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Topic:Physiology

The physiology of horses encompasses the study of the biological functions and processes that occur within the equine body. This includes the examination of various systems such as the cardiovascular, respiratory, musculoskeletal, digestive, and nervous systems. Understanding equine physiology is essential for comprehending how horses adapt to different environmental conditions, perform physical activities, and respond to health challenges. Research in this field often focuses on the mechanisms of energy metabolism, thermoregulation, and muscle function during exercise, as well as the physiological responses to stress and disease. This page compiles peer-reviewed research studies and scholarly articles that explore the diverse aspects of equine physiology, providing insights into the biological processes that support the health and performance of horses.
Effects of equine bone marrow aspirate volume on isolation, proliferation, and differentiation potential of mesenchymal stem cells.
American journal of veterinary research    May 1, 2013   Volume 74, Issue 5 801-807 doi: 10.2460/ajvr.74.5.801
Kisiday JD, Goodrich LR, McIlwraith CW, Frisbie DD.To compare the mesenchymal stem cell (MSC) yield and chondrogenic and osteogenic differentiation from 5- and 50-mL bone marrow aspirates from horses. Methods: Six 2- to 5-year-old mixed-breed horses. Procedures-2 sequential 5-mL aspirates were drawn from 1 ilium or sternebra. A single 50-mL aspirate was drawn from the contralateral ilium, and 2 sequential 50-mL aspirates were drawn from a second sternebra. The MSC yield was determined through the culture expansion process. Chondrogenesis and osteogenesis were evaluated by means of conventional laboratory methods. Results: The second of the 2 s...
Effects of hypoglossal nerve block and electrical stimulation of the thyrohyoideus muscles on position of the larynx and hyoid apparatus in healthy horses.
American journal of veterinary research    May 1, 2013   Volume 74, Issue 5 784-789 doi: 10.2460/ajvr.74.5.784
Zantingh AJ, Ducharme NG, Mitchell LM, Cheetham J.To determine the effects of hypoglossal nerve block and electrical stimulation of the thyrohyoideus muscles on position of the larynx and hyoid apparatus in resting horses. Methods: 16 healthy horses that underwent hypoglossal nerve block and 5 healthy horses that underwent electrical stimulation of the thyrohyoideus muscles. Methods: Horses underwent bilateral hypoglossal nerve block or electrical stimulation of the thyrohyoideus muscles. Positions of the basihyoid bone, ossified part of the thyroid cartilage, and articulations of the thyrohyoid bones and thyroid cartilage were determined in ...
Serum amyloid A (SAA) concentration after training sessions in Arabian race and endurance horses.
BMC veterinary research    May 1, 2013   Volume 9 91 doi: 10.1186/1746-6148-9-91
Cywinska A, Witkowski L, Szarska E, Schollenberger A, Winnicka A.Serum amyloid A (SAA) is the major acute phase protein in horses. Its concentration increases in various pathologies but also in response to prolonged, strenuous effort. The purpose of this study was to establish whether routine race and endurance training produces changes in the SAA level in Arabian horses. Additionally, the differences between SAA response in experienced endurance horses and endurance horses that were beginning their career were investigated. Results: There were no changes in SAA concentrations after race training and endurance training in experienced horses. In horses that ...
The art and the science of equine surgery.
The Veterinary record    April 30, 2013   Volume 172, Issue 17 442-446 doi: 10.1136/vr.f2555
Greet T.No abstract available
Identification of α-cobratoxin in equine plasma by LC-MS/MS for doping control.
Analytical chemistry    April 30, 2013   Volume 85, Issue 10 5219-5225 doi: 10.1021/ac4006342
Bailly-Chouriberry L, Cormant F, Garcia P, Kind A, Popot MA, Bonnaire Y.Cobra venom (Naja kaouthia) contains a toxin called α-cobratoxin (α-Cbtx). This toxin is a natural protein containing 71 amino acids (MW 7821 Da) with a reported analgesic potency greater than morphine. In 2007, in USA, this substance was found in the barns of a thoroughbred trainer and since then till date, the lack of a detection of this molecule has remained a recurring problem for the horseracing industry worldwide. To solve this problem, the first method for the detection of α-cobratoxin in equine plasma has now been developed. Plasma sample (3 mL) was treated with ammonium sulfate at ...
