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.
Ball BA, Gravance CG, Wessel MT, Sabeur K.A testis-specific isoform of angiotensin-converting enzyme (ACE) has been identified in a number of mammalian species. The purpose of this study was to characterize the activity of ACE in equine spermatozoa, seminal plasma, and testis. Activity of ACE was determined in seminal plasma, ejaculated and epididymal spermatozoa from mature stallions as well as from pre- and postpubertal testis. The effect of addition of angiotensin II on equine sperm motility was also evaluated. The activity of ACE in detergent extracted sperm plasma membrane was approximately 13-fold higher than that detected in se...
Brinsko SP, Blanchard TL, Rigby SL, Love CC, Varner DD.The aim of this study was to determine if dead spermatozoa reduced motility or membrane integrity of live spermatozoa in fresh and cooled-stored equine semen. Three ejaculates from each of three stallions were centrifuged and virtually all seminal plasma was removed. Spermatozoa were resuspended to 25 x 10(6) spermatozoa/ml with EZ-Mixin CST extender and 10% autologous seminal plasma, then divided into aliquots to which 0 (control), 10, 25, 50, or 75% (v/v) dead spermatozoa were added. Dead spermatozoa preparations contained 25 x 10(6) spermatozoa/ml and 10% seminal plasma from pooled ejaculat...
Love LB, Choi YH, Love CC, Varner DD, Hinrichs K.Two experiments were conducted to determine the effects of storage on equine ovaries or isolated oocytes. Ovaries were collected at an abattoir and were maintained at room temperature during collection and transport (3-9h total). After arrival at the laboratory, ovaries were divided into three groups: immediate oocyte collection (control), storage at room temperature overnight (15-18 h) before oocyte collection, or storage at 4 degrees C overnight before oocyte collection. Collected oocytes were cultured in maturation medium for 24h. There was a significant increase in the proportion of oocyte...
Dell'Aquila ME, Albrizio M, Maritato F, Minoia P, Hinrichs K.Follicle atresia and granulosa cell apoptosis may be related to oocyte meiotic and developmental competence. We analyzed the relationships among granulosa cell apoptosis, initial cumulus morphology, oocyte nuclear maturation in vitro, and pronucleus formation after intracytoplasmic sperm injection (ICSI) in the horse. For each follicle, the size was measured and granulosa cells were used for DNA laddering analysis. Oocytes were evaluated for cumulus morphology, cultured for in vitro maturation, and submitted to ICSI. Apoptosis was categorized as absent, intermediate, or advanced according to t...
Donahue JM, Williams NM, Sells SF, Labeda DP.Over the course of the past decade, actinomycetes have been isolated from the placentas of horses diagnosed with nocardioform placentitis. The incidence of this infection has generally been low, with typically no more than 30 animals affected in most years, but the incidence increased through 1999, with placentas from 144 mares found to be infected. Approximately half of the cases result in loss of the foal. A typical actinomycete with branching mycelium was isolated from placental lesions, and a comparison of the sequence of the 16S rDNA gene against the public databases indicated a relations...
Dirikolu L, Hughes C, Harkins D, Boyles J, Bosken J, Lehner F, Troppmann A, McDowell K, Tobin T, Sebastian MM, Harrison L, Crutchfield J, Baskin SI....The epidemiological association between black cherry trees and mare reproductive loss syndrome has focused attention on cyanide and environmental cyanogens. This article describes the toxicokinetics of cyanide in horses and the relationships between blood cyanide concentrations and potentially adverse responses to cyanide. To identify safe and humane blood concentration limits for cyanide experiments, mares were infused with increasing doses (1-12 mg/min) of sodium cyanide for 1 h. Infusion at 12 mg/min produced clinical signs of cyanide toxicity at 38 min; these signs included increased heart...
Squires EL, Carnevale EM, McCue PM, Bruemmer JE.Recent studies demonstrated that zwitterionic buffers could be used for satisfactory storage of equine embryos at 5 degrees C. The success of freezing embryos is dependent upon size and stage of development. Morulae and blastocysts <300 microm can be slowly cooled or vitrified with acceptable pregnancy rates after transfer. The majority of equine embryos are collected from single ovulating mares, as there is no commercially available product for superovulation in equine. However, pituitary extract, rich in FSH, can be used to increase embryo recovery three- to four-fold. Similar to human medic...
