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.
Pieńkowska-Schelling A, Becker D, Bracher V, Pineroli B, Schelling C.Numerical and structural aberrations of chromosomes may cause malformations, embryonal losses and reduced or missing fertility. In male horses the fertility is rather well controlled through their semen evaluation. For mares there are no mandatory regulations which specify their use in a breeding programme. Therefore, mares with chromosomal aberrations, which exclude reproduction success may be chosen for breeding. The present case describes a horse, offically registered as a female, which was presented for a breeding exam. On this occasion, small and inactive ovaries were diagnosed. Although ...
Cauchard J, Soldan A, Madeline A, Johnson P, Büscher P, Petry S.To evaluate the reproducibility of routine serological methods to detect Trypanosoma equiperdum antibodies in equine sera, two inter-laboratory ring trials were organized involving 22 European and 4 non-European reference laboratories for dourine. The serological methods were the complement fixation test (CFT; 25 laboratories) and the indirect fluorescent antibody test (IFAT; 4 laboratories). Three of the laboratories applied both these methods. The sample panels were composed of sera that were negative, positive or suspected for dourine. Of the negative sera, one was from a donkey naturally i...
Hanada M, Maeda Y, Oikawa MA.Uteri from 50 necropsied nonpregnant Japanese Thoroughbred brood mares (1-30 years of age) were investigated to clarify the histopathological characteristics of endometrosis in Japanese Thoroughbred mares and the distribution pattern of endometrosis lesions in the uterus as a whole. Endometrosis was observed in all animals over 6 years of age and in all of the 21 mares aged over 12 years of age. The affected mares showed elastofibrosis of arteries, veins and lymphatic vessels in the uterine wall, atrophy of the uterine smooth muscle layers and hyperplasia of collagen fibers among the smooth mu...
Vitte A, Rossignol F, Mespoulhes-Rivière C, Lechartier A, Röecken M.To report a combination of standing laparoscopic technique for intra-abdominal dissection of the mesovarial pedicle followed by a limited ventral median celiotomy under general anesthesia for removal of enlarged ovaries in mares. Methods: Case series. Methods: Mares (n = 20) aged 3-22 years with unilateral enlarged ovaries. Methods: Enlarged ovaries were confirmed by transrectal palpation and ultrasonography. After sedation, 3 laparoscopic portals were made in the paralumbar fossa. The mesovarium was desensitized and dissected using a vessel sealing device, and the ovary was left free in t...
Ali A, Alamaary M, Al-Sobayil F.The aim of this study was to evaluate the reproductive performance of the Arab mares in the Kingdom of Saudi Arabia (KSA). Methods: Managers of 26 studfarms containing 1014 Arab mares at five districts were asked to complete a questionnaire on the reproductive efficiency of their mares. Results: The mean age at first mating was 3.06 ± 0.5 years. Mares were mated equally in winter and all over the year. Natural mating was mostly used (76.9%). The means of estrus duration and estrus interval were 6.58 ± 1 days and 19.57 ± 1.8 days, respectively. The means of first service and over- all pregna...
Gallardo Bolaños JM, Balao da Silva C, Martín Muñoz P, Plaza Dávila M, Ezquerra J, Aparicio IM, Tapia JA, Ortega Ferrusola C, Peña FJ.To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h ...
Johannisson A, Lundgren A, Humblot P, Morrell JM.The decrease in foaling rates after artificial insemination with cooled semen warrants the search for new predictors of fertility. The objectives were to investigate levels of naturally occurring reactive oxygen species (ROS) in cooled, stored stallion semen doses for artificial insemination (AI), and their relationship with parameters of semen quality and with pregnancy rate. Semen was collected from warmblood stallions (n=15) and used to prepare commercial semen doses for AI. Sperm quality was evaluated after cooled transport to the laboratory overnight. The results were correlated with obse...
Mambelli LI, Mattos RC, Winter GH, Madeiro DS, Morais BP, Malschitzky E, Miglino MA, Kerkis A, Kerkis I.Mesenchymal stem cells (MSCs) due to their self-renewal potential and differentiation capacity are useful for tissue regeneration. Immunomodulatory and trophic properties of MSCs were demonstrated suggesting their use as medicinal signaling cells able to positively change local environment in injured tissue. Equine endometrosis is a progressive degenerative disease responsible for glandular alterations and endometrial fibrosis which causes infertility in mares. More precisely, this disease is characterized by phenotypic changes in the expression pattern of selected endometrial proteins. Curren...