Oxytocin stimulated release of PGF2α and its inhibition by a cyclooxygenase inhibitor and an oxytocin receptor antagonist from equine endometrial cultures.
Animal reproduction science    April 24, 2013   Volume 139, Issue 1-4 69-75 doi: 10.1016/j.anireprosci.2013.04.010
Penrod LV, Allen RE, Rhoads ML, Limesand SW, Arns MJ.Uterine inflammation results in a poor uterine environment and early embryonic loss in the mare due to an inhibition of maternal recognition of pregnancy caused from increased prostaglandin F2α (PGF2α). Oxytocin binds to endometrial cell receptors to activate prostaglandin synthesis. An oxytocin receptor antagonist (Atosiban) and a cyclooxygenase inhibitor (indomethacin) both decrease PGF2α production. The aim of this study was to evaluate the in vitro effects of Atosiban and indomethacin on equine uterine prostaglandin secretion. Equine endometrial explants were harvested on day two of beh...
Expression of enzymes and receptors of leukotriene pathway genes in equine endometrium during the estrous cycle and early pregnancy.
Theriogenology    April 24, 2013   Volume 80, Issue 2 145-152 doi: 10.1016/j.theriogenology.2013.03.025
Guzeloglu A, Atli MO, Kurar E, Kayis SA, Handler J, Semacan A, Aslan S.The aims of the present study were to elucidate the expression profiles of leukotriene (LT) pathway mRNA transcription and to determine the possible interaction of LT and prostaglandin (PTG) pathways genes in equine endometrium during the estrous cycle and early pregnancy. Endometrial biopsies were obtained from mares on the day of ovulation (d0), at late diestrous (LD, n = 4), and after luteolysis in the estrus phase (AL, n = 4) of the cycle. Biopsies were also taken on Days 14 (P14; n = 4), 18 (P18, n = 4), and 22 (P22, n = 4) during early pregnancy that were comparable days to cyclic sampli...
Stress modulates instrumental learning performances in horses (Equus caballus) in interaction with temperament.
PloS one    April 23, 2013   Volume 8, Issue 4 e62324 doi: 10.1371/journal.pone.0062324
Valenchon M, Lévy F, Prunier A, Moussu C, Calandreau L, Lansade L.The present study investigates how the temperament of the animal affects the influence of acute stress on the acquisition and reacquisition processes of a learning task. After temperament was assessed, horses were subjected to a stressor before or after the acquisition session of an instrumental task. Eight days later, horses were subjected to a reacquisition session without any stressor. Stress before acquisition tended to enhance the number of successes at the beginning of the acquisition session. Eight days later, during the reacquisition session, contrary to non-stressed animals, horses st...
Validation of the IDS Octeia ELISA for the determination of insulin-like growth factor 1 in equine serum and tendon tissue extracts.
Veterinary clinical pathology    April 23, 2013   Volume 42, Issue 2 184-189 doi: 10.1111/vcp.12038
Lygren T, Schjerling P, Jacobsen S, Berg LC, Nielsen MO, Langberg H, Thomsen PD.Insulin-like growth factor (IGF-1) is an important mediator of tissue repair in horses. Objective: The aim of the study was to evaluate whether IGF-1 could be measured reliably in equine serum and tendon tissue extracts, using an IGF-1 ELISA kit developed for human serum and plasma. Methods: A glycyl-glycine pretreatment protocol of samples was compared with the pretreatment procedure recommended by the manufacturer. Intra- and inter-assay imprecision were evaluated by repeated measurements of equine serum pools. Assay inaccuracy was determined based on the linearity of serially diluted equine...
Evaluation of intra-abdominal pressure in horses that crib.
Veterinary surgery : VS    April 22, 2013   Volume 42, Issue 6 658-662 doi: 10.1111/j.1532-950X.2013.12019.x
Albanese V, Munsterman AS, DeGraves FJ, Hanson RR.To measure intra-abdominal pressure (IAP) in horses that crib and compare it with IAP in horses that do not have this vice. Methods: Cohort study. Methods: Healthy cribbing horses (cribbing cohort, n = 8) and 8 healthy noncribbing horses (noncribbing cohort). Methods: A microsensor catheter was introduced into the peritoneal cavity through the right paralumbar fossa, using local anesthesia, for measurement of IAP. These pressures were recorded in 1-minute intervals for 2 hours, while the horses were standing tied in a stall. IAPs of cribbing horses were compared to the noncribbing cohort...
