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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.
Oral imipramine and intravenous xylazine for pharmacologically-induced ex copula ejaculation in stallions. This study is part of ongoing work toward developing pharmacological methods for enhancing and inducing ejaculation in stallions with ejaculatory dysfunction or disabilities that interfere with normal breeding behavior. The objective was to evaluate a treatment regimen involving oral imipramine followed by intravenous xylazine that, in uncontrolled field clinical trials, had shown promise for a higher rate of ejaculation and fewer side effects using a more easily obtained and administered form of imipramine. Eight stallions each underwent eight trials in which treatment consisted of imipramine...
Effect of homologous preovulatory follicular fluid on in vitro maturation of equine cumulus-oocyte complexes. The objective of this study was to test the hypothesis that incubating equine cumulus-oocyte complexes (COCs) in medium containing 50% or 100% homologous preovulatory follicular fluid would improve cumulus expansion and nuclear maturation. Oocytes were incubated in one of three media: 1) supplemented TCM-199 (control), 2) 50% (v/v) follicular fluid in control medium or 3) 100% follicular fluid. Cumulus expansion was evaluated subjectively, and nuclear maturation was evaluated by staining oocytes with Hoechst 33258. The hypothesis that incubating COCs in medium containing follicular fluid would...
Effect of periovulatory prostaglandin F2alpha on pregnancy rates and luteal function in the mare. The objective of this study was to determine whether periovulatory treatments with PGF2alpha affects the development of the CL, and whether the treatment was detrimental to the establishment of pregnancy. Reproductively sound mares were assigned randomly to one of the following treatment groups during consecutive estrus cycles: 1. 3,000 IU hCG within 24 hours before artificial insemination and 500 microg cloprostenol (PGF2alpha analogue) on Days 0, 1, and 2 after ovulation (n=8), 2. 2 mL sterile water injection within 24 hours before artificial insemination and 500 microg cloprostenol on Days ...
Embryo production by ovum pick up from live donors. Embryo production by in vitro techniques has increased steadily over the years. For cattle where this technology is more advanced and is applied more, the number of in vitro produced embryos transferred to final recipients was over 30,000 in 1998. An increasing proportion of in vitro produced embryos are coming from oocytes collected from live donors by ultrasound-guided follicular aspiration (ovum pick up, OPU). This procedure allows the repeated production of embryos from live donors of particular value and is a serious alternative to superovulation. Ovum pick up is a very flexible technique...
Regulation of ovarian follicular dynamics in farm animals. Implications for manipulation of reproduction. In this review, the main features of folliculogenesis are summarized and compared among species. In the past few years, ultrasonography has clarified follicle growth patterns, and our understanding of follicle maturation has improved considerably. As the follicles develop towards the ovulatory stage, three features appear to be highly conserved across all species: 1) the sequence of events (recruitment, selection and dominance); 2) the sequential need for gonadotropins (FSH for recruitment, LH for dominance) and 3) the large variability of numerical parameters (number of waves per cycle, numbe...
Embryo development rates after transfer of oocytes matured in vivo, in vitro, or within oviducts of mares. Objectives of the present study were to use oocyte transfer: 1) to compare the developmental ability of oocytes collected from ovaries of live mares with those collected from slaughterhouse ovaries; and 2) to compare the viability of oocytes matured in vivo, in vitro, or within the oviduct. Oocytes were collected by transvaginal, ultrasound-guided follicular aspiration (TVA) from live mares or from slicing slaughterhouse ovaries. Four groups of oocytes were transferred into the oviducts of recipients that were inseminated: 1) oocytes matured in vivo and collected by TVA from preovulatory folli...
Cryopreservation of equine embryos by open pulled straw, cryoloop, or conventional slow cooling methods. Cryopreservation of equine embryos with conventional slow-cooling procedures has proven challenging. An alternative approach is vitrification, which can minimize chilling injuries by increasing the rates of cooling and warming. The open pulled straw (OPS) and cryoloop have been used for very rapid cooling and warming rates. The objective of this experiment was to compare efficacy of vitrification of embryos in OPS and the cryoloop to conventional slow cool procedures using 0.25 mL straws. Grade 1 or 2 morulae and early blastocysts ( 0.1) in morphological grade or percent live cells among metho...