Curcio Bda R, Pereira GR, Antunes LI, Boff AN, dos Santos FC, Lucas T, Nogueira CE, Corcini CD, Liu I, Deschamps JC.In vitro fertilization (IVF) procedures are limited by the inability to mature equine oocytes on in vitro methods. Objective: The aim of this study was to evaluate structural integrity of equine oocytes subjected to vitrification with a synthetic polymer (PVA). Methods: The effect of eGH and its relationship with IGF-I on in vitro maturation (IVM) were evaluated. Compact cumulus oocytes complexes (n=122) were cultured in TCM-199 with eGH, IGF-I or eGH+IGF-I for 30h at 38.5C in air with 5 % CO2. Oocytes were fixed after IVM or subjected to the vitrification protocol. Cryopreserved oocytes were ...
Labitzke D, Sieme H, Martinsson G, Distl O.The objectives of this study were to show whether semen traits of 30 Hanoverian stallions regularly used in AI may be useful for breeding purposes. Semen characteristics were studied using 15 149 ejaculates from 30 Hanoverian stallions of the State Stud Celle of Lower Saxony. Semen samples were collected between 2005 and 2009. Traits analysed were gel-free volume, sperm concentration, total and motile sperm number and progressive motility. A linear multivariate animal model was employed to estimate heritabilities and permanent environmental variances for stallions. The same model was used to p...
da Silveira JC, Carnevale EM, Winger QA, Bouma GJ.Ovarian follicle growth and maturation requires extensive communication between follicular somatic cells and oocytes. Recently, intercellular cell communication was described involving cell-secreted vesicles called exosomes (50-150 nm), which contain miRNAs and protein, and have been identified in ovarian follicular fluid. The goal of this study was to identify a possible role of exosomes in follicle maturation. Methods: Follicle contents were collected from mares at mid-estrous (~35 mm, before induction of follicular maturation) and pre-ovulatory follicles (30-34 h after induction of folli...
Erol E, Jackson CB, Steinman M, Meares K, Donahoe J, Kelly N, Locke S, Smith JL, Carter CN.A comprehensive evaluation of the real-time PCR assay for leptospirosis in comparison with other diagnostic assays on a large-scale basis is fundamental in validating the assay and determining the causes of equine abortions. Objective: To compare and evaluate the diagnostic value of real-time PCR assay for leptospirosis with traditional methods in equine leptospiral abortions. Methods: Cross-sectional observational study. Methods: A Leptospira spp. fluorescent antibody test (FAT), microscopic agglutination test (MAT) and real-time PCR (targeting the LipL32 gene) were compared and evaluated in ...
Gallardo Bolaños JM, Balao da Silva CM, Martín Muñoz P, Morillo Rodríguez A, Plaza Dávila M, Rodríguez-Martínez H, Aparicio IM, Tapia JA....AKT, also referred to as protein kinase B (PKB or RAC), plays a critical role in controlling cell survival and apoptosis. To gain insights into the mechanisms regulating sperm survival after ejaculation, the role of AKT was investigated in stallion spermatozoa using a specific inhibitor and a phosphoflow approach. Stallion spermatozoa were washed and incubated in Biggers-Whitten-Whittingham medium, supplemented with 1% polyvinyl alcohol (PVA) in the presence of 0 (vehicle), 10, 20 or 30 μM SH5, an AKT inhibitor. SH5 treatment reduced the percentage of sperm displaying AKT phosphorylation, w...
Vackova I, Czernekova V, Tomanek M, Navratil J, Mosko T, Novakova Z.This study aimed to determine whether maternal cell contamination exists in cells derived from equine umbilical cord tissue, a perspective material for cell-based therapies in veterinary medicine. Potential maternal cell contamination was analyzed at DNA level via a set of 16 microsatellite markers in cells originating from the cord tissue of 22 foals. In these cells no maternal cell contamination was detected at a sensitivity level of 0.01%. Our results suggest that equine umbilical cord tissue-derived cells are entirely of fetal origin.
Leemans B, Gadella BM, Sostaric E, Nelis H, Stout TA, Hoogewijs M, Van Soom A.Sperm-oviduct binding is an essential step in the capacitation process preparing the sperm for fertilization in several mammalian species. In many species, capacitation can be induced in vitro by exposing spermatozoa to bicarbonate, Ca(2+), and albumin; however, these conditions are insufficient in the horse. We hypothesized that binding to the oviduct epithelium is an essential requirement for the induction of capacitation in stallion spermatozoa. Sperm-oviduct binding was established by coincubating equine oviduct explants for 2 h with stallion spermatozoa (2 × 10(6) spermatozoa/ml), during...