Abnormal plasma neuroactive progestagen derivatives in ill, neonatal foals presented to the neonatal intensive care unit.
Equine veterinary journal    April 22, 2013   Volume 45, Issue 6 661-665 doi: 10.1111/evj.12065
Aleman M, Pickles KJ, Conley AJ, Stanley S, Haggett E, Toth B, Madigan JE.Increased levels of pregnanes have been reported in foals with neonatal maladjustment syndrome (NMS). These steroids may cross the blood-brain barrier and have depressive effects in the central nervous system leading to behavioural abnormalities and altered states of consciousness in affected foals. Objective: The aim of this study was to determine the pregnane profile of foals with NMS and compare it with that of healthy controls and sick, non-NMS foals. Methods: Prospective-clinical study. Methods: Thirty-two foals with a clinical diagnosis of NMS, 12 foals with other neonatal disorders and ...
Effect of pressure support ventilation during weaning on ventilation and oxygenation indices in healthy horses recovering from general anesthesia.
Veterinary anaesthesia and analgesia    April 20, 2013   Volume 40, Issue 4 339-350 doi: 10.1111/vaa.12041
Ida KK, Fantoni DT, Souto MT, Otsuki DA, Zoppa AL, Silva LC, Ambrósio AM.To determine if pressure support ventilation (PSV) weaning from general anesthesia affects ventilation or oxygenation in horses. Methods: Prospective randomized clinical study. Methods: Twenty client-owned healthy horses aged 5 ± 2 years, weighing 456 ± 90 kg. Methods: In the control group (CG; n = 10) weaning was performed by a gradual decrease in respiratory rate (fR ) and in the PSV group (PSVG; n = 10) by a gradual decrease in fR with PSV. The effect of weaning was considered suboptimal if PaCO2 > 50 mmHg, arterial pH 50 mmHg or PaO2 10 L (p = 0.0001), fR /VT ratio ≤ 0.60 breaths ...
The effects of pulse-delivered inhaled nitric oxide on arterial oxygenation, ventilation-perfusion distribution and plasma endothelin-1 concentration in laterally recumbent isoflurane-anaesthetized horses.
Veterinary anaesthesia and analgesia    April 20, 2013   Volume 40, Issue 6 e19-e30 doi: 10.1111/vaa.12037
Grubb T, Frendin JH, Edner A, Funkquist P, Hedenstierna G, Nyman G.Anaesthetized horses commonly become hypoxaemic due to ventilation/perfusion (V·A/Q·) mismatch and increased pulmonary shunt fraction (Qs·/Qt·). Pulse-delivered inhaled nitric oxide may improve oxygenation but may increase plasma concentration of the potent vasoconstrictor, endothelin-1 (ET-1). Objectives: Study 1) compare arterial oxygen concentration (PaO2) and saturation (SaO2), calculated Qs·/Qt· and ET-1 concentration; and Study 2) assess V·A/Q· matching and measured Qs·/Qt· in isoflurane-anaesthetized horses in left lateral recumbency receiving pulse-delivered inhaled nitric ox...
Amino acid utilization by the hindlimb of warmblood horses at rest and following low intensity exercise.
The veterinary quarterly    April 19, 2013   Volume 33, Issue 1 20-24 doi: 10.1080/01652176.2013.775833
Peters LW, Smiet E, de Sain-van der Velden MG, van der Kolk JH.In particular branched-chain amino acids might limit muscle protein loss in pathological conditions. Little is known on basic amino acid utilization of muscle in horses. Objective: To assess amino acid utilization by the hindlimb of horses at rest and following low intensity exercise. ANIMALS & METHODS: Amino acid uptake by the hindlimb was investigated using the arteriovenous difference technique. Blood from six warmblood mares (mean age 12 ± 3 (SD) years and weighing 538 ± 39 kg) was collected simultaneously from the (transverse) facial artery and from the caudal vena cava. Food was wi...
Pre-selection by double layer density gradient centrifugation improves the fertilising capacity of frozen-thawed, capacitated stallion sperm.
