Fertility in horses encompasses the physiological processes and factors influencing reproductive success in equine species. It involves the study of reproductive anatomy, endocrinology, and behavior, as well as the management practices that affect breeding outcomes. Key aspects include the estrous cycle, conception rates, and factors impacting stallion and mare fertility. Reproductive technologies such as artificial insemination, embryo transfer, and hormonal therapies are also explored to enhance breeding efficiency. This page compiles peer-reviewed research studies and scholarly articles that examine the biological mechanisms, management strategies, and technological advancements related to equine fertility.
González N, Varela A, de Blas N, Gil L.At present, oviductal pathologies and their influence on mare fertility have not been clearly determined. Checking changes throughout the year in the oviductal structure may be relevant to understand the influence on the appearance of problems in oviductal functionality and, thus, the impact on fertility, which is the objective of this work. The oviducts of 56 mares were examined. Their anatomy, permeability, oviductal cellularity and the presence of pathologies were assessed to establish the correlations between the time of the year and each of these parameters. We cannot conclude whether the...
Latorre N, Gómez-Cuétara C, Cañizares E, Crespo F, Pérez-Aguilera V, Cuerda MI, Laborda-Gomariz JA, Soler AJ, Roldan ERS, Sanchez-Rodriguez A.Evaluating semen quality in Purebred Spanish Horses is essential to determine reproductive potential and optimize assisted reproductive techniques. Objective: This study compared several sperm analysis methods in Purebred Spanish Horses to standardize protocols suitable for both laboratory and field conditions. Methods: Fifteen stallions were evaluated through three ejaculates, analyzing sperm parameters using different techniques. Sperm kinematics was assessed by Sperm Computer Analyzer (SCA-CASA) or iSperm mCASA. Viability was analyzed by eosin-nigrosin-Giemsa (ENG) staining or Hoechst 33258...
Medica AJ, Gibb Z, Ortiz-Rodriguez JM, Swegen A.Heavy use of antioxidants in equine in vitro fertilisation (IVF) media may suppress the reactive oxygen species (ROS) signals stallion sperm need for capacitation, which depends on high mitochondrial activity and a well-maintained endogenous redox homeostasis. A species-specific approach balancing oxidative protection with controlled ROS signalling could improve fertilisation efficiency and reduce prolonged incubation times.
Griffin RA, Miller K, Colyvas K, Sheridan A, De Iuliis G, Aitken RJ, Baker MA, Gibb Z, Swegen A.Exposure to systemic heat stress in male mammals adversely affects sperm production, fertility, and DNA integrity. To date, few studies have investigated this phenomenon in horses, particularly in industry-relevant environments. Therefore, this study examined the relationship between ambient climatic conditions and fertility within a population of commercially fertile stallions. Post-coital semen samples ( = 804) were collected weekly from 46 Thoroughbred stallions during two successive breeding seasons (NSW, Australia; 22 weeks total). Semen samples were processed via single-layer colloid...
The ability of spermatozoa to withstand cryopreservation differs between stallions. While the underlying mechanisms of these differences are not fully understood, seminal plasma (SP) may play a crucial role in modulating sperm cryotolerance. Seminal plasma contains a heterogeneous population of extracellular vesicles (sEVs), nanometer-sized membrane particles that can transfer biomolecules to sperm modulating their function. This study aimed to isolate and characterize two sized-sEV subsets-small (S-) and large (L-)-from stallion SP, and to evaluate their involvement in sperm freezability. Sem...
Armangau M, Lurier T, Josson A.Offspring Sex Ratio (OSR) at birth in mammals is generally assumed to follow a 1:1 male-to-female distribution, yet deviations in OSR have been observed and may be influenced by maternal and paternal factors. In this study we have focused on the effect of paternal factors (stallion age and breed) and breeding methods on the OSR. Using a retrospective dataset of 251,783 foal births recorded in France between 2015 and 2024, we applied generalized linear mixed models to evaluate the effects of these variables on the likelihood of producing a female foal. Stallion identity was included as a random...
Tsopp E, Kilk K, Gambini A, Tvrdà E, Viljaste-Seera A, Tuppits U, Jaakma Ü.Equine in vitro embryo production (IVP) via ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) has gained prominence in sport-horse breeding due to high success rates, efficient semen utilization, and year-round embryo production, including from mares with fertility challenges or postmortem. However, assessing viability of early-stage ICSI-produced equine embryos remains challenging due to subjective morphological evaluations and limited visibility of developmental features. Non-invasive metabolomic analysis of culture media offers a promising alternative to enhance embryo selectio...
