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Topic:Reproduction
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.
Histomorphometric and Histochemical Changes in the Mare Tuba Uterina During the Oestrous Cycle and Follicular Cyst Condition. This study evaluated histomorphometric and histochemical changes in the mare tuba uterina during different reproductive stages and under follicular cyst conditions. Forty nonpregnant mares were classified into follicular, luteal, post-ovulation and follicular cyst groups (n = 10/group). Histological evaluation of the isthmus, ampulla and infundibulum was performed using haematoxylin and eosin staining, while periodic acid-Schiff (PAS) staining was used to assess epithelial secretory activity. Morphometric parameters included number of primary mucosal folds, fold length, epithelial height a...
Evaluation of a commercial multiplex immunoassay for assessing uterine inflammation in mares susceptible to persistent breeding-induced endometritis. Persistent breeding-induced endometritis (PBIE) is characterized by a prolonged and dysregulated uterine inflammatory response following breeding. Although inflammatory cytokines are central to the post-breeding immune response, their assessment in the equine uterus has historically relied largely on gene expression analyses. Multiplex immunoassays provide an opportunity to simultaneously quantify multiple cytokine proteins within uterine secretions; however, the application of equine-specific multiplex immunoassays utilizing the Luminex platform to low-volume uterine lavage (LVL) fluid remain...
Effects of the Freezing Diluent Tonicity and Prefreezing Cooling Rates on Post-Thaw Sperm Quality Markers in the Pure-Bred Arabian Stallion. Stallion semen cryopreservation remains limited by inconsistent post-thaw sperm quality and fertility. This study evaluated the effects of freezing extender effective tonicity and prefreezing cooling rate on post-thaw sperm characteristics in pure-bred Arabian stallions. In a two-phase design, ejaculates ( = 40) from 10 stallions were processed using extenders with different osmotic conditions (330, 390, and 450 mOsm/kg) and subjected to cooling durations of 45 min, 1.5 h, or 3 h. Sperm quality was assessed using CASA, functional assays, flow cytometry, transcriptomic analysis, and...
Extreme male reproductive skew limits effective population size in American Standardbred horses. Standardbred horses are widely used for harness racing in North America, Europe, Australia, and New Zealand. In the U.S., two gaits (Pacers and Trotters) are genetically differentiated at a level typical for different horse breeds. We used the complete pedigree for all registered American Standardbred foals born 1970-2014 (>200 K Trotters and > 500 K Pacers) to calculate annual and lifetime reproductive skew and its consequences for the effective number of breeders per year (Nb) and effective size per generation (Ne). With rare exceptions, females can only produce 0 or 1 foal per year, w...
The Effect of Different Extenders and Testicular Anaesthesia During Orchiectomy on Post-Thawed Spermatological Parameters of Stallion Epididymal Spermatozoa. Cryopreservation of epididymal stallion spermatozoa is a valuable tool for fertility preservation, particularly after orchiectomy. However, the potential interactions between local anaesthesia and extender composition on post-thaw spermatozoal quality remain poorly understood. This study investigated the effects of intratesticular lidocaine injection and the efficacy of various extender (egg yolk [EY] vs. lecithin [LC]) and cryoprotectant (glycerol [GLY] vs. dimethyl sulfoxide [DMSO]) combinations on stallion epididymal spermatological parameters. Using a paired factorial design, epididymal s...
The effects of clomiphene citrate on reproductive parameters in the mare. Clomiphene citrate (CC) is a longstanding human fertility drug that has yet to be investigated in horses. The objective was to evaluate effects of CC on mare reproduction. After follicle ablation on Day 0, treatment mares (n=5) received 2 grams of CC intravenously on Days 1-5 while control mares (n=4) received no treatment. Transrectal ultrasonography was performed on Days 1-5 and 8, and transvaginal oocyte aspiration occurred on Day 8. Hormones were analyzed on Days 0-5, 8, 11, and 15. Mares receiving CC exhibited higher uterine edema grades compared to control mares (P=0.009). Antral follicl...
Supplementation of IVM media with equine follicular fluid of large follicles increases blastocyst yield in equine ICSI programs. Despite advances in the use of intracytoplasmic sperm injection (ICSI) for the in vitro production of equine embryos, oocyte maturation and competence to sustain pre-implantation embryo development remain significant challenges. This study evaluated the supplementation of the in vitro maturation (IVM) medium with equine follicular fluid (eFF), obtained from young mares (2-15 years old) with oocyte-to-blastocyst conversion rates >22%, as a strategy to improve oocyte developmental competence. In the first experiment, IVM medium was supplemented with 10% eFF collected from ovarian follicles of di...
