Embryo transfer in horses is a reproductive technology that involves collecting a fertilized embryo from a donor mare and implanting it into a recipient mare, which then carries the pregnancy to term. This technique allows for the production of multiple offspring from a single mare within a breeding season. The procedure includes several stages: synchronization of the donor and recipient mares' estrous cycles, collection of the embryo, and its subsequent transfer to the recipient mare. Embryo transfer is utilized to enhance genetic dissemination, preserve valuable genetics, and manage breeding schedules. This page aggregates peer-reviewed research studies and scholarly articles that explore the methods, efficiency, and applications of embryo transfer in equine reproduction.
Coutinho da Silva MA.The use of assisted reproductive techniques (ART) has helped owners to produce offspring from valuable mares that were considered infertile using standard breeding techniques. Before referring a mare for an ART, the practitioner should be able to identify the underlying cause of subfertility of the mare. The objective of this review is to provide information regarding embryo transfer, oocyte transfer and intracytoplasmic sperm injection, the three most common ART used in equine practice. Knowing the complexity as well as the risks of these techniques, enables practitioners to refer a subfertil...
Scherzer J, Fayrer-Hosken RA, Ray L, Hurley DJ, Heusner GL.Embryo transfer has been an inherent part of cattle breeding for more than 35 years and has also gained remarkable interest from the equine industry after several breeds allowed registration of more than one foal per year. In both large animal species, non-surgical embryo recovery and transfer are well-established techniques. However, success rates after superovulation and cryopreservation of embryos in horses are still lagging behind those of cattle, and more research is needed to address these areas. To address the problem of freezing large equine embryos, we offer a preliminary demonstratio...
Bergfelt DR, Meira C, Fleury JJ, Fleury PD, Dell'Aqua JA, Adams GP.A regimen of progesterone plus estradiol (P&E) was used as a standard for ovarian synchronization to test the efficacy and evaluate the commercial application of ultrasound-guided follicle ablation as a non-steroidal alternative for ovulation synchronization in mares. Recipient mares at a private embryo transfer facility were at unknown stages of the estrous cycle at the start of the experiment on Day 1 when they were randomly assigned to an ablation group (n=18-21 mares) or to a P&E group (n=20-21 mares). In the ablation group, mares were lightly sedated and all follicles > or = 10...
Hinrichs K, Choi YH, Varner DD, Hartman DL.We evaluated the effect of different activation treatments on the production of blastocysts and foals by nuclear transfer. Donor cells were prepared using roscovitine treatment, which has previously been associated with increased production of viable offspring. All activation treatments were followed by culture in 6-dimethylaminopurine (6-DMAP) for 4 h. In experiment 1, blastocyst production after activation by injection of sperm extract followed by treatment with ionomycin was significantly higher than that for activation with a serial treatment of ionomycin, 6-DMAP, and ionomycin (12.5 vs 2....
Cran DG.Many tens of thousands of calves resulting from artificial insemination (AI) have been born worldwide after XY sperm separation and commercial production is underway in several countries. Accuracy of sex selection is some 90% and can be achieved both in research facilities and at AI studs in rural locations. Most facilities sort X- sperm which have also be utilised for superovulation and embryo transfer projects and for in vitro fertilisation (IVF) as well as AI. Sort rates of some 15 x 10(6) sperm/h are currently achievable and are used for low dose insemination, generally at 2 x 10(6) frozen...
Gandini G, Pizzi F, Stella A, Boettcher PJ.The aim of this work was to compare costs, in the horse, cattle, sheep, swine, and rabbit species, for the creation of gene banks for reconstruction of an extinct breed, using different strategies: embryos-only, embryos in combination with semen, and semen-only. Three cost measures were used: time required for population reconstruction, cost for creation of the gene bank, number of years-keeping-female to reach reconstruction. Semen costs were estimated across four scenarios: the presence or absence of a commercial market for semen, purchase of semen donors, and semen extracted from the epidid...
Hinrichs K, Choi YH, Walckenaer BE, Varner DD, Hartman DL.Viability of equine embryos produced by oocyte maturation, intracytoplasmic sperm injection and embryo culture to the blastocyst stage in vitro was evaluated after transfer of embryos to recipient mares. No pregnancies were produced after transfer of five blastocysts that had been cultured in G media. Transfer of 10 blastocysts cultured in modified DMEM/F-12 medium produced five pregnancies and three live foals; the two lost pregnancies developed only trophoblast (based on transrectal ultrasonography). To evaluate the status of the inner cell mass, equine blastocysts produced in vivo and in vi...
