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.
Ferreira JRM, Villela SB, Bianconi C, Ormieres M, de Melo GD, Pugliesi G, Gobesso AAO.Different approaches have been used to improve conception rates at foal heat. Omega-3 fatty acids family and derivatives have improved reproductive efficiency in ruminants, but literature lacks studies evaluating these components on equines. The objective of this study was to evaluate effects of mare dietary supplementation with microalgae rich in docosahexaenoic acid (DHA) during peripartum on follicular dynamics and uterine involution in early post-partum. Eighteen pregnant mares, no particular breed, 410 ± 39.5 kg body weight (BW), and 7.83 ± 2.01 yr old were used. Mares were randomly ass...
Viafara JAS, Portilho RV, Maculan R, Souza Júnior JF, Silva CM, Rodrigues PG, El Azzi MS, Souza JC.This study evaluated the presence of polyunsaturated fatty acids in circulating blood and in the ovarian follicular fluid of mares, after supplementation of the diet with linseed oil. Six Mangalarga Marchador mares, weighing 397.00±31.89 kg, were kept on native pasture, and assigned to the current study. In a switch over design, mares were randomly allocated to receive 150 ml of vegetable oil daily, containing polyunsaturated fatty acids n3 (62.23 g ALA, 20.34 g LA, 2.27 g EPA, 2.32 g DHA), (n=3) or no supplementation (n=3) in two replicates. Blood and follicular fluid samples were taken on t...
Requena F, Campos MJAPM, Martínez Marín AL, Camacho R, Giráldez-Pérez RM, Agüera EI.In equine reproduction, accurate and timely detection of the moment of ovulation is of great importance. Power Doppler ultrasound technology is a non-invasive method that enables to assess the morpho-echogenic features and blood flow changes during the estral cycle in mares. The objective of the present study was to evaluate the influence of age on ultrasonographic parameters (follicular diameter, follicular blood flow-FBF, corpus luteum (CL) area and corpus luteum blood flow-CLBF) and blood plasma progesterone concentrations in cycling Spanish Purebred mares (15 less than 8 years old and 15 e...
Ginther OJ.The equine embryo or embryonic vesicle on Days 11-15 postovulation travels with profound physiologic purpose throughout the lumen of the two uterine horns and uterine body making 12 to 22 trips between the two uterine horns per day. This phenomenon is termed embryo mobility and is unique in equids among domestic species. Apparently, the embryo first reaches the uterine body on Days 8 or 9. Mobility increases to maximum by Days 11 or 12 and continues until an abrupt cessation of mobility (fixation) on Days 15 (ponies) or 16 (horses and donkeys). The embryo is propelled by uterine contractions i...
Szóstek-Mioduchowska AZ, Shiotani H, Yamamoto Y, Sadowska A, Wójtowicz A, Kozai K, Hojo T, Kimura K, Skarzynski DJ, Okuda K.Glucocorticoids (GCs) are known to play an important role in maintaining basal and stress-related homeostasis by interacting with endocrine mediators and prostaglandins (PGs). Although a growing body of evidence shows that GCs exert their regulatory action at a multitude of sites in the reproductive axis through corticosteroid receptors, little is known about the direct role of cortisol, an active form of GCs, in the equine endometrium. Thus, the study aimed to determine the effect of cortisol on PGF synthesis in the endometrial tissue and cells in vitro. In Exp.1, the immunolocalization and ...
Beachler TM, Papich MG, Andrews NC, Von Dollen KA, Ellis KE, Withowski K, Bailey CS.Transcervical intrauterine infusion of antibiotics may more effectively treat pathogens associated with fetal and neonatal disease in pregnant mares than standard systemic routes. The objective of this study was to assess the safety of transcervical antibiotic infusion by characterizing the gestational outcome in nine healthy pregnant pony mares following a single transcervical infusion of 2.4 million IU of procaine penicillin and 200 mg of gentamicin in a 10 mL volume during late gestation. Assessment of fetal-placental health was performed through serial measurement of the combined thickness...
Becsek A, Tzanidakis N, Blanco M, Schmicke M, Bollwein H.The aim of this study was to investigate the effects of maternal, hormonal, and fetal factors on early fetal volume (FV) measurements in mares obtained by three-dimensional (3D) ultrasound. Furthermore, postpartum parameters were explored in regard to their association with early FV. For this purpose, 149 German warmblood mares that were artificially inseminated and confirmed to be pregnant between days 14-16 of gestation, were examined transrectally at day 45 ± 1 of gestation with the portable 3D ultrasound device Voluson® i (GE, Zipf, Austria). FV was calculated by using the extension so...
