the uterine bifurcations (Fig. 1). Embryos at the 2, 4, 8-cells and morula

recovered. In another study (Sumar and Franco,rovulation was induced in 8 donor alpacas usingG followed by 1000 U of hCG. Fifty six embryosted from the oviducts by laparotomy 3 daysg and 44 morulae were transferred into the44 recipients by laparotomy, resulting in 4 preg-

one live-born cria.t llama born by non-surgical collection andhnique was reported by Wiepz and Chapmanhe United States. In this study, synchroniza-ulation of the recipient was induced by GnRH,ion and transfer, of hatched blastocysts wass after GnRH treatment. One cria was bornr of two embryos to two synchronized females.l. (1991) reported that, in llamas, superovu-be achieved using eCG, and embryos can ben-surgically. Gatica et al. (1994) reported a suc-nancy through embryo transfer in a llama in

llent reviews about reproductive technologies South American camelids, have been publishedPugh and Montes, 1994; Del Campo et al., 1995;l., 2005; Miragaya et al., 2006; Vaughan and6).

n of males, donors, and recipients

thereare ptal papotennal g

Twgene(i.e., requihas ngrowof repdonosysteundemultdays 2010lactarecipsoft udates

4. Ti

Inmatintomealpacrespeova aafteruteruof alpeach Day 4, cos should emphasize the use of high ranking sires,ons of the external and internal genitalia, as

palpation and ultrasonography. The cost ofnsfer is high and will not pay dividends on off-ss they are from the meritorious females ande breed. Females selected as donors for the

nsfer should be well above the breed and herdenetic value. They should be of good conforma-e of heritable defects. Females with a history ofe problems do not make good donor animals;

to tubular bcysts on Dawere recov(Cardenas, et al., 2008)tocyst) doeand it devefrom Days 6

Results studies of in alpacas (Novoa and Sumar, 1968).

emales with a history of producing live criasble. Donors are further evaluated by transrec-n, ultrasonography and vaginoscopy to identifyngenital or acquired abnormalities of the inter-a.three recipients for each donor animal are

equired in single embryo collection programsut superovulation). The number of recipientsr multiple ovulation embryo transfer programs

been established. Recipients should be well-een 2 and 8 years of age, and have no history

tive failure. The same clinical approach used foruld be implemented for recipients. Other thanalth, recipients may be selected irrespective of

heritable or congenital defects (e.g., blue eyes,r coarse ber, prognathism), and should be 30artum. In a recent study in alpacas (Sumar et al.,pregnancy rate was higher (P < 0.001) for non-4%) than lactating (18.2%) recipients. An ideal

the time of the transfer will have a CL 12 mm, tone, a closed cervix, no vaginal or vulvar exu-

body condition score of 3 (out of 5).

embryo entry into the uterus

studies, ushing the oviducts 3 days afterlted in the collection of embryos from two blas-the morula stage (Novoa and Sumar, 1968). Inghtered at 4, 7 and 10 days after copulation,, Bravo et al. (1996) reported the collection oforula stage and compacted morulae by 7 days

g from the oviducts, and blastocysts from the days. Flushing of the excised reproductive tractfter slaughter at 215 days after mating (n = 34esulted in the collection of 24 cell embryos on

cell embryos on Day 3, early morulae on Day morulae and early blastocysts on Day 5, ovoid

lastocysts on Day 10, and lamentous blasto-

ys 1115, but for unknown reasons, no embryosered from alpacas on Days 69 after mating1997). Authors of a recent study (Cervantes

reported that the alpaca embryo (hatched blas-s not enter the uterus until 6 days after matinglops very rapidly to almost double in diameter

to 7 after mating.of embryo transfer studies are consistent withexcised tracts; i.e., embryos enter the uterus

172 J.B. Sumar / Animal Reproduction Science 136 (2013) 170 177

product

66.5 days(Bourke et study (Tayl55%, 79%, aning, respect

5. Single o

There arfor transfera single ovument. Dailyverify the p(i.e., betwe0). Some recafter matinCampo et adone on Da(i.e., that ovof ovulationtion at the out 7 daythe expandEmbryo colinconsistenafter matintion rate atare usuallyning to elonprostaglandand receptition cycle, athat time wopment andin healthy, 1972; Sumovulate evecol. A singlthese usheper year haovulation te

