Omneity® Pellets
All-In-One Vitamin & Mineral Pellet
Topic:Equid Semen
Equid semen refers to the reproductive fluid produced by male horses, which contains sperm cells necessary for fertilization. The quality and viability of equid semen are important for successful breeding programs and can be influenced by various factors, including genetics, age, diet, and environmental conditions. Key parameters used to assess semen quality include volume, concentration, motility, and morphology of sperm cells. Techniques for collecting, evaluating, and preserving equid semen, such as cryopreservation, are critical for artificial insemination practices. This page compiles peer-reviewed research studies and scholarly articles that explore the physiology, assessment, and management of equid semen in the context of equine reproduction.
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...
Metformin and rosiglitazone affect motility, lipid peroxidation and mitochondrial activity of thawed equine spermatozoa. Maintaining sperm energy homeostasis in vitro is very important to improve the efficacy of stallion sperm preservation. Equine spermatozoa preferentially utilize oxidative phosphorylation over glycolysis to generate ATP. Metformin and rosiglitazone are antidiabetic compounds that enhances metabolic flexibility and glucose utilization. The aim of this study was to evaluate metformin and rosiglitazone supplementation of the freezing medium on quality and oxidative status of thawed stallion semen. A total of 15 ejaculates from five horses were collected and supplemented before freezing with metfo...
Cryopreservation of Horse Sperm. Cryopreservation is currently the only strategy for long-term conservation of equine sperm. To get optimal post-thaw sperm survival, carefully following each step of the freezing protocol is crucial. First, one needs to obtain and exhaustively analyze an ejaculate of good sperm quality. Then, the seminal plasma is removed by centrifugation, and the resulting pellet is resuspended in a certain volume of the freezing medium to reach the right sperm concentration. Finally, sperm samples are packaged into 0.5-mL straws, cooled, and frozen using an automatic, controlled-rate freezer. Once the tempe...
Effect of pre-freeze sperm concentration, freezing extender, and epididymal flushing technique on post-thaw quality of cryopreserved epididymal stallion sperm. Cryopreservation of stallion epididymal sperm has become a common clinical procedure after routine castration, euthanasia, or acute death. Unique features of epididymal sperm compared to ejaculated sperm include the requirement to remove sperm from the cauda epididymis, lack of exposure to seminal plasma, and potential recovery of large sperm numbers. In this study, the effect of the flushing technique (Extender (INRA) vs. AIR), freezing extender (LE, CMLE, MFR5, CFR5, or BOTU), and the concentration at which sperm are cryopreserved (200, 400, or 800 × 10 sperm/mL) on post-thaw epididymal s...
Cryoprotective effect of zinc and gold nanoparticles during cooling and freeze-thawing on Marwari stallion sperm parameters and reactive oxygen species production. Sperm cryopreservation is one of the most important procedures in the development of biotechnologies for assisted reproduction. Cryopreservation of stallion spermatozoa tends to cause plasma membrane damage due to the low ratio of cholesterol to phospholipids. Different substances and compounds can be added to semen extenders to improve sperm quality. Objective: To investigate the effect of supplementing semen extender with zinc nanoparticles (ZnONPs) and gold nanoparticles (AuNPs) on cooled and frozen-thawed spermatozoa of Marwari stallion. Methods: A total of 20 ejaculates from four Marwari ...
Equine in vitro fertilization with frozen-thawed semen is associated with shortened pre-incubation time and modified capacitation-related changes. We recently reported successful equine IVF using fresh semen pre-incubated for a prolonged period (22 h) before co-culture with oocytes. In this study, we evaluated the feasibility of equine IVF with frozen-thawed sperm and evaluated capacitation-related changes in these sperm over the pre-incubation period. Sperm selected via a commercial sperm separation device (SSD) yielded significantly higher fertilization than did sperm selected by swim-up or by colloid centrifugation. Using the SSD method, fertilization rates with sperm pre-incubated for 15 min, 3 h, 6 h, and 9 h were 7.1, 22.2, 38...
Results of the “test-freeze” approach in a commercial program of stallion sperm cryopreservation and the relationship between pre-freeze sperm quality and “freezability.”. In the current study, we report the effect of different commercially available semen freezing extenders utilized for the "test-freeze" procedure for 13 years (2010-2023) as part of a commercial program of stallion sperm cryopreservation. Ejaculates obtained from sexually active and healthy stallions (n = 124) were cryopreserved using up to five commercially available semen freezing extenders (Lactose-EDTA [LE], MFR5, CryoMax LE [CMLE], CryoMax MFR5 [CMMFR5] or BotuCrio [BC]). Post-thaw sperm motility (total motility - TM [%]; progressive motility - PM [%]; and curvilinear velocity - VCL [μm...
