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.
Sertich PL.Pregnancy was established by transcervical transfer of embryos from performance mares into recipient mares. Estrus was synchronized between donor (n = 17) and recipient (n = 43) mares. After a greater than or equal to 25-mm follicle was detected, donor mares were bred artificially daily until ovulation. Day of ovulation was recorded. Uterine flushes (n = 111) were performed on donor mares 7 days after ovulation, and recovered embryos were transferred transcervically to recipient mares within 2 hours. Embryos were recovered from 40.5% of uterine flushes. Of transferred single embryos, 65.7% res...
Hinrichs K, Riera FL, Klunder LR.Embryo transfer was performed in three mares with gonadal dysgenesis. Karyotypes of the mares were as follows: Mare 1, 63,XX, 64,XX, 65,XX; Mare 2, 63,X; and Mare 3, 65,XXX. The mares were administered progesterone in oil, 300 mg intramuscularly daily, starting 1 or 2 days after donor mare ovulation. Embryos were transferred on day 7 after donor ovulation. Mare 1 became pregnant after the first embryo transfer and had a normally developing fetus on necropsy on day 45 of gestation. Mare 3 became pregnant after the third embryo transfer, but the embryo was lost between day 14 and day 18 of gesta...
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 ...
Pace-Owens S.Gamete intrafallopian transfer (GIFT), developed in 1984, was the result of further studies on in vitro fertilization (IVF). Since that time many nurses have worked in settings near in vitro fertilization centers and, therefore, have a basic understanding of the technology. An overview is given of the GIFT procedure to prepare nurses to advise and refer couples who may qualify for GIFT and to highlight the shift in the fertility program nurse coordinator's functions toward the positions of administrator and consultant.
Katila T, Oijala M, Kotilainen T, Väisänen K.Four subfertile mares (at least 2 barren years) were used as donors in an embryo transfer program. The embryo recovery rate was 50%, varying between 0-100. The transfer of 9 embryos resulted in the pregnancy rate of 44%. The efficiency of embryo transfer using subfertile mares is low. In this material 20% of flushings resulted in a pregnancy. Fyra subfertila ston (gatt galla i minst 2 är) användes som donatorer i ett program för embryot rans fer. Femtio procent av embryon tillvaratogs (varians 0–100%). Transfer av 9 embryon gav en dräktighetsresultat på 44%. Embryotransfer från subfert...
Ball BA, Little TV, Weber JA, Woods GL.The estimated embryonic loss rate between Days 4 and 14 after ovulation for young, normal mares (9%) was significantly lower (P less than 0.01) than the estimated embryonic loss rate for aged subfertile mares (62%). Fertilization rates, which were based on the recovery of embryos at Day 4 after ovulation, were 96% and 81% (P less than 0.1) for normal and subfertile mares, respectively. Day-4 embryos were collected from the oviducts of normal and subfertile donors mares. These embryos were transferred to the uteri of synchronized, normal recipient mares to test the hypothesis that the high inci...
Sirois J, Betteridge KJ.To recover intact Day-10.5 to Day-16.5 equine conceptuses (Day 0 = ovulation), a rigid catheter was used for 131 collections from donor mares diagnosed pregnant by ultrasonography. A total of 139 conceptuses were recovered, comprising 124 singletons, six pairs of twins and one set of triplets. Of these, 120 (86%) were intact after the collection, 14 (10%) had collapsed, and in five cases (4%), collapsed trophoblastic membranes were surrounded by an intact capsule. The recovery rate of intact conceptuses ranged from 99% on Days 10.5 to 12.5 to 40% on Day 16.5. More uterine flushes per recovery ...
Hinrichs K, Sertich PL, Solorzano NM, Caldwell LA.An immediate, qualitative enzyme-linked immunosorbent assay (ELISA) for progesterone was evaluated for use in determining the day of ovulation in an equine embryo transfer program. Plasma samples were collected from 27 mares from the third day of estrus to the second day of diestrus for 50 cycles. Ovulation was detected by ultrasound examination per rectum. Plasma progesterone concentrations were estimated using the qualitative assay to detect the time of the rise in progesterone after ovulation. Qualitative scores were compared to progesterone concentrations for the same samples as measured b...