Ellerbrock RE, Podico G, Scoggin KE, Ball BA, Carossino M, Canisso IF.The expression pattern and distribution of sex steroid receptors and steroidogenic enzymes during development of the equine accessory sex glands has not previously been described. We hypothesized that equine steroidogenic enzyme and sex steroid receptor expression is dependent on reproductive status. Accessory sex glands were harvested from mature stallions, pre-pubertal colts, geldings, and fetuses. Expression of mRNA for estrogen receptor 1 (ESR1), estrogen receptor 2 (ESR2), androgen receptor (AR), 3β-Hydroxysteroid dehydrogenase/Δ5-4 isomerase (3βHSD), P450,17α hydroxylase, 17-20 lyase...
Fernández-Hernández P, Marinaro F, Sánchez-Calabuig MJ, García-Marín LJ, Bragado MJ, González-Fernández L, Macías-García B.Equine fertilization cannot be performed in the laboratory as equine spermatozoa do not cross the oocyte's zona pellucida . Hence, a more profound study of equine oviductal fluid (OF) composition at the pre-ovulatory and post-ovulatory stages could help in understanding what components are required to achieve fertilization in horses. Our work aimed to elucidate the proteomic composition of equine OF at both stages. To do this, OF was obtained postmortem from oviducts of slaughtered mares ipsilateral to a pre-ovulatory follicle ( = 4) or a recent ovulation ( = 4); the samples were kept at -80°...
Benammar A, Derisoud E, Vialard F, Palmer E, Ayoubi JM, Poulain M, Chavatte-Palmer P.Although there are large differences between horses and humans for reproductive anatomy, follicular dynamics, mono-ovulation, and embryo development kinetics until the blastocyst stage are similar. In contrast to humans, however, horses are seasonal animals and do not have a menstrual cycle. Moreover, horse implantation takes place 30 days later than in humans. In terms of artificial reproduction techniques (ART), oocytes are generally matured in vitro in horses because ovarian stimulation remains inefficient. This allows the collection of oocytes without hormonal treatments. In humans, in viv...
Pennington PM, Splan RK, Jacobs RD, Wang Y, Wagner AL, Freeman EW, Pukazhenthi BS.The reproductive cycle and early pregnancy represent dynamic physiological states in mammals, but mechanisms involved in early pregnancy in the domestic horse remain poorly understood. Proteins in uterine secretions have been studied, but the proteome of peripheral serum during various reproductive states has not been investigated. This study characterized and compared the serum proteome in the domestic horse during various reproductive states. Serum was collected from three mares during: (1) estrus (day [d] -1; d 0 = ovulation), (2) diestrus (d 12.5, non-mated), (3) early pregnancy (d 12.5,...
Kútvölgyi G, Brabender K, Andersson M, Javkhlan A, Nagy S, Páble T, Egerszegi I, Hidas A, Soós I, Kovács A.The case of an 8-year-old, sexually active but infertile Przewalski's stallion (Equus ferus przewalskii) was studied. Besides the infertility, the stallion also showed permanent problems with its body condition, being obviously weaker than all the other group members. The horse was kept in a separate place for two years with 12 mares in its harem group (six mares had foals earlier); however, none of the mares covered got pregnant. Andrological and cytogenetic investigations revealed underdeveloped testes, arrested spermatogenesis, azoospermia, and XY/XXY/X0 mosaicism. We classify the case as a...
Vandenberghe E, Boshuizen B, Delesalle CJG, Goehring LS, Groome KA, van Maanen K, de Bruijn CM.In May 2018, Wolvega Equine Hospital (WEH) experienced an EHV-1 outbreak. This outbreak caused significant economic losses and negative publicity for the hospital. How should hospitals prepare themselves for these outbreaks and prevent shedding of the virus on multiple neighboring premises? The hospital transformed most of its activities into mobile practice and the entire infected hospital population was moved to a separate remote location. The hospital was cleaned and disinfected according to the latest recommendations before reopening. Four neighboring professional equine businesses and thr...
Medica AJ, Aitken RJ, Nicolson GL, Sheridan AR, Swegen A, De Iuliis GN, Gibb Z.Stallion sperm membranes comprise a high proportion of polyunsaturated fatty acids, making stallion spermatozoa especially vulnerable to peroxidative damage from reactive oxygen species generated as a by-product of cell metabolism. Membrane lipid replacement therapy with glycerophospholipid (GPL) mixtures has been shown to reduce oxidative damage in vitro and in vivo. The aims of this study were to test the effects of a commercial preparation of GPL, NTFactor® Lipids, on stallion spermatozoa under oxidative stress. When oxidative damage was induced by the addition of arachidonic acid to stall...
