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.
Effect of timing of frozen semen insemination on pregnancy rate in mares. 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 pregnan...
Recovery rate and quality of embryos from mares inseminated at the first post-partum oestrus. 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]. 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 ...
[Embryo transfer in horses–current status and future perspectives]. 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. 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. 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. 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 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. 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]. 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. 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. 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. 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. 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...
Cryopreservation of equine oocytes by 2-step freezing. Immature equine oocytes were frozen-thawed with ethylene glycol (EG), 1,2-propanediol (PD) or glycerol (GL) in PBS and cultured to assess the rate of in vitro maturation (Experiment 1). Compact-cumulus oocyte complexes were collected from slaughterhouse ovaries and equilibrated for 10 min in the freezing medium containing 10% (V/V) cryoprotectant and 0.1 M sucrose. The 0.25-ml straws, loaded with 10 to 30 oocytes, were seeded at -6 degrees C and cooled to -35 degrees C at 0.3 degrees C/min before being plunged into liquid nitrogen. The straws were thawed rapidly in a 37 degrees C waterbath for...
The dangers of disease transmission by artificial insemination and embryo transfer. This review summarizes the major infectious diseases of the three major agricultural species (cattle, sheep and pigs) and horses, and presents the evidence for and against the possibility of infectious agents being transmitted between animals via the venereal route or by the use of semen or early embryos in commercial artificial insemination (AI) or embryo transfer (ET). Cattle feature most prominently in the widespread distribution of frozen semen, and national and international organizations have set out guidelines to work towards disease-free bull studs with semen free from potential pathog...
Development to blastocysts of one- to two-cell equine embryos after coculture with uterine tubal epithelial cells. Development of 1- to 2-cell in vivo fertilized equine embryos cultured with or without uterine tubal epithelial cells (UTEC) was studied. One- to 2-cell embryos (n = 26) were collected surgically from the uterine tubes of pony mares 1 day after ovulation. Four- to 8-cell embryos (n = 9) were collected 2 days after ovulation. Presumptive zygotes and 2-cell embryos were cultured with (n = 17) or without (n = 9) UTEC, and all 4- to 8-cell embryos were cocultured with UTEC as positive controls. Uterine tubal epithelial cells were used as cell suspensions within 2 weeks after initiation of cultures...
[Early sonographic diagnosis of twin pregnancy in mares and its reduction to a single pregnancy]. The frequency of double ovulation and the chances of twin pregnancy in the mare are presented with reference to the relevant literature. A novel manual technique for reducing the embryo number from two to one is introduced and illustrated by ultrasonographic images.
Use of progesterone in microspheres for maintenance of pregnancy in mares. Administration of progesterone in poly(d-,l-lactide) microspheres was used to maintain pregnancy in mares after luteolysis was induced by treatment with prostaglandin F2 alpha at day 14 of pregnancy. Mares were given vehicle only (control, n = 6) or 0.75 g (n = 7), 1.5 g (n = 8), or 2.25 g (n = 5) of microencapsulated progesterone at days 12 and 22 of pregnancy. Serum progesterone concentrations were determined daily, and pregnancy was evaluated by transrectal ultrasonography on alternate days. Significantly (P less than 0.05) more mares given 1.5 or 2.25 g of progesterone (6 of 8 and 4 of 5 m...
Embryo-initiated oviductal transport in mares. The hypothesis that equine embryos initiate oviductal transport in mares was tested by placing day 6 uterine embryos in the oviducts of day 2 (n = 10) or day 5 (n = 10) recipient mares and attempting to collect the embryos from the uterus 48 h later. To determine whether the surgical transfer procedure initiated oviductal transport, medium alone was placed in the oviducts of day 2 (n = 10) inseminated mares (sham transfer), and uterine embryo collections were attempted 48 h later. Embryos were transported through the oviduct of day 2 recipients by day 4 (instead of day 5 to 6) in six of ten ma...
Relationships of age to uterine function and reproductive efficiency in mares. The uterine function and reproductive efficiency of 31 nonlactating pony mares were compared for two age groups: young (5 to 7 years, n=9) and old (>/=15 years, n=22). For pregnant mares, differences between age groups were not significant for the diameter of the largest follicle, cross-sectional area of the corpus luteum, growth profile of the embryonic vesicle or embryo mobility characteristics. Uterine contractility scores were lower (P<0.05), day of fixation of the embryonic vesicle was later (P<0.05), and uterine tone tended (P<0.10) to be lower in the old than the young mares...
Semen collection techniques. Semen collection techniques in the stallion have evolved considerably over the last 70 to 80 years and are used today primarily for artificial insemination. Semen can be collected from stallions that are otherwise unable to breed, allowing continued use of valuable animals. There are many options for collection of semen from stallions that present with ejaculatory dysfunction (see the article by McDonnell elsewhere in this issue.) Although there are many advantages to the use of artificial breeding, the collector must understand each step of the collection procedure as well as stallion prefere...
Artificial insemination and preservation of semen. Artificial insemination is an effective technique for improving utilization of stallions in breeding programs. When proper semen handling and insemination procedures are used, optimal pregnancy rates are attainable. When AI techniques are employed for mares and stallions with marginal fertility, pregnancy rates may be improved in comparison with natural mating. Preservation of stallion semen in the liquid or frozen state reduces the costs and potential health hazards incurred by transporting mares and provides easier access to genetic material that may otherwise be unavailable. Acceptable preg...
Common horse sense. This research article corrects a common misconception about the energy metabolism in horses during short sprinting and long-distance running events, emphasizing that short sprints are primarily powered by anaerobic activity, […]