Berghuis GA.The conception rates of semen intended for shipment and those of recently obtained semen are compared in the present paper. Conception rates using recently obtained semen were significantly superior to those obtained with semen intended for shipment. A number of factors to which this difference could be due are briefly discussed.
Carnevale EM, Squires EL, McKinnon AO.Forty equine embryos collected 7 d post-ovulation were stored at 5 C for 24 h in one of two culture media (n = 20/group): 1) Ham's F10 + 10% heat-treated fetal calf serum (FCS) buffered by gassing with 5% CO2, 5% O2 and 90% N2 and 2) Ham's F10 + 10% FCS with Hepes buffer (25 mM). Embryos cultured in Ham's F10 + CO2 maintained a better quality score and had a larger average increase in diameter (+34.8 micron) than embryos stored in Hepes buffered Ham's F10 (-10.2 micron). Embryos were transferred surgically into recipient mares that ovulated -3 to +1 d in relation to the donor mare. Twenty embr...
Sirois J, Betteridge KJ, Brault A.Embryo transfer was used in an equestrian teaching center in order to produce as many foals as possible from their preferred mares during a single breeding season. Embryo collection by uterine lavage was attempted in five donor mares on 25 occasions 6.5 days after ovulation. Sixteen of the collection attempts (64%) yielded a total of 17 blastocysts. Of these 17 embryos, 13 were immediately transferred transcervically into recipient mares that had ovulated within two days of the time of ovulation in the donors, three were frozen for later transfer, and one was lost. Eight of the freshly transfe...
Peyrot LM, Little TV, Lowe JE, Weber JA, Woods GL.Embryo autotransfer is defined as the collection of an embryo from and the transfer of this embryo into the same animal. The objectives of this study were to: 1) test the hypothesis that oviduct transport of the equine embryo from the oviduct into the uterus is not dependent on a unilateral embryo-corpus luteum interaction, 2) develop an embryo autotransfer technique for the mare and 3) compare the success rates of Day 4 embryos surgically autotransferred from the oviduct ipsilateral to ovulation to either the oviduct (n=10 mares) or the uterine horn (n=10 mares) contralateral to ovulation. Se...
Ball BA, Hillman RB, Woods GL.To test the hypothesis that an abnormal uterine environment was a cause of early embryonic loss in subfertile mares, morphologically normal embryos were transferred to normal mares (n = 20) and subfertile mares (n = 20), and embryo survival rates were compared. Embryos were recovered nonsurgically at Days 7 to 8 postovulation and transferred surgically to normal and subfertile mares that had ovulated on the same day or within 2 d after a donor. Survival of transferred embryos was monitored by ultrasonography of the recipient mare's uterus from Day 9 through Day 28 postovulation. There were no ...
Hinrichs K, Sertich PL, Palmer E, Kenney RM.Pregnancy was established and maintained after embryo transfer in 3 ovariectomized mares treated with progesterone only. Four ovariectomized mares were used as recipients, and 7 transfers were performed. Progesterone in oil, 300 mg i.m. daily, was given starting 5 days before transfer of a 7-day embryo. If the mare was pregnant at 20 days, progesterone treatment was continued to 100 days of gestation. The 3 pregnant mares carried to term and delivered live foals with normal parturition, lactation and maternal behaviour. No differences were seen between pregnant and non-pregnant ovariectomized ...
Summers PM, Shephard AM, Hodges JK, Kydd J, Boyle MS, Allen WR.Blastocysts were collected non-surgically from 2 Przewalski's horse and 2 Grant's zebra mares and transferred extra-specifically to domestic horse and donkey recipients. Nine Przewalski's horse embryos were transferred surgically, and 2 non-surgically, to domestic Welsh-type pony mares. After surgical transfer, 7 (77.8%) pregnancies were established and 4 foals were born. Twelve Grant's zebra embryos were transferred surgically to 5 pony and 7 domestic donkey recipients respectively and 1 non-surgically to a donkey; 3 (60%) zebra-in-horse pregnancies were established and 2 went to term. Only 2...