Blommaert D, Sergeant N, Delehedde M, Donnay I, Lejeune JP, Franck T, Serteyn D.The quality of fresh or thawed sperm in stallions has been generally determined by the viability and total and progressive motility of the sperm. Today, the expression of ProAKAP4, a protein present in the flagellum of spermatozoa, appears to be an innovative and relevant functional marker to assess semen quality and male fertility. This study aims to compare the concentration of ProAKAP4 in the semen from 5 stallions frozen with two different extenders immediately after thawing (T0) and 4 h post-thawing (T4). Viability, total and progressive motility were measured in parallel. Significant di...
Pinzón-Osorio CA, Cortés-Beltrán D, Sanchez-Isaza CA, Jiménez-Robayo LM, Lozano-Márquez H, Zambrano-Varón J, Jiménez-Escobar C.A 4-year-old Colombian Creole mare was presented for diagnosis because the external orifice of her cervix was not detectable when a uterine lavage as therapy for uterine fluid accumulation was attempted. Clinical and ultrasonographic evaluation of the genital tract revealed that ovaries were of normal size and showed structures suggestive of regular ovarian activity. However, granular free-floating fluid material distending the uterus was detected by ultrasound. Upon vaginal examination, the normal external cervical morphology was not evident. The vagina ended in a blind bag with a small papil...
Velloso Alvarez A, Boone L, Horzmann K, Hanson RR.To combine laparoscopic techniques with natural orifice transluminal endoscopic surgery (hybrid NOTES) to minimize number and enlargement of paralumbar laparoscopic portals for bilateral ovariectomy in standing, sedated mares. Methods: Case series. Methods: Six horses. Methods: Six mares with palpably normal ovaries were restrained in stocks, sedated, and had caudal epidural anesthesia performed. A 7.5 MHz ultrasound probe was used transvaginally to select placement of vaginotomy. An attempt was made to remove both ovaries with 70 cm esophageal forceps through the vaginotomy, while visualiz...
Rajabi-Toustani R, Watanabe H, Tsogtgerel M, Gao Y, Canbo L, Haneda S, Cheong SH, Nambo Y.The main difficulty of large equine embryo cryopreservation is the replacement of blastocoel fluid with cryoprotectant solution. The objective of this study was to improve the cryopreservation of large equine embryos with PMAP and/or LAP. Embryos were collected via the non-surgical transcervical procedure and divided into three groups based on their size (A ≤ 300 µm, 300 µm<B 300 µm). However, more research is required to find the best method for embryos ≥700 µm.
Kim SH, Lee JS, Lee JH, Kim YJ, Choi JG, Lee SK, Kim HJ, Yang SJ, Park T, Lee SK, Kang HE, Jeoung HY, Park JY.Venereal diseases caused by bacteria are important to the equine industry due to economic losses caused by decline of conception rate in breeding horses. Therefore, identification of infected animals as well as the implementation of appropriate managerial procedures based on accurate diagnosis is critical. In this study, two types of multiplex real-time polymerase chain reaction with high sensitivity and specificity were developed for the simultaneous detection and differentiation of five commonly associated bacterial pathogens of venereal diseases in horses, consisting of Taylorella equigenit...
Cooper HE, Davidson EJ, Slack J, Ortved KF.To report the clinical features, treatment, and outcome in horses with cellulitis and concurrent septic tendonitis and/or desmitis. Methods: Short case series. Methods: Medical records from 2000 to 2019 were reviewed, identifying horses with cellulitis and concurrent septic tendonitis and/or desmitis based on sonographic examination and positive bacterial culture. Signalment, ultrasonographic results, bacterial culture, treatment, duration of hospitalization, and complications were recorded. Long-term outcome data were obtained from follow-up examinations and/or telephone interviews. Successfu...
Ferrer MS, Canisso IF, Podico G, Ellerbrock RE, Hurley DJ, Palomares R.Different stallions exhibit a high level of variation in the ability of their sperm to survive cryopreservation. A large fraction of stallions show poor post-thaw sperm motility, and their semen is not suitable for commercial freezing. In this study, we hypothesized that the presence of sperm-bound antisperm antibodies (ASAs) was associated with poor cryosurvival of stallion sperm. Our objective was to assess the level of ASA binding to stallion sperm, and determine if it was associated with good or poor sperm cryosurvival. In Experiment 1, cooled shipped semen from 27 stallions was frozen usi...
Shang S, Jiang R, Luo R, Jia S, Irwin DM, Wang Z, Zhang S.Individual identification of horses for pedigree verification and registration is important for the sustainable development of the horse industry. Horse individual identification and parentage tests commonly use the 17 short tandem repeats (STRs) recommended by the International Society for Animal Genetics (ISAG) and the locus LEX33. While many multiplex STR typing systems have been established for the horse, a sex determining marker is usually absent, and none of them can simultaneously detect all 17 ISAG recommended loci and the locus LEX33. Here, we present a 19-plex STR typing system that ...
