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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.
Comparative Evaluation of Sperm Quality Assessment Methods in Purebred Spanish Horses. 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...
Changes in the Mare Oviduct Across Different Seasons Throughout the Year. 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...
Are antioxidants hindering stallion sperm capacitation? A perspective on hidden barriers to equine IVF success. 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.
Functional Characterization of Long Non-Coding RNAs Associated with Reproductive Fitness in Pura Raza Española Mares. Long non-coding RNAs (lncRNAs) are transcripts constituted of more than 200 nucleotides that have been associated with the regulation of different biological processes by modulating the expression of key genes. In horses, evidence suggests that lncRNAs play a role in female reproductive fitness, yet their functional implications remain poorly characterized. The objective of this study was to investigate potential DNA:RNA triplex interactions between the promoter regions of fertility-related genes and lncRNAs transcribed from non-coding loci located within ±50 kb of these genes. By doing so, w...
Deficiency in homozygous haplotypes reveals recessive lethal variants affecting fertility and viability in the Friesian horse. BACKGROUND: Recessive lethal alleles causing pre- or postnatal death in homozygous mutant animals, could lead to reduced fertility success. The Friesian horse breed has signs of reduced fertility and has faced high inbreeding rates in the past (∆F > 1%). Consequently, by genetic drift lethal alleles may have reached moderate to high frequencies in the population. Our aim was to identify lethal recessive alleles that — when homozygous — may cause pre- or postnatal death in the Friesian horse. RESULTS: We analyzed genotypes (70 K SNP) of over 8,000 Friesian horses, looking for hapl...
Gene Editing and the Future of Thoroughbred Breeding and Racing. Prohibited gene editing in horses (either in embryos or via cell culture and cloning) can result in both desired and undesired outcomes. If left undetected, changes can proliferate within the population in subsequent generations, posing a major threat to welfare and breed integrity.
Long-term micronutrient and amino acid supplementation increases embryo recovery per flush and ovarian responsiveness in donor mares. Background: Nutritional strategies may modulate ovarian function and embryo recovery in donor mares, but field evidence for long-term multinutrient supplementation remains limited.Aims/objectivesTo determine whether long-term micronutrient and amino acid supplementation increases embryo recovery per flush and ovarian responsiveness in donor mares.Methods: A total of 115 uterine flushes were performed under a crossover design across two consecutive breeding seasons in 12 multiparous, non-lactating crossbred donor mares (4-12 years). Mares received a basal diet or the same diet plus 40 g/day of ...
Characterization of isolated late preantral and early antral equine ovarian follicles. Understanding how to effectively isolate a large number of healthy follicles is essential for increasing success rates in ARTs, such as in vitro culture and cryopreservation. Late secondary (preantral) and early tertiary (antral) follicles isolated from the intermediary portion of equine ovarian tissue sliced at two different thicknesses (0.25 mm vs. 0.50 mm) were characterized according to regional distribution (dorsal, medial, and ventral), morphofunctional features, and epigenetic profiles. After isolation via microdissection, follicles were evaluated for membrane integrity (trypan blue), v...
Relationship between the duration of endometrial edema and reproductive parameters of estrus, diestrus, and fertility in mares: a field study. The duration of endometrial edema has been positively correlated with uterine receptivity and likelihood of pregnancy. Therefore, this study aimed to evaluate whether there is a relationship between the duration of endometrial edema (DEdema) and other key parameters of the estrous cycle: maximum edema (MEdema), largest follicle diameter (LPF), corpus luteum and uterine tone on day 5 post ovulation (CLD5 and TD5), progesterone concentration (P4) and fertility. A total of 88 cycles from 36 mares were evaluated and divided into Short Estrus (SE), Long Estrus (LE), Inseminated (AI) and Non-insemin...
Isolation and characterization of seminal extracellular vesicles subsets and their impact on sperm freezability in stallions. 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...
Effects of rectal examination on intraocular pressure and behavioral responses in pregnant and non-pregnant mares. 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...
Paternal determinants of Offspring Sex Ratio in horses: A retrospective analysis of stallion breed, age, and breeding method. 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...
Confocal laser endomicroscopy: exploring endometrial image quality in relation to the oestrous cycle of mares. Assessment of the endometrium is an essential component of a breeding soundness examination in mares. The current reference standard for evaluating equine endometrial health is histopathological assessment of an endometrial biopsy, however, inconsistencies may arise when a single blinded biopsy is used. Confocal laser endomicroscopy (CLE) is non-invasive and enables real-time in-vivo imaging of the endometrium across multiple locations within the uterine lumen. An initial study revealed variation in CLE image quality between mares. Objective: This study aimed to compare the quality of endometr...
Characterization of the Endometrial Microbiota of Healthy Mares Across the Estrous Cycle. 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...