Hebia I, Fiéni F, Duchamp G, Destrumelle S, Pellerin JL, Zientara S, Vautherot JF, Bruyas JF.The objective of this study was to determine whether the 10 wash cycles proposed by the International Embryo Transfer Society (IETS) for bovine embryos efficiently decontaminated equine embryos exposed to equine herpes virus 1 (EHV-1) in vitro. Donor mares and stallions were individually screened and shown to be negative for the virus by PCR detection of EHV-1 DNA in blood leukocytes, semen, and uterine lavages in which embryos were recovered. Twenty embryos were recovered and randomly assigned to one of two groups: 10 embryos were exposed for 24h to infectious EHV-1 at 10(6)TCID(50)/ml, and 1...
Lagutina I, Lazzari G, Duchi R, Turini P, Tessaro I, Brunetti D, Colleoni S, Crotti G, Galli C.Nuclear transfer (NT) is a complex procedure that requires considerable technical skills. Over the years attempts have been made to simplify the micromanipulations involved and to make the procedure more user-friendly. A significant step forwards has been the development of the zona-free NT methods. We have used zona-free NT with mechanical aspiration of the metaphase plate as a mean of enucleation, in a comparative approach with the conventional nuclear transfer zona-enclosed method in cattle, horse, sheep and pig. The absence of the zona considerably facilitates the enucleation step and sign...
Galli C, Colleoni S, Duchi R, Lagutina I, Lazzari G.Development of assisted reproductive technologies in horses has been relatively slow compared to other domestic species, namely ruminants and pigs. The scarce availability of abattoir ovaries and the lack of interest from horse breeders and breed associations have been the main reasons for this delay. Progressively though, the technology of oocyte maturation in vitro has been established followed by the application of ICSI to achieve fertilization in vitro. Embryo culture was initially performed in vivo, in the mare oviduct or in the surrogate sheep oviduct, to achieve the highest embryo devel...
Logan NL, McCue PM, Alonso MA, Squires EL.Superovulation could potentially increase embryo recovery for immediate transfer or cryopreservation. The objectives were to evaluate the effect of pretreatment with progesterone and estradiol (P+E) on follicular response to eFSH and compare doses of eFSH and ovulatory agents on follicular development and ovulation in mares. In Experiment 1, 40 mares were assigned to one of four treatment groups. Group 1 consisted of untreated controls. Group 2 mares were administered eFSH without pretreatment with P+E. Group 3 mares were administered P+E for 10 days starting in mid-diestrus followed by eFSH t...
Panzani D, Rota A, Pacini M, Vannozzi I, Camillo F.One year old fillies are able to conceive but, usually, not to give birth to a living foal. Although embryo transfer allows the production of foals from mature mares with repeated pregnancy losses, no reports are available on the use of one year old fillies as embryo donors. To evaluate this possibility, eleven 12-16 months old Haflinger mares were inseminated with fresh semen and subjected to embryo recovery. Some of the recovered embryos were non-surgically transferred into synchronized mature recipients. Pregnancies were terminated using PGF2alpha at day 25. Fillies' embryo recovery rate an...
Wilsher S, Kölling M, Allen WR.A level of synchrony between embryo and uterine environment is essential for the establishment of pregnancy when performing embryo transfer. The ability to extend the acceptable degree of asynchrony would allow more efficient use of recipient mares. Objective: To establish if administration of the prostaglandin synthetase inhibitor, meclofenamic acid, to asynchronous recipient mares could widen the acceptable window of asynchrony for embryo transfer. Objective: The prostaglandin synthetase inhibitory action of meclofenamic acid may act to suppress luteolysis and thereby allow for a greater deg...
Aurich J, Aurich C.The liberalization of European animal breeding legislation and an increasing diversity of equestrian sports have led to a constant rise in the number of horse breeds and breed registries. In addition to the trend towards more and smaller breed registries, there is another trend towards an international expansion of the bigger established sport horse breeds. Regional breeds, at least in smaller countries, may no longer be able to run an independent breeding programme. The typical horse breeder, in the future, will be a female and qualified in equestrian sports. Artificial insemination (AI) main...