Lu KG, Sprayberry KA.Selected conditions affecting broodmares are discussed, including arterial rupture, dystocia, foal support with ex utero intrapartum treatment, uterine prolapse, postpartum colic, the metritis/sepsis/systemic inflammatory response syndrome complex, and retained fetal membranes. Postpartum colic beyond third-stage labor contractions should prompt comprehensive evaluation for direct injuries to the reproductive tract or indirect injury of the intestinal tract. Mares with perforation or rupture of the uterus are typically recognized 1 to 3 days after foaling, with depression, fever, and leukopen...
Sprayberry KA, Lu KG.Selected emergency conditions of male and female reproductive tracts in horses are described, including injuries affecting the external genitalia of male horses and emergent conditions arising during gestation in mares. Conditions affecting male horses are discussed in the context of breeding stallions, but kicks or other mechanisms of trauma in the groin can also affect geldings. Priapism, paraphimosis, trauma to the scrotum and testicles, and penile injury are discussed. In mares, traumatic vestibular injury, placentitis, hydropsic conditions, prepubic tendon and abdominal wall compromise, a...
Podico G, Gray SM, Wang L, Canisso IF.An 8-y-old jenny was presented because of anorexia and mild depression. The jenny had weaned her colt 10 d before the admission. Upon arrival at the University of Illinois Veterinary Teaching Hospital, the heart rate was elevated, and the right udder was painful and swollen on palpation. Milk stripping of the affected side revealed purulent content; the contralateral udder had normal-appearing milk. Cytology of mammary gland secretions from the affected side revealed a large number of hypersegmented reactive neutrophils with phagocytized bacteria. Complete blood count, serum chemistry, and f...
Stefanetti V, Pascucci L, Wilsher S, Cappelli K, Capomaccio S, Reale L, Passamonti F, Coletti M, Crociati M, Monaci M, Marenzoni ML.Endogenous retroviruses (ERVs) are proviral phases of exogenous retroviruses, which have coevolved with vertebrate genomes for millions of years. The conservation of ERV genes throughout evolution suggests their beneficial effects on their hosts' survival. An example of such positive selection is demonstrated by the syncytin gene, which encodes a protein with affinity for various mammalian placentas that is involved in the formation of syncytiotrophoblasts. Although the horse has an epitheliochorial placenta, in which the fetal trophoblasts are simply apposed to the intact uterine epithelium, ...
Papas M, Govaere J, Peere S, Gerits I, Van de Velde M, Angel-Velez D, De Coster T, Van Soom A, Smits K.Anti-Müllerian hormone (AMH) reflects the population of growing follicles and has been related to mammalian fertility. In the horse, clinical application of ovum pick-up and intracytoplasmic sperm injection (OPU-ICSI) is increasing, but results depend largely on the individuality of the mare. The aim of this study was to assess AMH as a predictor for the OPU-ICSI outcome in horses. Therefore, 103 mares with a total follicle count above 10 were included in a commercial OPU-ICSI session and serum AMH was determined using ELISA. Overall, the AMH level was significantly correlated with the number...
Monteiro de Barros MR, Davies-Morel MCG, Mur LAJ, Creevey CJ, Alison RH, Nash DM.Persistent mating-induced endometritis is a major cause of poor fertility rates in the mare. Endometritis can be investigated using an ex vivo equine endometrial explant system which measures uterine inflammation using prostaglandin F2α as a biomarker. However, this model has yet to undergo a wide-ranging assessment through transcriptomics. In this study, we assessed the transcriptomes of cultured endometrial explants and the optimal temporal window for their use. Endometrium harvested immediately post-mortem from native pony mares (n = 8) were sampled (0 h) and tissue explants were cultured ...
Sampaio B, Ortiz I, Resende H, Felix M, Varner D, Hinrichs K.Exposure to the calcium ionophore A23187 may present a "universal" sperm treatment for IVF, as it bypasses capacitation pathways. However, success in utilizing A23187 is variable, especially in equine spermatozoa. Notably, albumin is used during A23187 treatment but paradoxically is thought to suppress A23187 action. Essentially no critical data are available on the effects of A23187 and albumin concentrations, ratios, or addition protocols on changes in intracellular calcium ([Ca] ) in any cell type. To determine factors that affect the action of A23187 on [Ca] in equine and murine spermatozo...