Measuring Noseband Tightness on the Lateral Aspect of the Horse’s Face. Measuring tools designed to objectively determine equine noseband tightness are inserted on the dorsal nasal planum in a rostro-caudal direction. The lateral aspect of the horse's head has several areas where minimal soft tissue intervenes between the skin and underlying bone, which makes them potentially useful sites for measuring noseband tightness. One hundred horses were fitted with a snaffle bridle with a cavesson, Swedish or dropped noseband in random order. The tightness of each noseband type was adjusted sequentially to 2.0, 1.5, 1.0, 0.5 and 0.0 finger-equivalents using an ISES Taper ...
The effect of resveratrol on the cryopreservation of Mongolian horse semen. Cryopreservation of semen has advanced significantly with the development of artificial insemination techniques, but post-thawed sperm often exhibit reduced viability, membrane integrity, and acrosome integrity compared to fresh sperm, leading to decreased fertilization capacity. Oxidative stress is a major concern during cryopreservation. This study investigated the use of resveratrol (RSV), a potent antioxidant, in the cryopreservation of Mongolian horse semen. Different concentrations of RSV were incorporated into semen cryopreservation extenders, and the morphological and antioxidant indic...
Ultrasensitive detection and quantification of bovine Deltapapillomavirus in the semen of healthy horses. BPV1, BPV2, BPV13, and BPV14 are all genotypes of bovine delta papillomaviruses (δPV), of which the first three cause infections in horses and are associated with equine sarcoids. However, BPV14 infection has never been reported in equine species. In this study, we examined 58 fresh and thawed commercial semen samples from healthy stallions. In 34 (58.6%), bovine δPV DNA was detected and quantified using droplet digital polymerase chain reaction (ddPCR). Real time quantitative PCR (qPCR) was able to identify bovine δPV DNA in 5 samples (8.6%). Of the BPV-infected semen samples, 15 were posi...
The use of sex-sorted semen in horses. The application of sex-sorted semen in horses has historically lagged the bovine industry due to differences in the reproductive physiology and grater variability in equine semen quality. Recent advancements, including SuperGen™ sorters and Ultraplus™ semen processing methods developed by STgenetics® (Navasota, Texas), have significantly improved the efficiency of the sex-sorting process and reduced the sperm damage previously reported. To facilitate the broader use sex-sorted semen in the equine industry, it is essential to address challenges such as shipping cooled semen to a central so...
Niosomal nanocarriers loaded with artemisinin has potential to enhance the functional characteristics of cryo-preserved equine spermatozoa. Cryopreservation is a crucial method for the preservation of genetic materials. Nevertheless, this technique can have adverse effects due to cryo-oxidative damage. The primary objective of this study was to investigate the cryo-protective effects of niosomal nanocarriers loaded with artemisinin (NN-Art) on the functional attributes of equine spermatozoa. The parameters examined included sperm motility and kinematics through computer-assisted sperm analysis (CASA), plasma membrane integrity via the hypoosmotic swelling test, viability using the eosin-nigrosin staining technique, morphology with...
Prolonged maintenance of stallion semen by optimization of cooling conditions. Bottlenecks to the success of equine assisted reproductive technologies (ART) include suboptimal conditions for prolonged storage of stallion sperm. Shipped stallion sperm are transported in cooling devices designed to maintain temperature for up to 48 h. Increasing the storage time of cooled semen while maintaining acceptable motility would relieve logistical ART challenges. Experiments were conducted to test the hypothesis that external regulation of shipment container temperature would prolong storage time of cooled stallion semen. Initial experiments determined the effect of pre-freezing c...
Evaluation of Platonia insignis Mart. (Bacuri Butter) and Biopolymers from the Puree of Allium cepa L. (Yellow Onion Bulb) for Wound Healing in Horses. Background/Objectives: Skin injuries are common in the equine clinical practice, requiring effective treatment to support natural healing. Bacuri butter is gaining attention for its potential in wound healing and its anti-inflammatory, antimicrobial, and antioxidant properties. Natural polymers such as onion (Allium cepa) bioplastics have been investigated for their potential as occlusive dressings and for tissue regeneration. Methods: This study evaluated the healing process of experimentally induced skin wounds on horses treated with bacuri butter, washed onion film, and unwashed onion film....
Optimizing equine sperm quality: an alternative to single layer centrifugation for sperm isolation. In vitro semen purification techniques have been developed that seek to mimic the in vivo selection process in order to generate the highest possible chance of oocyte fertilization following artificial insemination. Numerous methods have been developed to isolate functional spermatozoa for artificial insemination, yet only one method, single-layer centrifugation using commercial preparations like EquiPure, has been widely employed. In this study, we have introduced a novel approach for isolating spermatozoa and compared their quality to those isolated using EquiPure. The AI port system (Mempha...