Treatments resulting in pregnancy in nonovulating, hormone-treated oocyte recipient mares. Synchronization of follicle growth between oocyte donor and recipient mares is difficult. To avoid this, recipient mares in a clinical program were used during a period of low follicular activity, and were treated with estrogen before transfer and progesterone after transfer. Five pregnancies were established after oocyte transfer to nonovulating, hormone-treated recipient mares. One pregnancy was lost before 30 d gestation, and the other 4 foals were carried to term. One foal died at birth. Establishment and maintenance of pregnancy in these mares indicates that nonovulating, hormone-treated ...
Fertility of mares after unilateral laparoscopic tubal ligation. To develop a technique for laparoscopic tubal (oviductal) ligation and to evaluate pregnancy rates for mares that ovulated ipsilateral or contralateral to the ligated oviduct. Methods: Randomized prospective clinical trial comparing pregnancy rates after unilateral laparoscopic tubal ligation. Methods: Twelve mares of light horse breeds. Methods: One oviduct in each of 6 mares was surgically ligated with a laparoscopic technique; 6 other mares served as nonligated controls. Mares with unilateral tubal ligations (UTL) were inseminated with 500 million progressively motile sperm during 1 cycle w...
Comparison of culture and insemination techniques for equine oocyte transfer. This study was designed to test 3 approaches for insemination and transfer of oocytes to recipient mares. Oocytes were recovered transvaginally from naturally cycling donor mares 24 to 26 h after an intravenous injection of 2500 IU of hCG when follicles reached 35 mm in diameter. Multiple oocytes (1 to 4) were transferred surgically into the oviducts of 4 or 5 recipient mares per group. Three groups of transfers were compared: 1) transfer of oocytes cultured in vitro for 12 to 14 h postcollection with insemination of the recipient 2 h postsurgery; 2) transfer of oocytes into the oviduct within...
Factors affecting pregnancy rates and early embryonic death after equine embryo transfer. In the present study, 638 embryo transfers conducted over 3 yr were retrospectively examined to determine which factors (recipient, embryo and transfer) significantly influenced pregnancy and embryo loss rates and to determine how rates could be improved. On Day 7 or 8 after ovulation, embryos (fresh or cooled/transported) were transferred by surgical or nonsurgical techniques into recipients ovulating from 5 to 9 d before transfer. At 12 and 50 d of gestation (Day 0 = day of ovulation), pregnancy rates were 65.7% (419 of 638) and 55.5% (354 of 638). Pregnancy rates on Day 50 were significantl...
Vitrification of immature and mature equine and bovine oocytes in an ethylene glycol, ficoll and sucrose solution using open-pulled straws. Studies were conducted to compare viability of immature and mature equine and bovine oocytes vitrified in ethylene glycol. Ficoll using open-pulled straws. Oocytes from slaughterhouse ovaries (N=50/group) with >2 layers of compact cumulus cells were vitrified immediately after collection (immature groups) or vitrified after 36 to 40 (equine) or 22 to 24 (bovine) h of maturation (mature groups). Immature oocytes were matured after thawing. Before vitrification, oocytes were exposed to TCM-199 + 10 FCS + 2.5 M ethylene glycol + 18% Ficoll + 0.5 M sucrose (EFS) for 30 sec and then to 5 M ethyl...
Insemination of mares with low numbers of either unsexed or sexed spermatozoa. Two experiments were conducted to determine pregnancy rates in mares inseminated 1) with 5, 25 and 500 x 10(6) progressively motile spermatozoa (pms), or 2) with 25 x 10(6) sex-sorted cells. In Experiment 1, mares were assigned to 1 of 3 treatments: Group 1 (n=20) was inseminated into the uterine body with 500 x 10(6) pms. Group 2 (n=21) and Group 3 (n=20) were inseminated into the tip of the uterine horn ipsilateral to the preovulatory follicle with 25 and 5 x 10(6) pms, respectively. Mares in all 3 groups were inseminated either 40 (n=32) or 34 h (n=29) after GnRH administration. More mares ...
Echotextural changes in the follicular wall during follicle deviation in mares. The echotextural changes occurring in the follicular wall in association with deviation in diameters were studied in 8 pony mares. Echotextural changes could be useful as a reference point for future studies of the follicle-selection phenomenon. Follicles were examined daily by transrectal ultrasound from 3 d before to 3 d after the beginning of deviation (Day 0). The following echotextural end points were recorded based on a scoring or percentage system: 1) thickness of granulosa, 2) echogenicity of granulosa, 3) prominence of an anechoic layer located beneath the granulosa, and 4) extent of ...