Effects of L-selenomethionine supplementation on nutrient digestibility and metabolism, antioxidant capacity, hormone levels, and fecal microbiota diversity in pregnant Yili mares during mid- to late gestation. L-selenomethionine (L-SeMet), a highly bioavailable organic form of selenium, plays a critical role in maintaining antioxidant homeostasis, regulating reproductive hormone secretion, and improving intestinal microbial ecology. Previous studies have demonstrated that appropriate supplementation with L-SeMet can significantly enhance the production performance and health status of ruminants. However, the nutritional regulatory mechanisms and physiological effects of L-SeMet in monogastric herbivores, particularly horses during mid- to late gestation, remain inadequately understood. Therefore, th...
Comparative Evaluation of Follicular Flushing Frequency and Scraping Time During Ovum Pick-Up in Mares: Effects on Oocyte Recovery Rate and Technical Considerations. 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...
Differential profiles of GH, IGF-1, and fructosamine in follicular fluid and plasma of cyclic mares. 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...
Amniotic Mesenchymal Stromal/Stem Cell-Derived Extracellular Vesicles for Equine Chronic Degenerative Endometritis Treatment. 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...
Estrus synchronization and follicular development patterns in jennies. Seasonal variations and breed differences significantly influence reproductive efficiency in jennies, yet optimized, evidence-based estrus synchronization protocols for key Chinese breeds are lacking. Objective: This study aimed to compare the efficacy of three hormonal protocols on estrus synchronization, and to characterize follicular development patterns in DeZhou and Xinjiang jennies across different seasons. Methods: In two experiments (April-May, n = 94; July-August, n = 144), 238 non-pregnant jennies received one of three treatments: GnRH-PGFα with two injection timings (Treatment Grou...
Diagnosis of Equine Genital Leptospirosis (EGL) in mares with poor reproductive performance. 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...
Effective methods of horse reproduction in the production of mare’s milk. 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 - ...
Determining the timeline of gonadal and genital differentiation in male and female equine fetuses allows for early detection and intervention in malformations. 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...
GnRH Administration Increases Synchronisation of Ovulation in Mares. 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...
Infertility in an American Quarter Horse mare with cervical duplication. 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...
Doppler assessment of testicular arterial blood flow in stallions: influence of age and 180° spermatic cord torsion. Doppler ultrasonography is a valuable tool for monitoring testicular hemodynamics, offering improved diagnostic insight and supporting treatment monitoring. However, its use in equine andrology remains limited due to a scarcity of published data and reference values. Therefore, this study aimed to characterize the testicular arterial blood flow patterns in clinically normal stallions and stallions with 180° spermatic cord torsion. Spectral Doppler ultrasonography was performed on both testes of 48 Mangalarga Marchador stallions. Thirty stallions with normal testes were divided into three age ...
Unraveling ovarian histology: The key morphological aspects that spur the development of the Fossa ovarii in equine. The equine ovary exhibits unique structural and developmental features that distinguish it from those of other domestic species, including the presence of an ovulation fossa and an inversion of cortical and medullary layers. This study aimed to investigate the morphostructural development of the equine fetal ovary, with particular emphasis on the formation of the ovulation fossa. Fourteen female equine fetuses, ranging from 70 to 200 days of gestation, were collected from a commercial abattoir. Results revealed a progressive increase in ovarian volume and marked histological changes througho...
Evidence linking ambient air pollution to domestic animal health outcomes: a scoping review. Air pollution is a major global health threat that is expected to worsen, yet its effects on domestic animals remain poorly understood. The objective of this scoping review was to synthesize existing evidence on associations between ambient air pollutants and health outcomes in domestic animals and to identify gaps to guide future research. Unassigned: A scoping review was conducted following the Arksey and O'Malley framework and Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extension for Scoping Reviews guidelines with 2 search databases (PubMed and Web of Science) using ...
Investigation of gene stability in equine luteal tissue during mid-diestrus phase and early pregnancy – Research Article. Real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a technique that allows for the semi-quantification of mRNA transcripts present within a tissue of interest. Differences in the relative abundance of mRNA between samples detected by RT-qPCR require normalization with a reference gene or genes whose transcript abundance is stable within the tissue of interest independent of experimental conditions. In the field of equine reproductive studies, ACTB, GAPDH and B2M genes are the most widely used as reference genes for the normalization of RT-qPCR results. However,...
Assessing the Sperm Head DNA Damage in Frozen/Thawed Horse Spermatozoa via Xenogeneic ICSI. 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...
Impact of melatonin on inflammatory cytokine profiles in 24-hour cultured equine uterine explants’. Melatonin has demonstrated beneficial effects in regulating inflammatory processes in various tissues, but its role in mare uterine physiology remains unclear. Objective: To investigate the effects of melatonin on the gene expression of cytokines (IL-1β, IL-6, IL-8, IL-10, and TNF-α) in equine uterine explants cultured for 6, 12, or 24 hours. Methods: The study was divided into two experiments. Part 1 aimed to evaluate the cytotoxicity of melatonin at concentrations of 0, 0.5, 1, 2, 4, and 8 mM on endometrial cells. Part 2 aimed to determine the expression of inflammatory cytokines in uterin...