Squires EL, McCue PM.Embryo recovery from single ovulating mares is approximately 50 per cent per estrous cycle. Superovulation could be used to increase embryo recovery and provide extra embryos for embryo freezing. This review addresses some historical approaches to superovulation, as well as examines factors that affect the response of mares to equine FSH. eCG, GnRH and inhibin vaccines have been of limited success in stimulating multiple ovulation. Numerous studies have shown that injection of equine pituitary extract (EPE) will result in three to four ovulations per estrous cycle and two embryos. A purified, ...
Squires EL, McCue PM.Embryo recovery from single ovulating mares is approximately 50 per cent per estrous cycle. Superovulation could be used to increase embryo recovery and provide extra embryos for embryo freezing. This review addresses some historical approaches to superovulation, as well as examines factors that affect the response of mares to equine FSH. eCG, GnRH and inhibin vaccines have been of limited success in stimulating multiple ovulation. Numerous studies have shown that injection of equine pituitary extract (EPE) will result in three to four ovulations per estrous cycle and two embryos. A purified, ...
Hinrichs K, Choi YH, Love CC, Chung YG, Varner DD.We evaluated the effects of different donor cell treatments and activation methods on production of blastocysts after equine nuclear transfer. Nuclear transfer was performed by direct injection of donor cells, using a piezo drill, and standard activation was by injection of sperm factor followed by culture with 6-dimethylaminopurine. There was no difference in blastocyst development between embryos produced with roscovitine-treated or confluent donor cells (3.6% for either treatment). Addition of injection of roscovitine or culture with cycloheximide at the time of activation did not affect bl...
Choi YH, Love LB, Varner DD, Hinrichs K.Holding immature oocytes before the onset of maturation simplifies oocyte transport and aids in scheduling later manipulations. We report here a method for holding equine oocytes in the absence of meiotic inhibitors. In Experiment 1, immature oocytes with expanded cumuli were cultured at 38.2 degrees C in medium containing cycloheximide, or were held at room-temperature in M199 with Hanks' salts, for 16-18 h before maturation. Control oocytes were matured immediately after recovery. Oocytes were fertilized by intracytoplasmic sperm injection and cultured for 4d. Embryo development was not diff...
Lagutina I, Lazzari G, Duchi R, Colleoni S, Ponderato N, Turini P, Crotti G, Galli C.The objective of the present work was to investigate and clarify the factors affecting the efficiency of somatic cell nuclear transfer (NT) in the horse, including embryo reconstruction, in vitro culture to the blastocyst stage, embryo transfer, pregnancy monitoring and production of offspring. Matured oocytes, with zona pellucida or after zona removal, were fused to cumulus cells, granulosa cells, and fetal and adult fibroblasts, and fused couplets were cultured in vitro. Blastocyst development to Day 8 varied significantly among donor cells (from 1.3% to 16%, P < 0.05). In total, 137 nuclear...
Choi YH, Love CC, Varner DD, Hinrichs K.This study was conducted to evaluate the effects of thawing, division into aliquots and refreezing on fertilizing capacity (ability to support embryo development after intracytoplasmic sperm injection; ICSI) of frozen stallion semen. Frozen semen from a fertile stallion was thawed, diluted 1:100 with freezing extender, and refrozen (2F treatment). Control semen was frozen only once. In vitro matured equine oocytes were injected with: (1) motile control spermatozoa; (2) motile 2F spermatozoa; (3) non-motile 2F spermatozoa; or (4) non-motile 2F spermatozoa, followed by injection of sperm extract...
Allen WR.Although the horse was probably the first animal to experience and benefit from artificial insemination, it trailed the field somewhat with regard to the application of embryo transfer and other oocyte and embryo-related modern breeding technologies. But with a late run it is now back in mid-field and gaining fast on the other large domestic species in the application of the many technological advances of the past 20 years to sound breeding practice. Improvements in extenders and cryoprotectants have resulted in a veritable upsurge in the transport and insemination of cooled and frozen stallio...
Carnevale EM, Coutinho da Silva MA, Panzani D, Stokes JE, Squires EL.Oocyte transfer is a potential method to produce offspring from valuable mares that cannot carry a pregnancy or produce embryos. From 2000 through 2004, 86 mares, 19.2 +/- 0.4 yr of age (mean +/- S.E.M.), were used as oocyte donors in a clinical program at Colorado State University. Oocytes were collected from 77% (548/710) of preovulatory follicles and during 96% (548/570) of cycles. Oocytes were collected 21.0+/-0.1h after administration of hCG to estrous donors and cultured 16.4 +/- 0.2 h prior to transfer into recipients' oviducts. At 16 and 50 d after transfer, pregnancies were detected i...