Effectiveness of two systems for transporting equine semen. The storage and transport of cooled, liquid semen is an effective way of facilitating the use of desirable stallions for breeding mares located on distant farms. The Equitainer System is the most widely used transport container and it has been shown that it is possible to ship semen in this container and obtain good conception rates. However, the cost of Equitainers is high, and stud-farms that ship large quantities of semen have tended to rely on cheaper alternatives, even though little documentation exists concerning their reliability, especially under extreme temperature conditions. Two dif...
Effect of mare’s age and recovery methods on the recovery rate of equine follicular oocytes for IVM procedures. Mares (n = 39) were classified according to age as young (less than 1.5 yr, n = 17) or old (more than 1.5 yr, n = 22) and sacrificed. Ovaries were measured and weighed, and the number of follicles and CL were counted. Follicle size and distribution were recorded (external: > 20 mm, 5 mm, < 5 mm). External follicles were aspirated while internal follicles were sliced. The number and Type of oocytes recovered using each method were recorded. Oocyte recovery rates (oocytes/ovary) resulted in a mean of 0.92 oocytes by aspiration and 1.36 oocytes by additional slicing. The mean numbers of av...
Production of embryos by assisted reproduction in the horse. In vitro embryo production is not yet successful in the horse, largely due to low rates of fertilization in vitro. However, methods to produce embryos from isolated oocytes have been developed. Oocytes may be recovered from living mares by aspiration of the dominant preovulatory follicle by trans-abdominal puncture, and from both preovulatory and immature follicles by trans-vaginal ultrasound-guided puncture. Transfer of in vivo-matured oocytes to the oviducts of bred recipient mares has resulted in good pregnancy rates (75-85%). Little work has been done on transfer of horse oocytes matured i...
Effects of melatonin implants in pony mares. 2. Long-term effects. The effects of melatonin implant treatment over a 4 wk period at the summer solstice on the transition into and out of the following anovulatory season were evaluated in ovary-intact and ovariectomized mares. Melatonin implants tended to delay the timing of the final ovulation of the breeding season (P = 0.0797) in the ovary-intact mares. Although the decline in LH secretion associated with the end of the breeding season was parallel between treatments and ovarian statuses, the rate of LH secretion, as expressed by its mathematical accumulation, was lower in ovariectomized, melatonin-treated m...
Effect of insemination dose on pregnancy rate in mares. Different insemination doses have been used for artificial insemination(AI) in horses. Since the insemination dose can affect the pregnancy rate, it is important to ensure that an adequate dose be used regardless of the type of inseminationprotocol used. The aim of this study was to find out if it is possible to decrease the insemination dose from 500 x 10(6) progressively motile spermatozoa to 300 x 10(6) progressively motile spermatozoa and still maintain an acceptable pregnancy rate when using extended fresh semen. Thirteen stallions of known fertility and a well-defined group of 64 mares w...
Effect of spermatozoal concentration and number on fertility of frozen equine semen. Information on the number of motile spermatozoa needed to maximize pregnancy rates for frozen-thawed stallion semen is limited. Furthermore, concentration of spermatozoa per 0.5-mL straw has been shown to affect post-thaw motility (7). The objectives of this study were 1) to compare the effect of increasing the concentration of spermatozoa in 0.5-mL straws from 400 to 1,600 x 10(6) spermatozoa/mL on pregnancy rate of mares, and 2) to determine whether increasing the insemination dose from approximately 320 to 800 million progressively motile spermatozoa after thawing would increase pregnancy r...
Reproductive parameters of miniature stallions. Breeding soundness evaluation (BSE) of stallions is a routine component of stud farm practice. Guidelines for assessing satisfactory breeding potential have been developed using data derived from stallions of full-size breeds. In view of the increasing popularity of miniature stallions, knowledge of normal semen parameters of these stallions is important. Therefore, testicular measurements and semen parameters from 216 sexually rested miniature stallions were obtained. Semen was collected twice, 1.5 to 3 h apart, using an artificial vagina. Values were averaged over the 2 collections because o...
Changes in intrauterine pressure after oxytocin administration in reproductively normal mares and in those with a delay in uterine clearance. Intrauterine pressure was measured in 4 reproductively normal mares and 4 mares with delay in uterine clearance after administration of oxytocin to determine if intrauterine pressure varied between dosage and group. Changes in intrauterine pressure were measured during estrus, when a follicle was > or =35 mm, using a Millar "Mikro-tip" catheter that had 3 discrete pressure sensors/channels. Mares received 4 different treatments of 10, 5, 2.5 or 0 IU (vehicle) of oxytocin. The protocol for each treatment consisted of a 10-min baseline recording, administration of treatment and measurement of...