Tanaka Y, Taniyama H, Tsunoda N, Herath CB, Nakai R, Shinbo H, Nagamine N, Nambo Y, Nagata S, Watanabe G, Groome NP, Taya K.To clarify the source of inhibins in equine female fetuses, concentrations of immunoreactive (ir-) inhibin, inhibin pro-alphaC, and inhibin A in both fetal and maternal circulation and in fetal ovaries were measured. In addition, the localization of inhibin alpha and inhibin/activin beta(A), and beta(B) subunits and the expression of inhibin alpha(A) and inhibin/activin beta(A) subunit mRNA in fetal ovaries were investigated using immunohistochemistry and in situ hybridization. Concentrations of circulating ir-inhibin, inhibin pro-alphaC, and inhibin A were remarkably more elevated in the feta...
Martoriati A, Duchamp G, Gérard N.Paracrine factors have significant effects during folliculogenesis. Because of various morphological features, the mare is a convenient model to study in vivo the effects of factors involved in periovulatory events. In the present work, epidermal growth factor (EGF; experiment 1, n = 49 mares) and interleukin-1beta and interleukin-1RA (IL-1beta and IL-1RA, respectively; experiment 2, n = 80 mares) were injected intrafollicularly to evaluate the influence of these factors on in vivo maturation of equine preovulatory follicles. A transvaginal ultrasound-guided injection was performed when the di...
Guay KA, Brady HA, Allen VG, Pond KR, Wester DB, Janecka LA, Heninger NL.Matua bromegrass hay (Bromus willdenowii Kunth) is a high quality forage, but its value for mares during gestation and lactation is not well known. Intake, rate of passage, performance, and reproduction by gestating and lactating Quarter Horse mares fed the hay was investigated. In this experiment, 12, 2- to 12-yr-old gravid mares (mean BW = 553 kg, SD = 36) were fed Matua hay (CP = 11.5%) or alfalfa hay (Medicago sativa L.) (CP = 15.4%) for variable days prepartum (mean 59.9 d; SD = 23.5) and for 70 d postpartum. Matua and alfalfa hay were fed as the roughage portion of the diet with a grain ...
Diekman MA, Braun W, Peter D, Cook D.To determine whether secretory patterns of melatonin change throughout the seasons in mares, blood samples were drawn byvenipuncture from nine mares at noon and midnight for five successive days at monthly intervals from August through July at the University of Missouri in Columbia, MO. In addition, during September, December, March, and June, blood samples were drawn from indwelling catheters at 2-h intervals for 48 or 72 h. Mares were predominantly Quarter Horses weighing approximately 450 kg and ranged from 3 to 12 yr of age. Mares were housed in outdoor paddocks with three-sided run-in she...
Aurich C, Gerlach T, Aurich JE, Hoppen HO, Lange J, Parvizi N.In the non-breeding season, LH release is reduced via dopaminergic systems in the ram. On the other hand, our previous studies demonstrated an opioidergic inhibition of LH release in stallions outside the breeding season. Thus, in the present study we investigated the dopaminergic regulation of LH and prolactin secretion in stallions, considering interactions between dopamine and opioids. To achieve this, stallions (n=8) were treated with the dopamine antagonist sulpiride (0.6 mg/kg), the opioid antagonist naloxone (0.5 mg/kg), sulpiride plus naloxone or saline in December, March and June. Two...
Giese A, Jude R, Kuiper H, Raudsepp T, Piumi F, Schambony A, Guérin G, Chowdhary BP, Distl O, Töpfer-Petersen E, Leeb T.The cysteine-rich secretory protein (CRISP) family consists of three members called acidic epididymal glycoprotein 1 (AEG1), AEG2, and testis-specific protein 1 (TPX1), which share 16 conserved cysteine residues at their C-termini. The CRISP proteins are primarily expressed in different sections of the male genital tract and are thought to mediate cell-cell interactions of male germ cells with other cells during sperm maturation or during fertilization. Therefore, their genes are of interest as candidate genes for inherited male fertility dysfunctions and as putative quantitative trait loci fo...
Rosati I, Berlinguer F, Bogliolo L, Leoni G, Ledda S, Naitana S.It is clear that, in the horse, there are many weak links in the process of in vitro embryo production; an optimal culture system for equine oocytes does not exist, and related data are conflicting. Therefore, the ability of 3 different culture systems to support embryonic development of ICSI horse oocytes was examined. Oocytes (n = 261) suitable for culture were collected from 55 ovaries and divided, according to cumulus morphology, into 2 categories: expanded cumulus and compacted cumulus. Oocytes with expanded and compacted cumulus were cultured for in vitro maturation in TCM 199 + 10% FCS ...