Begg AP, Todhunter K, Donahoe SL, Krockenberger M, Slapeta J.A case of amoebic placentitis in a mare from eastern Australia was diagnosed postpartum by histopathological examination of the placenta. The identity of the etiological agent was confirmed as Acanthamoeba hatchetti by use of diversity profiling based on a next-generation sequencing approach.
Schulman ML, Becker A, van der Merwe BD, Guthrie AJ, Stout TA.Equine herpesvirus 1 (EHV-1) is one of the most common causes of infectious abortion in mares. Analysing the demography of outbreaks and detailing subsequent reproductive performance of affected mares will assist in the management of future (threatened) epizootics. Objective: To examine the epidemiology and reproductive outcomes of 2 EHV-1 abortion epizootics with very different patterns of morbidity. Methods: Epidemiological and reproductive data were analysed retrospectively following abortion epizootics associated with EHV-1, but initiated via different routes, among unvaccinated mares on 2...
Woodward EM, Christoffersen M, Horohov D, Squires EL, Troedsson MH.Research has shown that 6 h after breeding is a critical time during the uterine innate immune response, and the failure to respond appropriately will result in persistent breeding-induced endometritis. This presents a potential opportunity to modulate the course of inflammation towards a timely resolution. Objective: To evaluate the effects of immune modulation on endometrial mRNA expression of inflammatory genes in susceptible mares 6 h after breeding. The hypothesis was that immune modulation alters endometrial cytokine expression in susceptible mares. Methods: A randomised controlled s...
Choi YH, Ritthaler J, Hinrichs K.Cloned animals possess mitochondria derived from the host ooplast, which typically differ genetically from those of the donor. This is of special concern to horse breeders, as maternal lines are prized and athletic performance is a key factor in genetic value. To evaluate the feasibility of producing mitochondrial-identical cloned foals, we collected oocytes from immature follicles of two mares, BL and SM, maternally related to the donor stallion. In vitro matured, enucleated oocytes were treated with roscovitine-synchronized donor cells and blastocysts were transferred transcervically to reci...
Lund CM, Ragle CA, Lutter JD, Farnsworth KD.To describe the use of a motorized morcellator for elective bilateral laparoscopic ovariectomy in standing equids and to evaluate long-term outcome. Methods: Retrospective case series. Methods: 30 equids (20 horses, 9 mules, and 1 pony). Methods: Medical records of equids undergoing elective bilateral laparoscopic ovariectomy from 2007 to 2013 were evaluated. Cases were selected on the basis of use of a motorized morcellator for ovary extraction. Data collected included age, breed, reason for surgery, surgery date, surgical approach, intraoperative complications, surgery and morcellation times...
Balssa F, Fischer M, Bonnaire Y.5(10)-Estrene-3β,17α-diol is an essential reference material for doping analysis in horse-racing laboratories. It is used to detect misuse, for doping purpose, of the pregnancy status in the mare. Its stereoselective synthesis from 17β-estradiol-3-methyl ether (prepared from estrone or 17β-estradiol) was performed in four steps: (1) Mitsunobu inversion of the 17β-alcohol; (2) Birch reduction of the aromatic ring; (3) stereoselective reduction of the 3-ketone via Noyori asymmetric transfer hydrogenation; (4) chemoenzymatic purification.
Furukawa S, Kuroda Y, Sugiyama A.The primary function of the placenta is to act as an interface between the dam and fetus. The anatomic structure of the chorioallantoic placenta in eutherian mammals varies between different animal species. The placental types in eutherian mammals are classified from various standpoints based on the gross shape, the histological structure of the materno-fetal interface, the type of materno-fetal interdigitation, etc. Particularly, the histological structure is generally considered one of the most useful and instructive classifications for functionally describing placental type. In this system,...
Rodrigues MN, Carvalho RC, Franciolli AL, Rodrigues RF, Rigoglio NN, Jacob JC, Gastal EL, Miglino MA.Since the horse has a highly precocial reproductive strategy, most organs are functionally well developed at birth and thus, embryonic and fetal life is interesting. Data on the development of important organs are very limited. Here, we detailed macroscopically and histologically the equine digestive system, focusing on the first third of gestation. At 21 days, the oral cavity was an empty space, and the liver contained proliferating endodermal cells. At 25 days, a fusiform stomach and the pancreatic bud were present. At 28 days, a small tongue and the esophagus occurred. At 30 days, primary a...
