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Topic:Reproductive Technology

Reproductive technology in horses encompasses a range of scientific techniques and procedures aimed at assisting and enhancing equine reproduction. These technologies include artificial insemination, embryo transfer, and cryopreservation of gametes and embryos. They are employed to improve breeding efficiency, manage genetic diversity, and preserve valuable genetic material. Artificial insemination involves the collection and introduction of semen into the mare's reproductive tract, while embryo transfer allows for the harvesting and implantation of embryos from donor to recipient mares. Cryopreservation involves freezing and storing sperm, oocytes, or embryos for future use. This page compiles peer-reviewed research studies and scholarly articles that investigate the methodologies, applications, and outcomes of reproductive technologies in equine breeding and management.
Preliminary observations in in vitro development of equine embryo after ICSI.
Reproduction, nutrition, development    February 5, 1999   Volume 38, Issue 6 653-663 doi: 10.1051/rnd:19980607
Guignot F, Ottogalli M, Yvon JM, Magistrini M.The objective of this study was to perform intracytoplasmic sperm injection (ICSI) on in vitro matured equine oocytes and to improve in vitro embryonic development on Vero cells after activation of the microinjected oocytes with calcium ionophore. After maturation (23 or 40 h, 38.5 degrees C, 5% CO2), the cumulus-oocyte complexes were denuded, centrifuged and all oocytes exhibiting the first polar body were microinjected. ICSI was performed using fresh semen from three fertile stallions. Microinjected oocytes were activated with calcium ionophore A23187 (10 min, 10 microM) and cultured individ...
Equine Embryo Transfer IV. Proceedings of the 4th International Symposium on Equine Embryo Transfer. Reims, France, 16-18 January 1997.
Equine veterinary journal. Supplement    June 6, 1998   Issue 25 6-112 
No abstract available
Follicular fluid is not a compulsory carrier of the oocyte at ovulation in the mare.
Equine veterinary journal. Supplement    May 21, 1998   Issue 25 22-24 doi: 10.1111/j.2042-3306.1997.tb05094.x
Palmer E, Duchamp G, Cribiu EP, Mahla R, Boyazoglu S, Bézard J.The aim of this study was to test the possibility that ovulation can occur from a preovulatory follicle emptied of its follicular fluid. Transport of the oocyte into the oviduct and fertilisation in 29% of cases demonstrated that ovulation can occur in the absence of follicular fluid but the higher fertility achieved in control mares (62.5%) suggested that follicular fluid does serve a role during ovulation, fertilisation and oviductal transport. Injection of horse oocytes into preovulatory follicles in mules after removal of the follicular fluid, followed by insemination of the mules with hor...
The use of early pregnant mares as embryo recipients.
Equine veterinary journal. Supplement    May 21, 1998   Issue 25 77-79 doi: 10.1111/j.2042-3306.1997.tb05106.x
Camillo F, Cela M, Vannozzi I, Romagnoli S, Aria G.Fourteen normal, cyclic mares, treated to synchronise oestrus and ovulation and inseminated artificially with fresh semen, were assigned to a donor or a recipient group after ovulation, with the aim of obtaining a degree of synchrony of > or =2 days. Ten embryos, collected on Day 6 or 7 after ovulation (Day 0), were transferred nonsurgically to inseminated recipient mares (IRM) that had ovulated up to 5 days after the respective donors, or to pregnant recipient mares (PRM) that had ovulated 2-7 days before the donors. Embryonic size and development, as determined by ultrasound examination, ...
Treatment of equine oocytes with A23187 after intracytoplasmic sperm injection.
Equine veterinary journal. Supplement    May 21, 1998   Issue 25 51-53 doi: 10.1111/j.2042-3306.1997.tb05100.x
Kato H, Seidel GE, Squires EL, Wilson JM.In vitro matured horse oocytes with a first polar body (n = 68) were each injected with a single spermatozoon and divided into 2 groups: Group 1 oocytes were treated with 10 microM calcium ionophore A23187 for 5 min while Group 2 oocytes received no activation treatment. After culture in vitro for 2 days, significantly more oocytes treated with A23187 (5/24, 21%) cleaved than oocytes without activation treatment (2/44, 5%, P<0.05). All 7 cleaved zygotes from both treatment groups were transferred to recipient mares but no pregnancies resulted.
Cryopreservation of equine embryos with glycerol plus sucrose and glycerol plus 1,2-propanediol.