The Journal of physiologyNovember 15, 2002
Volume 547, Issue Pt 1 67-76 doi: 10.1113/jphysiol.2002.027409
Giussani DA, Forhead AJ, Gardner DS, Fletcher AJ, Allen WR, Fowden AL.This study used between-breed embryo transfer in the horse to investigate the effects of maternal size and uterine capacity on fetal growth and postnatal cardiovascular and neuroendocrine functions. Equine embryos were transferred to establish eight Thoroughbred-in-Thoroughbred (TinT), seven Pony-in-Pony (PinP), five Thoroughbred-in-Pony (TinP) and eight Pony-in-Thoroughbred (PinT), pregnancies. Maternal and foal weights and placental microscopic area were measured at birth. At 6 days of postnatal life, arterial blood pressure and heart rate were monitored and blood samples were taken for horm...
Rathi R, Colenbrander B, Stout TA, Bevers MM, Gadella BM.Progesterone (P(4)) is a physiological inducer of the acrosome reaction (AR) in stallion spermatozoa. However, the capacitation-dependent changes that enable progesterone binding, and the nature of the signaling cascade that is triggered by progesterone and results in induction of the AR, are poorly understood. The aim of the current study was, therefore, to investigate the protein kinase dependent signaling cascades involved in progesterone-mediated induction of the AR in stallion spermatozoa. In addition, we aimed to determine whether bicarbonate, an inducer of sperm capacitation, acted via ...
Giese M, Bahr U, Jakob NJ, Kehm R, Handermann M, Müller H, Vahlenkamp TH, Spiess C, Schneider TH, Schusse G, Darai G.Equine arteritis virus (EAV) is the causative agent of the equine viral arteritis. It is a small RNA virus with a linear, non-segmented plus RNA genome. EAV is a member of the Arteriviridae family that includes porcine reproductive and respiratory syndrome virus (PRSSV), simian haemorrhagic fever virus (SHFV) and lactate dehydrogenase virus (LDV). The viral transmission is via respiratory and reproductive routes. Clinical signs in horses vary, and severe infection can lead to abortions in pregnant mares or neonatal foal death. The aim of this study was to investigate the development of the imm...
Davies Morel MC, Newcombe JR, Holland SJ.In order to assist in the accurate prediction of the timing of parturition in the mare true gestation length, along with the potential effect of a number of factors, was investigated. Data from 433 Thoroughbred foal pregnancies were used. Sequential ultrasonic scanning allowed the true gestation length (fertilisation-parturition) to be ascertained, as apposed to previous work, which used the mating-parturition interval. An average gestation length of 344.1 +/- 0.49 days was evident. Colt foal pregnancies were significantly (P < 0.001) longer (346.2 +/- 0.72) than fillies (342.4 +/- 0.65). Mont...
Devireddy RV, Swanlund DJ, Alghamdi AS, Duoos LA, Troedsson MH, Bischof JC, Roberts KP.The effects of extracellular ice and cryoprotective agents on the measured volumetric shrinkage response and the membrane permeability parameters of equine spermatozoa have been reported previously. The volumetric shrinkage data were obtained using a differential scanning calorimeter technique that was independent of cell shape. The aim of this study was to examine the effects of collection and cooling conditions on the motility and the water transport parameters at subzero temperatures of equine spermatozoa. Stallion semen samples were collected using either a commercial lubricating agent, wh...
Gentry LR, Thompson DL, Gentry GT, Davis KA, Godke RA, Cartmill JA.An experiment was conducted to determine the effects of high vs low body condition scores (BCS) produced by restricted feeding on reproductive characteristics, hormonal secretion, and leptin concentrations in mares during the autumnal transition and winter anovulatory period. Mares with BCS of 6.5 to 8.0 were maintained on pasture and/or grass hay, and starting in September, were full fed or restricted to produce BCS of 7.5 to 8.5 (high) or 3.0 to 3.5 (low) by December. All but one mare with high BCS continued to ovulate or have follicular activity during the winter, whereas mares with low BCS...
Johnson CA, Thompson DL, Cartmill JA.The present experiment characterized the pituitary responsiveness to exogenous GnRH in the first 10 d after ovulation following commercially available deslorelin acetate implantation at the normal dosage for hastening ovulation in mares. Twelve mature, cyclic mares were assessed daily for estrus and three times weekly for ovarian activity starting May 1. Mares achieving a follicle at least 25 mm in diameter or showing signs of estrus were checked daily thereafter for ovarian characteristics. When a follicle >30 mm was detected, mares were administered either a single deslorelin acetate impl...
