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.
COHEN H, BATES RW.THE occurrence of substances, in the extracts of human urine and of testes, that are capable of augmenting the effects of estrogens or androgens has been postulated and demonstrated by various workers. Freud and co-workers (1933, 1935) obtained factors from extracts of testes and human urine that augmented the effects of androgens, although possessing no androgenic activity themselves. Emmens (1938) described the presence of substances in the phenolic fraction of normal human female urine which in themselves were non-estrogenic, but when given orally, increased the potency of estriol injected ...
Hannan MA, Watanabe H, Takeyama A, Yoshida S, Wudamu D, Lkhagvasuren N, Claes A, Stout TAE, Cheong SH, Haneda S, Nambo Y.This study evaluated the viability of in vitro embryo production using ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) as breeding techniques for pure and crossbred Hokkaido native ponies (n = 9). Oocytes were collected using transvaginal ultrasound-guided follicle aspiration. ICSI was performed on in vitro matured oocytes using frozen semen. Embryonic cultures were monitored using time-lapse cinematography. Blastocysts were cryopreserved and, after thawing, were transferred non-surgically into recipient mares. Over nine OPU sessions, the mean number of aspirated follicles was 2...
Renaudin CD, Picandet V, Kass PH.Using the equation of Hartwig et al. and the table established for light breed horses, day of gestation was predicted in 108 Warmblood (WB) and 59 racehorses (Thoroughbred [TB] and Standardbred [STB]) pregnant mares. In WB, gestational ages were predicted within 2 weeks from 100 to 200 days of gestation using biparietal diameter (BPD), aortic diameter (AortD) or femur length (FL); within 2 weeks from 100 to 250 days using BPD or AortD; within 2 weeks from 100 to 300 days using BPD alone; and within 3 weeks from 100 to 350 days using eye approximated volume (EyV) alone. In the ...
Baatar T, Song D, Weng Y, Wang G, Jin L, Guo R, Li B, Dugarjaviin M.The epigenetic regulation of gene expression through the covalent modification of histones is crucial for developing germline cells. To study the regulatory role of alternative splicing (AS) of euchromatic histone lysine methyltransferase 2 (EHMT2/G9A) in spermatogenesis in Mongolian horses, this study first examines the localization of the EHMT2 gene in testicular support cells and then predicts the higher-order structures of sequences with and without AS. Two types of lentiviral vectors for overexpression were subsequently constructed for the EHMT2 gene, one with AS and one without, to infec...
Warren AS, Werner LA, Thombs LA, Rodgerson DH, Spirito MA, Sandow CB, McCracken MJ.To report the impact of ventral midline colopexy on colic, recurrence of large colon volvulus (LCV) and time to next live foal. Methods: Retrospective case control study. Methods: A total of 188 Thoroughbred broodmares surgically treated for LCV (2016-2020). Methods: Horses treated for LCV were grouped based on whether they received a colopexy (CP) or did not (NCP). All broodmares that underwent a second LCV surgery were subsequently categorized into two groups: those that received a colopexy (RCP) and those that did not (NRCP). Differences in the number of horses with large colon compromise, ...
Ziadi C, Valera M, Laseca N, Perdomo-González D, Demyda-Peyrás S, de Los Terreros AR, Molina A. By simultaneously integrating both genotyped and non-genotyped animals into genetic evaluation, the single-step genomic BLUP method enhanced the accuracy of genetic assessments. This study aimed to compare the increase in prediction reliability (R) between restricted maximum likelihood (REML) and single-step genomic REML (ssGREML) in the Pura Raza Española (PRE) horse breed. The dataset comprised reproductive records for seven fertility traits from 47,502 females, with a total of 57,316 animals represented in the pedigree. A total of 4009 animals were genotyped using the EQUIGENE 90K SNP ar...
Medica A, Aitken RJ, Swegen A, Gibb Z.Context Equine reproductive technologies are crucial for overcoming challenges in natural fertilisation, particularly in sub-fertile stallions and breeding programs focused on genetic conservation and performance enhancement. Assisted reproductive technologies (ARTs), such as artificial insemination (AI), intracytoplasmic sperm injection (ICSI), and in vitro fertilisation (IVF), improve fertility outcomes and enable breeding across geographical distances. Aims This review examines sperm isolation techniques used in ART, evaluating their efficacy, limitations, and potential to enhance reproduct...
