Reproduction in horses encompasses the biological processes and mechanisms involved in the breeding and development of equine offspring. This includes the study of reproductive anatomy, physiology, and endocrinology in both mares and stallions. Key areas of interest include the estrous cycle, ovulation, conception, gestation, and parturition. Researchers also examine factors influencing fertility, reproductive technologies such as artificial insemination and embryo transfer, and management practices that impact reproductive success. This page compiles peer-reviewed research studies and scholarly articles that explore the physiological, genetic, and environmental aspects of equine reproduction.
Brück I, Hyland JH.The contributions of 2 biochemical pathways to the total metabolism of glucose (the Embden-Meyerhof pathway [EMP] and the pentose phosphate pathway [PPP]), were assessed for equine embryos recovered on Day 4.5, 7.5 and 11.5 post ovulation. At all developmental stages studied, glucose was metabolized through both pathways. Through the EMP, the amounts of glucose metabolized per nl embryo volume per hour were 4.0, 9.9 and 3.1 pmol, whereas via the PPP, amounts were 0.9, 1.7 and 0.07 pmol for Day-4.5, -7.5 and -11.5 embryos, respectively. The ratio of EMP:PPP with age was 9.7 for Day -4.5 embryos...
LeBlanc M, Ward L, Tran T, Widders P.A direct ELISA was used to measure immunoglobulin (Ig) isotypes G, Gt, A, and M recognizing Streptococcus zooepidemicus epitopes in uterine lavage fluids collected during the early post ovulatory period. A S. zooepidemicus isolate, used as the plate antigen in this assay, was inoculated into the uteri of 8 mares (3 resistant and 5 susceptible to endometritis) at oestrus prior to ovulation during Oestrous Cycles 1, 3 and 5. Resistant mares aged 2-5 years were nulliparous, with clinically normal reproductive tracts as determined by physical examination, bacteriological culture of the uterus, and...
Lucas Z, Raeside JI, Betteridge KJ.Field observations of 400 totally unmanaged feral horses on Sable Island, Nova Scotia, were complemented by oestrogen determinations in faecal samples from 154 identified females over a 4-year period (454 mare-years). Of mares that were sampled throughout the year and subsequently produced foals, 92.1% exhibited elevated faecal oestrogens between 15 October and 30 March. The results confirm that faecal oestrogens are a useful indicator of pregnancy after approximately 120 days gestation. Distribution of foaling resembled that seen in other feral populations, with 95% of births occurring from A...
Boyle MS, Skidmore J, Zhang J, Cox JE.The effect of long-term treatment of stallions with a powerful gonadotrophin-releasing hormone (GnRH) analogue was investigated. In the first part of the study, 9 sexually mature pony stallions were allocated to 1 of 3 groups, each of which was treated with 240 micrograms or 60 micrograms Buserelin per day administered by subcutaneously sited osmotic pumps or 30-50 micrograms Buserelin per day via solid, slow-release implants injected subcutaneously. Peripheral blood plasma and serum samples were collected frequently and assayed for follicle-stimulating hormone (FSH) and luteinizing hormone (L...
Tucker KE, Henderson KA, Duby RT.Twenty-three follicles were collected from 14 mares on specific days and grouped to represent follicles from early (Group 1; n = 6), mid (Group 2; n = 11) and late (Group 3; n = 6) oestrus, as described previously (Tucker et al., 1988). Isolated granulosa cells (GC) from each follicle were cultured in multiwell plates containing either Eagle's Minimum Essential Medium (MEM) alone, eLH (300 ng/ml), eFSH (300 ng/ml) or eLH + eFSH (300 ng/ml each), in the presence or absence of 0.5 microM testosterone. Media were collected and replaced at 24 h of culture, and 24 h later, media were again collecte...
Koskinen E, Kurki E, Katila T.Sixty-four mares (27 foaling, 37 barren or maiden), mainly Finnhorses, were subjected to treatment with 14.5 h of light and 9.5 h of darkness, starting at the beginning of December. The onset of cycling in non-foaling mares was estimated by weekly serum progesterone determinations. All of the non-foaling mares cycled in the middle of March. They started to cycle on average in the middle of February, 11.1 weeks after the beginning of the trial. There were statistically significant differences in relation to breed (Finnhorses started to cycle 2 weeks later than warm blooded, p less than 0.02) an...
Schwarzenberger F, Möstl E, Bamberg E, Pammer J, Schmehlik O.Faecal samples were collected at weekly intervals from pregnant Lipizzan mares during Weeks 7-16 following mating and from Lipizzan, Trotter and Thoroughbred mares during the last 3 months of gestation. After parturition, samples were taken daily from the Thoroughbred mares for another 6 days. Non-pregnant mares served as controls. The concentrations of unconjugated oestrogens (Eg), 20 alpha-OH-progestagens (20 alpha-G) and 20 beta-OH-progestagens (20 beta-G) were measured by enzyme immunoassay. In the faeces of Lipizzan mares, immunoreactive progestagens were significantly (P less than 0.01) ...