Cortez JV, Cervi D, Ruiz AJ, Grupen CG.Nicotinic acid (NA) treatment during in vitro maturation (IVM) has been shown to elevate nicotinamide adenine dinucleotide (NAD) levels and improve oocyte developmental competence. Suboptimal equine oocyte IVM systems currently limit the efficiency of viable embryo in vitro production. This study evaluated NA supplementation during IVM for cloned equine embryo production, using oocytes from abattoir-sourced ovaries and live mares via ovum pick-up (OPU). Abattoir-derived oocytes (n = 694) were treated without or with 50 or 200 μM NA during the 18 h holding period (Pre-IVM). Next, OPU-deriv...
Equine chronic degenerative endometritis (CDE) is a progressive process characterized by endometrial fibrosis that could be responsible for alterations of uterine environment and foetal-maternal communication. Objective: The aim of this study was to try to restore this communication by intrauterine administrations of amniotic cell-derived extracellular vesicles (AMC-EVs) in a case series. Methods: Twelve mares were selected on the basis of their reproductive history of early embryonic loss or abortion and clinical suspicion of CDE subsequently verified with histopathological examination of end...
Motta D, Aymée L, Roussouliéres I, Di Azevedo MIN, Jacob JCF, Lilenbaum W.Leptospirosis is a zoonotic disease caused by spirochetes of the genus Leptospira. Equine genital leptospirosis (EGL) has been described as a chronic and silent syndrome, presenting reproductive alterations such as abortion, stillbirth, placentitis, embryonic loss, repeat breeding syndrome, and subfertility. This study aimed to investigate the genital Leptospira infection in naturally infected mares with poor reproductive performance, as well as to genetically characterize the agents. A total of 41 mares with a history of poor reproductive performance were selected. Sera were collected for ser...
The research aims to study the dynamics of reproductive system pathologies in mares of Kazakh and Mugalzhar horse breeds and their impact on the reproductive traits of the breeding stock. The object of the research was horses of the Kazakh and Mugalzhar breeds. The formation of experimental groups was carried out using the pair-analog method. It was found that the frequency of detection of infertile mares is 13.8-25.0%. Subinvolution of the uterus from the total number of infertile mares is 8.3-10.7%, endometritis - 16.6-23.3%, atony and hypotonia of the uterus - 8.3-13.3%, ovarian diseases - ...
Cui B, Liu Y, Wu X, Li X.The objective of this study was to evaluate the effects of exogenous GnRH administration at the beginning of estrus synchronisation in mares during the spring transitional period. Estrus was synchronised using a progesterone releasing intravaginal device (PRID). The PRID was left in the vagina for 10 days, followed by an injection of 0.4 mg of cloprostenol at PRID removal. The GnRH group (n = 32) was subjected to intramuscular administration of 100 μg of the GnRH agonist triptorelin at PRID insertion, while the control group (n = 32) received 1 mL of sterile physiological sali...
Khalil WA, Sharf MI, Derbala MK, Hassan MAE, Abdelnour SA, El-Harairy MA.This study aims to investigate the cryoprotective effect of Omega-3 nano-emulsion (Omega-3 NE) on stallion sperm quality, kinematic parameters, acrosome status, subcellular ultrastructure, oxidative/antioxidant markers, and semen microbiota. Forty ejaculates were collected, extended, and cryopreserved from 5 fertile Pure Egyptian stallions (Equus caballus). The ejaculates were divided into five groups: a control group (without additive) and four groups supplemented with 25, 50, 100, and 200 μg of Omega-3 NE/mL. The Omega-3 NE exhibited an average particle size of 51-146 nm, a PDI of 0.58,...
Rychtarova J, Fulka H, Loi P, Fulka J.In the mouse, spermatozoa are highly resistant to DNA damage, even when frozen without cryoprotectants, and can produce offspring when subsequently used for ICSI (intracytoplasmic sperm injection). It is not known whether the same applies to other mammals as well. For example, in the horse, even conventional sperm freezing is still very problematic and frequently leads to sperm immobility. It has, however, never been tested whether sperm immobility also mirrors sperm head DNA damage, and if so, to what extent. In our study, we evaluated the damage to DNA in horse frozen and thawed motile and i...
Moroni R, Fanelli D, Maltinti S, Orefice M, Rota A, Camillo F, Melanie P, Cantile C, Miragliotta V, Pirone A, Lazzarini G, Passamonti F, Marmorini P....Equine persistent breeding-induced endometritis can lead to chronic uterine inflammation and fibrosis, reducing fertility. Intrauterine ozone (O₃) has recently been proposed as a potential treatment for mares unresponsive to conventional therapies. This case series describes O₃ treatment in eleven mares barren for at least one year despite multiple inseminations attempts. During the first oestrous cycle, low-volume uterine lavage and endometrial biopsy were performed, followed by 3 consecutive days of intrauterine O₂-O₃ gas insufflation. In the subsequent cycle, both procedures were re...