Barandalla M, Colleoni S, Perota A, Galli C, Lazzari G.The introduction of ovum pick-up, Intracytoplasmic Sperm Injection (ICSI) and in vitro embryo production in equine reproduction offers significant advantages, including overcoming infertility, high blastocyst yields, embryo cryopreservation, and preimplantation genetic testing (PGT) by collecting few cells extruded spontaneously during embryo growth using a mouth pipette. PGT enables embryo selection before transfer and is particularly useful in breeds with a preference for specific offspring genders, coat color, and detecting genetic diseases. Here, we present the first large-scale validation...
Knudsen AR, Marsh JS, Pye JL.To describe reproductive performance in Thoroughbred broodmares post-large colon resection and to compare to performance post-colopexy. Methods: A single-institution retrospective study. Methods: A total of 29 client-owned horses: post-large colon resection (LCR) n = 19, post-colopexy: n = 10. Methods: Thoroughbred broodmares that survived until discharge following LCR (Group R) or colopexy (Group P) between 2006 and 2023 were included. Clinical data were collected from clinical records. Breeding and foaling data were collected from the Australian and New Zealand Studbooks. Statistical...
Couto G, Grippo A, Ismer A, Hoogewijs M, Pedro B, Vasconcelos L, Santos G, Wilsher S.Vitrified in vitro-produced embryos can be successfully warmed in isotonic media at room temperature (RT; 22°C). However, this protocol has not been reported for in vivo embryos, which are more challenging to vitrify and warm. Study objectives were to see if vitrified in vivo embryos warmed in RT isotonic medium gave equivalent pregnancy rates to stepwise serial dilution warming, and if embryo size influenced the results. One hundred and seventeen embryos were divided into groups by size (G1:≤ 300 μm, n = 59; G2:> 300-400 μm, n = 33; G3:> 400-500 μm, n = ...
Hernández-Avilés C.Sperm quality analysis using computer-assisted sperm analysis (CASA) systems and fluorescence-based techniques has become common in the animal reproduction industry, particularly for large animals (i.e., bovine, porcine, equine). In this chapter, the methods commonly utilized in the author's laboratory to examine sperm motion characteristics via CASA and plasma membrane intactness by flow cytometry will be described. These include methods to properly dilute fresh (stallions, bulls, boars), cool-stored (stallions, boars), or frozen/thawed (stallions, bulls, boars) sperm for assessment of sperm ...
Murase H, Wachi S, Matsuyama R, Makita K, Sato F.Pituitary pars intermedia dysfunction (PPID) is an age-related disease considered to have a negative impact on fertility. To understand the true impact of PPID on fertility, the influence of age must be considered. Objective: This study aimed to assess the impact of PPID on fertility while accounting for the effect of aging. Methods: A total of 332 Thoroughbred mares aged 10-20 years and managed across 137 private farms in Hidaka, Hokkaido, Japan were classified as PPID, equivocal, or non-PPID based on plasma adrenocorticotropic hormone concentration in 2019 and 2020. The prevalence of PPID an...
Al-Kass Z, Morrell JM, Ntallaris T.Sperm quality is adversely affected by cryopreservation due to the increased production of reactive oxygen species, which affects the integrity of sperm membranes, motility, and DNA fragmentation. Three methods for removing seminal plasma, washing (centrifuging extended semen at 800× for 10 min) and Single Layer Centrifugation with high or low density Equicoll, were used to prepare 29 ejaculates from ten stallions for freezing. Sperm quality parameters (kinematics, plasma membrane integrity, superoxide and hydrogen peroxide production, mitochondrial membrane potential, and DNA fragmentation)...