Moussa M, Bersinger I, Doligez P, Guignot F, Duchamp G, Vidament M, Mermillod P, Bruyas JF.Vitrification using open pulled straw (OPS) has provided encouraging results with embryos from other species. The aim of this study was to compare the survival of 6.5- and 6.75-day-old equine embryos after OPS vitrification and slow-cooling. Eighteen embryos were frozen using a slow-cooling method. Embryos were placed in modified PBS with increasing glycerol concentration (2.5%, 5%, 7.5% and 10% (v/v) 5 min each). Embryos were loaded into 0.25 ml straws then placed in a programmable freezer and subsequently plunged into liquid nitrogen. After thawing, cryoprotectant was removed by five steps w...
Vanderwall DK, Woods GL, Aston KI, Bunch TD, Li G, Meerdo LN, White KL.The objectives of the present study were to: (1) clone horses using adult cumulus cells; and (2) determine whether the cumulus cell donor affected the outcome. In vivo-matured cumulus-oocyte complexes were obtained using transvaginal ultrasound-guided follicle aspiration; oocytes were used as cytoplasts, whereas cumulus cells (from one of three different mares) were used as donor cells. Immediately following nuclear transfer and activation procedures, cloned embryos were transferred surgically to the oviduct of recipient mares (n = 2-5 embryos per recipient) that had ovulated within 24 h prior...
Eldridge-Panuska WD, di Brienza VC, Seidel GE, Squires EL, Carnevale EM.Experiments were conducted to determine viability of equine embryos in vivo after vitrification. In a preliminary study (Experiment 1), embryos were exposed in three steps to vitrification solutions containing increasing concentrations of ethylene glycol and glycerol (EG/G); the final vitrification solution was 3.4 M glycerol + 4.6 M ethylene glycol in a base medium of phosphate-buffered saline. Embryos were warmed in a two-step dilution and transferred into uteri of recipients. No pregnancies were observed after transfer of blastocysts >300 microm (n = 3). Transfer of morulae or blastocyst...
Stout TA, Meadows S, Allen WR.Early embryonic development in the horse is characterized by the formation of an unusual acellular glycoprotein "capsule" between the trophectoderm and the overlying zona pellucida. This structure is first detected between days 6 and 7 after ovulation and completely envelops the spherical conceptus until as late as day 23 of gestation. In the present study, a micromanipulator was used to remove the capsule from 15 embryos on day 6-7 after ovulation. None of these denuded embryos developed into ultrasonographically detectable pregnancies after surgical transfer into recipient mares whereas four...
Pomar FJ, Teerds KJ, Kidson A, Colenbrander B, Tharasanit T, Aguilar B, Roelen BA.The occurrence of pregnancies and births after embryo transfer (ET) of in vivo produced embryos is generally more successful compared to that of embryos produced in vitro. This difference in ET success has been observed when embryos of morphological equal (high) quality were used. The incidence of apoptosis has been suggested as an additional criterion to morphological embryo evaluation in order to assess embryo quality and effectively predict embryo viability. In this study, equine, porcine, ovine, caprine and bovine in vivo and in vitro produced morphologically selected high quality (grade-I...
Carnevale EM, Coutinho da Silva MA, Maclellan LJ, Seidel GE, Squires EL.Parentage identification was used to test the developmental competence of oocytes cultured under different conditions and fertilized in vivo after oocyte transfer. Oocytes were collected transvaginally from follicles of estrous mares approximately 22 h after administration of human chorionic gonadotropin. Oocytes were cultured for approximately 16 h in one of three media, with or without addition of hormones and growth factors. Groups of three or four oocytes, cultured in different media, were transferred into the oviduct contralateral to a recipient's own ovulation. Recipients were inseminate...
Maxwell WM, Evans G, Hollinshead FK, Bathgate R, De Graaf SP, Eriksson BM, Gillan L, Morton KM, O'Brien JK.Sex-sorting of mammalian spermatozoa has applications for genetic improvement of farm animals, in humans for the control of sex-linked disease, and in wildlife as a captive management strategy and for the re-population of endangered species. Considerable research has been undertaken worldwide on the Beltsville sperm sexing technology, the only effective method for pre-selection of sex of offspring. The combination of this method with assisted reproductive technologies has resulted in the birth of offspring in a wide range of animals, including cattle, the only livestock species in which sperm ...