Animal reproduction science    April 18, 2013   Volume 139, Issue 1-4 62-68 doi: 10.1016/j.anireprosci.2013.04.005
Morató R, Soares JM, Orero G, Mogas T, Miró J.The effect of combining double layer density gradient centrifugation (DL-DGC) with different capacitation treatments on the fertilising capacity of frozen-thawed stallion sperm was examined via a heterologous assay involving in vitro-matured, zona pellucida-free bovine oocytes. In a first experiment, aliquots of frozen-thawed stallion sperm were subjected to one of five capacitation treatments without DL-DGC - ionomycin at 1.0μM, 0.1μM, 0.05μM or 0.01μM, or caffeine at 200μg/mL. The fertilising capacity of the semen was then assessed at 18h by staining the above oocytes with 4,6-diamidino...
Change of antibody levels to ferritin in the sera of foals after birth: possible passive transfer of maternal anti-ferritin autoantibody via colostrum and age-related anti-ferritin autoantibody production.
Animal science journal = Nihon chikusan Gakkaiho    April 18, 2013   Volume 84, Issue 12 782-789 doi: 10.1111/asj.12069
Numata M, Kondo T, Nambo Y, Yoshikawa Y, Watanabe K, Orino K.Antibody (immunoglobulin G (IgG), IgM or IgA) levels relative to ferritin in six foal sera (three male and three female) after birth (day 0 and 2, 6, 10, 20, 28, 36, 40, 52 and 56 weeks of age) were semi-quantitatively measured with normalization with antibody activity to ferritin in one adult horse serum. After addition of horse spleen ferritin to the serum sample, the complex formed between antibodies to ferritin in the serum and ferritin was co-immunoprecipitated using antibody to horse spleen ferritin. Antibody classes of the co-immnoprecipitate were detected with antibodies specific for h...
Sperm trajectories form chiral ribbons.
Scientific reports    April 17, 2013   Volume 3 1664 doi: 10.1038/srep01664
Su TW, Choi I, Feng J, Huang K, McLeod E, Ozcan A.We report the discovery of an entirely new three-dimensional (3D) swimming pattern observed in human and horse sperms. This motion is in the form of 'chiral ribbons', where the planar swing of the sperm head occurs on an osculating plane creating in some cases a helical ribbon and in some others a twisted ribbon. The latter, i.e., the twisted ribbon trajectory, also defines a minimal surface, exhibiting zero mean curvature for all the points on its surface. These chiral ribbon swimming patterns cannot be represented or understood by already known patterns of sperms or other micro-swimmers. The...
The role of polyphosphates in the sequestration of matrix metalloproteinases.
International wound journal    April 17, 2013   Volume 12, Issue 1 89-99 doi: 10.1111/iwj.12058
McCarty SM, Percival SL, Clegg PD, Cochrane CA.This study outlines the potential of a novel therapeutic dressing for the management of chronic wounds. The dressing incorporates polyphosphate, a non toxic compound with a number of beneficial characteristics in terms of wound healing, in a foam matrix. The aim of this study was to identify the potential of polyphosphate incorporated in the foam dressing to sequester the activity of matrix metalloproteinases (MMPs) and proteases derived from Pseudomonas aeruginosa. Methods used included gelatin zymography and milk-casein agar plate analysis. Results have shown that this dressing is effectivel...
Investigation of the solubility and the potentials for purification of serum amyloid A (SAA) from equine acute phase serum–a pilot study.
BMC research notes    April 16, 2013   Volume 6 152 doi: 10.1186/1756-0500-6-152
Christensen MB, Sørensen JC, Jacobsen S, Kjelgaard-Hansen M.Serum amyloid A (SAA) is useful as a diagnostic marker of systemic inflammation in horses, but only heterologous assays based on non-equine calibration and standardization are available for measurements of equine SAA. More accurate measurements could be obtained using purified species-specific SAA in native conformation for assay calibration and standardization. Further knowledge about the biochemical properties of SAA would facilitate a future production of native species-specific calibration material Therefore, the aim of the study was an investigation of the solubility and potentials for pu...
Effect of Treadmill Exercise and Hydrogen-rich Water Intake on Serum Oxidative and Anti-oxidative Metabolites in Serum of Thoroughbred Horses.
