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Topic:Fertility
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.
Phenylbutazone treatment of mares does not reduce embryo production via intracytoplasmic sperm injection. Production of embryos via intracytoplasmic sperm injection (ICSI) is a common clinical procedure in equine practice. A recent study indicates that treatment of mares with phenylbutazone paste significantly lowers blastocyst production after ICSI of recovered oocytes. The current study was conducted to determine the effect of different methods of phenylbutazone administration on in vitro embryo production of recovered oocytes in mares. Unassigned: In study 1 (July 29 through September 18, 2024), 14 mares received either oral liquid phenylbutazone once daily for 10 days or no treatment. On the l...
Clinical safety of intrauterine infusion of a uterine sanitizer in mares. 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...
Blastocyst production by conventional in vitro fertilization (cIVF) in horses: Effects of sperm storage method, incubation timing of cool-stored semen before gamete co-incubation, and comparisons between cIVF and intracytoplasmic sperm injection (ICSI). 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...
Natural Frequencies in Sexual Pelvic Thrusting. Seventy percent of mammals copulate using repeated pelvic thrusting, while the transfer of sperm requires just a single intromission. Why did thrusting evolve to be the dominant form of sexual intercourse? In this study, we investigate how the rate of sexual pelvic thrusting changes with body size. By analyzing films of copulating mammals, from mice Mus musculus to elephants Elephantidae, we find that bigger animals thrust slower. The rate of pelvic thrusting decreases from 6 Hz for the pocket mouse Pergonathus to 1.3-1.8 Hz for humans to an absence of thrusting for the rhino Rhinocerotidae an...
Influence of pituitary pars intermedia dysfunction on reproductive performance in Thoroughbred mares. 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...
Endocrine profile and OPU-ICSI outcomes in mares: a comparative study. Despite recent improvements in equine ovum pick-up (OPU) combined with intracytoplasmic sperm injection (ICSI), there is still significant inter-individual variability. In this study, serum concentrations of the oocyte-secreted factors growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), as well as the GDF9/BMP15 complex, anti-Müllerian hormone (AMH), progesterone (P4), and free and conjugated estradiol-17β (E2), were determined in 59 mares (3-24 years) at the time of OPU. Mares were classified retrospectively, based on the number of embryos obtained, into low or...
Comprehensive Integrated Analyses of Proteins and Metabolites in Equine Seminal Plasma (Horses and Donkeys). 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...
Direct Warming of Vitrified In Vivo Equine Embryos. 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 = ...
Reproductive success in Thoroughbred broodmares post large colon resection or colopexy. 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...
Influence of Cryopreservation on the Acrosome Reaction in Hucul Stallion Spermatozoa. 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...
A comparison of the efficacy of three commercial human embryo vitrification kits for cryopreservation of in vivo produced equine embryos. Different cryoprotectants can influence the ability of embryos to successfully survive vitrification and subsequent warming before transfer. Objective: To compare pregnancy rates for embryos ≤500 μm vitrified, without puncture or aspiration of the blastocoele cavity, with one of three commercial human embryo vitrification kits containing the same penetrating cryoprotectants (DMSO and EG) but varying in their non-penetrating cryoprotectants (NPCPAs; sucrose, trehalose, dextran serum supplement [DSS], and hydroxypropyl cellulose [HPC]). Methods: In vivo experiments. Methods: Embryos (n =...
Effect of Centrifugation of Stallion Semen Through a Low Density Colloid Prior to Freezing on Sperm Cryosurvival. 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)...
Two Cases of Chromosome 27 Trisomy in Horses Detected Using Illumina BeadChip Genotyping. Autosomal trisomy, a genetic disorder characterized by the presence of an extra autosome, is a rare but important chromosomal abnormality in horses, often associated with infertility, developmental abnormalities, and reduced life expectancy. This study represents the largest population-level screening for autosomal trisomy in horses; the analysis used single nucleotide polymorphism (SNP) panel genotype intensity data from 17,078 horses, 6601 of which were juveniles (i.e., ≤12 months of age) when genotyped. Using methodologies adapted from similar screening studies in cattle, the only aneuplo...