The secoof the topic

ill noperovamelidunpredps the

uperste folliubord, 1990affect , 2007o electrstimu

been asterond folli; Huanes in the suprstimupture

3). Mauperovlids, invs. sur

condirationn in dal tim

s. CattFig. 2. Single ovulation vs. superovulation protocols used for embryo

after mating at the hatched blastocyst stageal., 1991; Del Campo et al., 1995). In a llamaor et al., 2000), embryos were collected fromd 100% of donors at 7, 8 and 10 days after mat-ively; all were at the hatched blastocyst stage.

vulation vs. superovulation protocols

e two basic strategies for producing embryos (Fig. 2). The simplest involves collection afterlation, with no ovarian superstimulatory treat-

transrectal ultrasonography has been used toresence of a viable, mature dominant follicleen 7 and 12 mm) at the time of mating (Dayommend administration of GnRH immediatelyg to ensure ovulation (Bourke et al., 1991; Dell., 1995). Transrectal ultrasonography may bey 5 after mating to conrm the presence of a CLulation occurred). Alternatively, conrmation

may be done by ultrasonographic examina-time of ushing. Embryo collection is carrieds after mating, when embryos are generally ined blastocyst stage (Ratto and Adams, 2007).lection rate from the uterus is low at Day 6,t at Day 6.5 or 7, and optimal at 7.5 daysg (Sumar, 2008). Although, the embryo collec-

8 days after mating is very high, the embryos very fragile because they are large and begin-gate (Taylor et al., 2000). Donors may be givenin after uterine ushing to ensure luteolysis

and wto sucan cand perhaing sof ththe set al.cally et al.cols tsupehaveprogeguide2007studithat tsupeual ru(Fig. the scamecaya bodyprepamatiooptimcyclevity by the time of the next mating and collec-lthough luteolysis will normally be complete byithout treatment. Since ovarian follicular devel-

ovulatory capability are present year-aroundwell-nourished alpacas (Fernandez-Baca et al.,ar et al., 2010), a donor may be induced tory 1214 days using a single-ovulation proto-e embryo may be expected from about 85% ofs (Sumar, 2008), and 1218 embryos per donorve be produced using the continuous single-chnique (Taylor et al., 2000; Sumar, 2008).nd approach involves superovulation. A review

is provided in this publication by Ratto et al.

for three trembryo rec

6. Embryo

The nonused most wrequire spevention elimscarring. Ulthe non-surdiagnose prcervix (threion in domestic South American camelids.

t be recapitulated here. Briey, the responseulatory treatment protocols in South Ameri-s may be characterized as extremely variableictable. Although critical studies are sparse,

greatest variability is a consequence of initiat-imulatory treatment irrespective of the statuscular wave. The dominant follicle suppressesinate follicles within the same wave (Adams), an effect that has been shown to dramati-the superovulatory response in cattle (Mapletof) and in camelids (Ratto et al., 1997). Proto-ively induce follicular wave emergence so thatlatory treatment may be initiated at that timettempted using a combination of estrogen ande, GnRH or LH, and transvaginal ultrasound-

cle ablation (Ratto et al., 2003; Ratto and Adams,ca et al., 2009). In this regard, results of recenthe authors laboratory (unpublished) suggesterovulatory response is improved by initiatinglatory treatment (eCG) 2 or 3 days after man-

of the dominant follicle (via rectal palpation)ny issues remain to be addressed to optimizeulatory response in domestic South Americancluding species (alpaca vs. llama), breed (hua-i), age (yearlings vs. adults), lactational state,tion, environmental conditions, and hormones and protocols. Furthermore, there is no infor-omestic South American camelids about thee interval between successive superovulatoryle may be superovulated at 2-month intervals

eatments without an appreciable decrease inovery (Farin et al., 2007).

collection

-surgical method for collection of embryos isidely because it is non-invasive and does not

cialized equipment. Avoiding surgical inter-inates complications caused by adhesions and

trasonography has also been implemented ingical method to monitor follicular activity andegnancy. The anatomical characteristics of thee irregular annular or spiral folds) and rectum

J.B. Sumar / Animal Reproduction Science 136 (2013) 170 177 173

Fig. 3. Ovaries iddle anSuperstimulat to superovulatory resp r of larg

permit relaboth the esphases.