Segabinazzi LGTM, Oba E, Alvarenga MA.Equine practitioners often prescribe the combined use of hCG and GnRH to hasten ovulation due to presumed synergistic effects. Therefore, this study aimed to test whether the combination of hCG and deslorelin acetate to hasten ovulation in mares would show any effect in inducing ovulation more efficiently than when either drug is used by itself, and to verify whether this association would affect progesterone concentrations; corpus luteum (CL) diameter and blood flow; and pregnancy outcome in recipient mares after embryo transfer (ET). Seventeen mares had the ovulation hastened (≥35 mm folli...
Powell DM.Successful management of captive populations of wild animals requires effective control of reproduction. Contraception is one tool for controlling reproduction of animals in zoos; however, the options available to the animal manager are limited. Contraceptives vary in efficacy, reversibility, and side effects, and thus may not be suitable for widespread use. One consideration when selecting a contraceptive is its potential for side effects on behavior, especially given the fact that reproduction plays such a prominent role in the biology of any species. To date, there have been few evaluations...
Hendrickson D.Laparoscopic surgery has become commonplace in the field of equine urogenital surgery. As with most surgical procedures, the limiting factors in developing new surgical techniques are limited to the patient size and demeanor, the skills of the surgeon, and the available equipment. Some of the greatest benefits of laparoscopic surgery in the horse include better visualization of the important structures; tension-free amputation of the testes or ovaries,which generally leads to less postoperative pain; and the ability to evaluate the transected stump carefully to make sure there is no hemorrhage...
Ballagi-Pordány A, Klingeborn B, Flensburg J, Belák S.Primers and probes were selected from the gene encoding glycoprotein 13 (gp 13) of equine herpesvirus 1 (EHV-1). The polymerase chain reaction (PCR) was run on infected and noninfected cultured cells and on 63 specimens from 29 aborted equine fetuses. The results were evaluated by electrophoresis and dot-blot hybridization using an oligonucleotide probe labeled with biotin. In the infected samples electrophoresis showed a PCR product of about 280 base pairs. The dot-blot hybridization confirmed that this product contained EHV-1 DNA sequences. PCR took 4 h and hybridization another 14 h; the re...
Ginther OJ, Beg MA, Gastal EL, Gastal MO, Baerwald AR, Pierson RA.Changes in systemic concentrations of FSH, LH, oestradiol and progesterone during the ovulatory follicular wave were compared between 30 mares and 30 women. Based on a previous study, the emergence of the future ovulatory follicle was defined as occurring at 13.0 mm in mares and 6.0 mm in women, and deviation in diameter between the two largest follicles was expected to begin at 22.7 mm in mares and 10.3 mm in women. Mean FSH concentrations were high in mares during the luteal phase, resulting from statistically identified FSH surges occurring in individuals on different days and in different ...
Briski O, Salamone DF.During the past 2 decades, intracytoplasmic sperm injection (ICSI) has become a routine technique for clinical applications in humans. The widespread use among domestic species, however, has been limited to horses. In horses, ICSI is used to reproduce elite individuals and, as well as in humans, to mitigate or even circumvent reproductive barriers. Failures in superovulation and conventional in vitro fertilization (IVF) have been the main reason for the use of this technology in horses. In pigs, ICSI has been successfully used to produce transgenic animals. A series of factors have resulted in...
Muñoz PM, Ferrusola CO, Lopez LA, Del Petre C, Garcia MA, de Paz Cabello P, Anel L, Peña FJ.Stallion-to-stallion variability in the quality of cryopreserved ejaculates postthaw affects the commercial acceptability of frozen semen and thus is a major constraint for the equine industry. In recent years, the molecular mechanisms associated with sperm damage during cryopreservation have become better understood. Identification of the freezability of the ejaculates before the freezing process is initiated will have a major impact on the equine industry. We studied three markers of oxidative stress in sperm, including 8-iso-PGF2alpha, 8-OH guanosine, and 4-hydroxynonenal (4-HNE); the prese...
Johnson L, Blanchard TL, Varner DD, Scrutchfield WL.Spermatogenesis is a process of division and differentiation by which spermatozoa are produced in seminiferous tubules. Seminiferous tubules are composed of somatic cells (myoid cells and Sertoli cells) and germ cells (spermatogonia, spermatocytes, and spermatids). Activities of these three germ cells divide spermatogenesis into spermatocytogenesis, meiosis, and spermiogenesis, respectively. Spermatocytogenesis involves mitotic cell division to increase the yield of spermatogenesis and to produce stem cells and primary spermatocytes. Meiosis involves duplication and exchange of genetic materia...
Rizzo M, du Preez N, Ducheyne KD, Deelen C, Beitsma MM, Stout TAE, de Ruijter-Villani M.Aneuploidy of meiotic origin is a major contributor to age-related subfertility and an increased risk of miscarriage in women. Although age-related aneuploidy has been studied in rodents, the mare may be a more appropriate animal model to study reproductive aging. Similar to women, aged mares show reduced fertility and an increased incidence of early pregnancy loss; however, it is not known whether aging predisposes to aneuploidy in equine oocytes. We evaluated the effect of advanced mare age on (1) gene expression for cohesin components, (2) incidence of aneuploidy and (3) chromosome centrome...