Rieger D, Bruyas JF, Lagneaux D, Bézard J, Palmer E.The decrease in embryo viability caused by cryopreservation may be due, in part, to metabolic disturbances. To determine the effect of cryopreservation on metabolism, Day -6.5 horse embryos were either frozen and thawed using glycerol as the cryoprotectant, given only the glycerol treatment or washed an equal number of times in phosphate buffered saline (PBS). Before and after treatment, individual embryos were incubated with L-[14C(U)]-glutamine, to measure Krebs cycle activity, and D-[5-3H]-glucose, to measure Embden-Meyerhof pathway activity. Before treatment, glucose metabolism ranged from...
Cleaver BD, Grubaugh WR, Davis SD, Sheerin PC, Franklin KJ, Sharp DC.Melatonin is thought to play a role in relaying photic information to the central nervous system as part of the seasonal reproductive cycle of the mare. However, the mechanisms by which melatonin may act are unknown. Therefore, this study was designed to determine whether exposure to constant light would, by reducing circulating melatonin concentrations, have any effect on hypothalamic gonadotrophin-releasing hormone (GnRH) content and circulating levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Blood samples were collected for 12 h at 15-min intervals from 8 ovariect...
Tomanelli RN, Sertich PL, Watson ED.Receptors for progesterone and oestrogen were measured in cytosol extracted from the endometrium of mares. Samples of endometrium were obtained from mares during the oestrous cycle and during other physiological and pathological states, and from mares with persistent endometritis. The concentration of binding sites was high around oestrus and early in dioestrus and was low by mid- to late dioestrus. Numbers of receptors were low in mares sampled on Day 15 of pregnancy and in mares in prolonged dioestrus, whereas receptor concentrations in ovariectomized mares were high. In 2 mares with endomet...
Houghton E, Holtan D, Grainger L, Voller BE, Rossdale PD, Ousey JC.Radioimmunoassay (RIA) and gas chromatography-mass spectrometry (GC-MS) were used to determine plasma progestagen concentrations in the normal and premature foal. Radioimmunoassay provides a profile of plasma progestagens with respect to time but, due to the non-specific nature of the technique and without prior chromatographic purification, quantitative data based on RIA analysis must be interpreted with caution. In contrast, the greater specificity of GC-MS allows identification of specific plasma progestagens and measuring of multiple analytes in a single analysis. Both techniques demonstra...
Ball BA, Altschul M, McDowell KJ, Ignotz G, Currie WB.Research has indicated that trophoblastic vesicles (TV) formed from Day-14 equine conceptuses would prolong luteal maintenance in mares after surgical transfer to the uterus at Day 10 after ovulation. The current study assesses TV as a further model for maternal recognition of pregnancy in mares. The objectives of the study were to determine the ability of TV to prolong luteal maintenance in mares, their effect on endometrial production of prostaglandin F (PGF) in vitro, and their ability to secrete polypeptides in vitro. In contrast to our previous study (Ball et al., 1989b), transfer of TV f...
Varner DD, Forrest DW, Fuentes F, Taylor TS, Hooper RN, Brinsko SP, Blanchard TL.Eighteen fertile mares were used to determine the effects of the oestrous cycle and location in the reproductive tract on the amount and concentration of glycosaminoglycans in luminal fluids. Ovariohysterectomies were performed in 3 groups of 6 mares on Day 3 of behavioural oestrus, within 6 h after ovulation or on Day 8 of dioestrus. The lumina of the uterine horns and oviducts ipsilateral and contralateral to the active ovary were flushed and fluid was aspirated from the dominant follicle in the oestrous preovulation group. Glycosaminoglycans and protein concentrations were measured in these...
Rossdale PD, Ousey JC, Cottrill CM, Chavatte P, Allen WR, McGladdery AJ.Between 1985 and 1990, 25 Thoroughbred mares were classified as having an abnormal pregnancy on the basis of precocious signs of impending parturition. Throughout the remainder of their pregnancies, plasma progestagen and mammary secretion calcium concentrations were monitored. After parturition, evidence of placental pathology was recorded and their foals were assessed with regard to adrenocortical function and behaviour. Seven pony mares were used to establish a model of placental pathology and, following induced placental separation, were subjected to the same protocol. Control animals cons...