Hinrichs K, Kenney RM.Ovariectomized recipient mares were divided into two groups. Group A mares received 300 mg progesterone in oil i.m. daily starting 5 days before transfer of a 7-day embryo. Group B mares received the same dose of progesterone, but starting at least 4 days before donor ovulation. Presence of an embryonic vesicle was determined by ultrasonography; mares were considered to be pregnant if they had normal vesicle development to Day 18. Pregnancy rates were: Group A, 6/8; Group B, 1/12 (P less than 0.01). An additional 4 mares in Group B had a vesicle visible at 14 days which degenerated or did not ...
Falge R, Ehling C, Niemann H.The conservation of endangered breeds as live animals is at present the main national strategy of the government and breeding organizations to maintain genetic diversity. Fourty-three breeds and some old strains of cattle, pig, sheep, goat and horses are currently involved. Cryopreservation and banks for sperm, embryos or DNA are another type of genetic material which could subsequently be used for breeding and production in agriculture. Present semen banks involve 9 endangered cattle breeds and also a small amount of deep-frozen sperm of some endangered sheep and horse breeds. Only 2 embryo b...
Jeannerat E, Marti E, Thomas S, Herrera C, Sieme H, Wedekind C, Burger D.The major histocompatibility complex (MHC) influences sexual selection in various vertebrates. Recently, MHC-linked social signaling was also shown to influence female fertility in horses (Equus caballus) diagnosed 17 days after fertilization. However, it remained unclear at which stage the pregnancy was terminated. Here we test if MHC-linked cryptic female choice in horses happens during the first days of pregnancy, i.e., until shortly after embryonic entrance into the uterus and before fixation in the endometrium. We exposed estrous mares to one of several unrelated stallions, instrumentally...
Coutinho da Silva MA.The use of assisted reproductive techniques (ART) has helped owners to produce offspring from valuable mares that were considered infertile using standard breeding techniques. Before referring a mare for an ART, the practitioner should be able to identify the underlying cause of subfertility of the mare. The objective of this review is to provide information regarding embryo transfer, oocyte transfer and intracytoplasmic sperm injection, the three most common ART used in equine practice. Knowing the complexity as well as the risks of these techniques, enables practitioners to refer a subfertil...
Pérez-Marín CC, Vizuete G, Borge C, Galisteo JJ.While testing for uterine bacterial infection is usually performed prior to artificial insemination (AI), samples taken during or after embryo flushing are generally not assessed either in subfertile and old mares or in fertile mares, even though knowledge of the status of the uterine environment in which the embryo is to develop would help to predict the outcome of embryo transfer programmes. The presence of bacteria and inflammatory cells in the liquid retained in the filter after uterine flushing in donors was determined at the moment of embryo recovery. Primarily, a group of mares (n = 8) ...
Squires EL.Most equine embryos are collected from the donor mare and transferred immediately as fresh embryos or shipped cooled to a recipient station for transfer within 24 hours. Very few equine embryos are frozen despite the numerous advantages of embryo cryopreservation. There are 2 major hurdles: Only the small embryos (<300 μm) provide good pregnancy rates after freezing/thawing and transfer. Also there is no good procedure for superovulating mares; thus, extra embryos for freezing are not readily available. Using either a slow cool or a vitrification method, pregnancy rates of small equine e...
Rajabi-Toustani R, Tsogtgerel M, Gao Y, Li C, Sakato M, Haneda S, Cheong SH, Nambo Y.Japanese native horses, which consists of 8 breeds, are threatened with extinction. Embryo transfer (ET) is used to reproduce endangered animals in various mammalian species. We aimed to perform ET using native ponies from Kiso and Hokkaido as donors and recipients, respectively. ET operation included long-distance transport of non-cryopreserved embryos from Nagano Prefecture to Hokkaido. Embryos were transported 1500 km over 9 h in a container maintained at 22°C. After transferring two embryos to two recipients, one mare delivered a healthy live foal. These results demonstrated that reciproc...