Equine veterinary journal. Supplement    May 21, 1998   Issue 25 88-93 doi: 10.1111/j.2042-3306.1997.tb05109.x
Ferreira JC, Meira C, Papa FO, Landin e Alvarenga FC, Alvarenga MA, Buratini J.Six or 7-day-old equine embryos were divided into 4 groups; Group 1, n = 15, Day 7 embryos destined for immediate transfer; Group 2, n = 15, Day 6 embryos destined for deep-freezing with glycerol plus sucrose as cryoprotectant; Group 3, n = 10, Day 6 embryos destined for deep-freezing with glycerol plus 1,2-propanediol as cryoprotectant and Group 4, n = 3, fresh embryos destined for ultrastructural analysis. All the frozen/thawed embryos were transferred to recipient mares, except 3 embryos in Group 3 that were subjected to ultrastructural analysis. After thawing the cryoprotectants were remov...
Effects of follicular fluid supplementation of in-vitro maturation medium on the fertilization and development of equine oocytes after in-vitro fertilization or intracytoplasmic sperm injection.
Human reproduction (Oxford, England)    February 10, 1998   Volume 12, Issue 12 2766-2772 doi: 10.1093/humrep/12.12.2766
Dell'Aquila ME, Cho YS, Minoia P, Traina V, Lacalandra GM, Maritato F.The aim of this study was to compare the effect of the addition of follicular fluid (FF) collected from preovulatory follicles with that of oestrous mare serum (EMS) (acting as the control) to TCM-199 medium on the in-vitro maturation, fertilization and development of equine cumulus-enclosed oocytes. Oocytes (<30 mm in diameter) were obtained from the ovaries of slaughtered mares. After in-vitro maturation in the presence of the two supplements, their fertilization, cleavage and developmental potential were compared after conventional in-vitro fertilization (IVF) or intracytoplasmic sperm i...
Vestibulo-vaginal hypoplasia in a mare.
The Veterinary record    January 7, 1998   Volume 141, Issue 20 521-522 doi: 10.1136/vr.141.20.521
Freeman SL, England GC.No abstract available
Prostaglandin F2alpha release associated with an embryo transfer procedure in the mare.
Equine veterinary journal    July 1, 1997   Volume 29, Issue 4 286-289 doi: 10.1111/j.2042-3306.1997.tb03125.x
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.
Pregnancies from imipramine and xylazine-induced ex copula ejaculation in a disabled stallion.
The Canadian veterinary journal = La revue veterinaire canadienne    March 1, 1997   Volume 38, Issue 3 171-174 
Card CE, Manning ST, Bowman P, Leibel T.Breeding or semen collection was attempted using: natural cover, manual stimulation, artificial vagina, pharmacologic induction of ejaculation, and electroejaculation. Sperm cells were recovered from the ductus deferens and epididymides post mortem. Only semen collected ex copula by imipramine and xylazine treatment resulted in conceptions (4/5). This is the first report of pregnancies in horses from ex copula semen collection.
Cytogenetic analysis of horse oocytes matured in vitro for different periods of time.
Reproduction, nutrition, development    January 1, 1997   Volume 37, Issue 1 63-68 doi: 10.1051/rnd:19970107
Sosnowski J, Lechniak D, Brzozowska M, Switoński M.This paper presents the results of recovering horse oocytes by aspiration and maturation in vitro for 24, 30, 36 or 42 h. A total of 522 oocytes were recovered from 221 ovaries (2.4 per ovary) and 271 oocytes (51.9%) were selected for in vitro maturation (IVM). Oocytes were cultured in maturation medium (TCM 199 + estrus cow serum [ECS] + follicle-stimulating hormone [FSH] + 17 beta-estradiol + gentamycin). One hundred and seventy oocytes were cytogenetically analysed (68.3%). Cytogenetic analysis showed that the stage of maturation (first telophase-TI or second metaphase-MII) for fertilizatio...
Effect of GnRH and hCG on ovulation and pregnancy in mares.
The Veterinary record    August 3, 1996   Volume 139, Issue 5 119-120 doi: 10.1136/vr.139.5.119
Kilicarslan MR, Horoz H, Senunver A, Konuk SC, Tek C, Carioglu B.No abstract available
[Preservation of genetic variation in domestic animals using biotechnical methods].
DTW. Deutsche tierarztliche Wochenschrift    August 1, 1996   Volume 103, Issue 8-9 336-340 
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...
Cryopreservation of equine embryos.
The Veterinary clinics of North America. Equine practice    April 1, 1996   Volume 12, Issue 1 85-99 doi: 10.1016/s0749-0739(17)30296-1
Seidel GE.Principles and procedures for cryopreservation of equine embryos are described. Embryos less than 250 microM in diameter can be cryopreserved successfully if glycerol is used as the cryoprotectant. Cooling is takes place in such a way that most of the water leaves the cells before intracellular ice forms, and glycerol is removed after thawing without undue osmotic swelling of cells. Vitrification procedures also show promise for small embryos. Satisfactory procedures for cryopreserving embryos of more than 250 microM in diameter are not yet available.