Love CC, Garcia MC, Riera FR, Kenney RM.The semen of 26 stallions, 2-20 years of age, was collected once a day for 7 consecutive days to determine daily sperm output (DSO). After the last collection, 17 stallions were castrated. Testicular volume was estimated using two methods. Length, width, height, of each testis were measured by caliper. Length, width, height, cross-sectional area and circumference at the widest point of the testis were measured by ultrasonography. Both caliper and ultrasound measurements were first made in the live animal, and again in vitro. There were no differences in these measurements because of method or ...
Hofferer S, Duchamp G, Palmer E.Twelve anoestrous mares were treated with an intravaginal sponge containing 0.5 g allyl trembolone (Regumate; Roussel UCLAF, Paris) and 50 mg oestradiol benzoate for 7 days, followed by daily intramuscular (i.m.) injections of 25 mg crude equine pituitary extract (CEG), with (n = 6) or without (n = 6) 0.25 mg porcine growth hormone (pGH). No difference in ovarian response to this superovulation treatment was observed between the 2 groups (2.2 +/- 0.4 vs 2.3 +/- 0.4 ovulations per mare, respectively). CEG treatment was then combined with allyl trembolone (40 mg per os per day) and prolonged in ...
Ainsworth CG, Hyland JH.Continuous infusion of gonadotrophin releasing hormone (GnRH) has been shown to induce oestrus and ovulation in experimental mares in seasonal anoestrus. The present study sought to apply this form of treatment to anoestrous mares subjected to normal management procedures on 15 commercial Thoroughbred studfarms in Australia. The GnRH was delivered via subcutaneously implanted osmotic minipumps (OMP) at dose rates of 100 ng/kg/h and 200 ng/kg/h. The ovaries of treated mares were palpated on the day of OMP insertion (Day 0) and blood samples were collected twice weekly from a proportion of mares...
Davis SD, Sharp DC.This experiment investigated steroid production by ovarian tissues, in vitro, of pony mares during vernal transition from anoestrus to the breeding season. Follicular dynamics were monitored to detect the first, second, third or fourth transition follicle, greater than or equal to 30 mm diameter or the first large post luteal follicle of the breeding season. Twenty-four hours after a large follicle was detected, theca (T) and granulosa (G) tissues were harvested. Separate and co-incubations of these tissues were conducted to determine steroid production in early transition (ET), late transitio...
Kydd JH, Butcher GW, Antczak DF, Allen WR.The expression of class I Major Histocompatibility Complex (MHC) molecules by early trophoblast of intraspecific horse and donkey, interspecific mule and extraspecific donkey-in-horse conceptuses was determined using a rat monoclonal antibody (MAC 291) in a peroxidase anti-peroxidase immunohistochemical technique. Most non-invasive allantochorion of horse, donkey and mule conceptuses did not express class I MHC molecules at any stage of gestation except in small isolated patches of pseudostratified trophoblast lying adjacent to the openings of endometrial glands. In contrast, MHC class I molec...
Turner JE, Irvine CH.Gonadotrophin-releasing hormone (GnRH) was used in several regimens in anoestrous mares in an attempt to investigate the effects of frequency and dose on follicle-stimulating hormone (FSH) and luteinizing hormone (LH) release, and consequently on ovulation and corpus luteum establishment. Thrice daily injections of GnRH induced development of pre-ovulatory follicles, but hourly injections or constant infusions were required to induce ovulation. Hourly injections induced a much higher LH:FSH ratio in the presence or absence of ovarian hormones. When anoestrous mares were given an implant that r...
Weitkamp LR, Kenney RM, Bailey E, MacCluer JW, Brown JS, Blanchard TL, Sertich PL, Love CC, Hunt PR.Histological features of the endometrium, as assessed in biopsy samples, were related to Standardbred mare genotypes for transferrin, esterase (as a control) and equine leucocyte antigens (ELA). Pathological changes were found more frequently in each successively older age group of mares. Among mares aged 6-19 years, there were significant pathologic changes on first examination following an infertile breeding season for 46 of 90 (51%) of transferrin homozygotes and 50 of 146 (34%) of transferrin heterozygotes. The difference between the two groups was significant for the total data (chi 1(2) ...
Blanc G, Magistrini M, Palmer E.Semen from three ejaculates from each of 4 stallions was frozen in liquid nitrogen. Morphology was evaluated by coating the spermatozoa with fluorescein-labelled Concanavalin A (FITC-ConA2) and motility was measured by computer-assisted image analysis. Coating was performed at each step of the freezing procedure (dilution, cooling, addition of glycerol and freeze-thawing) and observations were made after each step, to evaluate changes, or after subsequent steps, to determine protection provided by the coating method. All the parameters showed progressive changes during the freezing procedure. ...