Follicular fluid secretome enhances the meiotic competence of in vitro matured equine oocytes, but not the developmental competence of zygotes after intracytoplasmic sperm injection (ICSI). In vitro embryo production is widely used in equine clinical reproduction; however, its efficiency remains suboptimal due to the undermined developmental competence of in vitro-matured oocytes. In this work, we evaluated the effect of mare preovulatory follicular fluid (PFF) secretome supplementation during in vitro maturation (IVM) of equine oocytes recovered post-mortem. Secretome was isolated from PFF and was added to the IVM medium at concentrations of 20 µg/ml (S20) or 40 µg/ml (S40). The percentage of mature oocytes, as well as embryo cleavage, blastocyst development, and gene expr...
Equine MicroRNAs: Performance, Reproduction, and Disease. MicroRNAs (miRNAs) are molecules that regulate gene expression by targeting the 3' untranslated region (UTR) of mRNAs. They are essential in numerous biological processes like growth, metabolism, and muscle development. miRNA research has become crucial in livestock breeding, offering solutions for improving animal health and productivity. This review focuses on miRNAs' roles in equine performance, reproduction, and disease, highlighting key findings and future applications in these areas. It discusses the use of circulating miRNAs (ci-miRNA) as biomarkers for athletic performance, particularl...
Catalase in Unexpected Places: Revisiting H2O2 Detoxification Pathways in Stallion Spermatozoa. Oxidative stress plays a critical role in regulating sperm function, yet species-specific antioxidant mechanisms remain poorly understood. This study compared hydrogen peroxide (H2O2) tolerance in horse and human sperm and investigated the roles of catalase and glutathione peroxidase (GPx) in horses. A H2O2 dose-response assay (0-2000 µM) showed that horse sperm were significantly more resistant to oxidative damage, with an IC50 for progressive motility over 14-fold higher than that of human sperm (391.6 µM vs. 27.3 µM). Horse sperm also accumulated more intracellular H2O2 without loss of m...
Seminal vesiculitis and blocked ampullae occurring simultaneously in a stallion. A Gypsy Vanner stallion presented with a history of subfertility and abnormal dense, grey ejaculate. Reproductive examination including ultrasound, cytology, and bacteriology confirmed the diagnosis of seminal vesiculitis. After evaluation of several ejaculates, bilateral blocked ampullae were confirmed by azoospermia and a low level of alkaline phosphatase (30 IU/L) in the filtered fraction. Continuous semen collections, along with oral antibiotics treatment, were performed at the owner's facility for five weeks. After readmission, a successful collection was obtained with a high concentratio...
Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development Without Inducing Genetic Aberrations. In vitro maturation (IVM) of oocytes retrieved from ovum pick-up (OPU) or ovarian tissue (OT) is a standard approach for patients with specific conditions where prior hormonal stimulation is contraindicated. However, the developmental competence of oocytes matured in vitro is still inferior to that of oocytes matured in vivo. Capacitation IVM (CAPA-IVM) includes an extra step of pre-maturation culture (PMC) with c-type natriuretic peptide (CNP) as a meiotic arrestor to better synchronize cytoplasmic and nuclear maturity in oocytes by allowing the cytoplasm additional time to acquire essential ...
Stallion spermatozoa express LDH isoforms A, B, and C, with LDHC playing a crucial role in sustaining sperm viability. Three isoforms of lactate dehydrogenase (LDH) - LDHA (cytoplasmic), LDHB (mitochondrial), and LDHC (flagellar) - have been identified and localized in stallion spermatozoa. Functional inhibition assays indicate that these three isoforms constitute a lactate shuttle of crucial importance for sperm function. Unassigned: Stallion spermatozoa use different energy sources; while oxidative phosphorylation predominates, glycolysis and beta-oxidation of fatty acids are also present. Glycolysis depends on the availability of NAD+ as an electron acceptor. During glycolysis, NAD+ is reduced to NADH. To e...
Advancements in microfluidic and electrophoretic techniques for stallion sperm isolation. 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...
Seminal plasma MicroRNA dynamics in stallion semen due to progressive motility and conception success. This study aims to investigate the expression patterns of miR-34b, miR-122, let-7a in seminal plasma of stallions in relation to progressive motility and conception success, and to evaluate their potential as biomarkers. Fifteen adult stallions were enrolled in study. One ml of seminal plasma was kept at -80 °C, 2 ml of fresh semen was sampled from each stallion for sperm analysis, and the rest were used for artificial insemination only for one mare. Two groups were formed as high progressive motility (HPM; n:8) and low progressive motility (LPM; n:7), based on progressive motility. Besides...