Carneiro GF, Liu IK, Hyde D, Anderson GB, Lorenzo PL, Ball BA.In vitro fertilization (IVF) is being routinely used in humans and several domestic species, however, limited success has been achieved in the horse. Although immature equine oocytes are capable of completing meiosis in vitro, subsequent fertilization, and embryonic development of those oocytes are questionable. The lack of development of these oocytes could be attributed to an impaired cytoplasmic maturation. In the horse, the study of oocyte cytoplasmic maturation and post-fertilization development has been hindered by the lack of progress in IVF. In mammalian oocytes, migration of cortical ...
Pommer AC, Rutllant J, Meyers SA.Phosphorylation of tyrosine residues on sperm proteins is one important intracellular mechanism regulating sperm function that may be a meaningful indicator of capacitation. There is substantial evidence that cryopreservation promotes the capacitation of sperm and this cryocapacitation is frequently cited as one factor associated with the reduced longevity of cryopreserved sperm in the female reproductive tract. This study was designed to determine whether stallion sperm express different levels of tyrosine phosphorylation after in vitro capacitation and whether thawed sperm display similar ph...
Mantovani R, Rora A, Falomo ME, Bailoni L, Vincenti L.The aims of this study were to compare glycerol (G) at customary concentrations and ethylene glycol (EG) as cryoprotectants for stallion semen in a skimmed milk (SM) extender, to test different EG concentrations and to compare the results of manual and computerized analysis with the hypoosmotic swelling (HOS) test. Ejaculates from two stallions were collected over 3 weeks (6 ejaculates per stallion), diluted in a SM based extender, divided into 4 fractions, centrifuged and diluted again to a concentration of 100 x 10(6) mL(-1) progressive motile spermatozoa (PMS) in addition with the cryoprote...
Guillaume D, Bruneau B, Briant C.The effects of two GnRH antagonists were tested in order to delay and/or synchronise ovulation in mares. Five mares received Antarelix (0.01 mg.kg(-1)), 5 mares received Cetrorelix (the same dose), 5 mares (control mares) received the vehicle intravenously, twice daily, for 8 days from the day the largest follicle reached 22 mm following prostaglandin administration. Ovulation was postponed in all mares injected with Antarelix (19.4 +/- 1.2 days after the beginning of the treatment) and in 2/5 mares injected with Cetrorelix (20 +/- 1 days) vs. 6.2 +/- 0.4 days in control mares. During the trea...
Stock AE, Bouchard N, Brown K, Spicer AP, Underhill CB, Doré M, Sirois J.In contrast to other species, the preovulatory rise in gonadotropins in mares causes a remarkable expansion of the entire granulosa cell layer in vivo, suggesting that hyaluronan (HA) synthesis may be regulated in mural granulosa cells in this species. The objectives of this study were to clone and characterize equine hyaluronan synthase 2 (HAS2) and investigate the regulation of its transcript and of HA synthesis in equine follicles during human chorionic gonadotropin (hCG)- induced ovulation. Results showed that the equine HAS2 cDNA contains a 5'-untranslated region of 436 bp, an open readin...
Pommer AC, Rutllant J, Meyers SA.Cryopreservation requires exposure of sperm to extreme variations in temperature and osmolality. The goal of this experiment was to determine the osmotic tolerance levels of equine sperm by analyzing motility, viability, mitochondrial membrane potential (MMP), and mean cell volume (MCV). Spermatozoa were incubated at 22 degrees C for 10 min in isosmolal TALP (300 mOsm/kg), or a range of anisosmolal TALP solutions (75-900 mOsm/kg), for initial analysis, and then returned to isosmolal conditions for 10 min for further analysis. Total sperm motility was lower (P < 0.05) in anisosmolal conditio...
Pozor MA, McDonnell SM.For the purpose of establishing clinical reference values, this paper reports results of ultrasonographic examination and measurement of accessory sex glands, ampullae, and the pelvic urethra of 102 mature, healthy breeding stallions (2-29 years of age) of various size types (7 Miniature Horses, 27 small ponies, 53 light horses and 15 heavy horses). Examinations were done per rectum in mostly unsedated stallions using an Aloka 210 scanner with a 7.5 MHz linear veterinary transrectal transducer (Corometrics Medical Systems, Inc., North Wallingford, CT, USA). Most measures of accessory sex gland...
Francl AT, Amann RP, Squires EL, Pickett BW.The effects of extender and storage at 20 degrees C on equine spermatozoa were evaluated in two experiments using embryo recovery as the end point. In both experiments, inseminations were every other day, starting on Day 2 or 3 of estrus or after a 35-mm follicle was detected, with 250 x 10(6) progressively motile cells (based on initial evaluation). In Experiment 1, semen from two stallions was used to compare the motility and fertility of spermatozoa maintained in a) heated skim milk extender at 37 degrees C with insemination in <1 h; b) E-Z Mixin extender at 37 degrees C with inseminatio...