Bugno-Poniewierska M, Bielecka M, Pietras N, Kij-Mitka B, Podstawski Z, Długosz B.The Hucul horse is a Polish primitive breed with a small population size, which highlights the importance of preserving the genetic resources. The cryopreservation of semen is essential for creating gene banks, but its effect on the acrosome reaction in Hucul stallions has not yet been investigated. The acrosome reaction is one of the most important physiological events associated with the fertilization process. Therefore, our goal was to determine the level of acrosome reaction in chilled and frozen/thawed Hucul stallion semen using the FluoAcro test and the SCA semen analysis system. We foun...
Cortez JV, Hardwicke K, Méndez-Calderón CE, Grupen CG.The asynchrony of cytoplasmic and nuclear maturation in cumulus-oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of pre-IVM treatment with cAMP modulators for different durations on the developmental potential of equine oocytes used for cloned embryo production. Collected COCs were transferred to cryovials filled with transport medium at 20-22 °C. Within the cryovials, the COCs were either untreated (Control) fo...
Wen X, Bou G, He Q, Liu Q, Yi M, Ren H.The reproductive ability of equine species is a critical component of equine breeding programs, with sperm quality serving as a primary determinant of reproductive success. In this study, we perform an integrative analysis of proteomics and metabolomics in seminal plasma to identify proteins and metabolites associated with sperm quality and reproductive ability in equine species. Methods: We utilized the CEROS instrument to assess the morphology and motility of sperm samples from three horses and three donkeys. Additionally, we statistically analyzed the mating frequency and pregnancy rates in...
Ramírez-Agámez L, Crowley JB, Love CC, Hernández-Avilés C.Currently, Intracytoplasmic Sperm Injection (ICSI) on in vitro-matured equine oocytes obtained by transvaginal oocyte aspiration (TVA) is the method of choice for in vitro production of equine blastocysts in a clinical setting. A protocol for conventional in vitro fertilization (cIVF) using either fresh or frozen/thawed stallion sperm incubated under capacitating conditions (Tyrode's Albumin Lactate Pyruvate medium + penicillamine, hypotaurine, epinephrine; FERT-PHE) has been recently reported. Several factors that may impact the clinical applicability of this cIVF protocol have yet to be st...
Segabinazzi L, Lisboa FP, Zutter B.Limited response to conventional therapies and rising antibiotic resistance emphasize the need for alternative treatments for infectious endometritis. A recent in vitro study showed promising antimicrobial activity of a commercial uterine cleaning sanitizer (UCS) against endometritis pathogens. However, its safety and efficacy in mares remain untested. Objective: This study aimed to assess the clinical safety of intrauterine infusion of UCS in reproductively sound mares. Methods: Exp. 1. Six healthy mares with endometrial biopsies classified as I or IIA had two intrauterine infusions of UCS pe...
Barbosa LA, Maran AP, de Almeida MER, Guimarães EC, Bringel B, Douglas RH, Lima TFG, da Silva ESM.A longer endometrial exposure to estradiol before progesterone has been shown to be beneficial in cyclic and acyclic recipient mares. Therefore, the selection of an estradiol ester that promotes longer endometrial exposure to estradiol using a single administration would be advantageous when preparing acyclic mares as embryo recipients. This study investigated plasma estradiol profiles in acyclic mares after a single administration of 17-β estradiol (17-β), estradiol benzoate (EB) and estradiol cypionate (EC), and the correlation between plasma concentrations and endometrial edema. Fifteen n...
Bailey VN, Gilbert BM, Vetter M, Oberhaus EL.The mechanism by which photoperiod influences the hypothalamic-pituitary-gonadal (HPG) axis and regulates seasonal reproduction in horses has yet to be fully elucidated. The hypophyseal pars tuberalis (PT) has been indicated as a critical site for the transduction of melatonin signals through melatonin-responsive, PT-specific cells that produce thyroid-stimulating hormone (TSH) in many mammalian species. However, this has yet to be investigated in horses. The objective of this study was to explore the interaction of melatonin and thyroid-stimulating hormone in the equine HPG axis. Pituitaries ...
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...