Thermal comfort and seminal traits in stallion reproduction: New methods to understand these relationships. Stallion breeding is mostly based on desirable phenotypic traits, with little consideration for semen quality. Objective: To identify relationships among semen parameters in stallions using a non-invasive, integrative approach across summer and winter seasons. Methods: Twenty-four stallions were evaluated, in which semen collections, testicular thermography, and Doppler velocimetry of the testicular artery by Doppler ultrasonography were performed, in addition to the measurement of physiological, environmental, and heat stress parameters, both in winter (coat weather) and summer (T-shirt weath...
Comparative Analysis of Proteomic Characteristics in Seminal Plasma Between Horses and Donkeys. Horses and donkeys, as integral members of the equine family, exhibit distinct reproductive capabilities and characteristics. Seminal plasma, the fluid component of semen, contains a variety of proteins that play critical roles in sperm function and fertility. This study aimed to systematically compare the protein profiles in the seminal plasma of horses and donkeys, thereby elucidating the molecular differences between these two species. The study utilized 4D-DIA proteomics technology to analyze seminal plasma from horses and donkeys and further validated key proteins through Western blot. Ou...
The relationship between post-thaw sperm quality parameters and blastocyst production following Intracytoplasmic Sperm Injection (ICSI) of in vitro-matured equine oocytes. In vitro production of equine embryos by Intracytoplasmic Sperm Injection (ICSI) is commonly utilized to maximize the availability of stallion sperm, particularly from stallions with limited semen supply. The relationship between standard measures of sperm quality in frozen/thawed semen (i.e., sperm motility, normal morphology, DNA quality) and embryo production after ICSI of in vitro-matured equine oocytes has not been thoroughly studied. In this study, frozen/thawed semen from 47 stallions utilized in a commercial ICSI program was analyzed for post-thaw sperm total and progressive motility (...
Comparison of RNA- and DNA-based 16S amplicon sequencing to find the optimal approach for the analysis of the uterine microbiome. Studies in humans and large animals indicate a relationship between the uterine microbiome composition and endometrial receptivity. Despite many studies have been performed, the analysis of the uterine microbiome remains challenging due to the very low microbial biomass. Studies in other biological systems showed that RNA-based microbiome analysis complements DNA-based results and provides information about active bacteria in a sample. Thus, the aim of this study was to establish a highly sensitive and specific 16S rRNA gene V3-V4 amplicon PCR from equine uterine cytobrush samples and to compa...
In vitro embryo production via ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) in pure and crossbred Japanese Hokkaido native ponies. 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...
Enhanced Reliability of the Evaluation of Fertility Traits in Pura Raza Española Horses Using Single-Step Genomic Best Linear Unbiased Prediction. 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...
Roles of GDF9 and BMP15 in equine follicular development: in vivo content and in vitro effects of IGF1 and cortisol on granulosa cells. In horses, the mechanisms behind ovarian follicle growth and oocyte maturation remain largely unknown. In other species, oocyte-secreted factors growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) have been related to the acquisition of developmental competence and to interaction with granulosa cells for the regulation of follicle development. This study assessed the expression and localization of GDF9 in the equine ovary, and its possible relationship with granulosa cell function. Results: Using custom-made antibodies, GDF9 protein was localized in oocytes from th...
Evidence of glycan mosaicism in the equine oviduct. We report a lectin histochemical study of oviductal ampulla and isthmus of thirteen mares euthanised at anestrus (1), estrus (5), 1-day post-ovulation (5) or diestrus (2). Staining with a panel of 5 lectins recognizing N-Acetylgalactosamine (GalNAc) residues revealed mosaicism in glycan expression in some specimens of ampulla and isthmus, peaking at estrus and 1-day post-ovulation (ov+1). In the ampulla, this presented as positively stained areas of both ciliated and non-ciliated cells in otherwise negatively-stained ampullae, ranging from small foci (diestrus) to areas up to 200 microns in le...
Sperm Vitrification in Horse and Donkey. Sperm vitrification is a novel-assisted reproductive technique that is increasingly gaining relevance in the last years. This technique allows to cryopreserve sperm from valuable stallions and donkeys without the exposure to permeable cryoprotectants, particularly toxic for the gametes of these species.This chapter aims to describe the current range of methodologies available that are key to ensure sperm quality after vitrification and warming of stallion and donkey sperm.