Francl AT, Amann RP, Squires EL, Pickett BW.The effects of extender and storage at 20 degrees C on equine spermatozoa were evaluated in two experiments using embryo recovery as the end point. In both experiments, inseminations were every other day, starting on Day 2 or 3 of estrus or after a 35-mm follicle was detected, with 250 x 10(6) progressively motile cells (based on initial evaluation). In Experiment 1, semen from two stallions was used to compare the motility and fertility of spermatozoa maintained in a) heated skim milk extender at 37 degrees C with insemination in <1 h; b) E-Z Mixin extender at 37 degrees C with inseminatio...
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 ...
Pool KF, Wilson JM, Webb GW, Kraemer DC, Potter GD, Evans JW.Two hormone regimens were utilized for recipient mares which were 2-14 days after ovulation at the time of non-surgical embryo transfer. In Exp. I, 20 embryos were transferred non-surgically into recipient mares which had been given 22 mg altrenogest daily starting the day of recipient ovulation. Higher (P less than 0.05) pregnancy rates (50% vs 0%) were obtained in mares which were 2-6 days after ovulation at the time of transfer compared with mares which were 7-12 days after ovulation. In Exp. II, on the day the donor mare ovulated (Day 0), 10 mg PGF-2 alpha were given to the recipient mare ...
Sirois J, Betteridge KJ, Goff AK.The outcome of 23 collections and reinsertions of conceptuses on Days 10.5-13.5, 4 transfers of Day-10.5, and 13 transfers of Day-6.5 embryos (ovulation = Day 0) was monitored in 30 mares. Blood samples were taken before and after each procedure to measure plasma 15-keto-13,14-dihydroprostaglandin F-2 alpha (PGFM), and then daily for progesterone determinations. Mares were also subjected to daily teasing for detection of oestrus, and to uterine ultrasonography for tracing the development of the conceptus. After the reinsertions, 12/23 conceptuses were detectable immediately after the procedure...
Allen WR, Kydd JH, Boyle MS, Antczak DF.Transfer of donkey embryos to horse mares provides a useful model of early fetal death. Endometrial cups do not develop in this one type of extraspecific pregnancy and 80% of donkey fetuses are aborted between Days 80 and 100 of gestation in conjunction with abnormal implantation and an intense accumulation of leucocytes in the endometrium of the surrogate mare. Treatment of mares carrying donkey conceptuses with progestagen (allyl trenbolone) or purified horse chorionic gonadotrophin does not prevent abortion. However, passive immunization with serum from mares carrying intraspecific horse fe...
Parry-Weeks LC, Holtan DW.Non-surgical embryo recovery attempts were done on Day 7 after ovulation. Embryo recovery rate from mares of varied reproductive histories was 57% (38/67). Non-surgical transfer of these embryos into altrenogest-treated recipient mares that ovulated between 3 days before and 3 days after the donor resulted in a 30-day pregnancy rate of 77% (10/13). Transfer of embryos into altrenogest-treated recipients that ovulated between 4 days before and 6 days after the donor resulted in an overall pregnancy rate of 64% (16/25) at Day 30 of gestation. No recipients that were in oestrus at the start of tr...
Squires EL, McKinnon AO, Carnevale EM, Morris R, Nett TM.For embryos collected from mares 7 days after ovulation, embryo recovery for single-ovulating mares was 53% compared to 106% for double-ovulating mares. Pregnancy rates 50 days after surgical transfer were 68 and 129%, respectively. Concentrations of LH were similar during the periovulatory period for cycles which included single or double ovulations. Horse pituitary extract given for 5.5 days resulted in greater than or equal to 2 ovulations (mean 3.8) in 26 of 28 mares and 2.0 embryos were recovered per donor compared to 0.65 for controls. Non-surgical pregnancy rates for embryos collected f...