Watson ED.Corpora lutea were recovered from mares either 4 to 5 days or 12 to 13 days after ovulation. Mixed populations of luteal cells were prepared by collagenase digestion and were incubated for 24 h in the presence or absence of prostaglandin (PG) F-2 alpha (250 ng/ml). PGF-2 alpha significantly (P = 0.03) reduced progesterone secretion by cells from late diestrous corpora lutea and tended (P = 0.06) to reduce secretion by early diestrous cells. PGF-2 alpha had no significant effect on leukotriene B-4 (LTB-4) production by cells from early diestrous corpora lutea, but significantly (P = 0.03) incre...
Cottrill CM, Jeffers-Lo J, Ousey JC, McGladdery AJ, Ricketts SW, Silver M, Rossdale PD.One hundred twenty-four horse placentas were evaluated according to gross appearance, weight and surface area. Grossly abnormal placentas were photographed, and histological sections were taken from abnormal appearing areas, and from representative areas (pregnant horn, non-pregnant horn, body and cervical pole) in both normal and abnormal placentas. Information about the resulting foals was obtained and correlations examined. Histological sections were reviewed and related to the resulting foals. Four normal fresh placentas were infused with dyes to ascertain the blood supply to various areas...
McCue PM, Troedsson MH, Liu IK, Stabenfeldt GH, Hughes JP, Lasley BL.Thirty-seven seasonally anoestrous mares were divided into treatment and control groups and given 10 micrograms of native GnRH (GnRH) per hour using a peristaltic pump, or 10 micrograms GnRH agonist (GnRHa) twice daily, beginning on either 13 January, 13 February or 14 March. Treatment with GnRH was equally effective in inducing ovulation in January (4/5), February (4/5) and March (3/4). GnRHa treatment was more effective in inducing ovulation in February (4/5) and March (4/4) than in January (2/8). Peak luteinizing hormone (LH) concentrations in mares induced to ovulate with GnRH (7.4 +/- 1.5...
Oriol JG, Donaldson WL, Dougherty DA, Antczak DF.Three monoclonal antibodies raised against equine trophoblast cells were tested to determine the characteristics of the identified molecules. First, the antibodies were used to precipitate molecules from radiolabelled equine trophoblast cells of the chorionic girdle. Antibody F71.1 precipitated a molecule of 115 kDa, whereas antibodies 71.8 and 71.10 precipitated a molecule of 66 kDa. Second, 2 of the antibodies were used in an indirect immunoperoxidase assay on frozen sections of equine conceptuses of different gestational ages beginning at Day 8. Antibody F71.1 labelled trophoblast cells fro...
Cope SE, Catts EP.Parahost behavior of adult Gasterophilus intestinalis (DeGeer) was observed during the summers of 1978 and 1979 near pastured horses on two farms near Newark, Del. Adult fly occurrence, although sparse in early summer, indicated a gradual increase, with three surges in emergence. In the laboratory, males lived 1-3 d and females 1-2 d. In the field, marked flies were not seen on days following their release. Adult behavior indicated an urgency in mating and oviposition dictated by a short life span. Male flight patterns were characterized by in-flight searches for females and intraspecific conf...
Baker CB, Adams MH, McDowell KJ.Horse conceptuses were collected on Days 13, 15, 20 and 25 after ovulation. Whole conceptuses (Days 13 and 15) or extra-embryonic membranes (Days 20 and 25) were homogenized and poly-adenylated RNA (poly A RNA) was isolated by binding to oligo (dT)-cellulose. Poly A RNA (1 microgram/well) was separated by size on a denaturing 1% agarose gel and blotted onto nitrocellulose filters (northern blotting). DNA probes were prepared from plasmids containing equine alpha 1, omega 1 and omega 2 interferons and human beta actin. The presence of messenger RNA (mRNA) was detected by specific hybridization ...
al-Bagdadi F, Hoyt P, Karns P, Martin G, Memon M, McClure R, McCoy D, Shoemaker S.Eleven unilateral cryptorchid stallions, two to three years old, were castrated at Louisiana State University Veterinary Teaching Hospital. Five of these cryptorchid cases were abdominal and the rest were inguinal. This study was initiated to document the differences between the abdominal and inguinal equine cryptorchid testes. Specimens were obtained from the abepididymal side of each cryptorchid testes and processed for light and electron microscopic study. The cryptorchid testes were smaller than the scrotal testes, with the abdominal testes being one-fourth the size of the scrotal testes. ...
Battut I, Bézard J, Palmer E.A culture for equine oviduct epithelial cells is described. Primary cultures reached confluence in 5-8 days, forming a monolayer of polygonal cells and remaining morphologically intact for about 20 days. Subcultures were obtained by collecting cells detached spontaneously from the monolayers, and confluence was reached again after 5-7 days. Cells frozen before primary culture were confluent 10-15 days after thawing. Dishes containing confluent cells also were frozen, and some cohesive monolayers formed after thawing. Equine embryos, collected 2 days after ovulation, were cultured alone or with...