6.1. Materi

The mat

i. A two-ii. A style

catheteiii. A sanit

inationvulva a

iv. A 10 mll the

v. An emcontainsure thuterus.

vi. Exit ansuggeshoses tpassag

vii. Petri dlter a

viii. A pipeembry

6.2. Donor

Collectiodonor in stsedated (AcIngelheim, and given 1 ml/100 kg

ped wwith ae pur

ve traas, them (Figmounpulatiperatothrougectumr, and ith theat theter. Thagina,ter tip of alpacas induced to ovulate a single follicle (left) or multiple follicles (mion was induced with 800 IU eCG. Ablation of the dominant follicle prior onse (right) compared with no ablation (middle, showing a large numbe

tively easy canulation of the cervix duringtrogen-dominant and progesterone-dominant

als for embryo collection

erials for embryo collection consist of:

way Foley catheter (1418 FR).t 6.2 in. length with a clip to hold it within ther.ary-plastic bag to prevent any possible contam-

during the passage of the catheter through thend the rst portion of the vagina.l syringe with sterile distilled water or saline to

Foley catheter balloon.bryo collection lter placed on a graduated

wrapsum For thductialpac20 cous amanithe osage the rwate

Wso thcathethe vcatheer. The container allows the collector to mea-e volume of ushing media recovered from the

d entry tubes for ushing medium. The authorts reducing the length of commercially availableo reduce the risk of losing the embryos duringe through the tubes.ishes into which the contents of the embryore emptied to search for the embryo.tte or syringe to rinse the lter in case theo is attached to the wall of the lter.

restraint and uterine ushing

n of embryos is done by maintaining theernal recumbency (Fig. 4). The donor may beepromazine, 0.020.05 mg/kg, IM, Boeringer-Germany; Promazil, Lab Chile, Santiago, Chile)caudal epidural anesthesia (Lidocaine 2%,

body weight; United Lab SA, USA). The tail is

vagina, therectum of tcervix and iushed at oan individuso that the tThe balloonor uterine os to securestylet is remted to the Fmedium. Wclamps on emptying ooverlling oan interval involves gethe uid. Atis deated, fully ushed right). Photographs were taken 8 days after mating/GnRH.stimulatory treatment appeared to induce a more uniforme non-ovulated follicles).

ith an elastic bandage and xed to the dor- clip to minimize contamination of the vulva.poses of transrectal manipulation of the repro-ct during embryo collection and transfer in

ideal circumference of the operators hand is. 4). Soft latex examination gloves and gener-

ts of lubricant are recommended for transrectalon. When introducing the hand into the rectum,r must gently rotate his/her hand during pas-h the anal sphincter. Feces are removed from

and the perineal area is cleaned with soap andrinsed with 70% ethanol.

help of an assistant, the vulvar lips are opened operator can introduce the uterine lavagee catheter is directed toward the dorsum of

after which the plastic cover that protects the is pulled back. Once the catheter is within the operator places a pre-lubricated hand into thehe donor and guides the catheter through thento the uterine body. The entire uterus may bence; hence, no attempt is made to catheterizeal uterine horn. The stylet is withdrawn slightlyip is caudal to the balloon of the Foley catheter.

is then inated with 510 ml of distilled waterush medium just cranial to the interval cervical

the catheter within the body of the uterus. Theoved from the catheter and a Y-junction is t-oley catheter to connect a bag of warm ushith the help of assistant controlling thumb-

the tubing, the operator guides the lling andf the uterus ush medium, being careful to avoidf the uterus. The uterus is ushed 3 times, with

of at least 30 s between each lavage. Each lavagentle massage of the uterine horns to evacuate

the end of the third uterine lavage, the balloonthe catheter is removed, and its contents care-d into the embryo lter. After uterine ushing,