Woods GL, Hillman RB, Schlafer DH.To evaluate embryo transfer as a possible method to circumvent infertility in mares, embryos from 14 normal and 14 infertile mares were collected three times and examined. Fewer flushes (p less than 0.05) from normal than infertile mares (1/42 vs 9/42) contained only abnormal embryos whereas more flushes (p less than 0.05) from normal than infertile mares contained one or more normal embryos (28/42 vs 8/42). More flushes (p less than 0.05) from normal than infertile mares contained embryos (29/42 vs 17/42). The embryo diameters (mm) at either day-7 or day-8 post ovulation were greater (p less ...
Woods GL, Steiner JV.The objective of this project was to produce multiple pregnant recipient mares during one year from each of four mares which were in athletic training and competition. Twenty-six embryos were recovered from the four mares in 38 collection attempts. Each embryo was surgically transferred to one of 20 recipient mares. Nine of 10 recipient mares which were transferred to on one occasion were detected pregnant at the one week post transfer pregnancy examination and six of eight mares which were transferred to on two occasions were detected pregnant at one of the one-week post transfer pregnancy ex...
Hyland JH, MacLean AA, Robertson-Smith GR, Jeffcott LB, Stewart GA.The removal of one of twin embryos was attempted by infusion of 24% (w/v) saline into the gestation sac in 2 mares by laparotomy. The treatment was successful in one mare (Case 1) and the untreated embryo remained viable. However, neither foetus survived in the second mare (Case 2). Plasma oestrone sulphate (E1S) concentrations fell immediately after treatment in both mares but recovered to approximately 50% of pretreatment levels in Case 1. In Case 2 plasma E1S concentrations declined steadily and were less than 1 ng/ml within 6 days of treatment. These preliminary results suggest that the me...
Iuliano MF, Squires EL, Cook VM.A 2 X 2 cross-classified experiment was conducted to investigate the effect of age of equine embryo (7 vs 8 d postovulation) and method of transfer (surgical vs nonsurgical) on pregnancy rates at 50 d of gestation. Embryos were recovered 7 or 8 d postovulation using a Foley catheter and 3 liters of modified Dulbecco's phosphate-buffered saline (PBS). Upon identification, the embryos were placed in millipore-filtered PBS containing 20% heat-inactivated steer serum and maintained at room temperature until transferred. At the time of recovery, embryos were randomly assigned to be transferred eith...
Allen WE, Goddard PJ.The uteri of 13 pony mares were examined daily by ultrasound during the first two months of gestation. The conceptus was first identified between 12 and 16 days after ovulation and the embryo was seen on the ventral surface of the conceptus after Day 21. The foetal heart could be visualised after Day 22 and spontaneous movement of the foetus occurred after Day 39. No consistent pattern was seen in the development of the foetal membranes, although attachment of the umbilical cord to the allantochorion was always on the dorsal aspect of the conceptus. Daily measurements were made of the diameter...
Sirois J, Betteridge KJ.To recover intact Day-10.5 to Day-16.5 equine conceptuses (Day 0 = ovulation), a rigid catheter was used for 131 collections from donor mares diagnosed pregnant by ultrasonography. A total of 139 conceptuses were recovered, comprising 124 singletons, six pairs of twins and one set of triplets. Of these, 120 (86%) were intact after the collection, 14 (10%) had collapsed, and in five cases (4%), collapsed trophoblastic membranes were surrounded by an intact capsule. The recovery rate of intact conceptuses ranged from 99% on Days 10.5 to 12.5 to 40% on Day 16.5. More uterine flushes per recovery ...
Gastal GDA, Scarlet D, Melchert M, Ertl R, Aurich C.In embryos subjected to assisted reproductive techniques, epigenetic modifications may occur that can influence embryonic development and the establishment of pregnancy. In horses, the storage temperature during transport of fresh embryos before transfer is a major concern. The aim of this study was, therefore, to determine the effects of two storage temperatures (5 °C and 20 °C) on equine embryos, collected at day seven after ovulation and stored for 24 h, on: (i) morphological development; (ii) expression of candidate genes associated with embryo growth and development, maternal recognitio...