Sayasith K, Sirois J.The interaction between stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) has been implicated in leukocyte attraction, tissue remodeling and angiogenesis. The objective of the present study was to characterize the expression and regulation of SDF1 and CXCR4 in equine follicles during the ovulatory process. Equine preovulatory follicles were isolated during estrus 0-39h after hCG treatment. Follicle wall preparations (theca interna with attached granulosa cells) and isolated preparations of granulosa cells and theca interna were obtained, and total RNA extracts wer...
Gaivão MM, Rambags BP, Stout TA.Experimental studies and field surveys suggest that embryonic loss during the first 6 weeks of gestation is a common occurrence in the mare. During the first 2 weeks of development, a number of important cell differentiation events must occur to yield a viable embryo proper containing all three major germ layers (ectoderm, mesoderm, and endoderm). Because formation of the mesoderm and primitive streak are critical to the development of the embryo proper, but have not been described extensively in the horse, we examined tissue development and differentiation in early horse conceptuses using a c...
Pérez-Marín CC, Requena FD, Arando A, Ortiz-Villalón S, Requena F, Agüera EI.There has been a lack of research into equine sperm vitrification to date, but studies of other species suggest it may have significant potential. To evaluate the impact of various cryoprotectant agents (CPA) and vitrification on equine sperm quality, a controlled study was carried out. A total of 12 ejaculates were subjected to exposure to CPA and vitrification. Sperm was diluted in a range of CPA: fresh, control (BSA), sucrose (0.15M, 0.3M and 0.5M), trehalose (0.15M, 0.3M and 0.5M) and the combination of sucrose and trehalose (M1: 0.15M sucrose+0.5M trehalose; M2: 0.5M sucrose+0.15M trehalo...
Mortensen CJ, Choi YH, Hinrichs K, Ing NH, Kraemer DC, Vogelsang SG, Vogelsang MM.The effect of exercise on mare reproductive efficiency was evaluated by comparing rates of embryo recovery from mares assigned to either an exercise regimen or a non-exercise (control) regimen. Exercised mares were worked daily for 30 min under average ambient conditions of >30 degrees C and >50% humidity. Mares were inseminated during estrus and subjected to uterine flush for embryo recovery on d 7 after ovulation for two consecutive cycles. After this, mares were allocated to the opposite group and allowed an estrous cycle without reproductive manipulation; then insemination and uterin...
Schofield WA.The goal of this review is to provide evidence supporting the physiologic basis for the use of acupuncture as a treatment in equine reproduction, and to discuss current uses of acupuncture for the treatment of anestrus, urine pooling, uterine infection and/or fluid, infertility, maintenance of pregnancy, cryptorchidism, musculoskeletal pain, and poor libido in stallions. The benefits achieved through acupuncture treatment are thought to be a result of hormonal regulation, altered smooth muscle motility, and general stress and/or pain relief from musculoskeletal or environmental conditions.
Al-Kass Z, Spergser J, Aurich C, Kuhl J, Schmidt K, Johannisson A, Morrell JM.Contamination of semen with bacteria arises during semen collection and handling. This bacterial contamination is typically controlled by adding antibiotics to semen extenders but intensive usage of antibiotics can lead to the development of bacterial resistance and may be detrimental to sperm quality. The objective of this study was to determine the effects of antibiotics in a semen extender on sperm quality and to investigate the effects of removal of bacteria by modified Single Layer Centrifugation (MSLC) through a colloid. Semen was collected from six adult pony stallions (three ejaculates...
Panzani S, Comin A, Galeati G, Romano G, Villani M, Faustini M, Veronesi MC.Thyroid hormones, insulin growth factor I (IGF-I) and non-esterified fatty acids (NEFA) represent important hormonal and metabolic factors associated with perinatal growth and maturation. Their action could be influenced by the type of parturition and the health status of the foal and therefore the aim of this work is to evaluate their plasma concentrations in newborn foals during the first 2 wks of life. Three groups of subjects were enrolled: 15 healthy foals born by spontaneous parturition, 24 healthy foals born by induced parturition and 26 pathologic foals. From each of the healthy foals,...
Fowden AL, Silver M, Ellis L, Ousey J, Rossdale PD.The factors influencing beta cell function in the foetal and neonatal foal have been investigated in chronically catheterised foetal foals and in newborn foals delivered either spontaneously at term or by induction at different gestational ages. Insulin was detected in the foetal plasma from as early as 150 days of gestation (term = 340 days) and during the last third of gestation the foetal beta cells responded to exogenous administration of glucose and arginine and to endogenous variations in the glucose level. Insulin secretion by the foetal beta cells was depressed by anaesthesia and surge...