174 J.B. Sumar / Animal Reproduction Science 136 (2013) 170 177

Fig. 4. Collect istant towall permit co e collecfor rectal palp

the donor mto prevent that the em

7. Embryo

The contthe lter is content of tthe ush mlocated anding to the (1990). Emin size fromexpanded bembryo diaafter matin(n = 8), respation in thedonors (Vas

A tubercfor embryoamount of mimize stick(Hyclone, Drate dishes dish. The eintroducingthe loss of tacteristics ohave not besystem for Embryo Traand Siedel, fer are idenmedium, encommerciaWA, USA), tcontaminatdish, at 37

3% BSA, unt

on-sur

oadin

e petrtereoscculin s, sucche strale, me5). Carof medol to fo

trans a fashansferip of threctal . Somee left r rightment ke et e reprnammnts ofion and transfer room: two chutes with one operator in each and an assntrolled access to the embryo manipulation laboratory, isolated from thation in alpacas is 20 cm.

ay be given a luteolytic dose of prostaglandinunwanted pregnancy in the donor in the eventbryo is not collected during ush.

evaluation and handling

ents of the lter are poured into a petri dish, andrinsed with ush medium using a syringe. If thehe lter is abundant and/or cloudy, a portion ofay be placed in a second petri dish. Embryos are

evaluated using a stereo- microscope, accord-recommendations of Springfellow and Siedelbryos collected 7.5 days after mating range

0.5 to 1.0 mm, and are usually found in thelastocyst stage. In another study, the averagemeter of llama and alpaca were similar at 8 daysg: 527.1 168.0 m (n = 6) and 534 151.4 mectively. There is, however, considerable vari-

size of embryos collected among and withinquez et al., 2007).ulin syringe attached to a pipette tip is used

handling, and care is taken to aspirate a smalledium before contacting the embryo to min-

ing and contact with air. Washing mediumPBS/Modied + Embiotic III) is placed in sepa-before the embryo is removed from the search

8. N

8.1. L

Ththe stuberstrawinto tbubb(Fig. umn alcohinto a

Inthe trThe ttranshornin thleft oplace(Bourof thand icontembryo is placed in the washing dish after the pipette tip into the medium, to preventhe embryo and air contact. Morphologic char-f embryo quality in South American camelidsen critically investigated, so the classication

bovine embryos published by the Internationalnsfer Society is used at present (Springfellow1990). Good quality embryos for embryo trans-tied and washed several times in the ushriched with fetal calf serum or in a maintenancel medium (Vigro Holding medium, Bioniche,o remove cervical mucus, debris, and bacterialion. Washed embryos are placed in a small petriC in the ushing medium supplemented withil loaded into a transfer straw.

and the tip plug effectinot leak bet

8.2. Recipie

Using a for single-oined ultrasgrowing dolation is indbe conrmeat the timehave been pbetween do record ultrasound ndings. The two windows in the backtion room. The ideal circumference of the operators hand

gical transfer to recipients

g transfer straws

i dish containing the embryo is placed underope to load in a 0.25 ml transfer straw. Using ayringe attached to the cotton-lled end of theeeding columns of medium and air are drawnw as follows: medium, air bubble, medium, air

dium with the embryo, air bubble, and mediume should be taken to ensure that the rst col-ium contacts the plug of cotton and polyvinylrm a water-proof seal. The straw is then loadedfer gun and covered with a transfer sheath.ion similar to that described for uterine ushing,

gun is introduced into the vagina of recipient.e transfer gun is guided through the cervix bymanipulation and positioned into the uterine

authors recommend placement of the embryouterine horn regardless of the side of the CL,

(Picha et al., 2010), while others recommendin the horn ipsilateral to the corpus luteumal., 1995; Trasorras et al., 2010). Manipulationoductive tract is minimized to avoid traumaation of the endometrium. After expelling the

the straw, the transfer gun is withdrawn slowly

of the gun is checked to conrm that the cottonvely expelled the contents and the contents didween the straw and sheath.

nt synchronization and pregnancy diagnosis

similar examination and treatment schedule asvulating donors (Fig. 2), recipients are exam-onographically to conrm the presence of aminant follicle between 7 and 12 mm, and ovu-uced by administering GnRH. Ovulation mayd by ultrasonography on Day 5 after GnRH, or

of embryo collection on Day 7. No reportsublished on the degree of synchrony requirednors and recipients for optimal embryo survival