Kot K, Tischner M.The aim of the experiment was to determine the effect of a year-to-year prolonged daylength on the patterns of equine reproductive activity and results of embryo recovery. Experiments using Konik Polski mares were conducted over four reproduction seasons. Five mares were exposed to a regimen of artificially prolonged daylength (APD) and another five mares in a control group were kept under conditions of natural daylight. Both the control and experimental groups were examined for appearance of estrus, ovulation and also for the state of their coats. A single stallion was used for breeding all o...
Rosas CA, Alberio RH, Barañao JL, Agüero A, Chaves MG.The efficiency of superovulating mares with an enriched fraction of equine follicle-stimulating hormone (feFSH) and an equine pituitary extract (EPE) with similar FSH content but differing in the LH amount was compared. Mares were randomly assigned to an feFSH (n = 5) or EPE (n = 5) treatment. The experimental period was of 2 successive estrous cycles, with the first cycle as the control. At Days 6 and 7 of the estrous cycle, the mares received 250 micrograms i.m. cloprostenol. The treatments consisted of daily injections of 25 mg feFSH or EPE beginning on Day 6 post ovulation. Mares were inse...
McKinnon AO, Lacham-Kaplan O, Trounson AO.The use of intracytoplasmic sperm injection (ICSI) for in vitro fertilization of equine oocytes and the developmental potential of these oocytes after transfer to the Fallopian tubes of synchronized mares were examined. Oocytes were aspirated from mature follicles 39 h after injection of a GnRH analogue and transported 190 km at 39 degrees C. Semen from a fertile and an infertile stallion was frozen and prepared for injection. Successfully injected oocytes were transferred surgically into the ampulla of the Fallopian tube either: (i) 4-8 h after semen injection; or (ii) after 24-48 h culture b...
Raz T, Carley SD, Green JM, Card CE.Reliable methods for regulating oestrus and superovulation in equine embryo transfer (ET) programs are desirable. The objective in this study was to compare two oestrus synchronization methods combined with equine follicle-stimulating hormone (eFSH) treatment in an ET program. In the progesterone and estradiol-17β (P&E) group, mares (n=12) were given progesterone and estradiol-17β, daily for 10 days, followed by prostaglandin (PG)F(2α) on the last day. In the PG group, mares (n=12) were given PGF(2α) 5 days post-ovulation. In both groups donor mares were allocated to eFSH therapy, and were...
Rodrigues LT, Segabinazzi LG, Oliveira SN, Frasson M, Papa FO.Pregnancy rates after embryo transfer (ET) are disappointing in donkey species. This study aims to report two successful ET of mini-donkey embryos using Brazilian Northeastern jennies as recipients. Eighteen embryo flushes were performed 9 days post-ovulation in two non-pregnant mini-donkeys jennies (11 and 7 cycles per jenny). Eleven embryos (61%, 11/18) were collected and transferred to Brazilian Northeastern jennies 4-6 days post-ovulation by conventional (n = 6) or an alternative (n = 5) technique. The alternative method consisted of inserting a Polansky equine vaginal speculum smear...
Silva ESM, Leite RO, Maciel LFS, Francia CCDA, Padovani CR, Oliveira-Filho JP, Meira C.Aiming to investigate the effects of different hormonal protocols on the endometrium morphometry of anestrous mares, 26 animals were assigned to four different treatment groups: (1) EB2.5LAP4: single dose of 2.5 mg of estradiol benzoate (EB); (2) EB5LAP4: 5 mg of EB in 2 consecutive days; (3) EB10LAP4: 10 mg of EB in three consecutive days, considering that all EB-treated groups received a single dose of 1,500 mg of long-acting progesterone (LA P4) after the single/last EB dose; and (4) LAP4: only 1,500 mg of LA P4. Results were also compared with those found in cyclic mares (control grou...