Jalim SL, McKinnon AO.To evaluate the fertility of mares bred at various intervals relative to surgical management of rectovaginal fistula (RVF). Methods: Surgical repair of RVF was performed in 28 mares at variable times relative to foaling (30 days to 24 months) and also relative to rebreeding (same cycle or delayed). Postoperative fertility was then evaluated. Results: Two mares were already pregnant at the time of surgery and 20 of 23 mares (87%) that were bred immediately prior to or following surgery conceived from their first service. When mares were bred in the same cycle as surgery, the next cycle followin...
Ginther OJ, Baldrighi JM, Castro T, Wolf CA, Santos VG.In experiment 1, daily blood samples were available from Days 0 to 20 (Day 0 = ovulation) in mares with an interovulatory interval (IOI, n = 5) and in mares that developed idiopathic persistent corpus luteum (PCL, n = 5). The PCL was confirmed by maintenance of progesterone (P4) concentration until end of the experiment (Day 20). Significant interactions of group and day revealed the novel findings that luteinizing hormone (LH) was lower (P < 0.05) in the PCL group than that in the IOI group on Days 0 to 4, and prolactin was lower (P < 0.05) on Days 1, 4, 6, and 7. In experiment 2, treat...
Campbell DL, Douglas LW, Ramge JC.Siliconized rubber tubes were used to cannulate one oviduct in 7 mares, and secretions were collected in a polycarbonate container located externally, in the region of the left paralumbar fossa. Secretion rates were recorded daily during the estrous cycle. Concentrations of calcium, magnesium, sodium, potassium, inorganic phosphorus and glucose were determined in the oviduct fluids secreted throughout the estrous cycle. Secretion rates were greatest during estrus (days 1-9), with a significant decrease (P<.01) noted during nonestrus (days 10-21). Concentrations of all constitutents measured...
Hoopes KH, Gradil CM, Vanderwall DK, Mason HM, Sarnecky BA, Davies CJ.There is an urgent need for practical methods of population control (i.e., contraception and/or sterilization) for free-roaming (i.e., "wild" or "feral") horses and burros on Western Public Lands in the United States. The objective of this study was to evaluate the contraceptive efficacy of a novel self-assembling three-part polymer-coated magnetic intrauterine device termed as an intrauterine POD (self-assembling; iUPOD) when there are natural breeding conditions when iUPOD use was managed by veterinary professionals with no prior experience with the device. Six mares were administered an iUP...
Smith IM, Girard A, Corner AH, Mitchell D.Using two known positive equine viral rhinopneumonitis (EVR) sera, conjugates were prepared with fluorescein isothiocyanate and tested for specificity using EVR infected tissue culture cells. The conjugate was then applied to selected tissues from 32 aborted fetuses and foals submitted during a natural outbreak of EVR. Antigen was detected in various tissues by immunofluorescence in 20 cases (62.5%). In 24 cases bovine fetal kidney cell monolayers were inoculated with a pool of lung and liver and EVR virus was isolated from 15 (62.5%). Histological examination of various tissues from 29 cases ...
Hannan MA, Murata K, Takeuchi S, Haneda S, Cheong SH, Nambo Y.The objectives of this study were to determine the plasma profile of equine chorionic gonadotropin (eCG) and its association with the formation of supplementary corpus luteum (CL) and plasma progesterone concentrations in embryo transfer Hokkaido native pony recipient mares. Blood samples and transrectal ultrasound examination of the reproductive tract were carried out weekly from the day of ovulation until week 32 of gestation (n = 4). Plasma concentrations of eCG and progesterone were measured by enzyme immunoassays. The eCG concentration was first detectable at week 5 for 2 mares and at wee...
da Silva CMB, Cano FEM, Gaitskell-Phillips G, Vega FJP.Flow cytometry is a powerful tool for the analysis of cell samples formed of multipopulations, such as spermatozoa. In recent years, multiparametric cytometers have evolved, allowing the study of different cellular characteristics, such as protein expression, DNA analysis, or mitochondrial activity. Whether using traditional fluorescent dyes or fluorophore-conjugated antibodies, each cell or cellular component is individually stained, the sample is analyzed at high velocities, and then is displayed and interpreted in a dot-plot. We hereby describe the procedure to perform a multiparametric flo...