Brown KA, Sayasith K, Bouchard N, Lussier JG, Sirois J.The type 1 form of 17beta-hydroxysteroid dehydrogenase (17betaHSD1) was the first isoform to be identified and is capable of converting estrone to 17beta-estradiol. This study was aimed at characterizing the molecular structure of the equine 17betaHSD1 gene and cDNA, as well as its molecular regulation during human chorionic gonadotropin (hCG)-induced follicular luteinization/ovulation in vivo. The equine 17betaHSD1 gene was cloned from an equine genomic library and shown to have a conserved genomic structure composed of six exons. Its cDNA sequence was also identified and coded for a 308 amin...
Duthie S, Mills H, Burr P.Infection with equine arteritis virus is a notifiable disease with sporadic occurrence in the UK. As stallions may harbour the virus after infection, horses are screened for exposure by serological testing prior to breeding. The virus neutralisation test is considered the 'gold standard' serological screening test, but it is time-consuming and labour intensive; consequently there is a move towards more rapid screening methodology. In this study, a commercially available EVA antibody ELISA is assessed. The ELISA performed poorly with a specificity [corrected] of 26% and a sensitivity [corrected...
Parlevliet JM, Kemp B, Colenbrander B.The semen characteristics and testicular size of 398 3-year-old maiden Dutch Warmblood stallions were studied during February and March. Mean values (+/- SD) of age (1030 +/- 88 days) and testicular size (9.8 +/- 0.9 cm) of the maiden stallions were determined as well as the following semen characteristics (mean of two ejaculates, taken 1 h apart): volume (65 +/- 26 ml), sperm concentration (2.061 +/- 1.685 x 10(8) ml-1), total number of spermatozoa (1.129 +/- 0.71 x 10(10)), percentage of progressively motile spermatozoa (68 +/- 9%), percentage of live spermatozoa with normal morphology (66 +...
Brinsko SP, Van Wagner GS, Graham JK, Squires EL.The aim of the present study was to determine whether there are characteristics of fresh, cooled and frozen-thawed semen samples that can be used to predict the suitability of stallion semen for preservation by cooling or freezing. Each of three ejaculates obtained from 12 stallions was divided into aliquots to be analysed for sperm motility, morphology and membrane integrity as fresh, cooled and frozen-thawed samples. The percentage of morphologically normal spermatozoa was similar in fresh and cooled samples and both were greater than in the frozen samples. There were no strong linear relati...
Ellerbrock R, Canisso I, Feijo L, Lima F, Shipley C, Kline K.Urospermia is known to affect semen quality in many mammals, including stallions. Determinations of semen pH and creatinine and urea concentrations have been used to diagnose urine contamination in raw stallion semen. Unfortunately, practitioners suspecting urine contamination in cooled-shipped samples have no proven means to confirm the presence of urine. Therefore, the objectives of this study were (1) to assess the effects of urine contamination on sperm motility of extended fresh and cooled-stored stallion semen, (2) to evaluate the usefulness of semen color, odor, pH, and creatinine and u...
Parlevliet JM, Colenbrander B.In the selection procedure to acquire a breeding licence, 3-year-old Dutch Warmblood stallions have to undergo a breeding soundness test It is questioned whether this evaluation is predictive of the stallion's fertility results in the first breeding season. Therefore, semen parameters at the beginning of their first breeding season were evaluated and correlated to nonreturn at first cycle and foaling rate of mares bred by stallions (n = 13). The total number of mares inseminated with chilled semen from those stallions was 1055. Semen parameters were recorded on 2 ejaculates, collected 1 h apar...
Moslemi S, Dintinger T, Dehennin L, Silberzahn P, Gaillard JL.The ability of human and equine placental microsomes to aromatize 7 alpha-methyl-19-nortestosterone (MNT) was studied. Kinetic analysis indicates that MNT shares the androgen-binding site of human and equine placental microsomal aromatases. Human placental microsomal estrogen synthetase had about a 2.5-fold higher relative affinity for MNT than the equine placental enzyme (KiMNT/Km androstenedione of 32 versus 87). However, MNT was not metabolized by human placental microsomes, whereas it was very actively metabolized by equine placental microsomes. Further studies using purified equine cytoch...