Prolonged incubation of frozen-thawed equine spermatozoa for in vitro fertilization: A preliminary study using low temperature and INRA96 medium. A protocol for conventional in vitro fertilization (IVF) in horses using fresh semen has been described, using a prolonged incubation in FERT-TALP medium (22 h) at 38.2°C in the presence of penicillamine, hypotaurine and epinephrine (PHE). Our work aimed to develop a protocol that maintains quality parameters in frozen-thawed equine spermatozoa incubated for 22 h in the presence of PHE using different media (FERT-TALP and INRA96) and incubation temperatures (30 and 38.2°C). Twelve frozen ejaculates from four stallions were thawed and then incubated in either FERT-TALP or INRA96 with PHE...
Effect of refreezing extender on stallion sperm quality and embryo production after intracytoplasmic sperm injection. Intracytoplasmic sperm injection (ICSI) is a valuable assisted reproduction technology in clinical practice, especially when semen availability is limited. Since the number of sperm required per ICSI cycle is much less than the number of sperm available in a standard straw of frozen semen, refreezing semen at lower sperm concentrations could yield multiple straws for ICSI use. However, there is little data on the effect of sperm refreezing on ICSI outcomes, especially on the effect of extender used for refreezing. The objective of the present study was to evaluate the effect of refreezing exte...
Noseband type and tightness level affect pressure on the horse’s face at trot. Poor noseband adjustment could create high pressures that may risk pain or tissue damage. Objective: To quantify sub-noseband pressures dorsally over the nasal bone and ventrally over the mandibular rami for a Cavesson, Swedish (crank), Drop and Flash noseband at five tightness levels (2.0 to 0.0 finger equivalents). Methods: In vivo experiments. Methods: Eight high-level dressage horses were ridden at the trot in a straight line by their usual riders. Two small pressure mats, attached to the noseband over the nasal bone and the mandibular rami, collected force (N) and pressure (kPa) data from...
Tracing horseback riding and transport in the human skeleton. Among the most widely used methods for understanding human-horse relationships in the archaeological record is the identification of human skeletal pathologies associated with mounted horseback riding. In particular, archaeologists encountering specific bony changes to the hip, femur, and lower back often assert a causal link between these features and prolonged periods of mounted horseback riding. The identification of these features have recently been used to assert the early practice of mounted horseback riding among the Yamnaya culture of western Eurasia during the third and fourth millenn...
Pre- and Post-Thaw Addition of L-Carnitine and Pyruvate: Effect on Stallion Sperm Parameters. The addition of antioxidants to cryopreservation media reportedly improves sperm post-thaw quality and reproductive performance after artificial insemination. Therefore, the objectives of this study were to evaluate if the addition of L-carnitine and pyruvate to freezing media, or their addition to samples after thawing, improves the post-thaw quality of equine spermatozoa. Thus, in Experiment 1, stallion semen samples were cryopreserved in: (1) EDTA-glucose-based extender with 20% egg yolk and 5% dimethylformamide (EDTA control); (2) skim milk-based extender with 20% egg yolk and 5% dimethylf...
In vitro, the aging of stallion spermatozoa at 22 °C is linked to alteration in Ca2+ and redox homeostasis and may be slowed by regulating metabolism. Conservation of equine semen in the liquid state is a central procedure in horse breeding and constitutes the basis of associated reproductive technologies. The intense mitochondrial activity of the stallion spermatozoa increases oxidative stress along the storage period, leading to sperm demise within 24-48 h of storage, particularly when maintained at room temperature. Recently, the relationship between metabolism and oxidative stress has been revealed. The study aimed to extend the period of conservation of equine semen, at room temperature through modification of the metabolites present i...
2D Kinematic Analysis of the Esbarrada and Volta Sobre Patas Manoeuvres of Criollo Breed Horses Competing in Freio de Ouro. The esbarrada and volta sobre patas manoeuvres are critical in Criollo horse equestrian activities, yet their kinematics remain largely unexplored. This observational study aimed to kinematically describe the manoeuvres in Criollo breed horses and explore the relationship between static goniometry and dynamics. A 2D kinematic analysis was conducted on 31 Criollo horses performing the manoeuvres using high-speed cameras. Joint angles were measured using static goniometry and analysed in relation to dynamic performance. The esbarrada covered a distance of 4.28 ± 0.99 m in 1.15 ± 0.11 s at a ve...