Journal of equine science    April 16, 2013   Volume 24, Issue 1 1-8 doi: 10.1294/jes.24.1
Tsubone H, Hanafusa M, Endo M, Manabe N, Hiraga A, Ohmura H, Aida H.The present study aimed to clarify changes of oxidative stress and antioxidative functions in treadmill-exercised Thoroughbred horses (n=5, 3 to 7 years old), using recently developed techniques for measurement of serum d-ROMs for oxidative stress, and BAP for antioxidative markers. Also, the effect of nasogastric administration of hydrogen-rich water (HW) or placebo water preceding the treadmill exercise on these parameters was examined. Each horse was subjected to a maximum level of treadmill exercise in which the horses were exhausted at an average speed of 13.2 ± 0.84 m/sec. Blood samples...
Endometrial inflammatory markers of the early immune response in mares susceptible or resistant to persistent breeding-induced endometritis.
Reproduction (Cambridge, England)    April 13, 2013   Volume 145, Issue 3 289-296 doi: 10.1530/rep-12-0452
Woodward EM, Christoffersen M, Campos J, Betancourt A, Horohov D, Scoggin KE, Squires EL, Troedsson MH.Transient endometritis after breeding is necessary for clearance of bacteria and spermatozoa; however, in a subpopulation of mares, the inflammation fails to resolve in a timely fashion. The objective of this study was to describe the uterine inflammatory response in mares susceptible or resistant to persistent breeding-induced endometritis (PBIE) during the first 24 h after induction of uterine inflammation.Twelve mares were classified as susceptible (nZ6) or resistant (nZ6) to PBIE. Mares were inseminated over five estrous cycles and endometrial biopsies were collected at one time point per ...
Serum anti-Müllerian hormone concentrations in stallions: developmental changes, seasonal variation, and differences between intact stallions, cryptorchid stallions, and geldings.
Theriogenology    April 13, 2013   Volume 79, Issue 9 1229-1235 doi: 10.1016/j.theriogenology.2013.03.019
Claes A, Ball BA, Almeida J, Corbin CJ, Conley AJ.Anti-Müllerian hormone (AMH), a homodimeric glycoprotein, is secreted early in fetal life when it exerts a crucial function in sexual differentiation. The secretion of AMH in male humans persists after birth and is characterized by high prepubertal concentrations followed by a significant decrease at the onset of puberty. The expression of AMH in the normal and cryptorchid equine testis is well characterized but data regarding circulating AMH concentrations are lacking. The objectives of this study were to determine serum AMH concentrations in neonatal colts and fillies, prepubertal colts, an...
Peeling back the evolutionary layers of molecular mechanisms responsive to exercise-stress in the skeletal muscle of the racing horse.
DNA research : an international journal for rapid publication of reports on genes and genomes    April 11, 2013   Volume 20, Issue 3 287-298 doi: 10.1093/dnares/dst010
Kim H, Lee T, Park W, Lee JW, Kim J, Lee BY, Ahn H, Moon S, Cho S, Do KT, Kim HS, Lee HK, Lee CK, Kong HS, Yang YM, Park J, Kim HM, Kim BC, Hwang S....The modern horse (Equus caballus) is the product of over 50 million yrs of evolution. The athletic abilities of the horse have been enhanced during the past 6000 yrs under domestication. Therefore, the horse serves as a valuable model to understand the physiology and molecular mechanisms of adaptive responses to exercise. The structure and function of skeletal muscle show remarkable plasticity to the physical and metabolic challenges following exercise. Here, we reveal an evolutionary layer of responsiveness to exercise-stress in the skeletal muscle of the racing horse. We analysed differentia...
The expression of mucin genes and the presence of mucin gene products in the equine endometrium.
Research in veterinary science    April 11, 2013   Volume 95, Issue 1 169-175 doi: 10.1016/j.rvsc.2013.03.012
Maischberger E, Cummins CA, Fitzpatrick E, Gallagher ME, Worrall S, Rousseau K, Thornton DJ, Meijer WG, Miranda-Casoluengo R, Duggan VE....In the equine reproductive tract, little is known about mucin gene expression and the role of mucins in barrier function and host-cell interaction. The aims of the study were to identify equine orthologs of mammalian mucin genes using available equine sequence data, to profile expression of equine orthologous mucin genes in the endometrium using reverse transcriptase polymerase chain reaction (RT-PCR), to determine spatial expression patterns of mucin genes using in situ hybridisation, and to confirm the presence of mucin gene products using Western blotting and equine-specific mucin antibodie...