Testicular Disorders in Equids: A Case Series. Equine testicular disorders represent a diverse group of conditions that impact the longevity of a stallion's breeding career. Stud farm veterinarians should be familiar with the variety of clinical presentations and diagnostic options for these conditions. This retrospective case series reports 57 cases of testicular abnormalities in stallions and donkeys presented to the clinic for further diagnostics and treatment. These disorders included neoplasia (three unilateral and one bilateral seminoma; two Sertoli cell tumors; two Leydig cell tumors; and one malignant mixed sex cord-stromal tumor),...
Genome-Wide Identification and Characterization of the Wnt Gene Family in Donkey (Equus asinus): Phylogenetic Analysis and Expression Profiling. The gene family encodes highly conserved signaling molecules involved in vertebrate development, tissue homeostasis, and cell fate regulation. However, the evolutionary characteristics and potential biological functions of genes in the domestic donkey () remain poorly understood. In this study, we performed the first genome-wide identification and comprehensive characterization of the donkey gene family. A total of 19 genes were identified, consistent with the conserved repertoire reported in other mammalian species. Phylogenetic and synteny analyses revealed strong evolutionary conservat...
Implications of dominance deviations for reproductive and morphological traits in the genetic management of the Pura Raza Española horse. Dominance is a non-additive genetic effect involving allele interaction at the same locus. In animal breeding, it has often been ignored due to its complexity and the low accuracy of pedigree-based estimates. This study analysed dominance variance components for reproductive traits [age at first foaling (AFF), reproductive efficiency (RE), interval between first and second foaling (I12)] and morphological traits [scapulo-ischial length (SIL), dorsal sternal diameter (DSD), knee circumference (KC)] in Pura Raza Española (PRE) mares. The aim was to assess the magnitude of dominance effects acro...
Russell’s viper envenomation in a 9-month pregnant mare saved with high-dose antivenom: a first-in-world case report. Snakebite during pregnancy is an uncommon but potentially catastrophic event, with maternal and foetal outcomes closely related to the severity of envenomation. While snakebite in pregnant women has been sporadically reported and is known to be associated with significant obstetrical complications, envenomation in animals, particularly in pregnant horses, is exceedingly rare. Only a few isolated cases of snakebite in horses have been documented, and to the best of our knowledge, snakebite in a pregnant mare has not been previously reported in the medical or veterinary literature. Russell's vip...
Body Condition, Metabolic Parameters and Semen Traits in Purebred Spanish Horses. The growing relevance of the Purebred Spanish horse has driven the extensive use of assisted reproductive technologies to preserve elite stallion genetics. Optimal reproductive performance is intrinsically linked to systemic health and high semen quality. This study aimed to evaluate the impact of body condition on the stallion's metabolic and endocrine profiles and their seminal parameters. Semen and blood samples were collected from 26 stallions (February-May in 2024-2025) whose body condition score (BCS) was assessed using the Henneke scale. They were classified into two groups, low BCS wit...
Somatic cell nuclear transfer as a complementary tool for genetic conservation of the Peruvian Paso horse. The Peruvian Paso horse, a breed of national significance in Peru known for its unique gait, has been selectively bred for nearly 500 years and now faces potential risks due to reduced genetic diversity. To investigate the potential of somatic cell nuclear transfer to enhance genetic conservation strategies, a program was initiated to produce cloned Peruvian Paso foals. Skin biopsies from a highly awarded stallion with exceptional breed-defining traits were used as the source of donor somatic cells. Of the 15 cloned embryos reconstructed, seven blastocysts were produced, and five were transfer...
Species-Specific Mechanisms of Sperm Transport: Predominant Roles of Motility in Ewes and Uterine Contractility in Mares. The factors controlling sperm transport within the female genital tract vary among species. The aim of this study was to quantify the respective roles of sperm motility and uterine contractions in passage of sperm within the reproductive tract of the mare and ewe. Sperm imaging was performed in vivo within the female genital tract of mares and ewes using probe-based confocal laser endomicroscopy (pCLE) after intrauterine insemination during two stages of the cycle characterized by high (oestrus) or low (luteal phase) uterine contractility. Fluorescent microbeads (2 µm in diameter) were insemi...
Combined effects of pre-freeze cholesterol-loaded cyclodextrin and post-thaw pentoxifylline on the functional longevity of cryopreserved stallion sperm. Cryopreservation causes structural and metabolic damage to stallion spermatozoa, impairing post-thaw functional competence. Objective: This study evaluated whether cholesterol-loaded cyclodextrin (CLC) supplementation before freezing, combined with pentoxifylline (PTX) after thawing, improves the functional quality, oxidative status, mitochondrial activity, and preliminary fertility outcomes of cryopreserved stallion semen. Methods: Semen from twelve stallions was assigned to four CLC concentrations (0, 1, 1.5, and 2 mg), frozen, thawed, and divided into subgroups with or without PTX supplemen...