Hinrichs K, Betschart RW, McCue PM, Squires EL.Mares with preovulatory follicles >33 mm in diameter were administered hCG and were randomly assigned for aspiration of the dominant follicle at 24 h or 35 h after hCG administration. Oocytes recovered at 24 h were cultured for 12 h before transfer and oocytes recovered at 35 h were cultured for 1 h. Oocytes were transferred by flank laparotomy to the oviduct of the same mare, or to the oviduct of another oocyte donor. Recipient mares were inseminated before and after transfer. The oocyte recovery rates at 24 h and 35 h after hCG administration were not significantly different (10/15 (66%) and...
Sirois J, Betteridge KJ, Brault A.Embryo transfer was used in an equestrian teaching center in order to produce as many foals as possible from their preferred mares during a single breeding season. Embryo collection by uterine lavage was attempted in five donor mares on 25 occasions 6.5 days after ovulation. Sixteen of the collection attempts (64%) yielded a total of 17 blastocysts. Of these 17 embryos, 13 were immediately transferred transcervically into recipient mares that had ovulated within two days of the time of ovulation in the donors, three were frozen for later transfer, and one was lost. Eight of the freshly transfe...
Heap RB, Hamon MH, Allen WR.Aromatase distribution in membranes of preimplantation horse and donkey conceptuses was compared by measuring the incorporation of [3H]androstenedione into oestrone and oestradiol-17 beta. In the donkey conceptus, aromatase activity was similar in all the tissues examined (yolk sac, chorionic girdle and allantochorion), whereas in the horse it was generally lower and showed the relationship chorionic girdle greater than yolk sac greater than allantochorion. A higher proportion of labelled precursor was incorporated into oestradiol-17 beta by extra-embryonic tissues of the donkey compared with ...
Lagneaux D, Pomarici AM, Sattler M, Bruneau B, Duchamp G, Camillo F, Palmer E.Day 6.5 equine embryos (n=30) were frozen in a medium containing glycerol (2.5-10.0%) supplemented with 0, 20 or 100 mmol L-glutamine 1(-1). After thawing, the embryos were tested individually, using 4',6'-diamidino-2-phenylindole (DAPI) staining to evaluate cell death. Three embryos (one frozen at each L-glutamine concentration) were transferred together into individual recipient mares. Pregnancy diagnosis was performed at day 12 (age of embryo). Embryos were collected at day 14 (age of embryo) and were identified by PCR amplified microsatellite analysis. Nine of ten recipient mares that rece...
Dordas-Perpinyà M, Pintart C, Terris H, Normandin L, Bruyas JF.Two cloned mares, produced from the same sample of skin fibroblasts, were bred during four breeding seasons from their second year of age, as embryo donors, in exactly the same conditions, using the same stallions for both cloned mares. The aim of this study was to test the embryo donor potential of cloned mares and to compare the results obtained from two cloned mares of the same mare with other embryo donor mares (n = 31-39 per breeding season) at the same stud. For both cloned mares, 19 embryos were recovered by 43 collection attempts (44%) (7/22 for one; 12/21 for the other), 16 (84%) pr...
Huhtinen M, Peippo J, Bredbacka P.Embryo biopsy has been used to detect inherited disorders and to improve the phenotype by analyzing of linkages between marker loci and the desired characteristics. Unfortunately, early procedures required the removal of a large portion (one-half) of the embryo for analysis, and the transfer of bisected equine embryos has not been particularly successful. Recent discovery of the polymerase chain reaction (PCR) has made possible the detection of specific DNA sequences from only a few cells. We investigated whether the removal of a small biopsy would allow for successful PCR and normal embryonic...
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...
Moussa M, Duchamp G, Mahla R, Bruyas JF, Daels PF.Equine embryos have been successfully transferred after 24h cooled storage in Ham's F-10. The aim of this study was to compare the viability of equine embryos in vitro and in vivo after 6 and 24h cooled storage using three media and to examine the relationship between embryo size and viability after 24h cooled storage. In Experiment 1, the viability of embryos was evaluated using DAPI-staining after 0, 6 or 24h in Ham's F-10, 24h in Emcare embryo holding solution (EHS) or 24h in ViGro holding plus (VHP) (n=10/group). The mean number of dead cells was similar for embryos stored in Ham's F-10, E...