Ginther OJ.Equine embryo mobility and cessation of mobility (fixation) provide explanations to several enigmas in reproductive biology of the pregnant mare and provide an efficient solution to the twinning problem, the bane of brood-mare owners. Embryo mobility is maximum on Days 12 to 15 (Day 0 = ovulation) while the spherical embryo is growing from 9 to 23 mm in diameter. During mobility, the embryo can be anywhere in the uterine lumen regardless of side of ovulation. Mobility solved the enigmas of how a small embryo can block luteolysis in a relatively massive uterus and why the side of ovulation do...
Alim S, Cheppali SK, Laitaoja M, Talluri TR, Jänis J, Swamy MJ.Fibronectin type-II (FnII) family proteins are the major proteins in many mammalian species including bull, horse and pig. In the present study, a major FnII protein has been identified and isolated from donkey (Equus hemionus) seminal plasma, which we refer to as Donkey Seminal Plasma protein-1 (DSP-1). The amino acid sequence determined by mass spectrometry and computational modeling studies revealed that DSP-1 is homologous to other mammalian seminal plasma proteins, including bovine PDC-109 (also known as BSP-A1/A2) and equine HSP-1/2. High-resolution LC-MS analysis indicated that the prot...
Holyoak GR, Ma A.Historical evidence of acupuncture predates written history. It has been a component of Traditional Chinese veterinary medicine for many generations and is officially recognized in recorded history for treating equine disease in the Zhou Dynasty, circa 1050 BC. Drawing from a range of searchable databases, we present the use of veterinary acupuncture related to theriogenology. We touch on human-based medicine only as an introduction to current uses within veterinary medical acupuncture. This review is confined to the use of acupuncture encompassing dry needle, electroacupuncture, aquapuncture,...
Atkins DT, Harms PG, Sorensen AM, Fleeger JL.5alpha-pregnane-3,20-dione was isolated from pooled pregnant mare serum using Sephadex LH-20 column chromatography and identified by the use of radioimmunoassay, gas-liquid chromatography and gas-liquid chromatography-mass spectrometry analyses. 5beta-pregnane-3,20-dione was not cross-reactive with the radioimmunoassay system and was not detected by gas-liquid chromatography. Peripheral blood levels of progesterone and 5alphs-pregnane-3,20-dione were determined by radioimmunoassay in four Quarter Horse mares for the first 150 days of gestation. Progesterone and 5alpha-pregnane-3,20-dione decli...
Cochran R, Meintjes M, Reggio B, Hylan D, Carter J, Pinto C, Paccamonti D, Graff KJ, Godke RA.In vitro fertilization in horses has been less successful than anticipated owing to: (i) the inability to collect large numbers of good quality oocytes; (ii) alterations in the zona pellucida that occur during in vitro maturation of equine oocytes; and (iii) inadequate preparation of equine sperm cells. In addition, studies in humans, mice and cattle have indicated that high concentrations of glucose in culture media may inhibit embryonic development in vitro and this may also be a problem for development of equine embryos in vitro. The aims of the present study were: (i) to achieve fertilizat...
Álvarez C, González N, Luño V, Gil L.The aim of this study was to evaluate the effect of trehalose and lactose extenders on ejaculated and epididymal stallion sperm vitrification. Ejaculated semen samples were collected from seven fertile stallions, and cauda epididymis samples were collected from ten stallion carcasses after slaughter. Both the ejaculated and the epididymis samples were diluted and vitrified using INRA 96® and bovine serum albumin as well as trehalose or lactose. As a control, ejaculated and epididymal samples were collected and frozen using the conventional method. Vitrification was performed by immersing sper...
The Kushum breed is one of the top breeds in the structure of horse breeds in the Aktobe region. In terms of numbers, it ranks third after the Kazakh and Mugalzhar breeds. The Kushum breed in the Aktobe region compares favorably with a number of horse-draft factory breeds in terms of early maturity, fertility, high meat, milk productivity, and economy of keeping. This article describes a new intra-breed type, the "Mamyr-Aktobe" type of the Kushum horse breed of the Aktobe population. The structure of the intra-breed type of Mamyr-Aktobe of the Kushum breed includes three factory lines of stall...
Mugnier S, Boittin S, Douet C, Monget P, Magistrini M, Goudet G.In human and rodents, sperm-zona pellucida binding is mediated by a sperm surface Galactosyltransferase that recognizes N-Acetylglucosamine residues on a glycoprotein ZPC. In large domestic mammals, the role of these molecules remains unclear: in bovine, they are involved in sperm-zona pellucida binding, whereas in porcine, they are not necessary. Our aim was to clarify the role of Galactosyltransferase and N-Acetylglucosamine residues in sperm-zona pellucida binding in ungulates. For this purpose, we analyzed the mechanism of sperm-zona pellucida interaction in a third ungulate: the horse, si...