Effect of Mitoquinone on sperm quality of cryopreserved stallion semen. This study aimed to investigate the effect of mitochondria-targeted antioxidants (Mitoquinone, MitoQ) on the quality of frozen-thawed stallion semen. Semen samples collected from three fertile stallions aged 10 - 13 years, were filtered, centrifuged in a skimmed milk-based extender, and diluted to a final concentration of 50 × 106 sperm/mL in freezing medium. Diluted semen was divided into five experimental groups supplemented with MitoQ at concentrations of 0 (control), 25, 50, 100, and 200 nM and then subjected to freezing after cooling and equilibration. After thawing, semen was evaluated ...
Impact of low-dose ozone supplementation on motility parameters and bacterial growth in horse cryopreserved semen. Two studies were conducted to evaluate the use of medical ozone (O) in commercial extenders for equine semen cryopreservation. In the first study (Study 1), 0, 5, and 15 µg/mL of O were added to diluents of refrigerated or frozen semen. Samples were evaluated for sperm kinematics at different time points for the chilled samples and after a thermoresistence test for the frozen/thawed samples. In the second study (Study 2), 0, 5, and 10 µg/mL of O were added to an antibiotic-free diluent for refrigerated semen for comparison with the control group in which semen was diluted in the same diluent...
Ex vivo biomechanical evaluation of tissue construct strength in an equine colopexy model. To compare strength of left paramedian colopexies using various techniques in equine ex vivo models. Methods: Experimental study. Methods: Equine cadavers euthanized for nongastrointestinal pathology (36 specimens derived from 9 horses). Methods: Colopexies were performed after euthanasia. Suture pattern (horizontal mattress vs. cruciate) and incorporation of dorsal sheath of the rectus abdominis (partial-thickness) versus incorporation of dorsal and ventral sheath of the rectus abdominis (full-thickness) were evaluated. Single cycle load to failure, work to peak load, stiffness, and mode of f...
16s gene metagenomic characterization in healthy stallion semen. Studies on the bacterial composition of seminal samples have primarily focused on species isolated from semen and their effects on fertility and reproductive health. Culture-independent techniques, such as 16S rRNA gene sequencing and shotgun metagenomics, have revolutionized our ability to identify unculturable bacteria, which comprise >90% of the microbiome. These techniques allow for comprehensive analysis of microbial communities in seminal samples, shedding light on their interactions and roles. In this study, we characterized the taxonomic diversity of seminal microbial communities in...
Effect of novel lyophilized extenders on stallion post-thaw sperm motility. The aim of this study was to assess the effect of lyophilized freezing extenders, which can be stored at room temperature, on stallion post-thaw sperm total motility (TM). Ejaculates of 28 stallions were frozen with four different extenders: two commercial freezing extenders offered worldwide and two novel lyophilized extenders (STAR and MX3), and two different cryopreservation protocols (CP1 with an equilibration period of 20 min. and CP2 with an equilibration period of 60 min.). The TM was assessed after thaw. Mean TM did not show significant differences between cryopreservation protocols wi...
The effects of oxidative stress and intracellular calcium on mitochondrial permeability transition pore formation in equine spermatozoa. The in vitro storage of stallion spermatozoa for use in artificial insemination leads to oxidative stress and imbalances in calcium homeostasis that trigger the formation of the mitochondrial permeability transition pore (mPTP), resulting in premature cell death. However, little is understood about the dynamics and the role of mPTP formation in mammalian spermatozoa. Here, we identify an important role for mPTP in stallion sperm Ca2+ homeostasis. We show that stallion spermatozoa do not exhibit "classical" features of mPTP; specifically, they are resistant to cyclosporin A-mediated inhibition...
Multiparametric Flow Cytometry for Determination of Viability, Caspase 3 and 7 Activity, and Lipid Peroxidation Adduct (4-Hydroxynonenal) in Equine Spermatozoa. Flow cytometry is a powerful tool for the analysis of cell samples formed of multipopulations, such as spermatozoa. In recent years, multiparametric cytometers have evolved, allowing the study of different cellular characteristics, such as protein expression, DNA analysis, or mitochondrial activity. Whether using traditional fluorescent dyes or fluorophore-conjugated antibodies, each cell or cellular component is individually stained, the sample is analyzed at high velocities, and then is displayed and interpreted in a dot-plot. We hereby describe the procedure to perform a multiparametric flo...
Rotation of liquid-preserved artificial insemination doses on roller benches affects sperm quality during storage in stallions. Appropriate stallion semen handling is of great importance in equine artificial insemination (AI) industry. Optimal treatment of AI doses is aiming for best sperm preservation by excluding strong environmental influences and adding media that favour sperm survival. One method widely used in stallion sperm handling is the rotation of liquid-preserved semen samples on roller benches during storage. As previous studies in boars give rise to the fact that rotation should not be considered beneficial for spermatozoa anymore, the present study investigated the influence of continuous rotation of dil...
