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Topic:Fertility
Fertility in horses encompasses the physiological processes and factors influencing reproductive success in equine species. It involves the study of reproductive anatomy, endocrinology, and behavior, as well as the management practices that affect breeding outcomes. Key aspects include the estrous cycle, conception rates, and factors impacting stallion and mare fertility. Reproductive technologies such as artificial insemination, embryo transfer, and hormonal therapies are also explored to enhance breeding efficiency. This page compiles peer-reviewed research studies and scholarly articles that examine the biological mechanisms, management strategies, and technological advancements related to equine fertility.
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...
Post-thaw motility and longevity of motility of imipramine-induced ejaculates of pony stallions. Imipramine-induced ex copula ejaculates (11) and fractionated in copula ejaculates were collected from each of 5 pony stallions for freezing in 5-ml straws (6), using a modified Kenney glucose skim-milk extender (2). Initial post-thaw total and progressive motilities and daily post-thaw total and progressive motilities, as well as the number of days to reach 0 progressively motile spermatozoa, were also similar for the 2 methods of collection. The percentage of morphologically normal spermatozoa both before freezing and after thawing were also similar for in copula and ex copula ejaculates. Co...
Preservation of ejaculated and epididymal stallion spermatozoa by cooling and freezing. The suitability of ejaculated and epididymal stallion spermatozoa for cooled storage (5 degrees C) and cryopreservation was examined in 5 ejaculates from each of 6 stallions and in spermatozoa recovered from the cauda epididymidis after castration of these stallions. The percentage of progressively motile spermatozoa, examined by subjective estimation (cooled samples) or by computerized analysis (frozen-thawed samples), was used as parameter. In ejaculated semen samples containing 5 and 25% seminal plasma in a skim milk glucose extender, the lower amount of seminal plasma supported spermatozoa...
Inhibin activity in the mare and stallion. An overnight double antibody RIA, employing a rabbit antiserum raised to bovine 31 kDa inhibin (rAs-#1989, NICHD) and purified bovine 31 kDa inhibin (bINH-I-90/1, NICHD) as trace and standard, was validated to measure immunoreactive inhibin (iINH) concentrations in equine peripheral plasma, follicular fluid (FF), ovarian vein (OV) plasma, testicular tissue extracts (TTE) and testicular vein (TV) plasma. The dynamic relationship of iINH and follicle stimulating hormone (FSH) was investigated during the estrous cycle of the mare and the annual reproductive cycle of the stallion. In the RIA, para...
Culture of equine embryos in media containing egg yolk, mare’s milk and saline: Preliminary results. A medium containing egg yolk, mare's milk and/or modified PBS was used to culture Day-8 to 8.5 equine blastocysts. Twenty-one variants of the medium containing different concentrations of the 3 components were prepared. Embryos were recovered nonsurgically and placed into the media at 37 degrees C for 24 h. A total of 45 embryos was cultured; of these 7 died in culture and 13 showed inadequate development at the onset, while 25 continued to grow in the media. It was established that embryos grew best in media containing 20 to 60% yolk, 20 to 60% mare's milk and/or 20 to 60% PBS. It was found e...
Fertilization rates in superovulated and spontaneously ovulating mares. Embryo recovery per ovulation has been shown to be lower in superovulated mares than in untreated controls. The objectives of this study were to 1) determine whether follicles stimulated with superovulatory treatment ovulate or luteinize without ovulation, 2) determine fertilization rates of oocytes in oviducts of superovulated and control mares, and 3) evaluate viability of early stage embryos from superovulated and control mares when cultured in equine oviductal cell-conditioned medium. Cyclic mares were randomly assigned to 1 of 2 groups (n=14 per group) on the day of ovulation (Day 0): Gro...
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...
[Cryopreservation trial with semen of purebred Arabian and Haflinger stallions in the Turkish national stud in Karacabey]. Within a German-Turkish university partnership deep freezing preservation of stallion semen was performed as a part project of the cooperation contract. In this study a modification of the introduced Makrotüb method was used for semen freezing. The investigated characteristics of fresh semen of the Arab stallions were in the normal range cited in the international literature. However, the semen data obtained from the Haflinger stallions were markedly and partially significantly in lower range than measured for the Arab stallions. This may reflect an incomplete adaptation process of the import...
Automated morphometric analysis of stallion spermatozoa. Tissue variation in microscope slides made for spermatozoon analysis and variation introduced by the subjective techniques used to analyze these slides reduce the statistical power of studies that seek to use spermatozoon morphology to predict fertility. A simple specimen preparation method was developed to standardize stallion spermatozoon morphologic smears, and a new, automated spermatozoa morphometry instrument was used to objectively analyze the efficacy of the specimen preparation technique. The method achieved a standard spermatozoon concentration and reduced field-to-field variation in...