Bueno VLC, Bastos HBA, Centeno LAM, Schmitt F, Larentis G, Mattos RC, Fiala-Rechsteiner S.The seminal plasma (SP), a fluid originating from the testes, epididymis, and accessory sex glands, represents 98 % of the volume of stallion ejaculate. Metabolomic analysis enables the identification of final products of metabolic pathways, providing insights into reproductive physiology. Objective: This study aimed to investigate the metabolomic profile of stallion seminal plasma and to investigate metabolic differences between stallions with high and low pregnancy rates, in order to identify potential biomarkers associated with fertility. Methods: Twenty-four Criollo stallions, were sampled...
Khalil WA, Sharf MI, Derbala MK, Alfattah MA, Hassan MAE, Alhujaili W, El-Harairy MA, Abdelnour SA.Oxidative stress is a major contributor to male infertility. Therefore, fortifying assisted reproductive technology with nanotechnology could enhance sperm preservation. Objective: This study aimed to examine the impact of myo-inositol nano-emulsion (MINE) supplementation in semen extender on sperm quality, redox balance, semen bacteriology, apoptosis, ultrastructure, and acrosome status of chilled stallion semen. Methods: Semen samples were collected and preserved with 0 (MINE0), 1 (MINE1), and 2 (MINE2) mg of MINE/mL of extender. Results: Results revealed that extender fortified with 1 or 2 ...
Vallejo-Soto P, Dorado J, Herrera-García R, Álvarez-Delgado C, Gómez-Laguna J, Santiago Á, Manrique M, González Ariza A, León Jurado JM....Assisted reproductive techniques are often extrapolated from horses to donkeys, despite poorer fertility outcomes in jennies. This issue has been attributed to unknown uterine species-specific differences. This study compared, through histomorphometry, the endometrium of jennies and mares. Endometrial biopsies (N = 12) were taken from reproductively sound jennies (n = 6) and mares (n = 6) in estrus. Histomorphometric analysis evaluated luminal (LE, µm) and glandular epithelium height (GE, µm), glandular lumen diameter (LD, µm), glandular area (GA, µm), the number of glands (#G), and glandu...
Jones CJP, Aplin JD, Wilsher S.There is little information on non-ciliated secretory cell numbers in the equine oviduct isthmus over the course of the estrous cycle and the effect, if any, of insemination. Lectin histochemistry was used on a series of 13 oviducts taken at different stages of the estrous cycle, with and without insemination, to monitor glycosylation changes and the distribution of secretory cells characterised by a prominent cytoplasmic accumulation of glycoconjugates. There appeared to be an increase in secretory cells expressing a wide range of glycans in all specimens at estrus. One day after ovulation, o...
Vilela VMT, Zutter BM, Gomes LLO, Silva DF, Diniz JHW, Sancler-Silva YFR, Ignácio FS, Papa FO, Monteiro GA.This study investigated the effects of cholesterol-loaded cyclodextrin (CLC) before cryopreservation and the use of FertTalp (FT) after thawing on the structural and functional quality of equine spermatozoa. Methods: Two ejaculates from twelve stallions were divided into four groups: G1 (0 mg CLC), G2 (1.0 mg), G3 (1.5 mg), and G4 (2.0 mg). Post-thaw semen quality was evaluated through computer-assisted sperm analysis and flow cytometry. Results: In Experiment I, supplementation with 1.5 mg (G3) significantly improved total motility (64.9 ± 10.1 %) and progressive motility (40.5 ± 8.8 %) com...
Pinto-Bravo P, Rebordão MR, Amaral AS, Szóstek-Mioduchowska A, Fernandes C, Galvão AM, Silva E, Alpoim-Moreira J, da Costa RPR, Skarzynski DJ....Oviduct glycoprotein 1 (OVGP1) is a key protein involved in oviductal functions. β-estradiol (E) and progesterone (P), oxytocin (OXT) and tumor necrosis factor-α (TNFα) modulate the equine oviduct function, through prostaglandin regulation. The objective was to evaluate OVGP1 expression within each equine oviduct segment (infundibulum, ampulla isthmus), throughout the estrous cycle. The in vitro effect of (i) E, P, OXT, TNFα; and (ii) spermatozoa, on oviduct OVGP1 transcription and secretion was studied. Gene transcription was assessed by real-time PCR; protein expression by western blot; ...