Age-related changes in the central nervous system in selected domestic mammals and primates.
Postepy higieny i medycyny doswiadczalnej (Online)    April 11, 2013   Volume 67 269-275 doi: 10.5604/17322693.1044490
Firląg M, Kamaszewski M, Gaca K, Bałasińska B.Aging is a process which operates at many levels of physiological, genetic and molecular organization and leads inevitably to death. Brain macroscopic changes by MRI investigation during aging were observed in humans and dogs but chimpanzees did not display significant changes. This suggestion led to the statement that brain aging is different in various species. Although human brain changes, e.g. β-amyloid storage, neurofibrillary tangle formation, lipofuscin, are relatively well known, we are still looking for a suitable animal model to study the mechanisms of aging and neurodegenerative di...
The absence of ER-β results in altered gene expression in ovarian granulosa cells isolated from in vivo preovulatory follicles.
Endocrinology    April 11, 2013   Volume 154, Issue 6 2174-2187 doi: 10.1210/en.2012-2256
Binder AK, Rodriguez KF, Hamilton KJ, Stockton PS, Reed CE, Korach KS.Determining the spatial and temporal expression of genes involved in the ovulatory pathway is critical for the understanding of the role of each estrogen receptor in the modulation of folliculogenesis and ovulation. Estrogen receptor (ER)-β is highly expressed in ovarian granulosa cells, and mice lacking ER-β are subfertile due to inefficient ovulation. Previous work has focused on isolated granulosa cells or cultured follicles and, although informative, provides confounding results due to the heterogeneous cell types present including granulosa and theca cells and oocytes and exposure to in...
Conceptualising the Impact of Arousal and Affective State on Training Outcomes of Operant Conditioning.
Animals : an open access journal from MDPI    April 11, 2013   Volume 3, Issue 2 300-317 doi: 10.3390/ani3020300
Starling MJ, Branson N, Cody D, McGreevy PD.Animal training relies heavily on an understanding of species-specific behaviour as it integrates with operant conditioning principles. Following on from recent studies showing that affective states and arousal levels may correlate with behavioural outcomes, we explore the contribution of both affective state and arousal in behavioural responses to operant conditioning. This paper provides a framework for assessing how affective state and arousal may influence the efficacy of operant training methods. It provides a series of three-dimensional conceptual graphs as exemplars to describing putati...
Getting a feel for equine colic.
The Veterinary record    April 9, 2013   Volume 172, Issue 14 354 doi: 10.1136/vr.f2119
No abstract available
Establishment and evaluation of a bead-based luminex assay allowing simultaneous quantification of equine IL-12 and IFN-γ.
Anticancer research    April 9, 2013   Volume 33, Issue 4 1325-1336 
Duran MC, Willenbrock S, Müller JM, Nolte I, Feige K, Murua Escobar H.Interleukin-12 (IL-12) and interferon gamma (IFN-γ) are key cytokines in immunemediated equine melanoma therapy. Currently, a method for accurate simultaneous quantification of these equine cytokines is lacking. Therefore, we sought to establish an assay that allows for accurate and simultaneous quantification of equine IL-12 (eIL-12) and IFN-γ (eIFN-γ). Methods: Several antibodies were evaluated for cross-reactivity to eIL-12 and eIFN-γ and were used to establish a bead-based Luminex assay, which was subsequently applied to quantify cytokine concentrations in biological samples. Results: ...
New seminal plasma removal method for freezing stallion semen.
Theriogenology    April 9, 2013   Volume 79, Issue 7 1120-1123.e1 doi: 10.1016/j.theriogenology.2013.01.014
Ramires Neto C, Monteiro GA, Soares RF, Pedrazzi C, Dell'aqua JA, Papa FO, Castro-Chaves MM, Alvarenga MA.Seminal plasma removal, an indispensable step in equine semen cryopreservation, is usually done by centrifugation, but this might cause mechanical damage to sperm. A new method for seminal plasma removal from stallion semen, namely a filter composed of a synthetic hydrophilic membrane (Sperm Filter, BotuPharma, Botucatu, Sao Paulo, Brazil), was recently proposed. The objective of this study was to test the use of the Sperm Filter in the removal of seminal plasma before freezing stallion semen. Ejaculates from 31 stallions were divided into two groups and cryopreserved. In group 1 (G1), seminal...