[Estrus synchronization and insemination results in Arabian mares in the Eskisehir stud]. An Arab broodmare herd of 30 mares age ranging from 4 to 12 years was grouped into 3 groups (group A, B, C) with 10 mares each and exposed to three different methods of cycle synchronisation. Group A mares received two PGF2 alpha injections 14 days apart. Group B mares got additionally 2,500 IU of hCG 5 days after second prostaglandin application. Group C consisted of mares which had foaled in a synchrony of one week and got a PGF2 alpha injection 3 weeks after parturition. All the mares responding to synchronisation were inseminated closely to ovulation detected by serial rectal palpations wi...
In vitro maturation of equine oocytes collected by follicle aspiration and by the slicing of ovaries. The aim of this study was to examine 2 techniques for oocyte recovery from equine ovaries at slaughter: by aspiration of follicles and by additional slicing of ovaries. The morphology and nuclear configuration of oocytes recovered with either technique, and the time course of nuclear maturation during in vitro maturation were evaluated. Recovery rates were 1.75 and 4.14 oocytes per ovary for aspiration and slicing (total 145 and 344 oocytes from 83 ovaries), respectively. The oocytes were classified according to their cumulus/ooplasm morphology into 4 groups: compact/circular(A), compact/semic...
Effect of constant administration of a gonadotropin-releasing hormone agonist on reproductive activity in mares: preliminary evidence on suppression of ovulation during the breeding season. During the breeding season, the effect of constant administration of an agonist analog of gonadotropin-releasing hormone (GnRH; goserelin acetate) on reproductive activity of mares was determined. Twenty-four mares undergoing estrous cycles were allocated at random to 6 groups (n = 4/group) and, on May 29 (day 0), received no treatment (group 1, controls), 120 micrograms (group 2), 360 micrograms (group 3), 600 micrograms (group 4), or 1,200 micrograms (group 5) of GnRH agonist/d for 28 days via a depot implanted subcutaneously. The final group of mares (group 6) was treated with 120 microgram...
Oviductal and uterine influence on the development of Day-2 equine embryos in vivo and in vitro. The objective of this experiment was to contrast the influence of the oviductal and uterine environments on development of Day-2 embryos. Embryos were transferred to oviducts or uteri of synchronous recipient mares, or were incubated in oviductal co-culture, in uterine co-culture or in defined culture medium. Significantly more (P < 0.02) embryos transferred to the oviduct versus the uterus survived until Day 11 after ovulation (5 7 vs 0 7 , respectively). Significantly more (P 0.1) in oviductal co-culture versus uterine co-culture (3 7 vs 6 7 , respectively), or in oviductal co-culture ve...
Effects of neodymium:yttrium aluminum garnet laser irradiation on endometrium and on endometrial cysts in six mares. Effects of neodymium:yttrium aluminum garnet (Nd:YAG) laser irradiation on equine endometrium were evaluated in vitro and in six mares with endometrial cysts. The Nd:YAG laser was applied to six endometrial sites, in each of five uterine specimens, with power densities of 5659 to 33,954 J/cm2. Depth of tissue ablation was measured and graded on histologic sections of the tissue lesions. Power density had a significant effect on the depth of tissue ablation (p < .001). Grade 3 lesions (full-thickness ablation of the endometrium) were created with energy densities of 16,977 to 33,954 J/cm2. Six ...
In vitro interaction between oviduct epithelial and equine sperm. Coculture of stallion sperm with monolayers of equine oviductal epithelial cells (OEC) was evaluated. Monolayers were obtained from frozen-thawed OEC. Live sperm attached to the OEC in vitro, whereas sperm killed by heat treatment or glutaraldehyde fixation did not. Sperm attached to OEC showed flagellar motion for 4 d in vitro, during which time they gradually became released. Scanning electron-micrographs showed an intimate association between the sperm and OEC. Incubation of sperm for 4 h with either control, heparinized or OEC-conditioned medium (Tyrode's albumin lactate phosphate) resulte...
Growth of small follicles and concentrations of FSH during the equine oestrous cycle. The temporal associations between increases in FSH and growth of small follicles (2-20 mm) were studied during one oestrous cycle (ovulation to ovulation) in 15 horse mares. Follicular diameters were obtained ultrasonically. For each day, follicles were combined for both ovaries, grouped from largest to smallest (excluding dominant follicles), and divided into three to five tiers of six follicles for each mare (for example: tier 1, six largest follicles; tier 2, next six largest follicles). A significant increase in mean diameters followed by a significant decrease was used to define a follicu...
Capacitation-like membrane changes and prolonged viability in vitro of equine spermatozoa cultured with uterine tube epithelial cells. Reliable capacitation of equine spermatozoa has been a major obstacle in the development of equine in vitro fertilization. Experiments were done to compare in vitro capacitation of equine spermatozoa by use of heparin/caffeine, calcium ionophore, uterine tube epithelial cell (UTEC)-conditioned medium, and direct culturing of spermatozoa with UTEC (coculturing). Capacitation-like changes, as determined by chlortetracycline membrane staining patterns, developed with UTEC-conditioned medium and coculturing, equivalent to that with calcium ionophore. Both of these treatments induced more (P < 0.05...