Joonè CJ, Schulman ML, Fosgate GT, Claes ANJ, Gupta SK, Botha AE, Human A, Bertschinger HJ.Circulating anti-Müllerian hormone concentration (AMH) is positively correlated to the number of small growing follicles in the mare and may reflect ovarian function. Dynamics of AMH during immunocontraception have not previously been investigated. This study aimed to compare serum AMH in mares following treatment with native porcine zona pellucida (pZP), recombinant pZP3 and pZP4 (reZP) or gonadotrophin releasing hormone (GnRH) vaccines, and saline-treated controls. Stored sera collected during two previous studies examining ovarian activity in mares during zona pellucida (ZP) or GnRH immuno...
Tarko A, Fabová Z, Kotwica J, Valocký I, Alrezaki A, Alwasel S, Harrath AH, Sirotkin AV.The influence of environmental contaminant toluene and of plant fennel (Foeniculum vulgare Mill.) on reproduction are reported, but the mechanisms of their action and the protective effect of fennel on contaminant influence remain to be elucidated. In this study, we hypothesized that toluene and fennel directly affects basic ovarian cell functions, and that fennel can be used as an appropriate natural protective agent against the potential adverse effects of toluene. This study aimed to examine the action of toluene (20 μg/mL) and fennel extract (0, 1, 10, 100 μg/mL), and assess their comb...
Argo CM, Turnbull EB.Spherical (35 mm diameter) glass intra-uterine devices (IUDs) are commonly understood to modify sexual behaviour in performance mares. The effect of IUDs on the endocrinology (luteinising hormone, oestradiol, progesterone, testosterone), ovarian and uterine function, sexual and social behaviours associated with the ovarian cycle were evaluated during the natural breeding season in nine, nuliparous pony mares (4.6±0.5 years old, 230±12 kg bodyweight). Age and weight-matched groups were allocated to sham-treatment (n=5) or IUD-implantation (n=4) groups. Fifty complete ovarian cycles were evalu...
Leadon DP, Jeffcott LB, Rossdale PD.Total calcium, total protein, albumin and globulin content were determined in 49 samples of mammary secretions from 37 crossbred and Thoroughbred mares. The mares were divided into three categories: Group 1--post partum samples from spontaneous full term Thoroughbred deliveries (n = 20); Group 2--pre- and post partum samples from spontaneous full term Thoroughbred deliveries (n = 6); Group 3--pre- and post partum samples from induced premature deliveries in crossbred mares (n = 11). Calcium concentrations of the mammary secretions proved useful in predicting full term and also in assessment of...
Dorado J, Bottrel M, Ortiz I, Díaz-Jiménez M, Pereira B, Consuegra C, Carrasco JJ, Gómez-Arrones V, Domingo A, Hidalgo M.Embryo transfer and the vitrification of embryos could be used for the conservation and recovery of endangered donkey breeds. It is important to develop techniques that optimize recovery rates and the cryotolerance of donkey embryos. This study evaluates factors affecting the recovery rate, quality, and diameter of embryos obtained from donor jennies as a starting point for the use of vitrification and embryo transfer in the conservation of the Andalusian donkey. A total of 100 embryos were recovered out of 124 estrous cycles (80.6%). The donor jenny affected the rates of positive flushings (P...
Wilsher S, Lefranc AC, Allen WR.During embryo transfer (ET) the equine embryo can tolerate a wide degree of negative asynchrony but positive asynchrony of >2 days usually results in embryonic death. There is still confusion over whether this is due to the inability of the embryo to induce luteostasis or to an inappropriate uterine environment. Objective: To assess embryo survival and development in an advanced uterine environment. Objective: Embryo-uterine asynchrony, not the embryo's inability to induce luteostasis, is responsible for embryonic death in recipient mares with a >2 days chronologically advanced uterus. Methods...
Paweska JT, Volkmann DH, Barnard BJ, Chirnside ED.Two in a group of five naturally seropositive donkey stallions were found to shed equine arteritis virus (EAV) in their semen as demonstrated by virus isolation. Direct intramuscular inoculation of sonicated semen from one virus-shedding stallion (S3) caused clinical disease in two donkeys from which virus was recovered and in which seroconversion was detected. Sexual transmission was confirmed in two mares mated to S3 when after a febrile response during which EAV was isolated from huffy coats and nasal and ocular exudates, both mares were found to have seroconverted. In-contact transmission ...