Poitras P, Guay P, Vaillancourt D, Zidane N, Bigras-Poulin M.The main objective of this study was to evaluate two freezing protocols and the effect of agar embedding on survival of day 6.5 equine embryos. A total of 133 embryos were used, in one group (n = 51), embryos were first embedded in agar before the freezing protocol was started. A freezing protocol to -30 degrees C or -33 degrees C was used before plunging embryos into liquid nitrogen (LN2). The embryos were thawed in water at 37 degrees C, evaluated and placed in culture. After 24 h culture, the embryos were evaluated for their morphology and development. No differences were observed between e...
Braun J.Although foals born after embryo transfer are eligible for registration in the majority of horse breeds, application of embryo transfer is still rare. This is mainly due to the lack of a possibility for superovulation. Uterine stage embryos can be recovered by a non-surgical flushing technique. Transfer can be accomplished by non-surgical as well as surgical methods. In contrast to the situation in cattle, most related technologies are scarcely available. Methods of cryopreservation as well as bisection of embryos are hampered by the fact that suitable embryos (morula) can be collected from th...
Peyrot LM, Little TV, Lowe JE, Weber JA, Woods GL.Embryo autotransfer is defined as the collection of an embryo from and the transfer of this embryo into the same animal. The objectives of this study were to: 1) test the hypothesis that oviduct transport of the equine embryo from the oviduct into the uterus is not dependent on a unilateral embryo-corpus luteum interaction, 2) develop an embryo autotransfer technique for the mare and 3) compare the success rates of Day 4 embryos surgically autotransferred from the oviduct ipsilateral to ovulation to either the oviduct (n=10 mares) or the uterine horn (n=10 mares) contralateral to ovulation. Se...
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...
Hinrichs K, Sertich PL, Solorzano NM, Caldwell LA.An immediate, qualitative enzyme-linked immunosorbent assay (ELISA) for progesterone was evaluated for use in determining the day of ovulation in an equine embryo transfer program. Plasma samples were collected from 27 mares from the third day of estrus to the second day of diestrus for 50 cycles. Ovulation was detected by ultrasound examination per rectum. Plasma progesterone concentrations were estimated using the qualitative assay to detect the time of the rise in progesterone after ovulation. Qualitative scores were compared to progesterone concentrations for the same samples as measured b...
Taveiros AW, Oliveira MA, Lima PF, Tenório Filho F, Bartolomeu CC, Santos MH, Oliveira LR, Iunes-Souza TC, Freitas JC.Ten pluriparous mares were used as donors to supply embryos which were transferred into 103 recipients, 31 of which were nulliparous, 34 were pluriparous and lactating, and 38 were pluriparous and non-lactating. The embryos were recovered eight days after ovulation and pregnancy was confirmed by ultrasound six days after the transfer; the length of the embryos was measured ultrasonographically on days 12, 14, 16, 18, 20, 25 and 30 after the embryo transfer. One hundred and fifteen of 200 flushes provided embryos, 12 being degenerate and 103 being viable embryos. From the 103 embryo transfers c...
Coutinho da Silva MA, Carnevale EM, Maclellan LJ, Seidel GE, Squires EL.The objective of the study was to compare embryo development rates after transfer of oocytes collected 22 or 33 h after hCG injection into recipients inseminated within the uterus or the oviduct. Oocytes were collected at approximately 22 or 33 h after hCG injections and incubated for approximately 16 or 1.5 h, respectively, before transfer. Intrauterine inseminations using 1 x 10(9) progressively motile sperm were done approximately 12 h before and 2 h after transfer. For intraoviductal inseminations (gamete intrafallopian transfer [GIFT]), semen was centrifuged through a Percoll gradient, an...