Reproductive performance of thoroughbred mares on six commercial stud farms. The records of 1630 mare years from 6 Thoroughbred stud farms in south eastern Australia were analysed for the years 1981 to 1986. Overall pregnancy and foaling rates were 83.9% and 69.3%, respectively. When calculated per served oestrous cycle, pregnancy and foaling rates were 54.7% and 43.1%, respectively. Pregnancy and foaling rates were higher (P < 0.001) for mares 3 to 10 years of age than for older mares. There was no difference in the pregnancy rates of maiden, barren and foaling mares. The foaling rate was significantly higher (P < 0.001) in mares that became pregnant during the ...
Steroid synthesis by equine conceptuses between days 7 and 14 and endometrial steroid metabolism. The objective of this study was to determine if changes in steroid synthesis occurred in the horse blastocyst about the time of maternal recognition of pregnancy. Embryos collected between days 7.5 and 14.5 were incubated for 8 hr in vitro in HAM's F10 containing radiolabelled pregnenolone. The steroid metabolites in the incubation medium were separated by reverse phase HPLC and the major peaks expressed as a percentage of total metabolites. It was found that there were no major changes in the profile of metabolites throughout the period of study, although there was increased conversion as the...
Ovarian follicles, ovulations and progesterone concentrations in aged versus young mares. The objectives of this study were: 1) to document age-related ovulation failure in mares and 2) to contrast the number of ovarian follicles, occurrence of ovulations, and postovulatory concentrations of progesterone in aged versus young mares. In Experiment 1, 4 of 10 aged (25- to 33-years-old) mares were anovulatory between July 1 and September 1, 1989. In Experiment 2, two of 25 aged (20- to 30-years-old) and none of 21 young (3- to 12-years-old) mares were anovulatory between February 1 and June 30, 1990. The average (+/- SEM) day of the first ovulation was later (P<0.05) for aged versus...
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...
Proceedings of the John P. Hughes International Workshop on Equine Endometritis. Davis, California, August 1992. The paper is a report from a workshop discussing equine endometritis, a condition affecting horse fertility. The event was held in honor of Professor John Hughes and his significant contributions […]
Binding of stallion spermatozoa to the equine zona pellucida after coculture with oviductal epithelial cells. The objective of this study was to determine whether coculture of stallion spermatozoa and mare oviductal (uterine tubal) epithelial cells induced sperm cell capacitation in vitro. Capacitation as determined by zona binding and chlortetracycline staining of the sperm cells was compared for stallion spermatozoa: (1) incubated with medium alone (negative control), (2) treated with calcium ionophore A23187 (positive control) or (3) cultured with mare oviductal epithelial cells (OEC) for 4 h. Chlortetracycline staining patterns of sperm cells bound to the zonae were used to group spermatozoa as un...
Serum levels of testosterone and oestrogens in normal and infertile stallions. The serum concentrations of testosterone and oestrogens were determined in stallions classified as geldings, normal (according to age) or infertile (azoospermic). There were significant differences in testosterone and oestrogen levels between the groups. Normal concentrations of testosterone and total oestrogens were attained after 16 months of age. Castrated and immature horses ( 4 years). The results suggest that serum levels of testosterone and total oestrogens may provide a sensitive index of the endocrine function of the testis in male horses.
Liquid storage and freezing of semen from New Forest and Welsh Pony stallions. Two experiments were conducted to examine the effects of liquid storage extender and of a modified freezing protocol on motility and morphology parameters of 3-year-old pony stallions. In experiment 1 ejaculates were diluted 1 + 1 (v+v) with glycine-egg-yolk extender (D11) or skim milk extender (SME), centrifuged, resuspended in the corresponding extender and kept at +5 degrees C. Concerning motion characteristics, both progressive motility and average path velocity of semen stored in SME was significantly superior to semen stored in D11 after 6, 18 and 42 hrs. However, over time of storage th...
Column separation of motile sperm from stallion semen. Subfertility in stallions is common, and methodologies are needed to increase the fertility in these animals. In other species, removal of the dead sperm from semen increases the quality and fertility of semen. With horse semen we evaluated 48 combinations of column separation techniques using micro-spin chromatography columns. The greatest improvement in motility was observed with glass wool, whereas glass beads exhibited the greatest recovery of motile sperm. Although centrifugation time did not influence recovery rate or percent motility, a column length of 2 cm was superior for recovery of...
Effect of surgical removal of endometrial cups on concentrations of chorionic gonadotrophin and subsequent fertility in the mare. Seven pregnant mares underwent general anaesthesia, laparotomy, hysterotomy and removal of a 50-day conceptus. Eversion of the uterine horn through the hysterotomy site allowed direct visualisation and electrosurgical removal of endometrial cup tissue from 5 randomly selected mares (Nos 1-5), while cup tissue in 2 mares (Nos 6 and 7) was left intact. Two pregnant mares served as unoperated controls (Nos 8 and 9). Efforts to re-establish pregnancy were initiated 20 days after surgery. Serum samples collected before surgery and during the post-operative period were analysed for concentration of ...