Mendoza Cerna MN, Anderson H, Avni G, Kahila Bar-Gal G, Juras R, Raudsepp T.Robertsonian translocations (ROBs) or centric fusions of acrocentric chromosomes are the most common structural chromosomal rearrangements in mammals. ROBs are of medical and veterinary concern because of associated subfertility and congenital disorders but are also of interest as a mechanism of chromosome and karyotype evolution. While ROBs are well documented in humans, mice, and cattle/bovids, they are extremely rare in horses, despite the 18 acrocentric chromosomes in the horse karyotype. Methods: We characterize the case using conventional and molecular cytogenetic approaches and DNA anal...
Otero JC, Neild DM, Ferrante AA, Gambarotta MC, Caldevilla ML.Glycerol, a penetrating cryoprotectant, is most employed for deep freezing spermatozoa. However, it can induce toxic, chemical, and osmotic effects, altering the lipid structure of the sperm membrane. Rapid glycerol addition has been demonstrated to have fewer toxic effects than its removal. Objective: To minimize both the toxic and osmotic effects of glycerol on equine sperm through post-thaw dilution using two different extenders. Methods: Samples of equine semen frozen with 5% glycerol from nine stallions were thawed, re-diluted 1:2, and incubated for 30 minutes at 22°C in Tyrodes (Tyr) an...
Fresa K, Catandi GD, Gonzalez-Castro R, Omar A, Whitcomb LA, Cheng MH, Chen TW, Carnevale EM, Chicco AJ.Advancing age is associated with a decline in fertility and functional capacity, which may result in part from suboptimal nutrition and impaired mitochondrial function. Dietary essential polyunsaturated fatty acids (PUFA) are broadly recommended to mitigate weight loss and reduce risk of chronic disease in aged populations, but their effects on mitochondrial function are less clear. The present study investigated the impacts of dietary supplementation with essential omega-3 PUFA (flaxseed oil; N3) or omega-6 PUFA (corn oil, N6) on blood, muscle and follicular cell fatty acid composition and mi...
Mules (Equus mulus), as sterile hybrids between mares and donkeys, present a unique uterine morphology and physiology that remains poorly characterized. This study provides the first histomorphometric evaluation and transcriptional profiling of the endometrium in adult mules. To achieve this, endometrial biopsies were analysed using quantitative stereology, histological classification (Kenney-Doig system), and qPCR of key immune-related and fibrotic genes. Histometric analyses revealed that the stratum spongiosum was the predominant component of the mule endometrium (mean volume density: 84.7...
Martin-Pelaez S, Orsolini M, Verstraete M, de la Fuente A, Takahashi K, Meyers S, Dini P.Equine in vitro embryo production (IVP) offers a valuable opportunity to enhance the efficient utilization of genetically and economically valuable horses. In cases of subfertility, sudden death, emergency euthanasia, or limited sperm availability, IVP becomes the only viable option to produce offspring. Due to steadily improving success rates, IVP has become one of the most sought-after assisted reproductive techniques. The rising global demand has created a clear need for veterinarians and stallion owners to provide IVP-suitable sperm, where the selection of intact sperm for intracytoplasmic...
Derar D, Ali A, Al-Sobayil F, Refaai W.Granulosa cell tumors (GCTs) represent the most common ovarian neoplasms in mares, yet data on their epidemiology, risk factors, and clinical outcomes in Arabian horses are limited. Understanding their prevalence and diagnostic features is crucial for improving fertility management in this breed. Unassigned: Clinical records from 807 Arabian mares housed on 35 stud farms in central Saudi Arabia were retrospectively reviewed to determine the prevalence of GCT and potential risk factors, including age, parity, and anabolic steroid use. Twenty-four confirmed cases were further investigated using ...
Solanki O, Talluri TR, Jhamb D, Kumar S, Pargi KK, Paul N, Kutty M, Dedar RK, Meena M, Gaur M, Bhattacharya TK.The current study evaluated the effect of L-Carnitine supplementation to the stallion semen extender on pre-freeze and post-thaw semen quality and oxidative parameters in Marwari breed stallions. Methods: For this purpose, thirty-six semen ejaculates (6 ejaculates from each stallion) were collected using an artificial vagina (AV) from six adult Marwari stallions. The gel free semen was extended with different concentrations of L-Carnitine (LC) i.e. 0 mM (control), 2.5 mM (T1), 5 mM (T2) and 10 mM (T3). Pre-freeze and post-thaw semen evaluations were conducted for various seminal attributes and...