Influence of epidermal growth factor on in vitro maturation of equine oocytes. The effect of epidermal growth factor (EGF) on the in vitro maturation rate of equine oocytes was examined. Oocytes were collected from an abattoir (Expt 1) or using ultrasound-guided follicular puncture in vivo (Expt 2). All oocytes with a compact or expanded cumulus at recovery were cultured for 30 h in: medium 1 (TCM199 + fetal calf serum (FCS) + crude equine gonadotrophin (CEG) + oestradiol + antibiotics); medium 2 (TCM199 + EGF); medium 3 (medium 1 without FCS + EGF); or medium 4 (medium 1 without CEG + EGF). In Expt 1, 84% (37/44) and 87% (40/46) cumulus expansion (P > 0.05), and 39% ...
Oviductal insemination of mares. A technique was developed for oviductal insemination of mares, in which a small number of motile spermatozoa are deposited directly into the oviduct. Pregnancy rates in mares inseminated by traditional intrauterine artificial insemination were compared with rates in mares inseminated by oviductal insemination. Fifteen mares were inseminated with 5 x 10(8) progressively motile spermatozoa by intrauterine artificial insemination, and 14 mares were inseminated with 5 x 10(4) progressively motile spermatozoa by oviductal insemination. Pregnancy rates in mares inseminated by intrauterine artificial...
Dopamine antagonist-induced reproductive function in anoestrous mares: gonadotrophin secretion and the effects of environmental cues. The effect of the dopamine antagonist sulpiride on FSH secretion and onset of reproductive activity in anoestrous mares under different environmental conditions was investigated. In Expt 1, sulpiride (0.5 mg (-)-sulpiride kg(-1) twice a day) had no affect on FSH pulse frequency, mean FSH concentration, basal FSH concentration or FSH pulse amplitude in anoestrous mares. These data do not support the hypothesis that dopamine inhibits reproductive activity by suppressing GnRH secretion, as it does in other species. In Expt 2, the interval to first ovulation (14.8 +/- 1.9 days; range 12-22 days) i...
Effect of L-glutamine for freezing equine embryos: evaluation by DAPI staining and transfer of multiple embryos to recipient mares. 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...
Ovulation synchrony after follicle ablation in mares. Two experiments were performed to determine the efficacy of ultrasound-guided transvaginal follicle ablation for synchronizing ovarian function in mares. The experiments were initiated at random stages of the oestrous cycle in control (nonablated) and follicle-ablated mares. On day 0, all follicles > or =10 mm in diameter were punctured, aspirated and curettaged in ablated mares, and, on day 4, two doses of PGF2alpha were administered with a 12 h interval between the doses to both ablated and nonablated (control) mares. In Expt 1, hCG was administered to the ablated mares on the first or se...
Effect of increased daylight during late pregnancy on the reproductive performance of mares after parturition. The aim of the present study was to determine the effect of prolonged photoperiod during late pregnancy on subsequent ovarian activity and fertility in mares. Pregnant mares (n=13) due to give birth in January and February were stimulated by a fixed photoperiod (16 h light: 8 h dark) from 15 November (during the last 2-3 months of gestation) until up to 1 month after parturition. A control group of mares (n=9) due to give birth at the same time were kept in the same stable and management regimen, but under natural light conditions. Light-treated mares ovulated during foal oestrus approximately...
Equine reproductive pharmacology. This article reviews therapies and strategies commonly used to treat diseases of the mare's genital tract and modulate the reproductive cycle of the mare. Many reproductive treatments are based on historical use and empirical evidence rather than well controlled clinical studies. This article attempts to present practical information in a summary form while highlighting the need for continued research documenting the efficacy and safety of reproductive therapies.
Molecular cloning of cDNA for equine follistatin and its gene expression in the reproductive tissues of the mare. A cDNA clone encoding equine follistatin was isolated from an equine ovarian cDNA library. Out of 1.2 x 10(5) independent clones screened, one positive clone was isolated and its cDNA sequence determined. The isolated clone, named EQ-FS-1, contained a complete open reading frame encoding 344 amino acid residues. The similarity of its deduced amino acid sequence to these of other mammalian species was greater than 95%. Although its expression level varied among the tissues examined, follistatin mRNA was detected in the equine uteroplacental tissues, follicles and corpora lutea by Northern blot ...