Toishi Y, Tsunoda N, Tagami M, Hashimoto H, Kato F, Suzuki T, Nagaoka K, Watanabe G, Tokuyama S, Okuda K, Taya K.Evaluation of a new chemiluminescent enzyme immunoassay, the PATHFAST assay system (PATHFAST), for measurement of circulating progesterone in mares was performed. Five mares at the mid-luteal stage were administrated a single i.m. injection of prostaglandin F2α analog (PGF2α; cloprostenol 250 μg/ml), and then blood samples were collected from the jugular vein at 0, 15, 30 and 45 min, at one-hour intervals until 24 and at 48 hr via a catheter in the jugular vein. To monitor the physiological changes in circulating progesterone in mares after induced luteolysis, concentrations of progesterone...
Johnson L, Amann RP, Pickett BW.Spermatozoa from four regions of the epididymis and from ejaculated semen were evaluated for their resistance to cold shock, progressive motility, and structural stability. Spermatozoa were incubated at 38 C and their percentage of eosinophilia was compared with that of spermatozoa cooled to 0 C in 2 minutes, 10 C in 12 minutes, or 4 C in 22 minutes. Spermatozoa motility was estimated visually under phase-contrast microscopy and was recorded by cinematography. Structural stability of spermatozoa incubated in 0.05 M sodium borate buffer, 0.035 M sodium dodecyl sulfate (SDS), 0.002 M dithiothrei...
Acland HM, Allen PZ, Kenney RM.After contagious equine metritis bacteria were inoculated into the uterus of mares, genital tract tissues were examined for presence of the organism by bacteriologic cultural technique and an indirect immunofluorescent staining technique. Up to 14 days after mares were inoculated, the organism was frequently in the lumen of the uterus and in the cervix and, less frequently, in the vagina, vestibule, clitoral fossa, clitoral sinus, and uterine tubes. After 21 to 116 days, the organism was occasionally found on the ovarian surface, in the uterine tubes, uterus, cervix, and vagina and more freque...
Zavodovskaya R, Stover SM, Murphy BG, Katzman S, Durbin-Johnson B, Britton M, Finno CJ.Osteoporosis has been associated with pulmonary silicosis in California horses exposed to soils rich in cytotoxic silica dioxide crystals, a syndrome termed silicate associated osteoporosis (SAO). The causal mechanism for the development of osteoporosis is unknown. Osteoporotic lesions are primarily located in bone marrow-rich sites such as ribs, scapula and pelvis. Gene transcription patterns within bone marrow and pulmonary lymph nodes of affected horses may offer clues to disease pathobiology. Bone marrow core and tracheobronchial lymph node tissue samples harvested postmortem from affected...
Troedsson MH, Liu IK, Thurmond M.The validity of measuring concentrations of immunoglobulins in undiluted uterine secretions was established. The concentrations of IgG, IgA, and cleavage factor C3 of the complement system in uterine secretions were compared in mares with different resistance to chronic uterine infection (CUI). The uteri of mares susceptible (n = 6) and resistant to CUI (n = 5) were inoculated with 5 x 10(6) Streptococcus zooepidemicus when the mares were in estrus. Uterine secretions were sampled, and sampling was immediately followed by a uterine lavage at 5 and 24 h after bacterial inoculation. During a sub...
Oquendo PS, Silva ESM, Oquendo FMG, Cuervo-Arango J, Beletti ME.The effect of three different hormonal protocols to prepare anestrous recipient mares on embryo survival was evaluated. The first group consisted of only progesterone administration (NE) 4 days before embryo transfer, while the recipients from the other two groups received a single administration of 2.5 mg of oestradiol benzoate (SE) 2 days earlier or 8 mg of oestradiol split in increasing doses for 5 consecutive days (LE) ending 3 days before progesterone treatment. The likelihood of recovering an embryo 2 days after transfer was 46.1% (6/13), 62.5% (5/8) and 85.7% (6/7) for recipie...
Ginther OJ, Gastal EL, Gastal MO, Siddiqui MA, Beg MA.The effects of ultrasound morphology, vascularity, and follicular-fluid hormones of the preovulatory follicle on oocyte recovery rate and on follicle and oocyte maturity rates were studied for 60 spontaneous and solitary preovulatory follicles in mares. An ovulation-inducing dose of hCG was given when the follicle was >or=32 mm (Hour 0), and a procedure for oocyte recovery was done 30 h later (Hour 30). Between Hours 0 and 30, diameter of the follicle increased less and circulating estradiol (E2) concentrations decreased more in groups with successful versus nonsuccessful oocyte recovery an...