Clulow JR, Evans G, Maxwell WM, Morris LH.The aim of the present study was to evaluate the potential oocyte binding ability and functional integrity of fresh or frozen-thawed, sex-sorted or non-sorted stallion spermatozoa. In the absence of effective IVF procedures in the horse, a heterologous sperm-binding assay was used as an indicator of fertilising capacity to assess differences in the ability of stallion spermatozoa to bind to bovine oocytes. The functional integrity of four treatment groups was assessed: (1) fresh non-sorted spermatozoa; (2) fresh sex-sorted spermatozoa; (3) frozen-thawed non-sorted spermatozoa; and (4) frozen-t...
Hendriks WK, Roelen BA, Colenbrander B, Stout TA.Equine embryos are cryopreserved by slow-freezing or vitrification. While small embryos (<300 μm) survive cryopreservation reasonably well, larger embryos do not. It is not clear if slow-freezing or vitrification is less damaging to horse embryos. Objective: To compare the type and extent of cellular damage suffered by small and large embryos during cryopreservation by slow-freezing vs. vitrification. Methods: Sixty-three Day 6.5-7 embryos were subdivided by size and assigned to one of 5 treatments: control, exposure to slow-freezing or vitrification cryoprotectants (CPs), and cryopreser...
Tsogtgerel M, Komyo N, Murase H, Hannan MA, Watanabe K, Ohtaki T, Tsumagari S, Kawate N, Nambo Y.Insulin-like peptide 3 (INSL3) is an important hormone for testicular descent during embryonic development and a factor for assessing functional status of Leydig cells of testes, but there is limited number of equine studies. Anti-Müllerian hormone (AMH) is a useful diagnostic marker for cryptorchidism in horses. This study aimed to compare serum concentrations and testicular expression intensity of INSL3 and AMH in intact and cryptorchid male horses. Serum INSL3 concentrations in intact (n = 9; mean ± SEM, 19.9 ± 5.9 ng/mL) and noncastrated unilateral cryptorchid (UC) male horses (n...
Li X, Morris LH, Allen WR.The influence of co-culture with either oviduct epithelial cells or fetal fibroblast cells on in vitro maturation of equine oocytes and their potential for development to blastocysts and fetuses after intracytoplasmic sperm injection (ICSI) was investigated. The oocytes were obtained from ovaries from abattoirs and were matured in vitro for 28-30 h in TCM-199 only, or in TCM-199 co-culture with oviduct epithelial cells or fetal fibroblast cells. Metaphase II oocytes were subjected to ICSI with an ionomycin-treated spermatozoon. The injected oocytes were cultured for 7-9 days in Dulbecco's modi...
Robinson SJ, Neal H, Allen WR.Equine embryos enter the uterus 144-156 h after ovulation, before which time the passage of embryos through the oviduct is halted in the region of the ampullary-isthmic junction. It is thought that further onward movement of embryos to the uterus is facilitated by secretion of prostaglandin E2 (PGE2) by the embryos, which relaxes the smooth muscle of the isthmus. In the present study, the effect of a single local application of PGE2 on oviductal transport was examined in 22 Thoroughbred and Pony mares that were inseminated on alternate days during oestrus. On day 4 after ovulation, the ipsilat...
Kimura Y, Haneda S, Aoki T, Furuoka H, Miki W, Fukumoto N, Matsui M, Nambo Y.The combined thickness of the uterus and placenta (CTUP) and ultrasonographic images of uteroplacental tissues were investigated in 35 pregnant heavy draft horses in Months 7-12 of pregnancy. The mares were divided into three groups: those pathologically diagnosed as placentitis (placentitis group, n=3); those who had abortion, premature birth, or fetal malformation (abnormal group, n=7); and those who had no abnormal findings (normal group, n=25). In the normal group, CTUP increased as pregnancy progressed from Months 7 (median, 7.08 mm; range, 5.68-11.27) to 12 (13.31 mm; 7.44-16.31 mm) (P<0...
Chavatte P, Holtan D, Ousey JC, Rossdale PD.Major progress on the endocrinology of the pregnant mare has been possible thanks to the catheterised equine fetal preparation developed by Marian Silver. In particular, these preparations led to the identification of the source of progestagens within the feto-placental unit and provided the impetus for further work on their biosynthesis and biological activities. The biosynthesis of the progestagens involves close interaction between the fetus, the endometrium and the placenta, and gives rise to some fundamental biochemical questions. The biological role of the progestagens is also discussed:...
Betteridge KJ.The developmental changes in the equine conceptus, its maternal environment and their interaction during the first 4 weeks following fertilization are reviewed. Attention is drawn to species-specific events to show why the horse is such a valuable model in which to study early pregnancy.