Gimeno BF, Bariani MV, Laiz-Quiroga L, Martínez-León E, Von-Meyeren M, Rey O, Mutto AÁ, Osycka-Salut CE.Cryopreservation by negatively affecting sperm quality decreases the efficiency of assisted reproduction techniques (ARTs). Thus, we first evaluated sperm motility at different conditions for the manipulation of equine cryopreserved spermatozoa. Higher motility was observed when spermatozoa were incubated for 30 min at 30 × 106/mL compared to lower concentrations (p < 0.05) and when a short centrifugation at 200× g was performed (p < 0.05). Moreover, because sperm suitable for oocyte fertilization is released from oviduct epithelial cells (OECs), in response to the capacitation process, we e...
Boyd EH, Allen WE.Plasma drug concentrations were measured after two commercially available potentiated sulphonamides, trimethoprim and sulfadoxine and trimethoprim and sulphadiazine, were infused daily for 2 and 3 days, respectively, into the uteri of pony mares which had been mated before ovulation. Intravenous administration of trimethoprim and sulfadoxine allowed uterine absorption of trimethoprim (23-43%) and sulfadoxine (29-34%) to be calculated. After intra-uterine administration trimethoprim and sulphadiazine were detected in the milk of a lactating mare. In order to maintain plasma concentrations likel...
Bergfelt DR, Meira C, Fleury JJ, Fleury PD, Dell'Aqua JA, Adams GP.A regimen of progesterone plus estradiol (P&E) was used as a standard for ovarian synchronization to test the efficacy and evaluate the commercial application of ultrasound-guided follicle ablation as a non-steroidal alternative for ovulation synchronization in mares. Recipient mares at a private embryo transfer facility were at unknown stages of the estrous cycle at the start of the experiment on Day 1 when they were randomly assigned to an ablation group (n=18-21 mares) or to a P&E group (n=20-21 mares). In the ablation group, mares were lightly sedated and all follicles > or = 10...
Blue HB, Blue MG, Kenney RM, Merritt TL.Forty uterine fluid samples were obtained from 4 mares classified as resistant to uterine bacterial infection. The uterus of each mare was flushed with 50 ml of saline solution during estrus and diestrus of successive estrous cycles. Bacteria or fungi were isolated from 4 samples, and 7 additional samples were obtained from a mare with active intrauterine infection. Fluid volumes obtained during estrus (means = 40.3 +/- 11 ml) tended to be greater than those recovered during diestrus (means = 36.8 +/- 7.9 ml), but the difference was not significant. Concentrations and yields of protein in reco...
Roels K, Smits K, Ververs C, Govaere J, D'Herde K, Van Soom A.In horse breeding, intracytoplasmic sperm injection (ICSI) has gained interest to obtain offspring from subfertile individuals. This paper presents a case report of a stallion with severe testicular degeneration. Semen analysis showed very low motility and 83.5% of detached heads. Histology of a testicular biopsy showed severely decreased spermatogenesis, while transmission electron microscopy of the sperm cells revealed no significant abnormalities. A total of 39 oocytes were fertilized by ICSI with frozen-thawed spermatozoa of this stallion: 25 oocytes with intact spermatozoa and 24 with det...
Linklater WL, Henderson KM, Cameron EZ, Stafford KJ, Minot EO.To investigate the utility of faecal oestrone sulphate (OS) concentrations for detecting pregnancy in mares during behavioural studies of feral horses, in which the collection and preservation of samples is not immediate. Methods: Oestrone sulphate concentrations were measured in fresh dung samples collected from 153 free-roaming Kaimanawa mares throughout the year. In addition, multiple samples were taken from the same pile to investigate the reliability of diagnosis from a single sample, as well as the influence of time until preservation on OS concentrations. Samples were also taken before ...
Brown-Douglas CG, Perkins NR, Stafford KJ, Hedderley DI.To test published models for predicting time of foaling using data derived from prepartum mammary secretions and to develop a new model based on concentrations of mammary secretion constituents that accurately predicts time of foaling in Thoroughbred mares. Methods: Concentrations of sodium, potassium, calcium, citrate and lactose were measured in prepartum mammary secretions of 20 Thoroughbred mares in the 2 weeks before expected date of foaling. Models to predict time of foaling were fitted to data based on absolute concentration, change in concentration, and percentage change in concentrati...
Dhakal P, Tsunoda N, Nambo Y, Taniyama H, Nagaoka K, Watanabe G, Taya K.Although equine gestation is unique from the standpoint of fetal gonadal enlargement and regression, the activator of this process is still unknown. The present study aimed to show a possible role of activin during equine gestation. In the first experiment, weekly plasma samples from six pregnant mares were used to measure activin A. In the second experiment, eight pregnant mares carrying female (gestational days 110, 140, 180, and 270) and male fetuses (gestational days 120, 180, 225, and 314) were used for immunohistochemistry of activin receptors (IA, IB, IIA, IIB), and their intracellular ...