Effects of a dimethylformamide-glycerol combination on freezing behavior, post-thaw sperm quality, and sperm-oocyte interaction in Arabian stallions. Although glycerol (GLY) and dimethylformamide (DMF) are widely used cryoprotectant agents (CPAs) for freezing equine semen, their combined use remains less characterized. Objective: To evaluate the effects of DMF, GLY, and their combination (DMF+GLY) on freezing behavior, sperm kinematics, membrane integrity, and IVF-related functional endpoints in Arabian stallion sperm. Methods: Twenty-four ejaculates from four stallions (six/male) were diluted in BotuSemen-Gold® extender supplemented with 5% DMF, 5% GLY, or 3% DMF+ 3% GLY, packed into 0.5-ml straws, and frozen. Sperm-oocyte binding and pro...
Efficiency of transvaginal oocyte aspiration (TVA), post-mortem ovary harvest (OH), and Intracytoplasmic Sperm Injection (ICSI) in a Quarter Horse mare population in North America. In a clinical setting, Intracytoplasmic Sperm Injection (ICSI)-derived equine blastocysts can be obtained either by transvaginal oocyte aspiration (TVA) or post-mortem ovary harvest (OH). Most peer-reviewed retrospective reports have described the suitability of these techniques for obtaining blastocysts, pregnancies, and live foals, particularly in Warmblood horse populations in Europe. In this study, retrospective data from Quarter Horse client-owned mares subjected to TVA-ICSI (n = 105; 151 cycles) or OH-ICSI (n = 42) at an academic institution in North America between 2023 and 2025...
Effects of equine sperm incubation under capacitation-supporting conditions on homologous and heterologous ICSI outcomes. Intracytoplasmic sperm injection (ICSI) in horses has become sufficiently efficient to be considered for clinical and commercial application; however, several factors influencing its efficiency remain poorly understood. This study evaluated whether incubating equine sperm under capacitating conditions induces capacitation-associated changes and affects embryo development following homologous equine ICSI, as well as oocyte activation, early cleavage, and 5-methylcytosine (5 mC) and 5-hydroxymethylcytosine (5hmC) levels following heterologous ICSI using bovine oocytes. Equine spermatozoa were ...
The effect of fenugreek (Trigonella foenum-graecum) on stallion spermatozoa motility and vitality in vitro. This study evaluated the effects of Trigonella foenum-graecum L. (fenugreek) seed extract on stallion spermatozoa quality in vitro, focusing on motility, vitality, and functional competence. In vitro spermatozoa analyses showed that fenugreek seed extract did not demonstrate overt cytotoxicity under the tested conditions: S1 = 1562 µg/mL; S2 = 781 µg/mL; S3 = 390.5 µg/mL; S4 = 195.25 µg/mL; S5 = 97.62 µg/mL; S6 = 48.81 µg/mL; S7 = 24.40 µg/mL. Computer-assisted semen analysis (CASA) revealed a concentration- and time-dependent Treatment × Time interac...
Development and validation of a liquid chromatography-tandem mass spectrometry method for the quantification of twenty-five steroids in equine serum. Steroids are potential biomarkers for monitoring equine pregnancy. However, immunoassays currently used for their quantification suffer from cross-reactivity and limited specificity, thus requiring more accurate methods. This study reports the development and validation of a robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of 25 steroids covering the main biosynthetic pathways of progestogens, corticosteroids, androgens, and estrogens. Steroids were extracted by protein precipitation followed by evaporation, derivatization, and reconstitut...
Seminal plasma exposure modulates peripheral T cell-associated transcription during early pregnancy in the mare. Successful establishment of pregnancy requires maternal immune tolerance to paternal antigens introduced by insemination. Regulatory T cell (Treg)-associated responses are thought to play a central role in this process, whereas pro-inflammatory T helper 1 (Th1) responses may disrupt pregnancy success. In other species, seminal plasma (SP) has been shown to modulate maternal immune adaptation; however, this relationship remains poorly defined in mares, where SP is frequently reduced during routine breeding practices. The objectives of this study were to (1) characterize temporal changes in syst...
Density gradient centrifugation preserves sperm kinetics and morphology in stallion ejaculates with severe urospermia. Urospermia is a frequent cause of subfertility in stallions due to the rapid deterioration of sperm quality. Objective: This study aimed to evaluate the effectiveness of different processing methods in minimizing the deleterious effects of urine on cooled equine semen. Methods: Ejaculates from 12 stallions were subjected to simple dilution, conventional centrifugation, and colloidal density gradient centrifugation, under challenges of 0%, 20%, and 50% urine, and stored at 5 °C for 24 h. Results: A severe dose-dependent effect was observed: increasing urinary concentration progressively reduce...