Hinrichs K.A research study is a product of not only a question and its pursuit but also the people, places, and facilities available at the time. My work in equine assisted reproduction has progressed from embryo transfer to oocyte maturation, oocyte transfer, intracytoplasmic sperm injection, embryo biopsy, embryo vitrification, and cloning, as a result of collaborations with an array of remarkable people. This is a summary of some of the stories behind the studies.
Meier HP, Gertsch U, Kohler S.Twin pregnancies are a serious problem in stud medicine as they terminate in most cases in abortion, stillbirth or the delivery of dead or weak and deformed foals. In recent years, the introduction of sonography has greatly improved the management of mares with twin conceptuses, in particular monitoring the phenomenon of spontaneous embryo reduction to a single vesicle. This allows supervision of pregnancy with relatively little expenditure and loss of time. We document the spontaneous reduction to single vesicles of unilateral twin pregnancies in a draught- and a warmblood-mare. In both mares...
Scherzer J, Davis C, Hurley DJ.The major difficulty in providing the benefits of embryo cryopreservation for equine agriculture is the mismatch between the optimal embryo age for collection from the mare (7-8 days after ovulation was detected) and the optimal age for freezing under current methods (6.5 days after ovulation). To overcome this limitation, we tested a method to enhance penetration of cryopreservative across the capsule and trophoblast of day 7 and 8 embryos combined with rapid freezing by vitrification. Six small embryos (<300 μm in diameter) were collected on day 6-7 after ovulation and twelve larger embryos...
Schmid RL, Kato H, Herickhoff LA, Schenk JL, McCue PM, Chung YG, Squires EL.In Expt 1, compact cumulus oocyte complexes (COCs) were matured in: (i) control medium (Hepes-buffered TCM-199 with 10% oestrous cow serum (OCS) + oestradiol, LH and FSH); (ii) Hepes-buffered TCM-199 with 20% follicular fluid; or (iii) control medium containing 250 ng progesterone ml(-1). Mature oocytes were collected by transvaginal aspiration as a positive control for the in vitro maturation (IVM) treatments. Oocytes were fertilized by ICSI and cultured in Menezo's B2 + 5% fetal calf serum (FCS). There were no significant differences among IVM treatments. In Expt 2, oocytes with expanded COC...
Blanco ID, Devito LG, Ferreira HN, Araujo GH, Fernandes CB, Alvarenga MA, Landim-Alvarenga FC.This study examined the effect of treating mares with equine pituitary extract (EPE) alone or in combination with hCG on the recovery rate of immature follicles by transvaginal follicular aspiration (ovum pick-up; OPU). Ten normally cycling crossbred mares aged 3-15 years and weighing 350-400 kg were subjected to each of three treatments in a random sequence with each exposure to a new treatment separated by a rest cycle during which a spontaneous ovulation occurred. The treatments were (1) superovulated with 25mg EPE and treated with 2500 IU hCG, (2) superovulation with 25mg EPE, and (3) cont...
McCue PM, Matthews PM, Prell MJ, Bellone RR, Allen H.Genetic testing is required for the registration of foals of most equine breeds. Objective: To describe two clinical cases of marked delayed embryonic development or delayed fertilisation in pregnancies generated by embryo transfer. Methods: Case report. Methods: Donor mares were inseminated with semen from one stallion during one oestrous cycle and semen from a different stallion on the subsequent oestrous cycle. Embryo(s) were collected 8 days after ovulation during the second oestrous cycle and transferred into synchronised recipient mares. Genetic testing was performed to determine paren...
Binder H, Jakob C, Bucher P.Successful application of embryo transfer (ET) has become common practice in cattle, horses, sheep, goats and a variety of other species held in captivity. Yet in cattle only has the technique been established commercially. In 1994 more than 100,000 bovine embryos have been transferred in European countries. Important progress in transvaginal ovum pick up (OPU), in vitro production (IVP) and cryopreservation have further improved the applicability of ET. Direct transfer simplifies the procedure considerably allowing individual transfers and eliminating the need of synchronizing recipients. In ...