Raz T, Card C.Reliable methods of regulating estrus and stimulating superovulations in equine embryo transfer programs are desirable. Our objectives were to investigate the efficacy of a progesterone and estradiol-17beta (P&E) estrus synchronization regimen in mares with and without subsequent equine follicle-stimulating hormone (eFSH) treatment and to examine the effects of eFSH on folliculogenesis and embryo production. Cycling mares were treated with P&E daily for 10 d. On the final P&E treatment day, prostaglandin F(2alpha) was administered, and mares were randomly assigned to one of two treatment group...
Ball BA, Miller PG.In this study we examined the ability of equine oviductal epithelial cells (OEC) to support the development of four- to eight-cell equine embryos in vitro and investigated the ability of co-cultured embryos to continue normal development after transfer to synchronous recipient mares. Equine embryos obtained at Day 2 after ovulation were cultured with or without OEC for 5 days. Those OEC co-cultured embryos that reached the blastocyst stage and embryos recovered from the uterus at Day 7 were surgically transferred to synchronous recipient mares. Co-culture with OEC improved (P < 0.01) develo...
Carnevale EM, Maclellan LJ, Coutinho da Silva MA, Squires EL.After euthanasia, ovaries were removed from 5 horses and shipped to a laboratory where 46 oocytes were collected. The oocytes were cultured for 24 to 30 hours, and 36 oocytes were transferred to 10 recipient mares via flank laparotomies. Recipient mares were inseminated with semen from various stallions. Sixteen days after transfer, 4 of the recipients were pregnant with at least 1 embryonic vesicle. Embryonic death occurred in 3 recipients, whereas a healthy live foal was born from 1 recipient. Ovaries from valuable mares can be a source of viable oocytes after death of the mare. For shipping...
Kask K, Odensvik K, Kindahl H.The pattern of the main metabolite of prostaglandin (PG) F2alpha was recorded following a nonsurgical embryo transfer technique in 9 mares under field conditions in Estonia. Three patterns were observed. Two of them were characterised by PG release, thereas the third was not. A tendency towards a shortened cycle was seen in 3 mares. Observations were made regarding the manipulation of the uterus as being normal or difficult to perform. In general, mares where the procedure was considered difficult were also found to have a PG release.
Ginther OJ.Recent findings on the origin and development of twins from ovulation (Day 0) to fixation (mean: Day 16) are reviewed. Available data show that almost all twins originate from multiple ovulations. Results of recent ultrasound studies indicate that the number of days between double ovulations does not affect the conception rate per ovum or embryo survival during the first 16 days after each ovulation. Embryo reduction is the natural elimination of excess embryos so that only one embryo enters the foetal stage. In two studies, embryo reduction before or on the day of fixation was not considered ...
Riera FL, Roldán JE, Gomez J, Hinrichs K.Transfer of donor oocytes to the oviducts of inseminated recipient mares (oocyte transfer, OT) presents a valuable method for production of foals from otherwise infertile mares. Little information is available, however, on factors affecting success of OT in a clinical setting. We report the findings over three breeding seasons in a commercial OT program developed at an equine embryo transfer center in Argentina. Overall, 25 mares were enrolled, and 197 follicle aspiration procedures were performed. The average mare age was 23 years. Follicle aspiration was performed with a needle placed throug...
Goretti RG, Araújo RR, Filho AN, Araújo GH, Lopes EP, Guimarães JD.The objective was to evaluate the effects of giving prostaglandin F₂(α) (PGF) to donor mares 48 h prior to embryo collection. Non-lactating donor mares (n = 20 estrous cycles in 10 mares), ranging from 2.5 to 10 y of age and 400 to 500 kg of body weight were used from September 2004 to February 2005 in the southern hemisphere (Brazil). Donor mares were randomly assigned in a cross-over design study. During a Treated cycle, 7.5 mg PGF was given 48 h prior to embryo collection, whereas in the Control cycle, 7.5 mg PGF was given at embryo collection. In Treated Cycles, serum progesterone conce...
Parker NA, Bailey TL, Bowen JM, Ley WB, Purswell BJ, Dascanio JJ.Like the human female, the mare experiences reproductive tract pathology that may sometimes be circumvented by the use of assisted reproductive technologies (ARTs). One such technology, gamete intrafallopian transfer (GIFT), may be used in mares that exhibit ovulatory, oviductal, or uterine abnormalities that limit the use of common ARTs, such as embryo transfer. Homologous GIFT has been successfully performed in the horse; however, the logistics, costs, and associated risks of surgically transferring gametes to the oviducts of a recipient mare are considerably high. Use of a less costly speci...