Carneiro JAM, Canisso IF, Bandeira RS, Scheeren VFC, Freitas-Dell'Aqua CP, Alvarenga MA, Papa FO, Dell'Aqua JA.This study aimed to evaluate the antioxidant properties of coenzyme Q10 (CoQ10) during cryopreservation of semen obtained from stallions having good and bad semen freezing ability (GFA vs. BFA, respectively). Forty ejaculates (n = 20 stallions) were split into five centrifugation and five freezing extenders containing different concentrations of CoQ10 (0, 25, 50, 75 and 100 μmols/L). If CoQ10 was added to the centrifugation extender, the freezing extender had no CoQ10 added; similarly, if CoQ10 was added to the freezing extender, the centrifugation extender had no CoQ10. Semen cryoprese...
Carnevale EM, Catandi GD, Fresa K.Numerous similarities in reproductive aging have been documented between the mare and woman. Aging is associated with a decline in fertility. In mares and women, oocyte transfer procedures were initially used to establish that oocyte donor age is associated with oocyte quality. Age-associated differences in oocytes include altered morphology, gene expression, and developmental potential. Reactive oxygen species and mitochondrial dysfunction are thought to be important contributors to loss of oocyte quality. In the woman, aneuploidy is a primary consideration with maternal aging. Although misal...
Johnson L, Thompson DL.Testes from 47 adult (4-20 years) stallions obtained in November-January (non-breeding season) and 41 adult stallions obtained in May-July (breeding season) were perfused with glutaraldehyde, placed in osmium and embedded in Epon 812. Percentage Leydig cell cytoplasm or nuclei in the testis was determined by point counting of 0.5 micron sections under bright-field microscopy. Testes from 6 randomly selected horses per season were processed for electron microscopy. The volume (ml) of SER/testis was calculated from the % SER in the cytoplasm % Leydig cell cytoplasm, and parenchymal volume. Numbe...
Pillet E, Labbe C, Batellier F, Duchamp G, Beaumal V, Anton M, Desherces S, Schmitt E, Magistrini M.Egg yolk is normally used as a protective agent to freeze semen of equine and other species. However, addition of egg yolk in extenders is not without disadvantages and the demand to find cryoprotective alternatives is strong. The objective of this study was to test the cryoprotective capacities of liposomes composed of egg yolk phospholipids. Two experiments were conducted: 1) the first to determine the optimal composition and concentration of liposomes to preserve post-thaw motility and membrane integrity of spermatozoa; 2) the second to assess in vivo the cryoprotective capacities of these ...
Spizziri BE, Fox MH, Bruemmer JE, Squires EL, Graham JK.Irreversible damage occurs to spermatozoal membranes, during the phase transition, when spermatozoa are cooled from room temperature to 5 degrees C. Some of this damage can be ameliorated by adding cholesterol to the membrane, thereby altering membrane lipid composition. Adding cholesterol-loaded cyclodextrins (CLCs) to stallion spermatozoa prior to freezing, increases cell cryosurvival. However, the fertilizing potential of CLC-treated stallion spermatozoa is unknown. To address this, experiments were conducted which evaluated the ability of CLC-treated stallion spermatozoa to capacitate, acr...
Andersson L, Sandberg K.In the present study an extensive amount of data, comprising more than 30,000 offspring in total, was analyzed to evaluate the influence of age and sex on the recombination frequency in the K-PGD segment of the equine linkage group (LG) I and the influence of age, breed and sex on recombination in the Al-Es segment of LG II. A highly significant sex difference is reported for both segments. Male and female recombination values in the K-PGD segment were estimated at 25.8 +/- 0.8 and 33.3 +/- 2.5%, respectively. Similarly, recombination was less frequent in the male (36.6 +/- 0.7%) than in the f...
Hidalgo M, Rodríguez I, Dorado J, Soler C.This work used semen samples collected from 12 stallions and assessed for sperm morphometry by the Sperm Class Analyzer (SCA) computer-assisted system. A discriminant analysis was performed on the morphometric data from that sperm to obtain a classification matrix for sperm head shape. Thereafter, we defined six types of sperm head shape. Classification of sperm head by this method obtained a globally correct assignment of 90.1%. Moreover, significant differences (p<0.05) were found between animals for all the sperm head morphometric parameters assessed.