Gabay R, Rozen S, Samokovlisky A, Amor Y, Rosenfeld R, Kohen F, Amsterdam A, Berger P, Ben-Menahem D.CGβ subunits comprise a unique carboxyl-terminal peptide (CTP) that has multiple O-linked glycans and extends serum half-life of the protein. It has evolved by incorporating a previously untranslated region of the LHβ gene into the reading frame. Although CTP-like sequences are encrypted in the LHβ genes of several mammals, the CGβ subunit developed only in primates and equids. To study this restriction in evolution, we examined whether the cryptic CTP decoded from the bovine LHβ gene (boCTP) possesses key characteristics of the human (h) CGβ-CTP. The boCTP does not impede several crucia...
Tyrnenopoulou P, Fthenakis GC.Antibiotic administration is a standard therapeutic practice for the treatment of reproductive disorders of equids. This might lead to undesirable microbial imbalance and could favour the acquisition of antibiotic resistance. Therefore, it is imperative for clinicians to understand patterns of antibiotic resistance when considering and developing treatment regimes. Continued engagement of clinicians with novel alternative approaches to treat reproductive infections would be essential in order to address this rising threat within the One Health perspective. The objectives of the present review ...
Riera FL, Roldán JE, Espinosa JM, Fernandez JE, Ortiz I, Hinrichs K.Embryo biopsy for fetal sexing has clinical application, but few reports are available of its use within an active embryo transfer program. We evaluated results on biopsy of 459 embryos over one breeding season. There were no significant differences in pregnancy rate between biopsied and non-biopsied embryos (72% vs 73%) or for biopsied embryos recovered at the centre (73%) compared with those shipped overnight (72%). However, the pregnancy rate decreased significantly in shipped embryos biopsied ≥20h after collection. Overall, 86% of biopsies provided a sex diagnosis. The likelihood of a po...
Rutllant J, Meyers SA.It is generally accepted that spermatozoa become functionally mature during epididymal transit. The objective of this study was to determine whether the cellular location of equine PH-20 is modified during epididymal transit and, if so, the mechanism for such modification. Sperm were isolated from caput and cauda epididymal regions from stallions undergoing castration (n = 7) and used as whole sperm cell or subjected to nitrogen cavitation for isolation of plasma membrane proteins. Both caput and cauda sperm and sperm protein extracts were subjected to N-deglycosylation, O-deglycosylation, or ...
Flood PF, Betteridge KJ, Irvine DS.Six samples of blastocoele fluid recovered between 10 and 22 days gestation were tested in human clinical radioimmunoassay systems measuring total oestrogens and total androgens. The results were erratic but in 5 cases measurements for oestrogen equivalent to between 1000 and 70,000 pg/ml and for androgen between 1000 and 85,000 pg/ml were recorded. Cells from two blastocysts were cultured in medium 199 with and without horse serum. When the used media were assayed, values equivalent to at least 8000 pg oestrogen/ml were obtained on 7 of 11 occasions. In 9 of 11 samples the androgen concentrat...
Pinto CR, Paccamonti DL, Eilts BE, Venugopal CS, Short CR, Gentry LR, Thompson DL, Godke RA.In the present study, follicular fluids of estrous mares treated with saline solution (Control) or nitric oxide synthase (NOS) inhibitors were analyzed for nitric oxide (NO), estradiol-17beta (E2) and progesterone (P4) concentrations before and 36h after administration of human chorionic gonadotropin (hCG). Follicular fluids obtained before (0h) hCG administration from control mares had lower concentrations of NO than those obtained 36h after administration of hCG (58.3+/-17.8 micromol versus 340.4+/-57.7 micromol; P<0.05). A similar pattern was also noted for intrafollicular P4 in control ...
Combarnous Y, Guillou F, Martinat N, Cahoreau C.The LH and FSH activities of equine choriogonadotropin (eCG) have been compared in several species with those of the highly purified homologous pituitary gonadotropins. The molar FSH/LH activity ratio of eCG determined by RRA is 0.20 in the pig, 0.25 in the rat and 0 in the horse. These data demonstrate the LH monospecificity of eCG in its own species as it is the case for hCG. We have also shown that equine LH exhibited a FSH-activity similar to that of eCG in the pig and in the rat but not in the horse. In the female rat, the binding activity to FSH receptors and the in vitro FSH activity of...
Kuiper H, Distl O.Intersexuality is a rare congenital anomaly of horses. Diagnosis of intersexuality is difficult because there are usually no specific changes in the reproductive tract visible. During a period of five years, ten patients with reduced fertility or suspected intersexuality respectively were investigated using cytogenetic, molecular genetic, histopathological and endocrinological methods. In one case a 64,XX/63,X0 mosaicism was found. In six cases male pseudohermaphroditism was verified. These patients showed a male karyotype, testes and rudimentary parts of a female reproductive tract were prese...
