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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.
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...
Effects of intrauterine ozone insufflation in eleven subfertile mares: a case series. 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...
Metabolomics of stallion seminal plasma: Identification of potential fertility biomarkers based on differences between high and low pregnancy stallions. 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...
Single-cell metabolic profiling of stallion spermatozoa by flow cytometry using NADH and FAD autofluorescence†. The metabolic activity of stallion spermatozoa was assessed using flow cytometry to detect NAD(P)H and FAD fluorescence without labels. Sperm were incubated with different energy sources-glucose, lactate, and pyruvate-individually or combined, and measurements of NAD(P)H, FAD, NAD(P)H/FAD ratio, and optical redox ratio (ORR) were taken. Additionally, a metabolic assay based on resazurin reduction and flow cytometry detection was developed. Changes in NAD(P)H and FAD fluorescence, the NAD(P)H/FAD ratio, and ORR were observed. The NAD(P)H/FAD ratio increased significantly, especially when glucos...
Histomorphometric Analysis of the Endometrium of Jennies (Equus asinus) and Mares (Equus caballus) in Estrus: Anatomical Differences and Possible Reproductive Implications. 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...
Transcriptome Sequencing and Differential Analysis of Testes of 1-Year-Old and 3-Year-Old Kazakh Horses. 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...
A pilot study to investigate the effect of insemination on secretory cell activity post-ovulation in the isthmus of the equine oviduct. 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...
Addition of cholesterol linked to cyclodextrin in processing of cryopreserved equine spermatozoa and addition of motility stimulants post-thaw. 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...
Seasonal effects on follicular dynamics and hemodynamics, ovarian hormone levels, nitric oxide levels, and lactate dehydrogenase levels in native mares. Summer heat stress affects the regularity of the estrous cycle in different animal species. Unassigned: This study aimed to investigate seasonal effects on follicular dynamics, hemodynamics, and associated biomarkers across the estrous cycle in native mares. Unassigned: Doppler ultrasound examinations and blood samples were collected along three sequential estrous cycles during the cold (November to March) and the hot months of the year (May to August). The diameter, antrum diameter, and color area of the dominant follicle (DF) were determined. The diameters, uterine horns, and corpus luteum (...
High glucose in the extender impacts the metabolic phosphoproteome and modifies the phosphorylation of AKAP4 in stallion spermatozoa. Commercial extenders for equine semen typically contain high glucose concentrations; however, recent research indicates that these supraphysiological concentrations of glucose may be detrimental. Spermatozoa lack translational activity and depend on post-translational protein modifications for their regulation. Supraphysiological glucose concentrations are known to modify cell regulation, and we hypothesize that post-translational modification deregulation associated with supraphysiological glucose concentrations occurs in spermatozoa extended in high glucose media; and increasing the concentr...