Artificial insemination: assessing stallion semen quality after freezing.
Equine veterinary journal    January 1, 1996   Volume 28, Issue 1 5-6 doi: 10.1111/j.2042-3306.1996.tb01580.x
Boyle MS.No abstract available
Effect of timing of frozen semen insemination on pregnancy rate in mares.
Acta veterinaria Scandinavica    January 1, 1996   Volume 37, Issue 3 361-365 doi: 10.1186/BF03548102
Katila T, Celebi M, Koskinen E.Thirty-four mares were inseminated with frozen semen from one stallion during 2 oestrous cycles, every 48 h until ovulation took place and within 12 h after ovulation. Semen was frozen using the Colorado method. The insemination dose was from 200 to 400 x 10(6) progressively motile spermatozoa. Ovaries were examined every 12 h to determine time of ovulation. Examination for pregnancy was carried out using ultrasonography, 15 days after ovulation. Thirty-five per cent of mares inseminated < 24 h and 23% of mares inseminated between 24-48 h before ovulation were pregnant (p = 0.388). The preg...
Recovery rate and quality of embryos from mares inseminated at the first post-partum oestrus.
Acta veterinaria Scandinavica    January 1, 1996   Volume 37, Issue 3 343-350 doi: 10.1186/BF03548100
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...
[Reproductive medicine in transition: new developments in embryo transfer].
Schweizer Archiv fur Tierheilkunde    January 1, 1996   Volume 138, Issue 5 245-250 
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 ...
Pregnancy diagnosis by ultrasound scanning.
The Veterinary record    April 8, 1995   Volume 136, Issue 14 371 doi: 10.1136/vr.136.14.371-a
Wooley P.No abstract available
[Embryo transfer in horses–current status and future perspectives].
Tierarztliche Praxis    December 1, 1994   Volume 22, Issue 6 558-566 
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...
Recent developments in cryopreservation of stallion semen with special emphasis on thawing procedure using thermal hysteresis proteins.
Zygote (Cambridge, England)    November 1, 1994   Volume 2, Issue 4 379-382 doi: 10.1017/s0967199400002264
Arav A, Carney JN, Pease GR, Liu KL.This research study explores the process of cryopreservation of stallion semen, focusing on improving the thawing procedures using thermal hysteresis proteins (THPs) from Antarctic and Arctic fish in order to [...]
In vitro development of day 2 embryos obtained from young, fertile mares and aged, subfertile mares.
Journal of reproduction and fertility    November 1, 1994   Volume 102, Issue 2 371-378 doi: 10.1530/jrf.0.1020371
Brinsko SP, Ball BA, Miller PG, Thomas PG, Ellington JE.This study was designed to investigate the development of day 2 embryos obtained from young and aged mares, co-cultured with oviductal epithelial cells obtained from mares in each age group in a 2 x 2 crossover design. Young, fertile mares (n = 19; 2-7 years of age) and aged, subfertile, mares (n = 16; 17-24 years of age) were used as embryo and oviductal epithelial cell donors. Embryos (n = 37) were collected from the oviducts 2 days after ovulation and were paired (embryos obtained from young mares with embryos obtained from aged mares) so that eight pairs were co-cultured with young mare ov...
In vitro viability of cryopreserved equine embryos following different freezing protocols.
Canadian journal of veterinary research = Revue canadienne de recherche veterinaire    October 1, 1994   Volume 58, Issue 4 235-241 
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...
Characteristics of cyclicity in maiden thoroughbred mares in the United Kingdom.
The Veterinary record    July 30, 1994   Volume 135, Issue 5 104-106 doi: 10.1136/vr.135.5.104
Watson ED, McDonnell AM, Cí·¯ord D.The characteristics of the cyclicity of 12 maiden thoroughbred mares kept in two groups were studied over a total of 58 cycles. On average, oestrus lasted 5.3 days and in 60 per cent of the cycles ovulation occurred in the last two days of oestrus. Oestrus and ovulation tended to be synchronised in each group of mares. The mean diameter of single-ovulating preovulatory follicles on the day before ovulation was 41.5 mm and during the seven days before ovulation they grew 2.5 mm/day. More than one follicle ovulated in 19 (33 per cent) of the cycles (seven double ovulations and 12 dioestrous ovul...
The equine embryonic capsule practical implications of recent research.