Gacem S, Valverde A, Catalán J, Yánez Ortiz I, Soler C, Miró J.This study aimed to characterize the sperm kinematic values with high frames per second, to define the subpopulation structure of a horse and a donkey and compare them. A total of 57 fresh semen ejaculates (26 Spanish and 16 Arabian horse breeds and 10 donkeys) were collected and subsequently analyzed for kinematic parameters using the Computer-aided sperm motility analysis ISAS®v1.2 system and using a Spermtrack® 10-μm depth counting chamber. Sequences were recorded at 250 frames per second, and eight kinematic parameters were automatically evaluated. All kinematic parameters showed signif...
Klewitz J, Struebing C, Rohn K, Goergens A, Martinsson G, Orgies F, Probst J, Hollinshead F, Bollwein H, Sieme H.Color Doppler sonography has become routine for the evaluation of high-risk pregnancies in human medicine. Previous studies documenting uterine blood flow parameters in the pregnant mare have found a decrease in peripheral blood flow resistance in the first pregnancy weeks and an increase in uterine blood flow, especially in the last trimester of pregnancy. However, these studies involved only a small number of mares. No naturally occurring pregnancy abnormalities occurred that would allow blood flow changes to be retrospectively examined and analyzed. The objective of the present study was to...
Love CC, Kenney RM.The relationship between fertility and susceptibility of sperm DNA to denaturation was determined in a group of 84 actively breeding, clinically fertile stallions. Susceptibility of DNA to denaturation was determined using the sperm chromatin structure assay (SCSA). The SCSA measures, mean of alpha-t (mean alpha t), standard deviation of alpha-t (SD alpha t), and the COMP of alpha-t (cells outside the main population)] were significantly correlated with the percentage seasonal pregnancy rate (SPR; mean alpha t, r = -0.24, P < or = 0.05; % COMP alpha t, r = -0.27, P < or = 0.05); percentage pre...
Centromeres exert an inhibitory effect on meiotic recombination, but the possible contribution of satellite DNA to this "centromere effect" is under debate. In the horse, satellite DNA is present at all centromeres with the exception of the one from chromosome 11. This organization of centromeres allowed us to investigate the role of satellite DNA on recombination suppression in horse spermatocytes at the stage of pachytene. To this aim we analysed the distribution of the MLH1 protein, marker of recombination foci, relative to CENP-A, marker of centromeric function. We demonstrated that the sa...
Aleksic D, Blaschke L, Mißbach S, Hänske J, Weiß W, Handler J, Zimmermann W, Cabrera-Sharp V, Read JE, de Mestre AM, O'Riordan R, Moore T....Pregnancy-specific glycoproteins (PSGs) are members of the carcinoembryonic antigen cell adhesion molecule (CEACAM) family that are secreted by trophoblast cells. PSGs may modulate immune, angiogenic and platelet responses during pregnancy. Until now, PSGs are only found in species that have a highly invasive (hemochorial) placentation including humans, mice and rats. Surprisingly, analyzing the CEACAM gene family of the horse, which has a non-invasive epitheliochorial placenta, with the exception of the transient endometrial cups, we identified equine CEACAM family members that seem to be rel...
Bromfield EG, Aitken RJ, Gibb Z, Lambourne SR, Nixon B.While IVF has been widely successful in many domesticated species, the development of a robust IVF system for the horse remains an elusive and highly valued goal. A major impediment to the development of equine IVF is the fact that optimised conditions for the capacitation of equine spermatozoa are yet to be developed. Conversely, it is known that stallion spermatozoa are particularly susceptible to damage arising as a consequence of capacitation-like changes induced prematurely in response to semen handling and transport conditions. To address these limitations, this study sought to develop a...
Ortega-Ferrusola C, Martin Muñoz P, Ortiz-Rodriguez JM, Anel-López L, Balao da Silva C, Álvarez M, de Paz P, Tapia JA, Anel L, Silva-Rodríguez A....We hypothesized that thiols and particularly glutathione (GSH) are essential for the regulation of stallion sperm functionality. To test this hypothesis, we initially investigated the relationship between sperm function and GSH content, revealing highly significant correlations between GSH, sperm viability, motility, and velocity parameters (P < 0.001). Furthermore, the deleterious effects of GSH depletion using menadione and 1,3 dimethoxy 1,4, naphtoquinone (DMNQ) were able to be prevented by the addition of cysteine, but no other antioxidant. Pre-incubation with cysteine prevented menad...
Kikuchi M, Nakano Y, Nambo Y, Haneda S, Matsui M, Miyake Y, Macleod JN, Nagaoka K, Imakawa K.A factor responsible for progression to pregnancy establishment in the mare has not been definitively characterized. To identify factors possibly involved in the establishment of equine pregnancy, the endometrium was collected from day 13 (day 0=day of ovulation) cyclic and day 13, 19 and 25 pregnant animals. From initial subtractive hybridization studies, a calcium regulating factor, Stanniocalcin-1 (STC1) mRNA, was found as a candidate molecule expressed uniquely in the pregnant endometrium. Endometrial expression of STC1 mRNA was noted on day 19 and was markedly increased in the day 25 grav...