J.B. Sumar / Animal Reproduction Science 136 (2013) 170 177 175

Fig. 5. Schematic drawing of a loaded embryo transfer straw.

in domesticis to have thTo minimizresulting frinhibitor (Frecipients recipients manesthesia mis wrappedthe rectumdescribed. Pultrasonogrence of an eof an embry(reviewed i

9. Cryopre

Cryoprelogistical anof embryosents, and it of embryosattempted ifreezing an

9.1. Conven

Very fewfrozen by costudied thedays) to pesolution oferol supplesodium hywas removeffect of supon trophobdifferencescontrol gropropylene progressiveture, but nore-expande

9.2. Vitric

Vitricadration of highly concfreeze thating the sostate. A lowcryoprotectcomparisontion, Lattan

ocyst umbryone serpansiond 57

frozendesign

pull ma ha). Embsed in ed dirpansioegnanved frescopelasm w surocyst, ients rose rec

ntersp

e rstican con Baa giva giv

(Sumlamas ferredients want ated at

were bht of 1

from a born tterpar

1 year (50%)ys afths of p

to alpa

utureas

ample llamales cofreein

embrcquire South American camelids. At present, the goale recipient ovulate within 1 day of the donor.e the luteolytic effects of uterine inammationom manipulations, a prostaglandin synthaselunixin meglumine) may be administered to3060 min before embryo transfer. Nervousay be tranquilized before transfer and epiduralay be induced, as described for donors. The tail

and tied out of the way, feces are removed from, and the perineal area is cleaned as previouslyregnancy diagnosis may be done by transrectalaphy by 1213 days after donor mating (pres-mbryonic vesicle), and conrmed by detectiononic heartbeat at 25 days after donor matingn Adams and Domnguez, 2007).

servation of embryos

servation of embryos offers several importantd economic advantages. It permits preservation

in excess of the number of available recipi-facilitates national and international movement. Two methods of cryopreservation have beenn South American camelids, conventional slow-d vitrication.

tional equilibrium or slow freezing method

studies are reported about alpaca or llamanventional methods. Palasz and Adams (2000)

exposure of llama trophoblastic vesicles (13rmeating cryoprotectant, exposing to a 10%

ethylene glycol, propylene glycol, or glyc-mented with 10% fetal calf serum or 0.1%

aluronate at 22 C for 15 min. Cryoprotectanted with 0.5 M sucrose solution. There was noplement or method of cryoprotectant removal

lastic vesicle survival in culture. There were no in trophoblastic vesicle survival after 24 h inups and those exposed to ethylene glycol orglycol. The embryos frozen in ethylene glycolly expanded to 90% of original volume in cul-ne of the embryos frozen in propylene glycold in culture.

ation

tion is a rapid process that consists of dehy-the embryo at room temperature by a veryentrated vitrication media and a very rapid

avoids the formation of ice crystals, allow-lution to change from a liquid to a glassy

toxicity vitrication solution consists of threeive agents (Vajta, 2000; Kassai, 1996). In a

of the slow freezing method and vitrica-

blasting. E(bovire-ex54% athosewas openof lla2002expomergre-exno prsurvimicrocytopto loblastrecipin th

10. I

ThAmerand va llama llamstudyand ltransrecippregnabortcriasweigborncriascounlar by3 of 645 damontborn

11. Fllam

Exfer infemathus more(2) azi et al. (2002) tested the viability of hatched be by-passsing two methods: vitrication and slow freez- viability was tested by culture in SOFaa + BSAum albumin) medium. After 48 h of culture,n of vitried and slow-frozen embryos was%, respectively, not signicantly different from

using the conventional method. Another studyed to determine the effect of vitrication bystraw (OPS) on the morphology and survivaltched blastocysts (Von Baer and Del Campo,ryos ranging from 300 m to 800 m wereone step to high 40% ethylene glycol and sub-ectly into liquid nitrogen. Results showed thatn of embryos after thawing was acceptable, butcies were obtained. Some of the embryos thatezing were examined by transmission electron