Preliminary Metabolomic Analysis: Serum Metabolomic Dynamics During Estrus Synchronization in Kazakh Mares. This study aimed to analyze the differences in serum metabolomes at different stages of estrus synchronization in Kazakh horses, to characterize dynamic changes in serum metabolites during estrus synchronization, and to screen potential metabolic markers and key regulatory pathways. Four sampling stages (M1: intravaginal device insertion; M3: device removal, labeled consistently with the pre-experiment timeline; P: 24 h post-PG injection; L: tertiary follicle emergence) were selected. The research results showed that 37 significantly different metabolites (DAMs) were identified in the M1-vs.-M...
Sequencing and Differential Analysis of miRNA in the Ovaries of Kazakh Horses During Estrus and Anestrus States. To reveal the regulatory role of miRNA in the estrous cycle transition of Kazakh horse ovaries and improve their reproductive efficiency, this study took the ovaries of Kazakh horses in the estrus period (DY group) and the quiescent period (DB group) as research objects. Small RNA sequencing and bioinformatics analysis were used to systematically analyze the expression characteristics and functional pathways of miRNA under the two physiological states. A total of 161 million high-quality miRNA tags were obtained through sequencing. Principal component analysis showed that the intra-group repea...
Pre-ovulatory follicles and ipsilateral ovarian stroma are cooler than non-dominant antral follicles and stroma on the contralateral ovary of the mare. Reports in several species have demonstrated that pre-ovulatory follicles (POFs) are cooler than neighbouring ovarian tissue, with cooling important for fertility. This study measured the temperature of POFs and compared it with other regions of the ovary, cervix, and rectum to determine if POF cooling occurs in mares. Fourteen oestrous cycles in 11 mares were monitored, and when an ovarian follicle ≥38 mm was present, deslorelin was given. Thirty hours later, rectal and cervical temperature measurements were taken. Mares were sedated and ovarian temperature measurements taken using a ther...
Sperm Energy-restriction and Recovery (SER) enhances equine sperm function and embryo development after ICSI. Assisted reproductive technologies (ART) are extensively used in domestic animal reproduction; however, improving their efficiency and consistency remains a major challenge for achieving reliable reproductive outcomes. The equine industry faces unique challenges related to sperm capacitation, which limited the success of conventional in vitro fertilization (IVF) for many years. Consequently, intracytoplasmic sperm injection (ICSI) became the most widely used technique for commercial in vitro embryo production. Even though a successful IVF protocol was recently developed, substantial opportunit...
Hair steroid hormones concentrations in jennies and their donkey foals from parturition to 6 months postpartum. The period after parturition/birth represents a delicate time for the dam and the offspring, during which several physiologic changes occur under finely tuned hormonal control, with the hypothalamic-pituitary- adrenal (HPA) axis playing a key role. This period, well studied in horses, received little attention in donkeys, despite the importance of species-specific knowledge for population preservation programs in many endangered donkey breeds. The present study aimed to investigate cortisol (C), DHEA(S), 17-β-estradiol (E2), progesterone (P4), testosterone (T), and allopregnanolone (AlloP) ha...
Platelet indices’ dynamic in broodmares: Effects of parity and lactation. Lactation and parity are physiological factors influencing platelet (PLT) indices, yet their combined effects in mares remain poorly characterized. As a dynamic process, lactation involves metabolic and endocrine changes that may influence thrombopoiesis; therefore, reproductive history and lactation stage may modulate PLT indices. Objective: To characterize longitudinal platelet (PLT) indices during early lactation in mares and evaluate the influence of parity on thrombopoietic responses. Methods: Fifty-five clinically healthy Spanish Purebred mares were included (20 primiparous and 25 multip...
Endometritis and endometrial fibrosis are associated with alterations in mare endometrial proteome. Equine persistent post-breeding endometritis may lead to endometrosis, characterized by excessive fibrosis, which is a major cause of subfertility/infertility. Endometrial proteomic changes induced by endometritis, when associated with fibrosis, were investigated to identify the biological processes (BP) involved. Cyclic mares´ endometrial biopsies were histologically classified (Kenney and Doig's) as: category IIA (n = 5), category IIA with endometritis (IIA-E; n = 4), category IIB (n = 5), or category IIB with endometritis (IIB-E; n = 5). Proteome was assessed by LC-MS/MS. I...
Evaluation of the Sidekick point-of-care progesterone lateral-flow assay for use in equine reproductive management. A point-of-care (POC; stall-side) test for progesterone that delivers rapid, accurate results has the potential to transform equine reproductive care. We evaluated the analytical and clinical performance of a POC device (Sidekick) that incorporates a progesterone lateral-flow assay (LFA) with a reader, and compared it with established methods (radioimmunoassay [RIA] and chemiluminescence immunoassay [CLIA]). Analytical performance was assessed using serially diluted mare plasma (0-8 ng/mL progesterone) to estimate precision and percent recovery. Clinical accuracy was assessed by comparison w...