Hemberg E, Lundeheim N, Einarsson S.During 1997-2001, a total of 430 Thoroughbred mares were mated by one of the two Thoroughbred stallions on a well-managed stud farm in the central part of Sweden. On arrival, a thorough gynaecological examination of all mares was performed. An early pregnancy examination was performed on days 14 and 15 after ovulation. The overall conception rates for the two stallions were 92.2 and 88.8%, and the corresponding live foal rates were 82.6 and 75.2%, respectively. The mean number of served oestrous cycles was 1.60 per mare per conception and 1.86 per live foal. The first and second cycle concepti...
Dorado J, Acha D, Ortiz I, Gálvez MJ, Carrasco JJ, Gómez-Arrones V, Calero-Carretero R, Hidalgo M.The main aim of this study was to evaluate the efficacy of two commercially available liquid stallion semen extenders for the preservation of Andalusian donkey semen at 5°C for up to 72h, and to evaluate the effect of amino acid addition on sperm quality of cooled donkey semen. In addition, this study investigated the effect of seasons on semen characteristics of Andalusian jackasses. Throughout a year, 50 ejaculates were collected from ten adult donkeys and a complete semen evaluation was performed immediately after collection. In Experiment 1, semen samples (n=32) were pooled, divided into ...
Hui A, Altan E, Slovis N, Fletcher C, Deng X, Delwart E.Circoviruses infect vertebrates where they can result in a wide range of disease signs or in asymptomatic infections. Using viral metagenomics we analyzed a pool of five sera from four healthy and one sick horse. Sequences from parvovirus-H, equus anellovirus, and distantly related to mammalian circoviruses were recognized. PCR identified the circovirus reads as originating from a pregnant mare with fever and hepatitis. That horse's serum was also positive by real time PCR for equine parvovirus H and negative for the flavivirus equine hepacivirus. The complete circular genome of equine circovi...
Baumann C, Daly CM, McDonnell SM, Viveiros MM, De La Fuente R.Pairing of the sex chromosomes during mammalian meiosis is characterized by the formation of a unique heterochromatin structure at the XY body. The mechanisms underlying the formation of this nuclear domain are reportedly highly conserved from marsupials to mammals. In this study, we demonstrate that in contrast to all eutherian species studied to date, partial synapsis of the heterologous sex chromosomes during pachytene stage in the horse is not associated with the formation of a typical macrochromatin domain at the XY body. While phosphorylated histone H2AX (γH2AX) and macroH2A1.2 are pres...
Hinrichs K, Choi YH, Varner DD, Hartman DL.We evaluated the effect of different activation treatments on the production of blastocysts and foals by nuclear transfer. Donor cells were prepared using roscovitine treatment, which has previously been associated with increased production of viable offspring. All activation treatments were followed by culture in 6-dimethylaminopurine (6-DMAP) for 4 h. In experiment 1, blastocyst production after activation by injection of sperm extract followed by treatment with ionomycin was significantly higher than that for activation with a serial treatment of ionomycin, 6-DMAP, and ionomycin (12.5 vs 2....
Brinsko SP, Love CC, Bauer JE, Macpherson ML, Varner DD.Semen samples were collected from six fertile stallions and seven stallions with unexplained infertility. Percentages of motile sperm (77.5 +/- 11.3 versus 67.5 +/- 12.2, P = 0.2), and progressively motile sperm (70.8 +/- 13.6 versus 60.7 +/- 14.0, P = 0.2) were similar between fertile and subfertile stallions, respectively. Morphologic characteristics in ejaculates of control and affected stallions (% normal: 60.2 +/- 18.2 versus 52.9 +/- 11.3, P = 0.4; % abnormal heads 7.3 +/- 4.8 versus 12.1 +/- 5.0, P = 0.11; and % abnormal acrosomes 1.6 +/- 2.1 versus 3.0 +/- 3.4, P = 0.4) did not differ....
Walter J, Neuberg KP, Failing K, Wehrend A.Aim of this study was to compare uterine smears made using the Knudsen catheter, the cytology brush and a uterine culture swab with regard to diagnostic usefulness and the occurrence of neutrophils. Additionally correlation between culture results and the occurrence of neutrophils in uterine smears was investigated. Samples were collected from 340 mares, 81.5% of which were in estrus. Smears made using the cytology brush yielded more endometrial cells per high-power field than those made using the other two instruments (p<0.0001), and a larger proportion had PMNs compared with smears made u...