Broothaers K, Jouneau A, Angel-Velez D, De Coster T, Festuccia N, Archilla C, Calderari S, Jouneau L, Van den Branden E, Peere S, Polfliet E....In vitro production of equine embryos has been performed using intracytoplasmic sperm injection (ICSI) for the last two decades. Since 2022, a repeatable protocol for conventional in vitro fertilization (IVF) provides a successful alternative. However, little is known about the influence of the fertilization method on embryo quality and the transcriptomic profile. In this study, we aimed to examine differentially expressed genes (DEGs) between ICSI and IVF embryos in the horse. Therefore, ten equine sibling blastocysts, produced in vitro by either ICSI or IVF from three different mares, were s...
Boerboom D, Paquet M, Hsieh M, Liu J, Jamin SP, Behringer RR, Sirois J, Taketo MM, Richards JS.Misregulation of the Wnt/beta-catenin signaling pathway is a hallmark of several forms of cancer. Components of the Wnt/beta-catenin pathway are expressed in ovarian granulosa cells; nevertheless, its potential involvement in granulosa cell tumorigenesis has not been examined. To this end, human (n = 6) and equine (n = 18) granulosa cell tumors (GCT) were analyzed for beta-catenin expression by immunohistochemistry. Unlike granulosa cells of normal ovaries, most (15 of 24) GCT samples showed nuclear localization of beta-catenin, suggesting that activation of the Wnt/beta-catenin pathway plays ...
Yánez-Ortiz I, Catalán J, Rodríguez-Gil JE, Miró J, Yeste M.Sperm cryopreservation is one of the most important procedures in the development of biotechnologies for assisted reproduction. In some farm animals, the use of cryopreserved sperm has so many benefits for which relevance has become more evident in recent decades. Values for post-thaw sperm quality, however, are variable among species and within individuals of the same species. There is no standardized methodology for each of the stages of the cryopreservation procedure (andrological examination, semen collection, dilution, centrifugation, resuspension of the pellet with the freezing medium, p...
LeBlanc MM, Causey RC.Endometritis, a major cause of mare infertility arising from failure to remove bacteria, spermatozoa and inflammatory exudate post-breeding, is often undiagnosed. Defects in genital anatomy, myometrial contractions, lymphatic drainage, mucociliary clearance, cervical function, plus vascular degeneration and inflamm-ageing underlie susceptibility to endometritis. Diagnosis is made through detecting uterine fluid, vaginitis, vaginal discharge, short inter-oestrous intervals, inflammatory uterine cytology and positive uterine culture. However, these signs may be absent in subclinical cases. Hyper...
Hahn S, Giaglis S, Hoesli I, Hasler P.The intention of this review is to provide an overview of the potential role of neutrophil extracellular traps (NETs) in mammalian reproduction. Neutrophil NETs appear to be involved in various stages of the reproductive cycle, starting with fertility and possibly ending with fetal loss. The first suggestion that NETs may play a role in pregnancy-related disorders was in preeclampsia, where vast numbers were detected in the intervillous space of affected placentae. The induction of NETosis involved an auto-inflammatory component, mediated by the increased release of placental micro-debris in p...
White IG.When sperm of the ram, bull, boar and stallion are cold-shocked by rapid cooling to near freezing point, motility and metabolic activity are irreversibly depressed and the acrosome and plasma membrane disrupted. Ram sperm become susceptible to cold shock in the proximal corpus region of the epididymis when the cytoplasmic droplet has moved backwards to the distal portion of the sperm midpiece. The membrane constituents phospholipids and cholesterol are important in cold shock which causes loss of lipid from sperm. The susceptibility of sperm to cold shock is linked with a high ratio of unsatur...
Gibb Z, Lambourne SR, Aitken RJ.The relationship between stallion fertility and oxidative stress remains poorly understood. The purpose of this study was to identify criteria for thoroughbred fertility assessment by performing a logistical regression analysis using "dismount" sperm parameters as predictors and weekly per-cycle conception rate as the dependent variable. Paradoxically, positive relationships between fertility and oxidative stress were revealed, such that samples that produced pregnancies exhibited higher rates of 8-hydroxy-2'-deoxyguanosine release (1490.2% vs. 705.5 pg/ml/24 h) and lower vitality (60.5% vs. 6...
Katila T.This review attempts to summarize the current knowledge on uterine inflammatory response after mating in horses, pigs and cattle. Post-mating endometritis has been extensively studied in horses as it has been considered to cause infertility. The inflammation is known to occur also in cattle, but it has not been investigated to a similar extent. There are a number of publications about mechanisms of post-mating uterine inflammation in pigs, which seem to resemble those in horses. The major focus of this review is the horse, but relevant literature is presented also on swine and cattle. Spermato...