Liu J, Yang Y, Wen L, Wen M, Zeng Y, Ren W, Yao X.The Kazakh horse is an outstanding dual-purpose dairy and meat breed in China, characterized by early maturity, tolerance to coarse feed, and strong stress resistance. Previous studies have examined gene expression patterns in the testicular tissues of Kazakh horses at different age stages, but the molecular mechanisms regulating testicular sexual maturation remain unclear. To address this gap, this study conducted HE staining and in-depth transcriptome sequencing analysis of Kazakh horse testicular tissue before and after sexual maturity. HE staining showed that the G3 group had well-formed s...
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...
Cortes D, Mendoza M, Raudsepp T, Ferrer M.Cervical duplication is a disorder of sexual development (DSD) that may be present in the absence of chromosomal aberrations. It is a rare cause of infertility in mares and is attributed to the failure of the Müllerian ducts to fuse properly, leading to the persistence of the median walls along the whole length of the cervix. This case report involves a 13-year-old American Quarter Horse mare with cervical duplication. Prior to this diagnostic, the mare was bred multiple times, but pregnancy was not achieved. The mare presented for a breeding soundness evaluation. During the examination an ab...
de Castro Sasahara TH, Gomes SP, Abdala FCM, Schimming BC, Miglino MA.Sexual differentiation in the equine fetus involves coordinated morphogenetic processes that shape both the gonads and the genital ducts. Although the formation of testes and ovaries has been relatively well documented, the temporal dynamics and morphometric patterns of the mesonephric (Wolffian) and paramesonephric (Müllerian) ducts remain insufficiently characterized. This study aimed to detail the timeline of gonadal and ductal differentiation in male and female equine fetuses during the critical developmental window between 60 and 80 days of gestation. Gross anatomical evaluation, histo...
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...
Satué K, Fazio E, Velasco-Martínez MG, Barbiera G, La Fauci D.Growth hormone (GH) influences follicular development mainly by stimulating insulin-like growth factor 1 (IGF-1), which acts systemically and locally within the ovary. However, the metabolic profile of follicular fluid (FF) in mares, particularly markers such as fructosamine (FRUCT), an indicator of glucose metabolism, remains poorly characterized. This study aimed to evaluate GH, IGF-1, and FRUCT concentrations in plasma and FF and their variation across follicle sizes. Sixty ovaries were collected during the breeding season from 30 clinically normal mares raised for meat production. Blood sa...
Márquez-Moya A, Carreras-Vico N, Sala-Ayala L, Martínez-Boví R, Cuervo-Arango J.Transvaginal ultrasound-guided follicular aspiration or ovum pick-up (OPU) has become the standard technique for oocyte collection in mares for intracytoplasmic sperm injection (ICSI). Although repeated follicular flushes and wall scraping are commonly used to improve oocyte recovery rate (ORR), the relative contribution of each remains unclear. This study aimed to compare the effects of multiple flushes versus controlled scraping time on ORR in mares. A controlled trial was conducted in two phases: (1) an ex vivo phase using slaughterhouse ovaries (n = 32), and (2) an in vivo phase in c...
Çortu A, Çınar H.Rectal examination is routine in equine reproduction but may trigger acute stress affecting welfare and safety. Intraocular pressure (IOP) responds rapidly to autonomic changes and may capture short-lived stress responses. Objective: To evaluate whether rectal examination acutely affects IOP and behavioral responses in mares, and whether reactions differ by reproductive status. Methods: Twenty Rahvan mares (10 pregnant, 30-40 days' gestation; 10 non-pregnant; 4-7 years) underwent transrectal examination without sedation. IOP was measured bilaterally at six time points: 2 min pre-exam (T1), dur...
Donato GG, Necchi D, Gionechetti F, Ala U, Nebbia P, Robino P, Stella MC, Vandaele H, Pallavicini A, Nervo T.In the past, bacteria detected in the mare's uterus were generally interpreted as a sign of endometritis, since the uterus was considered a sterile environment. This assumption has been challenged by the introduction of culture-independent molecular techniques, particularly 16S rRNA amplicon sequencing, which have demonstrated that healthy mares harbor an endometrial microbiota. The aim of this study was to characterize the endometrial microbiota of healthy mares and to determine whether microbial composition differs between estrus and diestrus. Endometrial samples were collected from eleven h...