Moussa M, Bersinger I, Doligez P, Guignot F, Duchamp G, Vidament M, Mermillod P, Bruyas JF.Vitrification using open pulled straw (OPS) has provided encouraging results with embryos from other species. The aim of this study was to compare the survival of 6.5- and 6.75-day-old equine embryos after OPS vitrification and slow-cooling. Eighteen embryos were frozen using a slow-cooling method. Embryos were placed in modified PBS with increasing glycerol concentration (2.5%, 5%, 7.5% and 10% (v/v) 5 min each). Embryos were loaded into 0.25 ml straws then placed in a programmable freezer and subsequently plunged into liquid nitrogen. After thawing, cryoprotectant was removed by five steps w...
Huhtinen M, Reilas T, Katila T.The pregnancy rate is lower in mares inseminated at the first post-partum (p.p.) oestrus (40-50%) compared with pregnancy rates in subsequent oestrous cycles (55-65%). The causes of the lowered pregnancy rate are not fully understood. The aim of the present study was to examine if embryonic defects could be one of the reasons for lowered pregnancy rate. A total of 23 p.p. and 14 non-lactating control mares were flushed 7 days after detection of ovulation. Embryo recovery rate was 48% and 71% in p.p. and control mares, respectively (p = 0.16). Embryos were photographed, measured, graded and sta...
Pérez-Marín CC, Vizuete G, Vazquez-Martinez R, Galisteo JJ.Few studies have been published about cryopreservation and embryo assessment in horses and donkeys. Objective: To evaluate the viability of embryos collected from mares and jennies that were cryopreserved by slow freezing or by vitrification. Methods: Randomised controlled experiment. Methods: Horse (n=19) and donkey (n=16) embryos (≤300 μm) were recovered on days 6.5-7.5 post-ovulation and assigned to control or cryopreservation protocols of slow freezing or vitrification. For slow freezing, 1.5 mol/L ethylene glycol (EG) was used. For vitrification, horse embryos were exposed to 1.4 mol/L...
Sales FABM, Azevedo MV, Souza NM, Ferreira-Silva JC, Chaves MS, Junior VR, Rocha JM, Dos Santos Filho JP, Freitas VJF, Oliveira MAL.The aim of this study was to evaluate the correlation between the corpus luteum vascularization with the concentration of progesterone and the fertility of embryo recipient mares. Mangalarga Marchador mares (n = 33) were distributed into groups according to the days (D) after ovulation, as follows: D3 (n = 8), D4 (n = 8), D5 (n = 9), and D6 (n = 8). The evaluations of the corpus luteum, endometrium, and blood collection to quantify the progesterone concentration were carried out on D3, D4, D5, and D6. Among the parameters evaluated, only progesterone concentration on D6 differed from the other...
Rigoleto Júnior WL, Dias de Camargo Neto W, de Paula Nogueira G, Ferrari TA, Bertan Membrive CM, Giometti IC, Castilho C.The objective of this study was to quantify serum progesterone levels, uterine features, and pregnancy rates in acyclic, embryo recipient mares using a bovine progesterone-releasing intravaginal device in a commercial embryo transfer (ET) program. The study included 73 recipient mares of unknown breed, aged 3-10 years, weighing 350-500 kg, and kept under an intensive management system on Tifton 85 (Cynodon spp.) pastures with water and mineral salt ad libitum. The horses were divided into two groups: a group with a progesterone-releasing intravaginal device (1 g progesterone, G-IVP4, n = 24...
Maciel LFS, Silva ESM, Oliveira-Filho JP, Fritsch SC, Rossi RS, Lourenção JAC, Meira C.Administration of progesterone (P4) after estradiol is usually performed to prepare non-cyclic mares as embryo recipients. However, there are successful pregnancy reports after embryo transfer in non-cyclic mares treated only with progestins. The objective of this study was to evaluate endometrial gene expression and immunostaining for estrogen receptor alpha (ERα), beta (ERβ) and progesterone receptor (PR) in seasonal anestrous mares treated with long acting P4 (LA P4). Endometrial tissue from eight seasonal anestrous mares was collected immediately before administration of 1.5 g of LA P4 ...