Aitken RJ, Gibb Z, Mitchell LA, Lambourne SR, Connaughton HS, De Iuliis GN.The prolonged incubation of human spermatozoa in vitro was found to induce a loss of motility associated with the activation of mitochondrial reactive oxygen species generation in the absence of any change in mitochondrial membrane potential. The increase in mitochondrial free radical production was paralleled by a loss of protein thiols and a concomitant rise in the formation of 4-hydroxynonenal, an electrophilic product of lipid peroxidation that was found to directly suppress sperm movement. These results prompted a search for nucleophiles that could counteract the action of such cytotoxic ...
Kenney RM.Normal histologic changes that occurred in concert with the ovarian cycle were evaluated. When combined with behavioral and physical findings, this evaluation enabled determination when there was synchrony between ovarian steroids (estrogen and progesterone) and their target tissues of endometrium. A system for classification as well as definitions of patterns types and degrees of severity of inflammatory and fibrotic changes was developed to facilitate a uniform system for description and communication. An attempt was made to relate the pathologic changes to their effect on fertility when fer...
Ball BA.Oxidative stress is an important component of the cytopathology of equine spermatozoa undergoing storage as liquid or frozen semen. Damage to chromatin, membranes and proteins of sperm are important components of oxidative damage to sperm. Similarly, sperm are exposed to a variety of osmotic stresses during storage that result from exposure to hypertonic media or result as a consequence of osmotic changes induced during freezing. A number of changes induced during processing and storage of equine sperm also appear to induce apoptotic-like changes which may adversely affect sperm survival and f...
Su TW, Choi I, Feng J, Huang K, McLeod E, Ozcan A.We report the discovery of an entirely new three-dimensional (3D) swimming pattern observed in human and horse sperms. This motion is in the form of 'chiral ribbons', where the planar swing of the sperm head occurs on an osculating plane creating in some cases a helical ribbon and in some others a twisted ribbon. The latter, i.e., the twisted ribbon trajectory, also defines a minimal surface, exhibiting zero mean curvature for all the points on its surface. These chiral ribbon swimming patterns cannot be represented or understood by already known patterns of sperms or other micro-swimmers. The...
Riddle WT, LeBlanc MM, Stromberg AJ.Endometrial cytology and culture specimens (n=2123) were collected concurrently with a guarded uterine culture instrument from 970 mares (738 barren, 1230 foaling and 155 maiden mares) during three breeding seasons (2001-2004). Results were compared to the 28-d pregnancy rate for the cycle from which the samples were taken. Cytological smears were evaluated for inflammation at x100 and graded as: not inflammatory (0-2 neutrophils/field), moderate inflammation (2-5 neutrophils/field), severe inflammation (>5 neutrophils/field), or hypocellular (scant epithelial cells and no neutrophils). Ute...
Janečka JE, Davis BW, Ghosh S, Paria N, Das PJ, Orlando L, Schubert M, Nielsen MK, Stout TAE, Brashear W, Li G, Johnson CD, Metz RP, Zadjali AMA....Dynamic evolutionary processes and complex structure make the Y chromosome among the most diverse and least understood regions in mammalian genomes. Here, we present an annotated assembly of the male specific region of the horse Y chromosome (eMSY), representing the first comprehensive Y assembly in odd-toed ungulates. The eMSY comprises single-copy, equine specific multi-copy, PAR transposed, and novel ampliconic sequence classes. The eMSY gene density approaches that of autosomes with the highest number of retained X-Y gametologs recorded in eutherians, in addition to novel Y-born and transp...
H A Morris L, M McCue P, Aurich C.Endometritis in the mare begins as a normal physiological inflammatory response to breeding that involves both a mechanical and immunological response pathway activated to rid the uterus of semen and bacteria. With successful resolution of this inflammation, the mare's uterus will provide a hospitable environment for the development of the semi-allogenic conceptus. If the mare fails to resolve this inflammatory response within 48 h of breeding, she will become susceptible to persistent breeding-induced endometritis (PBIE) which will have detrimental effects on her fertility. This condition can...