Equine veterinary journal    May 1, 1994   Volume 26, Issue 3 184-186 doi: 10.1111/j.2042-3306.1994.tb04366.x
Oriol JG.In most domestic animals, the zona pellucida is the outermost extracellular layer that covers the blastocyst before implantation. However, in the horse, an acellular membrane, the capsule, replaces the zona pellucida and envelops the developing conceptus during the 2nd and 3rd weeks of gestation. Although this structure was first described by Bonnet in 1889, it received little attention until the 1970s when its rediscovery by Marrable and Flood (1975) led to a series of reports (see review by Betteridge 1989). Nevertheless, until recently the structure, origin, and function of the capsule have...
[The effect of the covering of mares during the postpartum heat on the pregnancy rate].
Berliner und Munchener tierarztliche Wochenschrift    February 1, 1994   Volume 107, Issue 2 48-49 
Sönmez C, Eroglu A.The present study was carried out to investigate the pregnancy rate after covering in the foal heat (Group I), in the subsequent spontaneous heat (Group II), and in the induced heat (Group III) after administration of 7.5 mg of the prostaglandin F2 alpha analogue Luprostiol (Reprodin, Bayer) between the 20th and 22nd day post partum. Breeding during foal heat resulted in a pregnancy rate of 43.9% compared to 48.6% in the subsequent spontaneous heat post partum. Of 18 mares in group III, 14 mares had a foal heat, whereas 4 others had not shown a foal heat. 1-8 days after treatment, 14 mares (77...
Ovarian follicular growth and development in mammals.
Biology of reproduction    February 1, 1994   Volume 50, Issue 2 225-232 doi: 10.1095/biolreprod50.2.225
Fortune JE.Evidence from several species indicates that the initial stages of follicular growth proceed very slowly. In contrast, the stages after antrum formation are much more rapid. Atresia seems to be most prevalent as follicles approach the size at which they could be recruited for potential ovulation. Although most follicles become atretic around that stage, a few are recruited into a cohort or wave of follicles that continue to grow beyond the stage at which atresia normally occurs. Next, a species-specific number of follicles is selected for dominance. In some species (e.g. rats, primates, pigs),...
Insemination results with slow-cooled stallion semen stored for approximately 40 hours.
Acta veterinaria Scandinavica    January 1, 1994   Volume 35, Issue 3 257-262 doi: 10.1186/BF03548330
Heiskanen ML, Huhtinen M, Pirhonen A, Mäenpää PH.Semen from 3 stallions was extended using 2 methods (Kenney extender and a modified Kenney extender), slowly cooled, and stored for 41 +/- 6 (s.d.) h before insemination. An insemination dose (40 ml) contained 1.5-2 billion spermatozoa. In the experiment, 26 mares were inseminated in 30 cycles. The pregnancy rate per cycle obtained with sperm stored in the Kenney extender was 87% (n = 15). When the semen was extended with the modified extender, centrifuged and stored, the pregnancy rate was 60% (n = 15). Inseminations were done every other day until ovulation was detected. If a mare ovulated m...
Pregnancies following transfer of equine embryos cryopreserved by vitrification.
Theriogenology    January 1, 1994   Volume 42, Issue 3 483-488 doi: 10.1016/0093-691x(94)90686-d
Hochi S, Fujimoto T, Braun J, Oguri N.The objective of this study was to investigate the in vitro and in vivo developmental abilities of equine embryos cryopreserved by vitrification. Twenty-eight embryos were recovered from Native pony and Thoroughbred mares at Days 5 to 7 by nonsurgical uterine flushing (detection of ovulation=Day 0). The vitrification solution contained 40% ethylene glycol, 18% Ficoll, and 0.3 M sucrose in PBS. The embryos were placed for 1 to 2 min in vitrification solution (Group 1) or following exposure to 20% ethylene glycol in PBS for 10 to 20 min (Groups 2 and 3). Single embryos were loaded in 0.25-ml str...
Freezability and fertility results with uncentrifuged stallion semen.
Acta veterinaria Scandinavica    January 1, 1994   Volume 35, Issue 4 377-382 doi: 10.1186/BF03548311
Heiskanen ML, Hilden L, Hyyppä S, Kangasniemi A, Pirhonen A, Mäenpää PH.The first (1 to 3) sperm-rich fractions of the ejaculate were collected from 4 stallions using an open-ended vagina. The volume of the collected fractions was 12 +/- 8 ml with a density of 475 +/- 200 million spermatozoa/ml. Before freezing, the semen was diluted with a skim-milk based extender 1:1 to 1:8 (volume of semen: volume of extender), depending on the initial sperm concentration to achieve a final concentration of 100 million/ml. The total number of spermatozoa in an insemination dose ranged from 0.7 to 1 billion spermatozoa. Within 12 h after ovulation, 48 mares were inseminated in 7...
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