(TEM), revealing a high lipid content in theof llama oocytes and embryos may contributevival after vitrication. With expanded llamaAller et al. (2002) reported 50% of pregnancy ineceiving vitried embryos (2/4) and 33.3% (2/6),eiving fresh embryos.

ecies embryo transfer

two reports of interspecies transfer in Southamelids were published by Taylor et al. (2001)er (personal communication). In the rst case,es birth to an alpaca cria. In the second case,es birth to a guanaco cria. In a more recentar, 2008), embryos were recovered from alpacasby non-surgical ush 7.5 days post-mating and

to recipients of the opposite species. In llamaith alpaca embryos, 4/7 (57%) were detected

15, 25 and 35 days after donor mating, but one6 months of pregnancy. Three healthy alpacaorn from llamas (Fig. 6), with an average body

0.5 kg; i.e., 3.5 kg more than that of alpaca criaslpacas (7.0 kg). At 6 months of age, the alpacao llama recipients gained 12 kg more that theirts born to alpacas, but body weights were simi-

of age. In alpaca recipients with llama embryos, alpacas were detected pregnant at 15, 30, ander donor mating, but one alpaca aborted at 4.5regnancy. The body weight of the 2 llama criascas was about 8.0 kg (Fig. 6).

of the embryo transfer in alpacas and

s of the potential application of embryo trans-s and alpacas are: (1) embryos from valuable

uld be transferred to less valuable recipients,g the valuable animal for the production ofyos than would otherwise be possible (Fig. 6),d infertility problems in valuable animals may

ed by embryo transfer, (3) preservation of

176 J.B. Sumar / Animal Reproduction Science 136 (2013) 170 177

Fig. 6. Top (frdonor alpaca, eMiddle: Threeof the HuacayAlpaca recipie

endangered(4) geneticAmerican cthe last 30 y

Despite in domesticnot been cspecies. In aanisms invthe uterus, nature of ecies. Develosplitting, gedepend on technique.

Conict of

No con

Acknowledgement

I am grateful to Dr. Gregg Adams for the critical readingof the manuscript.

ences

, G.P., Dacas. In

rge AnimA, pp. 8, G.P., Sd reproma). J. R.F., Rebu

of vitr1127.e, D.A., An-surgice, D.A., Aulation ., Maythternatio. 1831e, D.A., Kt synchreriogen

P.W., MRefer

AdamsalpLaUS

Adamsangla

Aller, Jfer12

Bourkno

BourkovA.JInpp

BourkenTh

Bravo,om left to right): Elite breeding male alpaca, elite embryombryo recipient alpaca, and an elite cria from the recipient.

llama recipients with alpaca crias. The cria in the center isa breed, and the other two are of the Suri breed. Bottom:nt with a llama cria.

camelid species (vicuna and guanaco), and improvement may be accelerated in Southountries that have suffered genetic erosion inears through indiscriminate exportation.the commercial potential of embryo transfer

South American camelids, the technology hasritically and systematically studied in theseddition research is needed regarding the mech-olved in the time of entry of the embryo toembryo migration to the left uterine horn, thembryonic loss, and the loss of twin pregnan-pment of in vitro embryo production, embryone transfer, and embryo sexing early will all

successful development of the embryo transfer

interest

ict of interest.

spermatoz173179.

Cardenas, H.,1embrionartca AnuaNacional A

Cervantes, M.,ine ushinpacos). In:Reproduct

Correa, J., Rattoand equintion. Anim

Del Campo, MThe appliccamelids.

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Embryo transfer in domestic South American camelids1 Introduction2 History of embryo transfer in domestic South American camelids3 Selection of males, donors, and recipients4 Time of embryo entry into the uterus5 Single ovulation vs. superovulation protocols6 Embryo collection6.1 Materials for embryo collection6.2 Donor restraint and uterine flushing

7 Embryo evaluation and handling8 Non-surgical transfer to recipients8.1 Loading transfer straws8.2 Recipient synchronization and pregnancy diagnosis

9 Cryopreservation of embryos9.1 Conventional equilibrium or slow freezing method9.2 Vitrification

10 Interspecies embryo transfer11 Future of the embryo transfer in alpacas and llamasConflict of interestAcknowledgementReferences