Ambruosi B, Uranio MF, Sardanelli AM, Pocar P, Martino NA, Paternoster MS, Amati F, Dell'Aquila ME.Phthalates are ubiquitous environmental contaminants because of their use in plastics and other common consumer products. Di-(2-ethylhexyl) phthalate (DEHP) is the most abundant phthalate and it impairs fertility by acting as an endocrine disruptor. The aim of the present study was to analyze the effects of in vitro acute exposure to DEHP on oocyte maturation, energy and oxidative status in the horse, a large animal model. Cumulus cell (CC) apoptosis and oxidative status were also investigated. Cumulus-oocyte complexes from the ovaries of slaughtered mares were cultured in vitro in presence of...
Weems CW, Weems YS, Randel RD.Prostaglandins impact on ovarian, uterine, placental, and pituitary function to regulate reproduction in female livestock. They play important roles in ovulation, luteal function, maternal recognition of pregnancy, implantation, maintenance of gestation, microbial-induced abortion, parturition, postpartum uterine and ovarian infections, and resumption of postpartum ovarian cyclicity. Prostaglandins have both positive and negative effects on reproduction; they are used to synchronize oestrus, terminate pseudopregnancy in mares, induce parturition, and treat retained placenta, luteinized cysts, ...
Das PJ, McCarthy F, Vishnoi M, Paria N, Gresham C, Li G, Kachroo P, Sudderth AK, Teague S, Love CC, Varner DD, Chowdhary BP, Raudsepp T.Mature mammalian sperm contain a complex population of RNAs some of which might regulate spermatogenesis while others probably play a role in fertilization and early development. Due to this limited knowledge, the biological functions of sperm RNAs remain enigmatic. Here we report the first characterization of the global transcriptome of the sperm of fertile stallions. The findings improved understanding of the biological significance of sperm RNAs which in turn will allow the discovery of sperm-based biomarkers for stallion fertility. The stallion sperm transcriptome was interrogated by analy...
Kirkpatrick JF, Lyda RO, Frank KM.Wildlife, free-ranging and captive, poses and causes serious population problems not unlike those encountered with human overpopulation. Traditional lethal control programs, however, are not always legal, wise, safe, or publicly acceptable; thus, alternative approaches are necessary. Immunocontraception of free-ranging wildlife has reached the management level, with success across a large variety of species. Thus far, the immunocontraceptive research and management applications emphasis have been centered on porcine zona pellucida and gonadotropin-releasing hormone vaccines. Contraceptive succ...
Rathi R, Colenbrander B, Bevers MM, Gadella BM.The primary aim of this study was to establish a flow cytometric technique for determining the capacitation status of stallion spermatozoa. To this end, a flow cytometric technique that demonstrates changes in plasma membrane fluidity; namely, merocyanine 540 staining, was compared with the more conventional Ca(2+)-dependent fluorescence microscopic technique, chlortetracycline (CTC) staining, for assessing capacitation status. In addition, the effect of bicarbonate/CO(2) on the progress of capacitation and the acrosome reaction (AR) and on temporal changes in sperm motility, with particular r...
Binder AK, Rodriguez KF, Hamilton KJ, Stockton PS, Reed CE, Korach KS.Determining the spatial and temporal expression of genes involved in the ovulatory pathway is critical for the understanding of the role of each estrogen receptor in the modulation of folliculogenesis and ovulation. Estrogen receptor (ER)-β is highly expressed in ovarian granulosa cells, and mice lacking ER-β are subfertile due to inefficient ovulation. Previous work has focused on isolated granulosa cells or cultured follicles and, although informative, provides confounding results due to the heterogeneous cell types present including granulosa and theca cells and oocytes and exposure to in...
Galli C, Duchi R, Colleoni S, Lagutina I, Lazzari G.Assisted reproductive techniques developed for cattle in the last 25 years, like ovum pick up (OPU), intracytoplasmic sperm injection (ICSI), and somatic cell nuclear transfer, have been transferred and adapted to buffalo and horses. The successful clinical applications of these techniques require both the clinical skills specific to each animal species and an experienced laboratory team to support the in vitro phase of the work. In cattle, OPU can be considered a consolidated technology that is rapidly outpacing conventional superovulation for embryo transfer. In buffalo, OPU represents the ...
da Silveira JC, Winger QA, Bouma GJ, Carnevale EM.Age-related decline in fertility is a consequence of low oocyte number and/or low oocyte competence resulting in pregnancy failure. Transforming growth factor (TGF)-β signalling is a well-studied pathway involved in follicular development and ovulation. Recently, small non-coding RNAs, namely microRNAs (miRNAs), have been demonstrated to regulate several members of this pathway; miRNAs are secreted inside small cell-secreted vesicles called exosomes. The overall goal of the present study was to determine whether altered exosome miRNA content in follicular fluid from old mares is associated wi...