Lorenzo PL, Liu IK, Illera JC, Picazo RA, Carneiro GF, Illera MJ, Conley AJ, Enders AC, Illera M.Epidermal growth factor (EGF) has been reported to promote different functions in mammalian ovaries, including oocyte maturation. The aim of the present study was to establish: that EGF influences oocyte maturation in ovine and equine, that a tyrosine kinase-dependent intracellular mechanism mediates EGF effect and, that EGF-R receptor is detectable in ovarian follicles by immunohistochemistry methods. Selected ovine and equine oocytes were aspirated from 2-5 mm (ovine) or 25 mm (equine) follicles and cultured in TCM 199 for 22 (ovine) or 36 hours (equine). They are then subjected to culture w...
Foster SJ, Marshall DE, Houghton E, Gower DB.In on-going studies of 'classical' and ring B-unsaturated oestrogens in equine pregnancy, the products of metabolism of [2,2,4,6,6-2H(5)]-testosterone and [16,16,17-2H(3)]-5,7-androstadiene-3 beta,17 beta-diol with equine placental subcellular preparations and allantochorionic villi have been identified. Using mixtures of unlabelled and [2H]-labelled steroid substrates has allowed the unequivocal identification of metabolites by twin-ion monitoring in gas chromatography-mass spectrometry (GC-MS). Two types of incubation were used: (i) static in vitro and (ii) dynamic in vitro. The latter invol...
Magalhaes HB, Canisso IF.This study aimed to establish normal ranges and determine variables affecting the duration of gestation, placental and fetal eyeball measurements in donkeys. Jennies (n = 121) were submitted to transrectal ultrasonography at 15-day-intervals from 120 days of gestation until parturition. During each examination, variables assessed included: the combined thickness of uterus and placenta (CTUP), fetal eyeball diameter, amniotic membrane thickness, allantoic and amniotic fluids echogenicity, and chorioallantois edema. Parametric data were analyzed with t-test, linear regression, mixed models, AN...
Song X, Jiang H, Lv P, Cui K, Liu Q, Yin S, Liu H, Li Z.Oocyte in vitro maturation is necessary for the study and application of animal-assisted reproduction technology in animal reproduction and breeding. The comprehensive transcriptional profile of equine oocyte maturated in vitro has not been fully mined yet, which makes many key transcriptional events still unidentified. Here, Smart-seq2 was performed to analyse the gene expression pattern and the underlying regulatory mechanism of horse germinal vesicle (GV) and in vitro metaphase II (MII) oocytes. The results showed that 6402 genes (2640 up-regulated and 3762 down-regulated in MII samples com...
Silberzahn P, Dehennin L, Zwain I, Reiffsteck A.Follicular fluid was obtained from equine follicles at different stages of development as determined by ultrastructural study. Gas chromatography-mass spectrometry associated with stable isotope dilution permitted the demonstration of high levels of 4-estrene-3,7-dione and 17 beta-hydroxy-4-estren-3-one, 17 beta-hydroxy-4-estren-3-one levels often being about 10 times higher than those of testosterone. These findings suggest that in the mare ovary, an aromatizing pathway may proceed using these 19-norsteroids as intermediates. As a consequence of this high level of 19-norsteroids, testosterone...
Cheng FP, Fazeli AR, Voorhout WF, Tremoleda JL, Bevers MM, Colenbrander B.The aim of this study was to investigate whether mare follicular fluid (FF) induces the acrosome reaction (AR) in stallion spermatozoa and, if so, to identify the component in FF responsible for it. Furthermore, the effect of this component on sperm-zona binding and the subsequent AR was studied. Pooled FF, aspirated from the preovulatory follicles of mares in oestrous, was used and aliquots of the fluid were treated with charcoal to remove steroids (CFF). Charcoal treatment reduced the progesterone concentration in FF from 153 to < 2 ng/mL. Spermatozoa from fertile stallions collected by a...
Alvarenga MA, McCue PM, Bruemmer J, Neves Neto JR, Squire EL.Equine pituitary extract (EPE), has been reported to induce multiple ovulation in mares, however ovulation rates are poor in comparison to those obtained in other species. Attempts to improve the effectiveness of EPE for induction of superovulation in cyclic mares has focused on daily frequency of EPE treatment. Two experiments were performed to compare the ovarian response of cyclic mares given EPE once or twice-daily. Mares were assigned to one of two treatment groups 6 to 8 days after ovulation: prostaglandin was given once and EPE (25 mg) was given once daily (Group 1) or twice daily (Grou...