Lopes Ede P, Siqueira JB, Pinho RO, Guimarães JD, Rocha AN, de Carvalho GR, Torres CA.The objective of this study is to evaluate the reproductive efficiency in donors and recipient Mangalarga Marchador mares in commercial programmes of embryo transfer (ET) and the effects of some reproductive characteristics and ET methodology on conception rates in the recipient mares. A total of 1140 flushing procedures were performed and 830 embryos (72.8%) were recovered. There were no differences between the rates of embryonic recovery in the different breeding seasons (p > 0.05) and 92.8% of the recovered embryos were 8-9 days old. There was no difference in the embryonic recovery regardi...
Sitzenstock F, Rathke I, Ytournel F, Simianer H.A reference horse-breeding programme with 13500 foals each year was modelled with ZPLAN+. This new software for the optimization of the structures in breeding programmes is based on ZPLAN. In two scenarios, the implementation of a rigorous selection of mares was implemented. In scenario I, the mare performance test was the point of selection, while in scenario II, further information on 20 competitions in two more years is available. These selected mares were used for embryo transfer (ET), partly in combination with multiple ovulation (MOET). The selection intensity and the number of foals out...
Douet C, Parodi O, Martino NA, Lacalandra GM, Nicassio M, Reigner F, Deleuze S, Dell'Aquila ME, Goudet G.Most wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a breed. In the equine, embryo production in vitro would allow the production of several embryos per cycle. Intracytoplasmic sperm injection (ICSI) is used to generate horse embryos, but it requires expensive equipment and expertise in micromanipulation, and blastocyst development rates remain low. No conventional in vitro fertilization (IVF) techn...
Enders AC, Meadows S, Stewart F, Allen WR.The mature preinvasive chorionic girdles of horse, mule, donkey and extraspecies donkey-in-horse conceptuses, and the very young endometrial cups on d 37 of gestation in mares carrying horse, mule and transferred donkey-in-horse conceptuses, were compared histologically and ultrastructurally to determine possible mechanisms underlying failure of endometrial cup development in the donkey-in-horse model of equine abortion. The progenitor chorionic girdle from the failing donkey-in-house pregnancy was similar in size to the normal donkey chorionic girdle but the trophoblast cells within the forme...
Aurich C, König N, Budik S.In mares, repeated embryo collection in successive oestrous cycles is necessary if a greater number of foals should be produced. We investigated effects of repeated embryo collection in fertile donor mares on embryo recovery rates. In addition, an influence of the individual mare and season on embryo recovery rates was studied. In nine mares, a total of 153 embryo collections were performed during 30 months (17 ± 2.2 embryo collections per mare). The overall embryo recovery rate was 64% and did not differ among mares. Between successive embryo collection procedures, recovery rate varied signi...
Segabinazzi LGTM, Andrade LRP, Alvarenga MA, Dell'Aqua JA, Canisso IF.The objectives of this study were: (1) to assess uterine features and serum progesterone concentrations of acyclic mares synchronized and resynchronized with intravaginal progesterone release device (IPRD), and (2) to compare pregnancy rates and losses of cyclic and acyclic embryo recipient mares treated with different synchronization protocols. In Experiment 1, mares (n = 12) received estradiol for 3 days (E2-3d), and then 24 h after the last injection, an IPRD was inserted and kept in place for 9 days. Three days after IPRD removal, mares were treated with E2-3d, and then a new IPRD was inse...
Tischner M.In the present study the growth and development of three pairs of matching gender foals from birth to maturity were compared. One Polish Pony embryo of each pair was transferred from a donor mare (mass 387-400 kg) to a much larger heavy type recipient mare (mass 561-780 kg). The other embryo of each pair underwent normal gestation (control). The transferred and control horses were examined at 9 and 13 years of age, and X-ray pictures were taken of their front legs to measure the length of the bones. On the basis of the results of this study, it is concluded that the processes of growth and dev...
Morelli KG, Lourenço GG, Marangon VR, Feltrin IR, Imura Oshiro TS, Rodrigues da Silva IV, Pugliesi G.We aimed to evaluate the impact of corpus luteum (CL) and uterine characteristics accessed by B-mode and Color-Doppler ultrasonography in recipient mares at the time of embryo transfer (ET) on pregnancy outcomes. Recipient mares (n = 110), between days 3-9 after spontaneous ovulation, received a fresh embryo. Immediately before ET, the reproductive system was assessed by transrectal palpation for the following parameters: uterine tone (0-3), CL echogenicity (0-6), CL type (homogeneous, trabecular or anechoic center), luteal area (cm2), uterine echogenicity (0-3), uterine edema (0-3), luteal ...