Muller B, Lewis N, Adeniyi T, Leese HJ, Brison DR, Sturmey RG.Mitochondria provide the major source of ATP for mammalian oocyte maturation and early embryo development. Oxygen Consumption Rate (OCR) is an established measure of mitochondrial function. OCR by mammalian oocytes and embryos has generally been restricted to overall uptake and detailed understanding of the components of OCR dedicated to specific molecular events remains lacking. Here, extracellular flux analysis (EFA) was applied to small groups of bovine, equine, mouse and human oocytes and bovine early embryos to measure OCR and its components. Using EFA, we report the changes in mitochondr...
Loux SC, Crawford KR, Ing NH, González-Fernández L, Macías-García B, Love CC, Varner DD, Velez IC, Choi YH, Hinrichs K.In vitro fertilization does not occur readily in the horse. This may be related to failure of equine sperm to initiate hyperactivated motility, as treating with procaine to induce hyperactivation increases fertilization rates. In mice, hyperactivated motility requires a sperm-specific pH-gated calcium channel (CatSper); therefore, we investigated this channel in equine sperm. Motility was assessed by computer-assisted sperm motility analysis and changes in intracellular pH and calcium were assessed using fluorescent probes. Increasing intracellular pH induced a rise in intracellular calcium, w...
Yang F, Fu B, O'Brien PC, Nie W, Ryder OA, Ferguson-Smith MA.We have made a complete set of painting probes for the domestic horse by degenerate oligonucleotide-primed PCR amplification of flow-sorted horse chromosomes. The horse probes, together with a full set of those available for human, were hybridized onto metaphase chromosomes of human, horse and mule. Based on the hybridization results, we have generated genome-wide comparative chromosome maps involving the domestic horse, donkey and human. These maps define the overall distribution and boundaries of evolutionarily conserved chromosomal segments in the three genomes. Our results shed further lig...
Cheng FP, Fazeli A, Voorhout WF, Marks A, Bevers MM, Colenbrander B.Peanut agglutinin (PNA) was used to assess the sperm acrosomal status and the acrosome reaction during gamete interaction in the equine species. PNA exclusively binds to the outer acrosomal membrane of stallion spermatozoa, as was established by transmission electron microscopy. Fluorescein isothiocyanate-PNA (FITC-PNA) labeling was used to monitor sperm acrosomal changes during a prolonged incubation period of 24 hours and during a 2-hours incubation in the presence of 5 microM calcium ionophore A23187. In addition, after a 4-hours preincubation in SP-TALP medium, sperm samples were incubated...
Carnevale EM.Reproductive aging and assisted reproduction are becoming progressively more relevant in human medicine. Research with human subjects is limited in many aspects, and consequently animal models may have considerable utility. Such models have provided insight into follicular function, oocyte maturation, and reproductive aging. However, models are often selected based on factors other than physiological or functional similarities. Although the mare has received limited attention as a model for reproduction in women, comparisons between these species indicate that the mare has many attributes of a...
Loomis PR, Graham JK.One of the challenges for those attempting to cryopreserve stallion spermatozoa is dealing with the stallion to stallion variability in the cryosurvival of their semen. In the dairy industry, each bull stud, essentially utilizes a single cryopreservation technique, and bulls that produce sperm that do not cryopreserve well using that technique are replaced by other bulls. However, replacing stallions is unlikely to prove acceptable to the equine industry, where specific genotypes are desired. Instead, to increase the number of stallions that can be effectively utilized for cryopreserved semen ...
Morris LH, Allen WR.The findings of a retrospective survey of 1393 Thoroughbred mares visiting 22 studfarms in the Newmarket region of the UK during the 1998 mating season were compared with those of a similar study undertaken in 1983. The effects of mare age and status, stallion, month of mating, application of uterine treatments and other parameters on the rates of singleton and twin conception and subsequent pregnancy losses were analysed. Mare age and status significantly affected the per cycle pregnancy rate and the incidence of pregnancy loss. Overall, the mean number of matings per oestrus was 1.12 and the...
Gravance CG, Garner DL, Baumber J, Ball BA.The fluorescent carbocyanine dye, JC-1, labels mitochondria with high membrane potential orange and mitochondria with low membrane potential green. Evaluation of mitochondrial membrane potential with JC-1 has been used in a variety of cell types, including bull spermatozoa; however, JC-1 staining has not yet been reported for equine spermatozoa. The aim of this study was to apply JC-1 staining and assessment by flow cytometry or a fluorescence microplate reader for evaluation of mitochondrial function of equine spermatozoa. Six ejaculates from four stallions were collected and centrifuged thro...
